fflONAL INSTITUTES OF HEALTH
NATIONAL INSTITUTE OF NEUROLOGICAL
DISEASES AND BLINDNESS
HATHONAi INSTITDTES OF HEALTH
PUBLIC HEALTH SERViCE :
U.i DEPARTMENT OF HEtfffl, EDUCATION, AiSDWELFAB
;iatio!i£! ;;u. ^ Health
BuilcJing 10
Batiiesda, Maryland 20014
MNUAL RSPOE!r - 1958
NIKDB Office of the
Director Project Reports
Title Chief Investigator
The Director's Report--- — Bailey
Direct Training------ ■-- — Wipf
Collaborative Research Masland
Epidemiology Branch — Kurland
Biometrics Branch — -- Goldstein
Extramural Progrsns- — Seger
lo Research Grants — — — --■ Seger
IIo Field Investigations and Pilot Projects Seger
III. Graduate Training Grants „_»„-— .^ »- Seger
IV« Special Traineeship Program ■- Harttnan
Vo Research Fellowships — — — Hartman
VI. Review and Approval of Grants — .---—-- — Seger
EC
clinical Research - 1958
Clinical Director's Reporfc-— — — — Shy
Services Given by the Clinical Investigations Unit — Shy
Blectroeacephalography and Clinical Neurophysiology
SrancE
Summary- ■ »---- — =. . Ajxaone-Marsan
Individual Project Report —
Serial No. Title Principal Investigator
1(c) /inaiytical study of focal cerebral
seizures--- — — ---- — • — Ajmone-Marsan
2(c) Depth electrography in epileptic
patients — — ■— Ajmone-Marsan
3(c) Electrocorticographic studies in
temporal lobe epilepsy and in focal
cerebfal seizures--- — ----- ~— . Ajcione-Marsan
4(c) Epileptic activation of unitary
elements of the cat cerebral cortex
and their relationship with EEG
discharges— — "- — -- — ■-■ — — — — — Enonioto
5(e) Unit analysis of the responses
elicitable in the visual corten — • Ajtoone-Marsan
Widen
6(c) The modification of sensory mechanisms
by subcortical structures — - — ■ Long
7(c) EEG changes induced with photic
stimulation in patients treated
with ACTH and adrenal corticoids- — Abraham
8(c) Bibliography on Electroencephalography
and Clinical Neurophysiology,
1948-1958—-—-- — — --— «— - — - — -- Ajmone-Marsan
Henry
9(c) EKCltation in medullated nerve -■ del Castillo
Serial No. Title Principal Investigator
10 (c) Study of Mechanisms of Transmitter
Liberation at presynaptic Nerve
Endings- — — -- - del Castillo
Medical Neurology Branch
Sutmnary-- -■- — - — ■ — ^
Individual Project Report
Serial No„ title Principal Investigator
11(c) The Use of a Monomaine Oxidase Inhibitor
(JB-516) as an Anticonvulsant Medication
for Centrencephalic Seizures-- Prockop
Smith
12(c) Clinical Pathological Correlative Study
of the Nervous System in Orthorostatic
Hypotension-- • ■ Drager
13(c) The Histopathological and Chemical
Investigations of Neuromuscular Disorders Shy
Wanko
14(c) Pathological Study of Intramuscular Motor
and Sensony Nerve Endings in the Normal
and in Neuromuscular Diseases Haase
15(c) The Physiology and Pharmacology in
Myasthenia Gravis— — • _— ghy
16(c) A Study of Progressive Parenchymatous
Degeneration of the Central Nervous System Smith
Drager
17(c) Spirochetes and viral antigens and their
relation to the spinal fluid and blood
of multiple sclerotic patients Korengold
18(c) Investigations in Localization of
Cerebral Neoplasia by Isotopic Detection- Shy
19(c) Metal Chelates as Possible Contrast Media
for Myelography — - — - — Ci Chiro
Rubin
20(c) Blood and tissue cholinesterases in
neuromuscular blockade-- =--- .- — Irwin
Serial No. Title Principal Investigator
21(c) The action of neuromuscular blocking
drugs on directly stimulated inner-
vated and denervated xnuscle ■ — -- Irwin
22(c) Study of muscle cholinesterase
and its inhibitors Irwin
23(c) A study of naturally occurring
choline esters. Irwin
24(c) A study to determine the effects of
depolarizing drugs on muscle
enzymes --^ — Irwin
25(c) Electrolyte and Energy Metabolism in
Normal and Epileptogenic Cerebral
Cortex in Vitro — . Tower
26(c) Comparative Biochemistry of Smooth
ly&iscle and Striated 14iscle Horvath
27(c) Amino Acid Metabolism in Normal and
Epileptogenic Cerebral Cortex
in Vitro — Tower
28(c) Clinical Evaluation of Various
Amino Acids and Related Compounds
in Control of Seizures including
Studies of their Metabolism in Vivo----- Tower
29(c) The Relation of Pyridoxine (Vitamin
B6) to Certain Seizure States McKhaan
30(c) The Metabolism of Y-Aminobutyric Acid
in Neural Tissue-- • McKhann
31(c) Microchemical Determination of
Acetylcholine Curtis
32(c) Physico-chemical studies of Human
Cerebrospinal Fluid--------- — - Curtis
33(c) The Surface- Chemical Behavior ©f
Urine in Relation to its Surface-
Active Macromolecular Constituents Curtis
Serial No. Title Principal Investigator
34(c) Cerebral Protein Metabolism and
Turnover in Tissue Slices incubatea
in vitro, -• — - — --— — ---- Wherrett
35(c) Distribution of Actin and Tropomyosin
in Normal and Diseased Muscle Horvath
36(c) Alterations of Actomyosin Tensile
Strength and ^aiscle Proteins in
Neuromuscular Diseases ^-— Proctor
Shy
Ophthalmology Branch
Summary — ■ — von .Sallman
Individual Project Report
Serial No» Title j:'rincipal Investigator
37(e) Studies on Diet and Drug Induced
Experiment*! Cataract von Sallmann
38(c) Study of Submicroscopic Structures
of Lens Tissue Componets by Phase
Contrast Microscopy ■ von Sallmann
Caravaggio
Aronson
39(c) Studies on Central Nervous System
Control of Intraocular Pressure,,
(Anatomy of Posterior Ciliary
Nerves)-- ~ — - — ---» von Sallmann
Grimes
40(c) Study of Afferent Electric Impulses
Induced by Intraocular Pressure
Changes-- .— — — Lele
41(c) Effects of Muscle Relaxants on lOP ■
and Extraocular Striate Mascles— BJacri
42(c) Study on the Pharmacodynamics of
Various Agents Affecting Intraocular
Pressure-----—-------------—"—' — -— --- Macri
Serial Ro., Title
43(c) Clinical Glaucoma Study — — — —
44(c) The Ocular Diagnosis and Treatment of
Ocular Complications of Primary
Familial Amyloidosis- — - — ■ — — — — —
45(c) Study of Toxoplasmosis and Its
Therapy- ■ _-_- — -
46(c) Thyroid Hormone Turnover in Uveitis
47(c) Detection of Ocular Tumor by Isotope
Tracer Methods — - — ■—
4S(c) Immunological Relations in Ocular
Tissues ■ ----
49(c) Basic Factors in Refraction Anomalies— •—-
50(c) Intraocular Angiography — »---—» — -»—-.—
51(c) Electron Microscope Studies on Epithelium,
Capsule and the Fibers ef the Lens and on
the Ef^ithelixim of the Ciliary Body and
the Optic Herve-------- — ~- ■
52(c) Electron Microscope Studies on Biopsies
of Human Muscle Diseases--- —
53(c) Study of Submicroscopic Structures of
Ocular Pigment Gellso (Staining of
the Living Tissue Culture Figment
Cell by Acridine Orange) — -- •-
54(c) A Study of the Proteins of the Lens--— — -
55(c) An Investigation of the Enzymatic Systems
Present in the Lensj Cornea and 4<|usous
Humor and their Relation to ^ vivo
Tissue Metabolism-—- -—
56(c) Electrophysiology of the Eye— — --— — —
57(g) erg Reactions of Pure-Cone Mammalian
Retinae—- — „-_.
Paton
von Sallmann
Kaufman
van Alphen
Kaufman
O'Rourke
O'Rourke
van Alphen
van Alphen
Cohan
Wanko
Wanko
Shy
Wolf
Aronson
Resnlk
Safelman
Fuortes
Tans ley
Copenhaver
Serial No. Title Principal lavestigator
58(c) Functional Studies in Retinal
Anomalies and Diseases
(Electsoretinography, Adaptometry,
and Perimetric Light Sense Studies)-^— Copenhaver
Gunkel
59(c) ERG Spectral Sensitivity Curves
on Caucasians^ Negroes, and Albinos — Dodt
60(c) Design and Construction of
Ophthalmic ingtruments — -- — ---—._„_ Gunkel
Surgical Neurology Branch
Summary- — - — — — — • Baldwin
Individual Project Report
Serial Title Principal Investigator
61(c) Epileptogenic Mechanisms in the
Brain of Man — — Baldwin
62(c) Functional Representation in the Temporal
Lobe of Man and Higher Primates-- — - Baldwin
63(c) Effect of Tumors upon the Central Kexvous
System Function and Structure Van Buren
64(c) A Study of the Functional Anatomy and
Pathology of the Human Visual System-- ■ Van Buren
65(c) Studies of Involuntary Movements- — ■ Van Buren
-66(c) Pain Mechanisms-- — ■ — ----- yan Buren
67(c) Study of Cortical Intracellular
Potentials „ — i^i
': 68(c) Factors Determining the Discharge of a
Motor Neuron in Cerebral Cortex i^i
69(c) The Problem of Synchronous Activity
of Nerve Cells in Cerebral Cortes - Li
71(c)
72(c)
73(c)
74(0)
75(c)
76(c)
77(c)
78(c)
79(c)
80(c)
81 (c)
82(c)
Title Principal Investigator
Neuromuscular Transmission in
Hypothermia— — ~ Li
Effect of Cooling on Conduction of
Iiiipulses in Cranial and Peripheral
Nerves——— — — — — .__„„„ Ortis
Study of Pharmaceutic Agents Acting on
Various Cortical and Subcortical
Structures of the Brain — — Ortiz
Properties of Cultured Nerve and Muscle
Ceils — — — — — " — Li, Klatzo
and Baldwin
Pinocytosis of Labelled Proteins in
Tissue Culture Klatzo
The Localisation of Myosin in Human
Striated Muscle by Fluorescent
Antibody-—-- • — - Klatso
Study of Pathology of Kuru Disease — Klatso
Study of Regeneration in the Central
Nervous System — — — - — Ortiz-Galvan
Histochemical and Electrophysiological
Observations on the Muscle Fibers Gro^m
in Vitro — =>-- --«=-_---_ Engel
A New Method for Quantitative Study of
Precipitin Reaction— — — — - — ---»---- Miquel
The Relationship between Edema j Blood-
Brain-Barrier and Tissue Elements in
Experimental Brain Injury- .— » Klatzo
Study of the Effects of Hypothermia
on Injured and Normal Brain Tissue- Laskowski
The Investigation of the Site, TjTpe and
EKtent of Lesions Involving the CNS in
Cerebral Palsy and Allied Conditions- Dekaban
Serial Mo. Title Principal Invesfcigafcoi
83(c) Mafcemal Condition Busing Pregnancy
and the Course of Birth in Relation
to Neurological Abnormalities in the
Infants and Pathologic Lesions in
Products of Abortion-— -------- — —- — Dekaban
84(c) Pathological Lesions in the Central
Hsrvous System Occurring During
Prenatal, Intranatal and Early
Early Postnatal Life • --■ Dekaban
85(c) The Incidence and the Type of the
Central Nervous System Abnormalities
Encountered in Offspring Bom to
Diabetic Mothers------ -— --— Dekaban
Baird
86(c) Measurements of External and Internal
Orbital Distance in Males and Females
from Birth to Adulthood -— — — Dekaban
87(c) Preparation of the Horizons of the
Normal Development of the CNS in Mice
and Experimental Production of Congenital
Malformations of the CNS— — — ■?— ?-ri- Dekaban
88(c) Effect of "fear-provoking" stimuli
on visual discrimination in primates — Lansdell
89(c) Psychological Evaluation of Temporal
Lobe Disease— — — — -- — ^~-.=.-----.---»--- Lansdell
90(c) Body Temperature in Chimpansees with
Bilateral Tssnporal Lobe Damage- — — — <. Blevins
91(c) Fiuothane Studies — -=- Hall
92(c) Hypothermia in Neuroanesthesiology- — • — -— Hall
93(c) Succinyl Choline in Awake Craniotomy——--- Hall
94(c) The Effect of Hypertonic Urea Solution
on Intracranial pressure—————— — ' Pritchard
Basic Research
Introduction------
Laboratory of Neuroanatomical Sciences
Summary" — —
Individual Project Report
Serial No. Title
Livingston
Windle
Principal Investigator
NiI?DB-KA-DR-l Development of Intrinsic Structures
of the Human Brain
NINDB-WA-DR-2 Histogenesis of normal and dystrophic
retinas in mice-- — ------- — . .
•NA-DR-3 Histogenesis in the embryonic
mammalian nervous system-- — -
NIM53-NA-DR-4 Regeneration in the central nervous
system ■ — . — — — -.
NINDB-NA-DR-5 Functional and Structural Changes in
Reserpinised Animals— — ---
NINDB-NA-DR-6 Neuronal specificity in the autonomic
nervous system- —
NIHDB-NA-DR-7 Heterogeneous Reinnervation of the
Diaphragm — — ■
HIi©B-NA-DR-8 Experimental Analysis of the nerve
fiber-taste bud relationship-^ — -
HIi©B-NA-DR-9 Hervous System Pathology in Macaca
Mulatta after Asphyxia Neonatorum—
NIKDB-NA-DR-IO The significance of the acridine .
orange staining of neurons iii vitro
and in vivo------- — •--- — -
NIHDB-NA-DR-ll Structure and chemistry of photoreceptor
cells — ———-—-.—«—--—
Windie
Guth
Sidman
Sidman
Windie
Windie
Guth
Guth
Guth
Ranck
Windie
Wolf
Sidman
Serial No. Title Principal Investigator
NII3DB-NA-DR-12 Development of new histcchsmical
methods Sidman
Feder
NINDB-NA-DR--3 Behavior and social organisation of
rhesus monkeys on Cayo Santiago,
Puerto paco— "— — — — Altraann
HIM)B~NA-ri-14 Physical measurements of rhesus monkeys
from birth to old age»- Altmann
KINDB-N/-DR-15 Technique of neurological examination of
the monkey (Macaca mulatta) Ranck
NIHDF,-NA-DR-16 Normal reproductive function in the
rhesus monkey — -— — Jacobson
NIT/;B-NA-DR-17 Maturation in infant rhesus monkeys;
and care required for rearing them— Jacobson
'/-NBB-NA-DR-IS The intrinsic nerve supply to the endometrium
in cat, guinea pig, monkey and man Jacobson
:^INDB-NA-DR-19 Neurological deficits of asphyxia
neonatorum in macaca mulatta — - Windle
Ranck
Combs
Jacobson
I3INDB-NA-DR-20 Psychological effects of asphyxia
neonatorum in rhesus monkeys-- — ----- — __=-_ Bailey
Saxon
NIM>B-NA-DR-21 Psychological and histopathological
deficits of asphyxia neonatorum in
guinea pigs — Bailey
NINDB-NA-DR-22 Centers and pathways involved in induced
cerebellar seisures— — „.——>--. combs
NINDB-NA'NG-1 Ultrastructure of the nervous system — Palay
NISDB-NA-NC-2 Ensymatic reactions of gamma- aminobuty rate
(V-AB) catalyzed by brain tissue — - — Albers
- .-.atitative histocheaiic . ,_.
o£ glutamic decarboxylase in the
ner\'ous system -„_»--.-...—» „
HII''iI)S-RA-NC-4 Micro- radiometric Eisasurairieiit of
dec£i-boxyiase reactions -»--- < -_ Albers
A fluroliQetric micromethod for the
determinatio-n of succinic samialdehyde Albirs
A-NG-6 Heurroyacration in the rodent ------> . — Bv:
ITIHDB-NA-HG-7 ,::.ai cholinesterase oS the
vertebrate central nervous systam B
I-IiroS-l: I'; Pathogenetical factors in the
clavelo/jviiarit o:" ni-^.^eonstbies--------- — •-■ — ■ Caiornan,, ./^ .
. ■'":"';'- ■:'\ , .,„___.._____.. _:. „.:._ _,. ihe epidural
space in msKnnals--- -__«-, — ___«„__„__ q-,
l'£l'lDB**NA-EP-3 Structure of brains of iconkeys in which
the pituitary gland had been irradiated
with high-energy deuterons .----» Canscasr;
KIIILly-KA-'J-^ - :tudy of the auditory afferent sad
..- .-.;:.- .- ,.-.;,... ..'_:.._ L_„::; ..„:.:._1.^^..._ snd genetic
sf:.;j.dj' of ths hearing raechanism in a strain
of coBgential deaf guinea pigs-- — ■ — -- — "-- Rsi ......
.;:1DB-1TA-FH~3' An experimental study of the niediai
longitudinal fasciculus of the brain stem
and spinal cord---- ,..,„„_„„«„ — — ^„_..„ jjassopust
IIXELI^-r. Neuronal connec . .3 functional
significance of the iiii:a';:.p£duacular
i>iIl\DE-l:!A-FN-5 :;tive anatomy of the efferent
cohlsi.:; iundle in selected subraiaalian
vertebrates; and experimental study"---"—"- Ecord
Se!;ial No. Title Principal lavaetigafcor
K'JM)B-NA-FN-6 A study of an efferent component
of the vestibular nerve arising
from the medulla oblongata------ Rasmussen
Lab oratory of Biophysics
Sun .mary--- ----.-.- — «»----»« — „_ »„__.^ — -,„___> -_ cole
individual Project Report
j Serial No. Title Principal Investigator
SIlTOB-B-l Ionic Permeabilities of the Squid
Giant Axon Membrane — Cole
'. TINDB-B-2 Ionic Permeabilities of Nerve Membranes
Theoretical Investigations — — — FitzHugh
■ iINDB-B-3 Correlation of Acetylcholinesterase
Inhibition with Nerve Action- — — Tiihitcomb
ilIWDB-B-4 Membrane Potentials of a Lobster
Giant Asron ^— — «-.— — Dalton
V3INDB-B-5 Ionic Permeabilities of Nodal Membrane-- Moore
del Castillo
I/iboratory of Neurophysiology
fjuimnary- —-»--—--...---- — -— — --_- — .--.__ — — Marshall
Individual Project Report
Serial No. Title Principal Investigator
NINDB-NP-SS-la The chemistry of neural activity — Tasaki
Spyropoulos
NINDB-NP-SS-3 Investigation of the sensory mechanism——— Tasaki
NlNDB-NP-SS-4 Physiological studies on the nervous
elements in tissue culture---- ■- Chang
Tasaki
KINDB-NP-SC-3 Generation of impulses in spinal
motoneurons---- — - — • — —-—--—■— — « — - Frank
Fuortes
NIM>B-NP~S2-4 Effects of locally applied drugs on
single spinal niotoneurons-— ------ — »------. Frank
Paton
Serial i-iO. Title Principal Investigator
NINDB-IIP-SC-5 "Direct" contralateral inhibition- — Frank
Sprague
Laboratory cf Neurochemistry
Summary — Livingston
Section oa Lipid Chemistry —----, — _--- »«- Brady
Individu/I g3:oject Report
Serifl Ko. Title Principal Investigator
HIt®'j-NC°l Biosynthesis of Sphingoiips— — Brady
Nli/iB-NC-S Bios3mthesis of. Aromatic Compounds Brady
TiM!0}B-NC-7 Metabolism of Inositol- — Agranoff
.IIHDB-WC-13 The Effect of ^^phingosine on Blood
Co agul at ion- "-----—— — - Hecht
NIM)B~NC-i4 Ensymatic Synthesis of Fatty Acida — Brady
NIKDB-NC-15 Biosytithesis of Cholesterol Brady
NINDB-NG-12 Visuo-Itotor Coordination in a Lower
Vertebrate-- -— — » — Livingston
Iranyi
Nn®B-KC-13 Vestibular Influences on Spinal
Mechanisms- — --- — ---- — -_»>— — » Geraandt
m^cmL JSSWSSS& of msssasuxsssuL cibsasbs abd wsjem^ss
Office of ^36 IMjFectar
BBtJaated OMLJgatione « F. Y, 1959
XtLreet BeiiAaraaBSBt Votal
AdBiaiBtraUoa $1S8»000 0 $128,000
Faibliest&as^ end Beports 79^000 $16,000 95,000
(Xttfoneatioa Office)
Msroct Ts^iaias 39>500 0 39*000
AMUAL REPORT
Calendar Year 1958
National Institute of Neurological Diseases and Blindness
National Institutes of Health
The Director's Report
The 1958 annual report of the National Institute of Neurological
Diseases and Blindness contains the Director's Report; the Reports of
the Chief of the Extramural Programs Branch, Dr. Gordon H. Seger; the
Annual Report of the Clinical Director, Dr. G. Milton Shy; and the
Introductory Annual Report of the Director of Basic Research, Dr. R. B,
Livingston. It also contains summary reports of branch and laboratory
chiefs and the Report of Dr. Richard L. Masland, Assistant Director of
the National Institute of Neurological Diseases and Blindness, who
heads the Institute's program in collaborative research.
Since the reports by Drs . Seger, Shy, Livingston, and Masland
represent comprehensive commentaries on their 1958 scientific activi-
ties, they shall not be referred to further except when they relate
to new trends or changes in program emphasis. The Director's Report,
therefore, shall confine itself ..r^ the main to new trends and
developments in program emphasis and to special important events
of 1958 which are relevant to the Institute's overall mission.
DIGEST OF EXTRAMURAL PROGRAMMING HIGHLIGHTS
At the end of the calendar year, 1958, there were Qk-^ active
research grants as compared to 67O at the end of 1957^ or an in-
crease of 26.8 percent. Of the total number of research grant
awards, 26 percent were devoted to sensory disorders. Two thirds
of this 26 per cent were related to studies of vision, and one
third to studies of speech, hearing and equilibrium, and to
studies of smell, taste, touch, and pain. A noteworthy change
in programming was the lifting by the National Advisory Council
of the arbitrary five-year ceiling for the support of research
grants .
The program in field investigations and pilot project grants,
though only initiated in 1957^ now has 6k active projects, totaling
$^,329^196. Ninety two percent of the funds expended in this program
support multi-institutional, collaborative and cooperative studies.
The largest of these is the collaborative study of cerebral palsy,
mental retardation, and other neurological deficits of infancy and
childhood. In this study, the NINDB functions as a coordination
- 2 -
center and central lalDoratory for 1.6 other institutions, in a study
Involving the examination of 4o,000 pregnant mothers and infants.
The cooperative study of intracranial aneurysms reached a full
complement in 1958. Twenty cooperating institutions are in the study
at an annual cost of $200,000. The cooperative anticoagulant
therapy study in cerebrovascular disease must "be conducted one
more year before complete data will be available . This involves
six institutions at the cost of $5^,000 per annum.
A geomedical collaborative epidemiological study of selected
neurological disorders in South Carolina, Nova Scotia, and Japan
will be completed in 1959* New collaborative enterprises scheduled
for 1959 include a glaucoma detection evaluation study in collabo-
ration with the Chronic Disease Division of the Bureau of State
Services, and a project of the Institute's Biometrics Branch with
scientists of the American Academy of Ophthalmology and Otolaryn-
gology.
Closely associated with the support of research programs
within the extramural branch is NIKDB's program for the development
of future investigators. Early in the development of the research
grants programs, it became evident that the lack of trained
investigators was a bottleneck in our research effort. As initial-
ly set up, the training program concentrated on the training of
clinical scientists and academicians in neurology and ophthalmology.
This was important since it was recognized that research in the
neurological sciences would require a "core of clinicians" well
versed in the basic sciences who could serve in several capacities:
as coordinators for clinical research, as teachers within
university centers, and as leaders in neurology and ophthalmology
to recruit and stimulate both basic and clinical research relating
to neurological and sensory disorders .
At the end of the calendar year, 195°^ the number of grant-
supported programs for graduate training in the field of neurological
disorders was 55 • These programs were supported at a level of
approximately $1,600,000 or at an average of $29,091 per program.
One hundred and ninety-six trainees were in training for a career
in clinical neurology during the period ending June 30, 1958' At
present, 2^9 individuals are in training and 61 are expected to
graduate in June, 1959- According to the present rate, 80 to 90
trainees will complete their training as specialists in clinical
neurology each year.
The sharp research focus on neurologic disorders of early
life has revealed the dearth of well-trained pediatric neurologists .
The ITINDB program for the training of pediatric neurologists is
- 3 -
still in a cradling stage . During 1958, one new training program was
organized, bringing the total active programs in this area to three.
These programs are being supported at the level of $63,768, or an
average of $21,256 per program. At present there are 10 trainees in
the program of pediatric neurology. At least 20 new programs are
needed in this field.
In 1958^ a greater program emphasis was given to the training
of scientists in the neurologic basic sciences. These programs
increased from seven in 1957^ to 25 in 1958. This increase included
eight new programs in neuropathology, three in neuroanatomy, three
in neurophysiology, one in neuropharmacology, and three in neuro-
chemistry. These grants now total 25 in number- -in the amount of
$675,^58.
In 1958, the graduate training programs for developing
future investigators in ophthalmology saw an increase of programs
from 35 to 38. These programs provide a total of 296 trainees with
approximately 87 potential tepcher-investigators finishing residency
training every year. Training programs in otolaryngology also
increased from six to I8. The present number of trainees in this
program total II6, of whom 25 are in the third year, nine in the
fourth, and one in the fifth. It is expected that this program
will produce between 20 and 30 potential teacher-investigators
each year.
Another new area of program emphasis in trainin-g is the
development of a training grant program in sensory physiology.
This is a postdoctoral program designed to train basic scientists
in the physiology of the special senses. Four applications for
training programs in this area, in the amount of $2^9,^75 will be
submitted for review by the National Advisory Council at their
March meeting.
In 1958, there was a substantial advance in the development
of the program of Special Trainee ships . A total of $905^750 was
awarded to 125 trainees, an increase of approximately 50 percent
over 1957- In the face of an acute need for neuropathologists,
neurochemists, and those trained for research in the neurological
deficits of the young, a special tralneeship program contributed
materially to training in these gap areas . The 1958 support in
these areas was twice that of 1957 with I6 awards in neuropathology,
seven in neurochemistry, and I8 in pediatric neurology.
An appraisal of the year's training activities indicates
that the number of clinical training programs directed to neurologic
disorders has reached a temporary plateau which will remain fixed
- k -
until more trainees under the program mature Into teachers. Meanwhile,
program emphasis is turning to training teacher -investigators in
pediatric neurology, a serious gap area, and in the sciences haslc
to neurologic disorders . There also is an opportunity for the
directors of clinical programs to bring their activities in closer
contact with the basic sciences by developing clinical basic
divisions under their own auspices, such as in clinical neuro-
pathology, clinical EEG and neurophysiology, clinical neuro chemistry,
and related specialties .
The training programs directed toward ophthalmic disorders are
progressing slowly without indication of a precipitous rise. The
development of a genuine research interest on the part of ophthal-
mologists, with notable exceptions, has been comparatively slow.
Programs in otolaryngology are on the march but of limited potential
at present. The program in sensory physiology is in an advanced
planning stage .
The multidisciplinary approach of the NIWDB's collaborative
projects has created a need for additional training for clinical
specialists in their own specialty, or in related clinical
specialties, or in the basic sciences. This important need is
being met by the availability of special traineeships which has
been one of the crowning achievements of the NINDB's training
program .
INTRAMURAL GUIDELIHES
During 1958^ "the National Institute of Neurological Diseases
and Blindness continued its multidisciplinary clinical and basic
project studies .
The clinical program advanced considerably in its research
on neiiromuscular, physiological, chemical, ophthalmolcgical,
pathological, and surgical problems in the field of neurological
and sensory disorders . The clinical area has always operated
under the guiding principle that increased knowledge of diseases
of the nervous system depends upon the application of basic
techniques to the study of disease . Accordingly, the personnel
of this program have been trained not only in the clinical
sciences, but also have had additional postgraduate training in
the basic sciences, such as chemistry, mathematics, biophysics,
anatomy, pathology, and other basic disciplines.
Another basic premise of this program is that the investi-
gator must be free to shift his activities in any direction in
which his research carries him. This gives to the program a
multidisclplinary trend. A similar trend also may be obsei'ved on an
international scale. Here^ even among scientists trained as physiologists
and chemists, the newer techniques of electron microscopy and isotopic
work is being applied.
The NIEDB's clinical investigative program is reaching an
optimal level of functioning within the present limits of the NIH.
It is believed, however, that a clinical program in otology must be
developed at some time in the future .
The basic research program is a segment of an established and
powerful world eehter in scientific disciplines basic to neurology.
Their guiding principle is to create and manipulate those concepts
which will lead to a more fundamental understanding of the nervous
system and its functions. During the next ten years, the biophysics,
physical chemistry, and chemical structure of nerve membranes,
axonal and synaptic, will undoubtedly be much better understood.
Included in its present program are biophysical, neuroanatomical,
neurophysiological, and neurochemical research. Predicted develop-
ments for the future lie in the fields of physical biochemistry and
sensory mechanisms . The first of these will be devoted to cogent
areas of genetics, cellular physiology, and theoretical chemistry;
the second will pursue the analysis of sensory systems with special
emphasis on mechanisms involved in the central control of perception.
SCIENTrFIC COUNSELORS
In October, 1958, the NIWDB's Board of Scientific Counselors
held its second meeting in Bethesda. The purpose of "this group is to
advise the Director, NINDB, and the Director, NIH, in matters pertaining
to the intramural programs in both clinical and basic research.
Another important function of the counselors is to monitor and make
periodic reviews of the policies and activities of the intramural
programs. Dr. Eichard L. Masland, Assistant Director of NINDB,
serves as a point of contact between the Board of Counselors and
the Institute Director. The members of the NIHDB's Board of
Scientific Counselors are: Hallowell Davis, Chairman of the
Central Institute for the Deaf j Raymond D. Adams, Chief of Neurology
Service, Massachusetts General Hospital; Howard Curtis, Brookhaven
National Laboratory; Algernon B. Reece of the Institute of Ophthal-
mology, College of Physicians and Surgeons; Roger J. Rossiter,
Biochemist, University of Ontario; and A. Earl Walker, Professor
of Neurological Surgery, Johns Hopkins University.
In their deliberations, the counselors indicated that the
NINDB had two especially unique opportunities for justifying its
existence as a national institute . The first is its program of
collaborative projects and collaborative research in which the
Institute serves as a coordination center and a central laboratory
for miiltldiscipllnary and multi-institutional enterprises. It is
the only Institution in the country that could fill this role.
The second is the Institute's program of clinical investigations^
through which an eill-out effort is being made to explore the
potentialities of a particular technical method in relation to a
definite clinical problem. NINDB alone, according to the Board,
had the facilities for prosecuting research of this type.
The counselors also observed that the Bayne-Jones recom-
mendation of no further expansion of the National Institutes of
Health intramurally was instituted at an unfortunate time for
NINDB. In their opinion, a new building would be most desirable
to give adeqixate facilities for the present intramural staff.
It would also allow a more efficient arrangement of space
presently assigned to other activities in order that these could
be more efficiently concentrated.
Among the several program gap areas discussed by the group,
the most apparent and widest gap concerned studies in otology and
the auditory and equilibratory fimctions of the inner ear. The'
counselors took full cognizance of the sterling basic research
now being conducted by Drs = Tasaki, Rasmussen, and Gernandt,
but strongly recommended the organization of a clinical division
in otology as soon as adequate space and competent personnel
became available .
Finally, the counselors were interested in the question of
adequate representation of neuropathology as an essential foundation
of clinical investigation at NIH. The opportunity of NINDB to
provide a central neuropathological laboratory, to assist all
institutes having an interest in systemic conditions involving,
the brain, and to service certain cooperating institutions in
their collaborative projects, was considered both unique and
challenging. Consequently, the counselors recommended that a
discussion of the place of neuropathology in the organization and
operation of the Institute be surveyed and made a topic of dis-
cussion at their next meeting.
LANDMARKS IN COLLABORATIVE RESEARCH
Since the activation of the Institute in 1951, there has
been a trend toward more collaborative and cooperative types of
research programs to meet certain special problems. The solution
of these problems called for a greater collaboa"ation between the
Institute ' s intramural and extramural programs as well as greater
cooperation between institutions on mialtidisciplinary levels .
These problems arose because many neurological and sensory
- 7 -
syndromes appear to stem from a multiplicity of causes, giving rise
to the problem of procuring adequate samples. Tlie variation in the
geographic and climatic prevalence of certain neurological and sensory
disorders emphasizes the need of collahorative studies in geographic
neuropathology, both national and international. The Institute's
program is related to several specialized medical disciplines which
operate as distinct specialties as a matter of practical expediency
in medical practice, hut which, from a research standpoint, overlap
to such a degree that they can "be regarded as merely different points
on the same assembly line.-'- The rapid postwar development of older,
and the introduction of newer disciplines, in basic and neurological
sciences, add to these Institute problems.
These special problems gave rise to a need for a cooperative
plan to establish coordination centers and to consolidate these
program aims on institutional, geographic, and disciplinary levels.
The earliest examples of Institute cooperative projects on an
institutional level came in 1952 . These included tjie cooperative
project in retrolental fibroplasia, the collaborative project for
the evaluation of glutamine and asparagine in epileptic sei.2ures,
and the epidemiological studies in amyotrophic lateral sclerosis
on Guam. More recent examples of cooperative projects on a
mult i- institutional level are the evaluation of therapy for
intracranial aneurysms and the evaluation of the administration
of anticoagulants in cerebrovascular disease.
The RIlSDB's largest collaborative project is the one in
cerebral palsy and other neurological and sensory deficits of
early life. The present status of this mult i- institutional and
multidisciplinary project is described by Dr. Masland in his
section of the annual report. It has become increasingly evident
that in this type of research the Institute can plan an important
role, if not its most important role, by serving as an integrating
force in these collaborative projects. More specifically in this
respect, it serves as a focal point for the planning and mapping
of collaborative projects as well as a central laboratory for
the biostatistical collation of data and for the examination and
correlation of pathological specimens .
This developing trend in collaborative research has influ-
enced the relative amount of funds expended in 1958 on individual
research projects, as compared to the amount expended on collabo-
rative and cooperative field investigations. As Dr. Seger points
out, 51 percent of the research budget in 1957 "was used for the
support of individual research projects as compared to ^7 percent
of the total research budget in calendar year 1958- This shifting
emphasis raises the question as to whether or not the growth of
collaborative and cooperative field investigations, or the planned
type of research, is being achieved at the expense of the individual
1. Referred to our clinical disciplines of neurology and neuro-
surgery, ophthalmology and otolaryngology, and their respective
counterparts in the basic sciences.
initiative and creativity of the individual project type of research.
I think not . There is evidence at hand that planned collaborative
research is providing leads for the development of more individual
research projects. In other words, the collaborative and cooperative
programs serve as a supplement to, and not as a replacement of,
individual research projects. Serendipity, therefore, will be
preserved.
INTERNATIONAL RESEARCH
An unmistakable trend in medical research is a growing
gravitation toward greater international cooperation. This was
evident over the past two years in the programs of international
scientists assemblies. More recently, it has been expressed along
political lines. A section of President Eisenhower's January 9^
1958^ message to the Congress on the State of the Union, emphasized
the value and opportunities of international cooperation in medical
research. Later in the spring of the same year. Congressman John
E. Fogarty and Senator Lister Hill presented a joint resolution to
both Houses calling for greater cooperation in international
research to provide more building blocks for the accumulation of
scientific knowledge and to serve as an instrument of peace.
In December, 1958^ Senator Hubert Humphrey (D-Minn.)
visited Western Europe, and the USSR principally, as Chairman
of a Government Operations Subcommittee, to make a study of
problems pertaining to international health, research, assistance,
and rehabilitation. During his visit to Moscow, the Senator
and Soviet Premier Nikita Khrushchev disagreed on many things
but they agreed that "the world is hungry for some evidence of
Soviet -American collaboration" and the best way to start is with
medical research. Highlighted by Senator Humphrey is the need
for international cooperation to conquer cancer, heart disease,
and the killing and crippling neurological disorders of early
life, arising from disease or injury during the perinatal period.
In Senate Resolution 3^1, Senator Humphrey invited
President Eisenhower to extend to other nations of the world,
through the World Health Organization, an invitation to meet and
discuss the feasibility of designating an International Health
and Medical Research Year.
GEOGRAPHIC AND CLIMATIC DISTRIBUTION OF DISEASE
International studies in the geographic and climatic
distribution of disease often provide clues of a general nature
which lead to further and more fundamental investigations of
- 9 -
the disease process involved. The value of such studies as those
of yellow fever in Central America^ and pellagra in the Southeastern
United States, recently have "been accentuated by the dramatic
discovery of Allison regarding the significance of sickle cell
anemia with relation to malaria in the tropical regions of Africa
and Asia.
The increasing emphasis on chronic diseases of the nervous
system has stressed the need for world epidemiological research
programs in chronic neurologic and sensory disorders „ Recent
studies, for example, of geographical variations in the frequency
of multiple- sclerosis show that this disease is more prevalent in
temperate than tropical climates, but the clinical manifestations
are the same in all climates . Recent Institute studies in the
distribution of a new disease called "kuru" among a tribe of
savages in New Guinea afford an opportunity to study this type
of neurologic disease under control conditions .
GEOGRAPHIC NEUROPATHOLOGY
These types of studies may be included under the title of
"geographic neuropathology" or "geomedicine" which refers to the
evaluation of the frequency of diseases, their pathology, and
their relationship to associated genetic and environmental factors
in diverse geographic regions and populations. It includes
geographic variations in the manifestations of illness as well
as total incidence and the prevalence figures.
THE WORLD FEDERATION OF NEUROLOG-Y
Symbolic of international cooperation in medical research
was the formation of a World Federation of Neurology during the
First International Congress of Neurological Sciences, held in
Brussels in July, 1957- This newly created body is a federation
of national neurological societies throughout the world. The
chief objectives of the Federation are; (l) the dissemination and
exchange of new scientific knowledge in clinical neurology and
neurological science on a world-wide basis; (2) the stimulation and
encouragement of international cooperation in neurologic research;
(3) the organization of international congresses and symposia; and
(h) the development and exchange of fellowships in neurology and
neurological science.
Most important of all, however, are the Federation's
functions as a coordinating mechanism and central clearing
station for the stimulation, formulation, and integration of
international collaborative research projects. In this respect.
- 10 -
the Federation Is uniquely equipped. It is a Federation of national
professional societies representing the academic pinnacles of neurology
throughout the world. The Federation is nongovernmental and does not
depend on fund raising for the administration of its routine affairs
which are underwritten with the proceeds of annual dues from the
national delegates of the many societies throughout the world, of
which it is composed. Because of these factors, communications are
free and easy between the delegates when the question of organizing
symposia or collaborative projects is involved. The Federation
hopes, however, to work in close liaison with the World Health
Organization and other government agencies for the advancement of
medical research .
Suggested among the primary problems to be attacked by the
World Federation of Neurology are: (l) the establishment of an
international reporting system and standards of nomenclature and
classification of disease processes; (2) the identification of
situations in various geographical regions which would lead to
study problems of international geographic pathology on a world-
wide scale. This preliminary planning involves the detection and
classification of population isolates--classification of climatic,
cultural, and economic factors within the regions to be studied;
and (3) the establishment of geomedical studies to attack the
major neurological disorders of mankind such as studies of perinatal
morbidity similar to the present collaborative investigations of
NIWDB in this field.
The research program of the WFN has, in jrelil^oy^ already
begun. A grant has been awarded the Federation to be used in the
planning and conduct of an international symposium on the neuro-
pathology of the encephalitides. This symposiiom, to be held in
Antwerp early in 1959^ will serve as a precursor for the establish-
ment of an international reporting system and the development for
standards of nomenclature and classification of disease processes.
A grant also has been made to the Federation for the
implementation of research, especially along the lines of collabo-
rative and cooperative studies in the cerebrovascular diseases and
perinatal morbidity. The project in cerebrovascular disease is
based on the preliminary findings of Baker and his associates at
the University of Minnesota. These illustrate that the nature,
frequency, and severity of degenerative changes in cerebrovascular
disease differ in cerebral arteries of different sizes. For
example, the larger arteries of the circle of Willis, the small
intracerebral (15O-5OO microns in diameter), and the intracerebral
arterioles (15O microns or below), differ in the type of degenerative
changes they undergo in cerebrovascular disease. Moreover, the
involvetnent of arteries of a certain caliber is not necessarily
- 11 -
associated with the involvement of the arteries of a different caliber.
Finally, the degree of correlation between the arteries of different
sizes varies with a niMber of factors: the pathological changes of
underlying brain parenchyma, the age of the patient, the presence or
absence of hypertension, and other factors. These variations in the
reaction of cerebral arteries of different sizes provide a unique
opportunity to establish leads by means of comparatiAre studies in
different geographic regions throughout the world. Thus, the effects
of climate, race, cultural environment, diet, somatic disease and
other factors can be measured.
We believe that this project can be easily and quickly
developed through several selected centers throughout the world,
particularly in Belgium, Brazil, Argentina, France, Japan,
United States, Australia, and Mexico. We anticipate such a
study, similar to the cooperative aneurysm study now underway in
the United States, could be completed in a period of two years.
Geomedical studies directed toward discovery of geographic
distribution of perinatal morbidity as it relates to the central
nervous system are already underway in Ireland, the Netherlands,
and Sweden. The protocol of the WIKDB's collaborative cerebral
palsy project is being used as a guide in the further organization
of these studies and as a guide by the European group for the
collection of their data. Eventually, the findings of the European
scientists will be collated and correlated with those of the WIITOB's
collaborative project through the coordinating mechanisms of, the
World Federation of Neurology.
At present, about ^0 major nations of the world have joir^ed
the World Federation of Neurology. Its current officers are:
Drs. Ludo van Bogaert (Belgium), President j Macdonald Critchley
(United Kingdom) and Auguste Tournay (France), Vice Presidents;
Pearce Bailey (United States), Secretary-Treasurer General; and
Richard L. Masland (United States), Assistant Secretary-Treasurer
General. The Federation's executive office is at the Institut
Bunge, 59; rue Phillipe Williot, Berchem-Anvers, Belgium. The
administrative offices are at the National Institute of Neuro-
logical Diseases and Blindness, Bethesda ik, Maryland.
COMPARATIVE NEUROPATHOLOGY OF MAN AND ANIMALS
A generally conceded weakness in a comprehensive attack
on neurologic disorders is the present lack of a systematic
comparative neuropathology of spontaneous diseases of animals,
both domesticated and wild. Further, what sparse literature
there is on the subject is widely scattered and difficult to
- 12 -
find. At the University of Bern in Switzerland, a research and
coordination center is in the making to bridge the gap between
veterinary and human neurology and to establish an information
exchange center of animal neuropathology. Heading this enterprise
at the University of Bern, are R. Fahkhauser, Professor of Animal
Neuropathology, and E. Frauchiger, Professor of Comparative
Neurology and alternate delegate (Switzerland) to the World
Federation of Neurology. Backed by fifteen years of experience,
these two Swiss investigators are org:anizing a project which
calls for the protracted clinical studies of domesticated-wild
animals suffering from neurological disorders . These studies
include complete postmortems with the application of modern
techniques for the examination of cerebrospinal fluids, and
investigations in neuroradiology and electroencephalography.
Their research also extends to studies of the embryology of the
central nervous system and spontaneous congenital cerebral
malformations arising in the perinatal period. The Institute
of Comparative Neurology at the University of Bern also is
tooling up as an information exchange center for the review
and distribution of publications, histologic sections or
paraffin blocs, photographs and slides.
The Bern project furnishes another example of international
cooperation through neurologic research. Here it concerns the
United States and Switzerland through the sponsorship of the
project by the National Institute of Neurological Diseases and
Blindness and the Swiss National Fund for the Advancement of
Scientific Research.
MEETINGS AND SHViPOSIA
During the calendar year, 1958^ several important meetings
and symposia were held in Bethesda under NINDB auspices. Among
these was a sym.posium on the Electrophysiology of the Visual
System, organized by Dr. M. G. F. Fuortesj a meeting of the
Eastern Section of the Association for Research in Ophthalmology
(January 17 - l8)> a conference on Graduate Training in Clinical
Ophthalmology (January 26); and a symposium on the History and
Prospects of Neurochemlstry (April 19); in cooperation with the
National Institute of Mental Health.
The proceedings of the symposium on the Electrophysiology
of the Visual System, edited by Dr. M. G. F. Fuortes, has been
published as a supplement to the September, 1958^ issue of the
AMERICAN JOURNAL OF OPHTHALMOLOGY. The proceedings of the other
meetings ar^ in press.
- 13 -
The proceedings of two symposia held in 1957 were published in
1958 as special supplements to MUEOLOGY: "The Sequelae of the
Arthropod-Borne Encephalitldes," edited by Drs . Pearce Bailey and
A. B. Baker J and "A Classification and Outline of Cerebrovascular
Disease," a report by an Ad Hoc Committee established by the
National Advisory Council of NINDB, Clark Millikan, Chairman.
In addition, in I958, five books were published by members of
the NINDB staff and collaborators. These are: "Temporal Lobe
Epilepsy," "New Research Techniques of Neuroanatomy/' "Biology of
Neuroglia," "Neurological and Psychological Deficits of Asphyxia
Neonatorum," "External Gollimation Detection of Intracranial
Neoplasia with Unstable Nuclides," and "The Epileptic Seizure."
In addition to the meetings and symposia held in Bethesda,
members of the NINDB staff participated in many important meetings
held elsewhere in the United States and abroad. Some mention of
these has been made in other parts of the total annual report.
This section will mention only important meetings abroad in
which the Office of the NINDB Director was officially concerned.
NINDB MISSION TO RUSSIA
At its June, I958, meeting, the National Institute of
Neurological Diseases and Blindness Council endorsed a proposal
by the Director, NINDB, to organize a neurologic mission to
Russia and authorized him to take the necessary administrative
steps .
By November, the mission had been formed and on November
18, they left Washington by air for Moscow via Paris . The
members of the NINDB mission were; Drs. Francis M. Forster,
Council Member j Clinton N. Woolsey, Council Member^ Louis S.
Goodman, former Council Member; Henry W. Woltmani Paul I.
Yakovlev; and Karl Frank. The purpose of the mission was to
observe the nature and conduct of research activities in the
physiology and pharmacology of the nervous system in the U.S.S.R.
The mission organized in collaboration with the Public Health
Service and the Department of State was the first of its kind
in the field of neurology and was made possible by a January,
1958, agreement between the United States and i he Union of
Soviet Socialist Republics to exchange missions in various
fields .
Dr. Forster, Chairman of the mission, already has
submitted a preliminary report of impressions perceived in the
Soviet Union (November I8 - December I8, 1958)- The report
concerns training of Soviet physicians and scientists; the
- Ik -
organization and orientation of Soviet research^ physical facilities
and research equipment; types of research personnel; and a consider-
ation of Soviet research and development. Among other things, Dr.
Forster saids "Neurological research is given the highest priority
"because of the Soviet concept that the central nervous system is
the central control of all bodily processes and must be regarded
as the ultimate target in biologic research."
Among the Soviet methodologies which impressed the U. S.
delegates were: "the application of descriptive anatomy to
research;, active studies in developmental morphology and the
phylogeny and ontogeny of the central and autonomic nervous
systems^ the preservation of a historical approach in the
training of scientists which provides a stimulating background
for further achievements, and the acute awareness by Soviet
scientists of things going on elsewhere and their determination
to assimilate and Improve them." Near the end of the report,
Dr. Forster recommended: "that research in the United States
in neurophysiology and neuropharmacology be supported even
more generously than it has been; that scientific literature
in English translation be made more available to American
scientists; that the program of scientific exchange missions
be promoted further; and that a program of exchange fellowships
be developed."
Dr. Forster adds a final note when he writes, "If one
appreciates that the majority of Soviet research institutes a:e
postwar in origin, and that their application to modern research
techniques is relatively recent, one cannot be complacent about
their potential for rapid progress in the future."
A return Russian mission will visit the United States
early in 1959- The members of the visiting Soviet team are:
S. V. Anlchkov (Pharmacologist and Physiologist specializing In
the study of chemoreceptors); V- S. Ruslnov (Physiologist,
specializing in KEG and electroretlnography); and V. V. Zakusov
(pharmacologist and Chemotherapist ) .
BABINSKI CEMTENARY
The NIWDB Director was appointed by the American Neurological
Association and the American Academy of Neurology to represent
the United States at the celebration of the centenary of the
famous French neurologist, Joseph Bablnskl (1857-I932). A
delegate from the Municipal Council of Paris opened the Inaugural
sessions of the centenary in the amphitheater of the Ecole des
Inflrmieres of the Salpetriere, where In l882 the world's first
professorial chair in neurology was created for J.-M. Charcot.
- 15 -
After the opening remarks ^ there were presentations by Professor
Raymond Gareln, President of the French Society of Neurology,
and by official representatives of l8 countries on the life and
works of Babinski. These papers have been published in the
Revue Neurologique .
On June 3^ the day following the centenary, the French
Society of Neurology conducted its Twenty-second International
Neurological Reunion at the Salpetrlere, with President Garcin
and A. Tournay presiding. Still in tribute to Babinski, the
morning sessions of the reunion were devoted to a symposium on
the cerebellum. The International Reunion continued on June ^,
with a program of free communications, interrupted at noon for
a convocation at the Hopitaux de la Pitie', to affix a medallion
of Babinski on the outside wall of his old neurologic service
(le Pavilion Benjamin Dalessert). The ceremonies were conducted
by A. Tournay, who espied and singled out from the audience, in
a moving scene, Mademoiselle Alips, Babinski 's faithful and
devoted chief nurse .
Both the Babinski Centenary and the International
Neurological Reunion were organized by the French Society of
Neurology and held under the patronage of the French Minister
of Foreign Affairs, the French Minister of Public Health, and
the President of the Municipal Council of Paris.
NEUROPHYSIOLOGICAL WEEK IN PARIS
Again In France, the NINDB Director had the privilege
to preside at one of the sessions of the annual neurophysiological
week of the Salpetri^re in Paris, under the Presidency of
Professor Th. Alajouanine and the Secretary-Generalship of
Antoine Remond. The central theme of the scientific sessions
(October 20 - 25, I958) was "Sensory Integration." "After an
introduction by Professor Alajouanine on the evolution of
ideas on sensory integration in neurology, some 28 neurologists
and neurologic scientists from France and other countries
participated in the presentations and discussions. Some of
the contributors had just returned from a series of neuro-
physiological meetings in Moscow, Kiev, fep''.'3' Ia^IIs.
NEUROLOGY UNIT INAUGURATED IN MEXICO CITY
On November I8, 195 8, the NINDB Director travelled as
an official guest of the Mexican Government for the ina'^juration
of a Neurology-Neurosurgery Unit of the Mexico City General
Hospital. The official ceremonies were opened by the President
- 16 -
of the Mexican Republic^ Don Adolfo Ruiz Cortlnes^ accompanied by
Dr. Ignacio Morones Prieto^ the Incumbent Secretary of Health and
Public Welfare. The scientific program began with an address by
Dr. Clemente Robles^ the head of the nev neurology unit, who
traced the history of Mexican medical advances from the time of
the first field hospital established by the Conguistador, Hernan
Cortes, through the development of the present General Hospital
which was founded by Profirlo Diaz in 1911- He concluded by
stating that the opening of the new Neurology-Neurosurgery Unit,
the only one of its kind in the country, represented an important
forward step in the study and treatment of neurological diseases
in Mexico. Following other speeches by Dr. Leonides Guadarrama,
Director of the General Hospital, and by President Cortines him-
self, the President and his retinue toured the Neurology-
Neurosu: I'.r-^- Unit. The new unit has k2. beds for adult patients,
two small pediatric wards, and surgical, EEG, and x-ray suites.
It is furnished with new and modern equipment and ready for
occupancy by patients. The cost of construction was 3 million
pesos. Present at the inaugural ceremonies were official guests
from other countries. Others from the U.S.A. were Derek Denny-
Brown, John F. French, and Webb Haymaker; from France, Professor
and Madame Paul Dell and Dr. Antolne Re'mondj from Chile, Raul
Hernandez Peon; and from Cuba, Dr. C M. Ramirez Corrla. The
day following the Inaugural ceremonies (November 19), the
visiting neurologists and neurosurgeons joined their Mexican
colleagues in a three-day scientific program held in the
General Hospital and at the National University. Among the
Mexican participants were Drs. Luis Saenz -Arroyo, Manuel
Velasco-Svarez (President of the Mexican Society of Neurosurgery),
Hernando Guzman West (President of the newly formed Mexican
Society of Neurology and Psychiatry), Roberto Gamboa Acosta,
Ramon del Cueto, Guillermo Santln, Mariano Vazquez, and
Armando Ortlz-Galvan, currently a Research Associate of
Maltland Baldwin at NINDB, who, with Jose Humberto Mateos,
were of great help in guiding the visiting groups.
THE INFORMATION OFFICE
An important arm of the Director's Office is the
Information Office, headed by Ruth Dudley. This unit functions
as a general Information and distribution center for the
dissemination of NINDB news and reports to the outside world.
The work of the Information Office this past year has
included the preparation and distribution of brochures, reports,
articles, speeches, exhibits, and press releases. The office
has also arranged press conferences, press interviews, and
picture stories . It has edited many manuscripts and has
answered many inquiries of all types.
- 17 ~
Mong the brochures completed in 1958 are: The Research
Attack Against Cerebral Palsy, iVfultlple Sclerosis — Hope Through
Research, NIKDB Conference Programs, Highlights of Progress in
Research on Neurologic Disorders, and Who's Who in NIEDB. Nearing
completion are three other brochures: Little Strokes, Parkinsonism,
and a brochure about the Institute.
The Information Office also prepares reports, articles, and
speeches to fill requests from individual Congressmen, Congressional
Committees, the Department, Public Health Service, the National
Advisory Council, voluntary health agencies, and World Health
Organization. In 1958^ these included: Highlights of Research
and Program Developments, Past Foreign Contributions to the Field
of Neurology and Blindness, NINDB's Puerto Rico Project, Progress
Reports on Electromyography and the Structure of the Synapse for
use of the NINDB Advisory Council, NINDB portions of the NIH and
PHS booklets, research accomplishments of NINDB for the Voice of
America, WHO, and UNESCO.
Numerous interviews vere arranged for press and magazine
science writers, including the Associated Press, United Press,
International, Scripps -Howard, Medical News, New York Times,
and such magazines as Fortune, Time, Scope, Farm Journal, and
Parents Magazine. Seventeen general and three individual press
releases were prepared and distributed to the scientific press,
wire services, and other science outlets. Background materials
were made available by the Information Office for the production
of two films (Conquest and Year of Birth) .
During 1958, the Information Office replied to 788 letters
of inquiryj mailed out 15OO individual pamphlets (bulk orders not
included) J and replied to I7OO telephone inquiries from the public.
Finally, the Information Office edited and cleared many articles
and scientific manuscripts, and gave assistance in the preparation
and distribution of three scientific exhibits and accompanying
literature for NINDB investigators .
AHillAX, HEPCra
(Silettdsr fear^ 1958
Direct Sraiaiag
wmmAL wsswjm cf heurolosical diseases ahd HasDis^'
Ftmds under ■^bils aefclirlty C$^,OT©) proffflde fcac the migRport
of t&@ ixiosesnrie^ trainios progrsm of tl3;@ Institute. IMs jpx'QgX'am
mak^s it possiM.e for tli© Institute to seeua?« iguallfied stafT for
scone of its e^ratlons by tralnixig youn^^^c- scientiots in pm^tieulsar
^ills Dee@BMary to owexy on eertaia prc^ram ap®ar®,tioaas.
Is eoop@rati(m -ultli ths CcBsnuialsabl^ Bis@a&@ Ceister 12;. Atlasitoj
Georgia, the Ii:mtitute is presetSEtly suspcrtix]^ traloixsg In t3s«» broad
eone^ts of ^id;^iiolo£^. Basis traizdsg sM. e3tsp@ri6B.ee in tl;^ £i@ld
of iitjs Bsore acute smd id.Sespresd ccBimimiea.ble dlsi^seis offers sa:^
iotxxt^etioa to tfee mare c»k^1@3£ smd p3:^traet«d epideosiolosieal
proM@iKS of nmxeologlcBl aM simsor^ disorders aiod dirmide di@@a0®B<.
In additiott, tte Institute ia supportissg tJi® tralaiiag of an
officer wbo l%as gea® to the institut® for Cellulsr (Sieralstry la
£&s!^c^j Gexraasisr to studjr the medtmiiisms in^olTed ia the fon^tiossi
of aeefeyldaoliae and 1& the relationship between aeetyldjollis® ^sd
esrtsia iiaportant lipid factioas of th® nerve aj^nlaraa®. He wiH &lso
te \fc>rkizig (m ths oxieiation of lipids 1;^ means of @@rtain 6.yws aM
tecteifues avaHsble oaly in that Institute in Mmidi.
Another officei* is undergoing traiaiag in tiourolc^ifflal
disorders st the Colusabia-Prestsffeerlan Bfedl@al Ceat®r, in Sfew taxfk,
so that h@ will beeoae a ajor® effee-fcl?® ®pidsaaiologistc AdditlOTal
traiBlng for a Mopbysieist ®M a physiologist is bssiag ptexmedo
PurUifir staff training in blOTtetxy and ©pidaBiology is coj3,t®Hiplst«i=
Siorfc^tQrjia courses, generally of & ^>eelfla t««toieaa. smture^
aa?e also undertaken ia this activity.
An Institute Coraaittee has b@@n foriaed-to ds^velop a 3C£s^et@
pro-am ia in-servlee training^ uadsr the a©"5r Seaisijsis Aet ptMs«d
l^ the Cssigress djjring I958.
mnnomL w&srmnE of keueolcsicm. dissasis asd BLWsmss
Ckjllaborative a^d Cooperative Projects
Cinc]luding Project Services ^fcra.acla)
Estiioated ObU^tioas for Fi 1955
Ibtal: $576,000
Direct: 478, 200
S^iisburseizsatB : 97,800
AISUAL RSPCSE
CslenSm* Year, 1958
Collaborative Heseai?da
latiffisal Xs^titute of Isurologlcal
Mseases end BLlMness
^tioaaX Institutes of le<li
Sie laost recent pliase of tlie de'vteloipm^nt of the ^stitut^'s
total piTC^raai has "be^n tfee creatioa of tli© eollalKsrative rweeardfe
srea. ^le respcaasibill'^ of tMs area is tl^e coordLoation of eertalia
types of resear^ whi^ aar@ carried on with difficulty withia a siagle
limtitutiosa. Most basic labxratop;^ ras@arch and imi^ cliMeal reeea^eh
eaa "be carried on effsctlveljf in isolated laboo^torles ae clinics.
lowe-^sr, research wMcli invol^«s tfee collation of data frcm large
mmba?s of individuals, 0£ tJ&e com^risem of health or sickness in one
ssssmxsaity as ece^mred to another; requires a cc^rdinatcd progrem, snd
f^pogu^rtly involves t^fe© <K>llateor®tion of a number of institutions.
Sie CQStSuct of suoji vos^ 03:^narlly requires t&e skill of tto
epidaElologist and tlie stsitistlcimn. ^erefore, wi'^ain the past fev
7^irs, the Institu^t® has leunc&ed a Isroad program tjhos® objective is
to provide the necessery coordination for programs of ^id^alological
immsti^tlosi® and f&r ot&er areas of resesreh vMch easmot be cazried
out wil^iout diffleulty vi-^ia any aiagle research cearfcer* Because of
•yi® "srery nature of sueih (^idaaiological investigatioas which presently
iK^mlv® wide areas of td^ UMted States ^ it is evid^st that their
fuUeat egression eventually vill require -^e extension of tliese
activities t© a worls^ride "basis.
tee of tiae first investigations carried on within this program
was the stu^ of the cause of retroleatal filsropliasia. Within a sh^rfc
period of time, mi investlgsti^a eonceraisjg preasatvires, carried on
siasult^aaeously witMa a number ©f resesreh centers, wss ahle to veri^
the proMffia as th© aasainlstration of o3^g@n and -aie duration of the
infteats. At -^e presaet time, iw^xtmxb, epidsnlological investigations
of nmltiple sclerosis ©re being coadueted ty this program. Saese have
ia"TOlved i^e colle<^ion of dst®, frcsa ^nada. United States, and laore
recently, J&pasi. Imgortaat sidles on the island of Guam, also, serve
to danonstrat® the genetic "basis of one fofsa of aasyoftroigMe lateral
sclerosis. Wqt assisting ia the conduct of such iavesti^tloas, the
eolla"b©rative resear^ area has developed an epidaaioloiy teaseh and
a "biostatistics branch.
ISi© mosrt recent prograai ^ow de'^loping within this area is the
eoilB^Ksrativ^e project f«r the -study of psrlnatal |»thology. ©ils
progr^B, res^ulring -m® collah^^tion of 15 dlffexsait institutions, is
relying osi the intranairal progrem to provide the central statistical
and eoordiaiating servi^ss. In order to achieve uniformity, both for
procedure and for the type of information being ©"btained, a eoordlna-
tiag staff is being devieio^ed in a project services branch. It is
the resj^s&sibility of ^bls ip?c%^ to establish training programs for
- 2 ~
eoilabsratffis-s, to pire|>ar« t&e prtjcedimal laaaualB, to visit tfe®
soliaboratifflg iastitutioas foo? the purpose of assistissg in the
egtabiislssisat of stsadard proosdures, and flsaUy, to colleet,
asE®iab3.e, sad process ^bjs data forwarded to -^e eentral office.
la tMs study, we have been coacemed ia tdae past year with
two lE^KStrtant ste^sj aasaely, the rBer«itm©at and traiaing of pes^osmel
for 1^© conduct of -ase steidy vithia &e collaborating iastitutioas
®3Ed ia tfe® ceatral offlcej end the develcpaeat of a defiaitive
ja?otocol for ezasaiaation of mother aad <aiild, recoarding the observa-
tione, aad p3P04^ssins t&e recorded data. Baese rcqair^aents have
feeea ®®t by a '^pretest period", during vMch over 12C0 voaea have
teen examiaed ia t&e etudj, and tlirou^ which persoisael have becoae
espss-ieaeed, methods of esEe^iaatiwi have he^a developed, and
knosfledge has beea gaiaed as to the tgrpes of iafocnaaticaa 'whiefe caa
1:^ oMaiaed.
^e Bisjor probiffla ensouatered duriag this year stesas feaoi
tk@ fast there ere aot available -^e basic methods of intervieviag
and esBBdoatioa ^^aic^ are rehired if oam is to obtain valid and
reliable ^ta in a vast study of this sort, ^lese methods had to
be developed befor© the study could be laua^ed, yet dtsriag this
recruitEeat ^ase, the personnel w®re aot available within the
central office. For this reasoa, service eontreets with established
groups have been utilised as a means of rapid developient of specific
phases of the study. largely through this means, as of Jajsuary 1,
19^t t£>s actual study Qf cases aeeor&ing to a sound protocol can
T^ uadeptakea. IKsAs protocol will include a thorou^ socio-economie
aM genetic history of Idie gravida, data froa her initial and return
prenatal e:!@2ainati<me, and detailed observaticms during "^e eovirse
of lalxxs' @nd deliv&^. War the <&ild there has been developed a
series of nietieulous neonatal esamlnations, including a special
neurolog?.cal e^amioatios^; a developmental essfsinatioa at ei^t months
and a repeat ne^trological at 12 months. Subsequesat examinations
remain to be developed. Special studies will include serological
esaminaticMa of i^e gravida for virus, t&eee studies to be conducted
within t^e laboratories of ^AH). ^sci®l eaabryological studies of
abortuses will be coodueted at Harvard, J^ins Hog^lsis, and Brown
ISaiversity vhere special labooratories Mve been establish^ for this
pu^^ose. @ie leuropat^iologieal escsminatlosss will b@ conducted at
the Warren Ifoseum, jfervard Ifeiversity, ustll such time as central
ffecilities can be developed at Bsthesda.
At the pres^it time the collaborativs project has a<ad.eved
an encouraging degree of coiiesiveness sad mutual understanding,
^irou^ whldi g?oup action has l^ecBae very effective. "Bm basis for
proceoclizg actively with t&e study has be@a established. Its
productivity will depend to a large extent oa our ability to bring
into the central grcazp the odditioixal investigators of vision and
imagis^tioa to fie-^^lop tfee fullest explol-featioa of tSie treaeadous
ogpcartimity vbldx t^e new research facility greseats.
The 'O0llSLbosnt±v9 resesr^ area apinraprifttely provides &
lirif]^ bet:w^sQi the esctraimiral aad i&e intrearural pxo&rtma. A slisilar
situation esistB la respect to t!» laboratory of p«Pisatai patliolosy
located in Puerto Rico. T^ wfisarfc of tMs laboratory, coaeemed with
the r^srodueti-^ I&ysiolo^ of -Sae Fiiesus mcaikey, paralleXs very
closely the int^^et of ttee «>ll&borative project ea perinatal
pa'i^iolegy in humaas. Is. ^is inetanee, the l&'borateo^ is related
directly to fa.6 1:^sic ^eseare^ lalsoaratories of ttis latrasmral program,
"but Tiecause of its relatlonaMp to t&e IMiversity of Puerto Hieo, it
is p?ope?ly re^rded as a eolla.'texrative tmdsrtafa^iisg. SSsre agais^
t^ IntraiBural px^ogran is pirovldistg -t^e a^ryices and coordination
required "i^ an outside reseasrdft orgesidsiatiozi.
Pathological studies of the bsiains of |«tieBte dying ia. Hev
Guinea fsoBi "Ku.?u" aay provide information regarding the nature of
the process responsible for i^sXe rqysterlous laalsdy. Reports from
the field indicate ^at vitMn a small area i^ ineid«nee of this
disorder is extr&m'iy hi^. It is still not ksimm vke^isit there
is a genetic Isasie for this disorder, ^Aether it represents an
infectious disease, or is the result of seme toasie agent to vhidh
the natives of '^Is particular area are suhijeeted. Definite changes
in the nervous systen have "teen demonstrated toy pathological means.
Extensive investigations vill be required to detersaine ^e nature
of -^tilB ptrocess dad vhether it has iiq^li cations in other less caescxm.
disorders ses^ ia this country, dimerous aspects of the disease,
including its peculiar age end sex predilection, its t^adency to
unifue paljwlogioal reaetica in -yse liarain, indicate that solution
of -^iie 'iKXiTu" preldea vould provide a eoasiderahle advance in our
vusderstanding of otdber disease processes in tSte nervous ^st€si.
Bstimte^ Qbllgp-t^Lcfes tow fff 1959-
Beijabusseffieatsi 18,500
CRP-2
AMUAL REPCSS' - 1958
PHS - NIH
NINBB - EPIDEMIOLOGHr BRIKCH
Reorganization diiring the year restated in the transfer of the
Epidemiology Branch to the new Collahorative Research Program. This
is in keeping with the cooperative services and collaborative type of
research in which the Branch has been active. Our present program
aims to develop a nucleus of highly trained and experienced personnel
both in epidemiology and in genetics \fb.o can function well within the
clinical categories for which the Institute is responsible. These per-
sonnel iriLll operate primarily on a cooperative basis with other Insti-
tutes and agencies or, when invited, in collaboration with extramural
projects.
Organization of the Branch into Sections on Genetics, Neurologic
Epidemiology, Ophthalmologic and Otologic Epidemiology has proceeded
slowly due to lack of personnel trained in both epidemiolo^ and the
clinical specialties; provision of limited clinical training for physi-
cians oriented in epidemiologic me-aiodology appears to be the practical
solution to this problem.
The recruitment of Robert S. Krooth, M. D-, Ph. D., represents
an important step in the development of the proposed Section of Genetics
within the Branch. Tlie proposed Head for the Section in Neurologic
Epidemiology will complete his training in neurology in 196O and in the
interim the Branch Chief will continue in his activity. The planned
cooperative studies on phl^ctenulosis and mastoiditis with the Arctic
HeaLLth Research Center and on a uveitis and strabismus collaborative
study with the Biometry Branch have provided further stimulus for
recruitment of trainee epidemiologists for the proposed Ophthalmologic
Epidemiology Section.
The Epidemiology Branch of the Coiamunicable Disease Center and
this Branch continue in a successful cooperative effort. Dr. Sledler,
on assignment to oxsr Branch, has participated in several of our research
projects while at the same time remaining available for Epidemic Intel-
ligence Service calls. (See project descriptions.) The Asian Influenza
collaborative study which he supervises is nearing completion; prelimin-
ary restilts fail to show any effect of the virus on the incidence of
prematurity, abortions or neonatal deaths. The teratogenic effect, if
any, must await refined statistical analysis of data which are still
being collected.
The Guam Field Station continues to represent a low-cost, highly
productive research operation and an excellent training facility for our
medical officers . The clinical type of studies of the highly prevalent
neujpological disorders as approached by our clinical ly-oriented
epidemiologist (Dr. Pieper who left the Service after a successful tour
on Guam to conqolete his training in neurology) are giving vTay to a series
of genetically-oriented mathematical and laboratory type of field
investigations under Br. Krooth. The population on Guam and neighboring
islands still provides a wealth of clinical information and e3q)erienee.
At the same time, pathological and othar specimens are available on
short notice for cooperative research activities at NIE, CDC, and for
collahorators in various universities in the United States and abroad.
The island is at a crossroads of the Western Pacific and provides
unusual opportunities in standard epidemiologic practice for our field
workers as well. The excellent cooperation of the Government of Guam
and the Havy Hospital facility continues.
During the year several major population surveys were completed.
The results of the intensive survey in Rochester, Minnesota, in collabora-
tion \rith the Mayo Clinic has been published. This report is expec'sed
to provide baseline incidence and prevalence statistics for numerous
neurological disorders for many years to come- The local population sur-
veys under Dr. ^Llter, in collaboration with staffs at the Medical College
of South Carolina, in Charleston, and Dalhousie University, Halifax,
Nova Scotia, have been completed and the reports are being readied for
publication. Dr. Alter, after two years in the field, has been assigned
for a year of training at the Nevirological Institute in Hew York.
Several other projects of limited scope were completed and
reported. These included the study of Sequelae of Japanese B and Mumps
Encephalitis on Guam, a review of the Epidemiologic and Genetic Factors
of Parkinsonism, and a genetic and physiologic report on Mirror Movements
with Dr. Robert Cohn of the Naval Hospital, 5ethesda. The progress of
the long-term cooperative study on the Natural History of Multiple
Sclerosis and Retrobulbar Neuritis ^Tlth respect to Multiple Sclerosis is
continuing \rith the National Research Council and Veterans' Administra-
tion. This study should be completed next year.
Dr. t5yrianthopoulos, geneticist, has aided in the completion of
a study of skeletal deformities in motor system disease and continues in
his extensive twin study on mxiltiple sclerosis. He has developed a new
program aimed at clarifying genetic factors in parkinsonism and some of
the genetic factors in cerebromacular degenerative disorders. In coopera-
tion with the Laboratory of Blood Products, NIH, the association of
blood groups with amyotrophic lateiral sclerosis is being evaluated.
Several other studies of more limited scope on Huntington's chorea and
peroneal muscular atrophy are also being pursued.
Hew projects cover various fields, particularly in chronic
neurologic disease. One of the isKJst encouraging steps forvTard in our
struggle \T±th amyotrophic lateral sclerosis (AIS) resulted from the
detection of a previously unrecognized amorphous intracutaneous sub-
stance in AIS patients by Dr. Harold Fullmer, of HIDR. Atten?)ts at
histochsmical identification, if successful, promise to open an entirely
new field of exploration in the study of the degenerative lower motor
neurone disorders. The availability of specimens from our field station
on GuaxD. has been particularly valuable in this project.
- 3 -
^EMs Bs'snc^ has & T^sponsibllity in '^veloping staoslas^dlzei
p^^oceterea ajad ^^portitig astlsoSs x^aicJi will allow for ^3Poa<a popula-
tion GQs^?^l&ona in seves^ geaB^dical ?esa®?eli p3?o@?aBss io »a«yolo^.
fhese iuGluSs p^epayatiosas for tl^ forfchccgaing sysaposixaa at tfee
In-^roatioaal Confess of Sfemrology (1961) sjai advisory ©tatus to
susv^jrs now undej? way ia several o^teef coua-fe^iea.
Psoblems of xioiBsaclatur© axigl efforts to revise classification
of neurologic disorSss^ for hospit&l use as^ for intsmatioaal ssortal-
ity eca^arieoas ase expected to bectaae sn important paxigrasa aeat yaa?.
Ebe Branch is sacw represents on tifcs Hsurology Section of tte Aasiriean
ttedical Association '*Staad&?d Nosienclaturs."
&3ie? activities vMch are not strictly of sa iatramaral
r^s^ir^ ssatursF are s«rvie®8 to the MeSical Advisory Bos3?as of t^
CffiWMtlan ant the National Multiple Seleroais Societies ; and tl^
Washington, B. C Chapter of ■thQ W^S; !£he Cosaaittsse on Sfozeenclatwre
and BioiKlaclcs of th© Aissricon Aea«3aay of Keurologyj aisa liaited
teat^dJBg responsibilities by staff msmb&sQ in tte lo<^l univiersiti@s .
Br. HyrianlJiopoulos has continued in instruction anS. eonsvLLtation
of genetics for clinical assoeiatsa at the Clinical Center aM for
studsnts at George Washington University. The B. C Heredity Coijnseling
Clinic -uhich he established at Georgs Washington University has mat
viih increasing success and serves as a \iseful cas@ naterial source for
res^trch ani clinical experience az^ at the saats tijize supplies a
herstofor© unavailable service for "Uiis region.
There are two factors at pres^at serving to intes^ea?© wi-^ tSse
full developssnt of ovr progs^a which is nesdsd in the Institute.
The first is %hs lade of trained Manpower referred to at ^t® begin-
ning of this report. The second is the shortage of satisfactory
space whi^ has resulted in frequent changes in office assignment
(now totaling 7 iQOves in k2 laonidis). These moves have been so
disruptive that we have not bean able to us®, with wsximm efficiency,
ths few hi^ily traiseS specialists available to us in this f i@Mj nor
are ^Sm latest assignissnts to off ice-typ© facilities sway fr?aa HSH
likely to iii5>rove our opporfemitiea f<» recrui'^^snt of scajree
professional psrsoasel or young physicians who would o-aisrtdse be
well inclined to-^ards our pro^;^3i. The contlnuisd location of l^ds
Branch in a building geogretphically r^scved from the Clinical Center,
where laboi^tories, lilscrary ftoilititss and patients are availsbls,
will seriously reduce -^n^ efficiency aod preductiv© capabilities of
this ES'suach.
ORP -2 AETOAL REPORT - 1958
PBS • T?TH
StMiARg
Suring the calendar year 1956^ the Bi*£mch becaiae paart of the nev
Collaborative Research program. One of Its major activities in descriptive
epidemiology has Toe&a cozapleted ^th a series of reports on the incidence
aod prevalence of neurological disorders in the tinited States. A series of
genetic and epideoiologlc projects of limited scope in neurology and
ophthalnology has also been completed. Collahorative projects on the
teratogenic effect of Asian influenza and the natural history of miltiple
sclerosis are expected to be completed during the next calez^ar year.
The Quasi Field Stati<m continues as an importfmt activity in the Branch
and has pz^vided numerous opportunities for epidemiologic, genetic,
clinical and pathological investigations of neurological, ophthalmological
and other disorders. The Joint project with BWR, begun late this year
following the detection of a previously unrecognized Intracutaneous
substance in ALS patients, promises to be a major program in forthcosilng
months as efforts are made to Identify the material, to determine its
significance in the pathogenesis of ALS aod to evaluate its specificity
and sensitivity as a diagnostic aid.
The genetics research program is now veil established and an in-
creasingly successful genetic counselling program has been developed in
cooperation >flth George Washington ^oiversity. The organization of a
Section of Senetics is e^^pected In the near future.
Other responsibilities of the Branch Include a program concerned
vlth international geomedlcal research of diseases of the nervous system
and forQicQmlzig revision by the section of neurology of the AMA Standard
SomeBClature .
PrellminflTy steps were taken to develop a nev activity on
phlyctenulosis az^ deafhess vlth the Arctic Health Research Center
in Anchorage.
Repeated moves (average 1 each 6 months 1) continue to disrupt
the operation of the program. Space poorly suited to the needs of the
Branch im|K>ses a serious problem for recruitment of capable personnel.
aRP-2 mWB - Spidealology Branch
HJBLIC/\TIOHS
1. Kxirland, Leonard T., Sachs, David, Kerpelman, I^rry C, and Davis,
F. Sterling, Jr.: Evaltiation of the "Phosphenator" Device: For
the Detection of Increased Intraocular Pressure. /\nierican Journal
of Ophthalmologir, Vol. k^, No. 2, February, 1958-
2. Cohn, Robert, and Kurland, L. T-: Synkinesia, Transaction of
Aiaerican Neurologic Association, June 1958.
3- Siedler, Howard D., Nicholl, Willard, and Kurland, Leonardo?.: OHie
Prevalence and Incidence of Multiple Sclerosis in Missoula County,
Montana. The Journal-Lancet, Vol. 78, No. 8, pp 358-360., Avigust
1958. ;
k. Pleper, Samuel J. L., and Kurland, L. T.: Sequeleie of Japanese B
and Mumps Encephalitis. Amer. J. of Tropical Medicine and Hygiene,
Vol. 7, No. 5, pp kQl'k90, September 1958.
5. Kurland, L. T., and Myrianthopoulos, N. T.: Skeletal Abnormalities
With Motor System Disease . Neurology, Vol. 8, No. 10, pp 727-733*
October 1958.
6. Kurland, Leonard T.: Descriptive Epidemiology of Selected Neurologic
and Myopathic Disorders With Particular Reference to A Survey in
Rochester, Minnesota. Journal of Chronic Diseases, 8:378-^18,
October 1958.
7. Kurland, Leonard T.: The Frequency of Intracranial and Intjraspinal
Neoplasms in the Resident Population of Rochester, Minn. Journal of
Neurosurgery, Vol. XV, No. 6, p 627, November 1958.
8. Parkinsonism - William Fields, Editor. Kurland, L. T.: Chapter I -
Parkinsonism. Epidemiology: Incidence, Geographic Distribution and
Genetic Considerations. Charles C Thomas, Springfield, Illinois,
1958.
9. Mackay, R. P. and Myrianthopoulos, H. C: Mxiltiple Sclerosis in
T\7ins and Their Relatives. Preliminary Report on a Genetic and
Clinical Stu^. Trans. Am. Neurol. Assn., I958. Also accepted for
publication by the AMA Arch. Neurol. Psychiat.
REPCBT3 (Unpublished data)
10. Alter, Milton., Allison, R. S., Talbert, R., Godden, J., and Kurland,
L. T.: Epidemiologic Investigations of Multiple Sclerosis and other
Neurological Diseases in Charleston County, S. C, and Halifax, N. S.
Series of three reports.
11- Alter, Bfilton and Talbert, R.: Myasthenia Gravis in Twins.
12. Siedler, Howard D-: Paralytic Poliomyelitis and Aseptic Meningitis
Syndrome in Washington, D. C, and Surrounding Counties During 1957*
OHP-2 fflMIB - Epidemiology Branch
13. J^yriautlaopouios, H., Pieper, S. J, L., Kiaxland, L. T.: The ASO
aod Bh Blood Groups Amoxsg the Chamorros of Guam.
ih. I^yriaathopoulos, 1., Rowley, P,, aod Kurlaad, L. T.: BmtiagtoB's
Chorea la Monozygotic Ti«ins .
13. FallsBSTf Harold M., Kurl£ao&, L. *£., and Siedler^ S. D.: A
Cutaneous Mucopolysaccharide in Amyotrophic L&teral Sclerosis «
16. Kurla&d, Leooard T.: !I!he loeideztce axd. Frevalezsee of Convulsive
Disorders in A Small Urban Comounity.
17. Pi€^r, S. J. L., Fields, W. S., and Kurlaod, L. T.: Failure of
Aaiyotrophic Lateral Sclerosis to Respond to Intrathecal Steroid
euad Vltaiain B12 Therapy and to the Use of Poly-alcoholic
Hydrocarbons .
ELmTm TO MME^)SBIP
Sntenaatlonal Society of Geographic Patholo©^, October, 1958.
Dr. lairl^nd.
Comaittee on Sbmenclataire, Beurolo^ Section, AHA. Standard
Nomenclature, September 1958. Br. Kurland.
Neurolo^ Instructor (Genetics), @eorge Washington University, I958-59,
Evening Classes. &r. Itiyriaathopoulos .
OEP-£ NIBDB - Bpideniology Branch
PHS-KH
IndivldTial Pro^lect Report
Calendar Year 1958
PARg A.
1. Project Title: Epidemiologic Investigations of Hevirological and
Ophthalmologic al Disorders and Other Conditions of
Unusual Prevalence in Guam and Other Islands of
Micronesia
Principal Investigator: Leonard T» Kurland
Other Investigators.: Rohert S. Krooth, Geneticist, HIHDB
SsBHuel J. L. Pieper, Jr., Epidemiologist, NOTDB
Nathan Malamud, Professor of Neurology, Langley
Porter Heuropsychiatric Institute, San
Francisco, California
William Fields, Professor of Meurology, Baylor
University School of Medicine, Hovtston, Texas
Robert J. Huebner, Chief, Lab. of Infectious
Diseases, MPiW
Leon Jacobs, Head, Section on Protozoal Diseases,
Laboratory of Tropical Diseases,
Cooperatirag Units: Division of Medical Services, Govenoment of Guam
Laboratory of Infectious Diseases, end Laboratory
of Tropical Diseases, KIAIB
Man-Yetars: Total - 1 3A
Professional -^/h
Other - 1/2
Project description:
a. Jtoyotrophic Lateral Sclerosis; A general point survey is underlay
to ascertain all affected in a specified population on the Island of
Guam to determine the extent of familial aggregation of cases. The
completion of a registry and the preliminary analysis may clarify the
extent of genetic factors and reasons for the high iacidence in the
population. The survey will determine the feasibility of the proposed
long term anterospective program.
Other studies on the ALS patients and comparisons vith members of the
unaffected population include cutaneous histochemistiy (see project 8)
and studies almet at identifying genetic markers. These include urine
and spinal fluid chromotography, serum protein studies and blood typing
of the native population (see project 12).
Therapeutic studies have been completed on intrathecal steroid and
Vitamin B12. These served the purpose of forestalling the publication
of a preliminary report by ©r. Fields on the supposed value of this
- 2 - WHIDB ~ Epidemiology Braaeh
treatment. Our study demonstrated the treatment as valueless among
the patients on Guam and enabled Dr. Fields to reevaluate the status
of his patients in Houston and to redirect his report,
A study of Inositol and Sorbitol failed to demonstrate any therapeutic
value by these drugs in ALS,
b. Parkinsonism; This disorder appears to be more prevalent in this
population than in corresponding populations of the Continental United
States. The mean age at onset is less than that in the United States.
There is a preponderance of males, there is a frequent association
with ALS in the same patient or in family members, and there is evidence
of appreciable diffuse brain damage and intellectual dysfunction in the
patients. Post encephalitic disease has been suspected as well as a
variant of amyotrophic lateral sclerosis. Serological, virus isolation,
and further clinical pathology, genetic and epidemiologic stx:^ULes are
now xmder way to clarify these Issues,
c. Sequelae of Japanese B and Mumps Encephalitis; A follow-up nine
years after the epidemic has been completed. Forty-six persons
(most infants and children) were known to be affected; there were ^3
survivors of the epidemic. Ho parkinsonism was observed. 7.^ per cent of
the patients with JBE died and hO per cent (11 per cent severe - mental
retardation or paralysis) had sequelae. 22 per cent of these with ME
had evidence of slight neuirological damage.
d. Toxoplasmosis and Chorioretinitis; A survey of the incidence of
chorioretinitis has been completed for a sample of the population on
Truk and Guam, Blood from humtans and animals has been collected for
toxoplasmosis antibody titration. The resvilts are now being analyzed.
e. Diaphyseal Aclasia; This rare disease is also called multiple
exostoses. The present plan is to study frequency at post mortem or
frequency among orthopedic admissions among the natives. One of the
most recent projects on Guam, the high incidence observed by ©r. Krooth,
will be followed by appropriate genetic analysts,
PART B.
Publications other than abstracts from this project;
Pieper, Samuel J. L., and Kurland, Leonard T«: Sequelae of Japanese B
and Mumps Encephalitis. Amer. J. of Trop. Med, and Hygiene, Vol. 7,
No. 5, September 1956
OKP-2 KBTDB - Epidemiology Braach
PHS-WIH
Individual Project Report
Calendar Yeax I958
PART A
2. Project Title: Epidemiologic Investigations of Multiple Sclerosis
and Other Neurological Diseases in Cliarleston County.,
South Carolina and Halifax, Nova Scotia
(Collahorative Project)
Principal Investigator: Milton Alter
Other Investigators: E. S, Allison, Neurological Dept., Royal Victoria
Hospital, Belfast Irelemd
Rhett Talbert, Professor of Neurology, Medical
College of South Carolina
John Goddard, Professor of Neurology, Dalhoi^ie
University, Halifax, N, S.
Leonard T. Kurland
Cooperating Units: University of South Carolina, Charleston, S« C.
Balhousie University, Halifax, K. S.
Man Years: Total - 1 I/2
Professional - 1 1/4
Other - 1/4
Project Description:
The incidence and prevalence of multiple sclerosis and other diseases of
the nervoiis system was deterniined for the populations of these two com-
munities. Data are being analyzed aaad reports are being prepared on the
frequency of multiple sclerosis, myasthenia gravj.s and other diseases of
the nervous system in the communities and for subgroups of their
respective populations. Clinical reports based on uni^ual cases observed
in these coimssunities are also being prepared. These inclvide a study of
myasthenia gravis in twins and an hereditary cerebellar ataxia vith
cataract formation.
PART B, Nona
KIUDB ~ Epidemiology Branch
ESS-NIH
Individual Project Report
Calendar Year 1958
P/^RT A
3. Project Title: The Prevalence and Incidence of Multiple Sclerosis
in Missoula County, Montana, and DuxlDury, Massachusetts
(Collahorative Project)
Principal Investigator: Howard Siedler
Other Investigators: Walter Deacon, Ihixhury, Ma8sach\isetts
Leo Alexander, Boston, Massachusetts
Willard Nicholl, Western Montana Clinic,
Missoula, Montana
Leonard T. Kurland
Ifen Years: Total - 3/8
Professional - l/k
Oth«r - 1/8
Project Description:
Missoula County, Mon-bana, A study of the frequency of multiple sclerosis
in Missoula County, Montana was conducted to determine whether the
clinical impression that nniltiple sclerosis was unduly prevalent in this
area was valid. The average annual incidence rate for Missoula County
was found to correspond to rates for other cities of comparahle climate.
It is concluded that the prevalence and incidence of M.S. in Missoula
County, Montana are consistent \Ttth the pattesn of rather uniform fre-
quency rates of the disease in widely sepsirated populations living in
comparatle regions of climate in the tenrperate zone of North Ainerica.
Dxixbury, Massachusetts. The prelitainary investigation of the frequency
of multiple sclerosis in Dxixbury, Massachusetts was determined because
of the suspected high frequency of M.S. among its residents. Althou^
the incidence and prevalence are high, the factor of chance in selection
rather -Oian local environmental situation cannot be ruled out. Further
studies in the vicinity of Duxbury and some additional genetic investi-
gation in this region are indicated.
PART B.
Publieations oldaer than abstracts from this project:
Siedler, Howard D., Mcholl, Willard, and Kurland, Leonard T-: The
Prevalence and Incidence of Multiple Sclerosis in Missoiila County,
Montana. The Journal Lancet, Vol. 78, No. 8, August 1958.
QSP-2 NIHDB - Epidemiology Brancb
PHS-HIH
Individual Project Report
Calendar Year 1958
PART A
k. Project Titl®: The Effect of Climate and Other Environmental Factors
in the Prognosis of Multiple Sclerosis. One Aspect
of A Broader Investigation of the Natural History of
Multiple Sclerosis ( Collahorative Project)
Principal Investigator: Leonard T. Kurland
Other Investigators: Gilbert Beehe, FoUow^ijp \gency, National Research
Council
J. p. Kurtzke, Chief, Neurology Service, V.A.
Hospital, Coatesville, Pennsylvania
Thomas Auth, Neurology Departiaent, Veterans'
Administration, Washington, B. C
Benedict Nagler, fomierly VeteKms' Administration
(now Lynchburg gaining School and Hospital,
Colony, Virginia)
Coopei^ating Units: Follow-up Agency, National Research Council,
Washington, D. C.
Veterans' Admi.nistration, Washington, D. C
Man Years: Total - l/lv
Professional - I/8
Other - 1/8
Project Description:
Retrobulbar neuritis has been reported to progress to multiple sclerosis
in k5-5<yfo of affected individuals within 10 to 15 years . It had been
hoped that correlation of residence with RBW might show whether some
climatic or residential status influenced prognosis with respect to
multiple sclerosis.
In the large population of military personnel with RBN, 19'*0-19U5,
only about 8 par cent have developed M. S. to date. This finding will
be reported J the main objective of the investigation cannot be achieved
In view of tAiis low incidence of M. S.
The project to analyze other factors in the Natural History of Multiple
Sclerosis is continuing.
PART B. None
CKP-2 HIM3B - Epidemiolog;^'^ Branch
PHS-NIH
Individtjal Project Report
Calendar Year 1958
PABT A
5. Project Title: Evaluation of Possible Teratogenic Effect of Asian
Influenza Virus (Collaborative Piroject)
Principal Investigator: Ho^mrd B. Siedler
Other Investigator: Leonard T. Kurland md H. Goldstein, Biometrics Branch,
NXHDB
Cooperating Units: 15 Cooperating Clinics, Universities and Health
Bepartments .
Man Years: Total - 1/2
Professional - l/k
Other - l/k
Project Description:
Certain virus infections -diar-Ang early pregnancy are known to have en
adverse effect on the fettis. The <Vsian influenza epidemic offered aJi
unususil opportimity to assess the teratogenic effect, if any, of this
strain of influenza virus. Histories have been collected and serological
specimens have been obtained from about 8,000 women in 15 centers. Sera
from mothers of affected offspring euad controls will be evaluated. A
consolidated report of the participating groups ^rlll probably be made
next yesa:'.
PABa? B. None
(S(P-'^. NIMDB - Epidaniology Bx'anch
PHS-H2E
Individual Project Report
Calendar Year 1958
PARC A
6. Project Title: Descriptive Epidemiology of Selected Nexsrologlcal
and Myopathic Disorders With Particiaar Reference
to \ Starvey in Rochester, Minnesota
Principal Investigator: Leonard T. Kurland
Cooperating Units: Sections of Hevirology and Biometry, Mayo Clinic,
Rochester, Minnesota
Man Years: Total - 1/2
Professional - 1/4
Other - 1/U
Project Description:
Statistics from a nmiber of selected sources, particularly those
sxirveyed by memhers of the Epidemiology Branch, NIHDB, were coraplled
and presented; the need for further descriptive and definitive
epidemiologic or other investigations were also considered. New data
trcm a recent survey in Rochester, Minnesota were also presented. Tha
discussion for each of the subjects covered In the paper was oriented
to^-raand the use of the available statistical and genetic data to provide
some foundation for further clinical, laboratory or definitive type of
epidemiologic research.
PART B.
Publications other than abstracts from this project:
Kurlemd, Leonard T.: Descriptive Epidemiology of Selected Keurologic
and Myopathic Disorders with Particular Reference to A Suarvey in
Rochester, Minnesota. Joiimal of Chronic Diseases 8: 378-.U18, Ctetober,
1958.
Kurland, Leonard T .: The Frequency of Intracranial and Intraspinal
Neoplasms in the Resident Population of Rochester, Minn. Journal of
Neurosurgery, Vol. XV, No. 6, p 627, Novaab«r 1958 •
ORP-2 NIKDB - Epidemiology Branch
PES -HIE
Individual Project Report
Calendar Year 1958
PABT A
7. Project Title: Mirror Movements and the General Phenomenon of
Synkinesia
Principal Investigator: Robert Cobn, U. S. Naval Hospital, Bethesda,
Md.
Other Investigator: Leonard T. Kvirland
Man Years : Total - I/16
Professional - I/32
Other - 1/32
Project Xtescription:
Mirror movements and other aberrant synkinetic actions may give Insight
into the normal functional organization of the motor system. This
survey was undertaken -wiiexi. four members of a single family who show
mirror movements were observed. An isolated case of "spontaneous"
mirror activity, one ease of acquired mirror movements and one case of
acquired synkinesia WBre also stxidied. These uncontrollable movements
appear to be the result of a functionally decreased control of a motor
system which noimally operates in parallel with the pyramidal tract
system. It is suggested that the inherited defect observed in the first
four cases was due to a developmental defect in the reticular system.
PART B„
Publications .
Cohn, Robert; Kurland, Leonard T.: Mirror Movements and the General
Phenomenon of Synkinesia. Presented at the American Neurological
Association, Atlantic City, June 1958.
ORP-2 iniDB " Epidemlolosr Branch
PES-NIH
Individual Project Report
Calendar Year 195^
PART A
8. Project Title: A Cutaneous Mucopolysaccharide in Amyotrophic
Lateral Sclerosis
Principal Investigator: Harold M. Fullmer, HIDR
Other Investigators: Leonard T. Kurland
Howard D. Siedler
Man Years: Total - 1/4
Professional - l/8
Other - 1/8
Project Description:
The papillary layer of the dermis in formalin- fixed sections of
abdominal skin in amyotrophic lateral sclerosis patients has been
found to contain an extracellular amorphous material which stains
with aldehyde fuchsin after paracetic acid oxidation and with the
Hale stain (Rinehart modification) . The material remains unstained
■*Tith azure A at pfi*", it is digested by testicular hyaluronidase
and by glucuronidase. On the basis of these findings it is believed
to be a neutral mucopolyseiccharide, or mucoprotein. It is possible
that the substance represents a heretofore unrecognized step in the
abnormal metabolism which is believed to be present in motor neurone
disease.
PART B„ Kone
Cgp-2 HIKDB ' Spifigmiology Branch
PHS-mS
liidividual Project Report
Calendar Year I958
PART A
9. Project Title: Paralytic Poliomyelitis and Aseptic Meningitis
Syntoome in Washington, D. C, and Surroimding
loTinties Boring 1957
Principal Investigator: Hotmra D. SiefflLer, on assignnsent to NHJIffi
from CTC. (Cooperative Project)
Cooperating Agencies: D. C- Health Bepartsnent
Man Years: Total - l/2
Professional - l/k
Other - l/k
Project Description:
A report of studies concerning paralytic poliomyelitis and aseptic
meningitis synSrome in Washington, B. C and surrounding counties in
1957. The outbreak originated in a small Southwest quadrant of the
city where the population is predominantly negro and of lower socio-
economic standing. Paralytic disease laanifested a marked selectivity
for preschool age children from this group, 'S&e same pattern observed
in the Chicago I956 epidemic. Aseptic meningitis syndrome In the
Washington ^ovip was believed to be associated in the naj owelty of
patients with poliovirus. This syndrome in the county ^?oup tj&b
associated with poliovlrus in only 1 of 11 instances where viral
cultures were positive.
PARI? B. Kone
^vaienflar Year 3.95o
pA'Tr /^
10. Rfojc ■ Pa3f.iinsoi>.is!H - Epideztdology: xncidence;.
Geo.g-jfapMc Distriljution said Genetic Considaratio:.
Frincip;sl Inyestigato:p; Laonard T. Kurland
;i|aa Yea?s: a?otal - i/4
Professional - 1/8
Other - l/S
Pyo,1ect Description;
TMs spideflil,ologic . ^' v;' parkliasonisia incl-udsd an analysis or
mortality statistics in the United States scn.d Canafe, a mDrbidi'fcy
survey in So Chester , Mimieaota, .aad an evaltiation of several, repoits
on possible etiolcgic msehanissss. Prevalence and incidence «3a.ta
s*sveal tJiat paufki53,0on:lsm is one of tb© ssost pi*evai.ent of tha e"h iikIc
nsurologifi diBordsrs. About 300? 000 cases are be].ie-ved to c
in taa OnJ-tea. States at px-eaent. Studies of cosv'sordancs foz:
par-ki/jisonisra in twins an& of tJi® possible coastitfiticasJL- fact,ora In
jfaactiosas to ataraxie cimpoiond^ were s-uggesteit as steps in deifinixig
fixe r^'elativs roles of g«5n.«tics aad exogeaous factors in 'the various
foams of p££=;xiB3oai8si. Ultijcate prevention aad control of the
"ideopatbie" forsm of parkinscnism await fcLe xGeatification of a
"chronic e?;? -,':.-)Tit' '"i>feot" wMch is believed to be pvre^eat a?:'' i-blc^i
is likely t-iCtic csture.
Publieatiora other than abstracts farom thia project;
Km^l£j:i&., L. T. Chapter X -> Parkinsonism. IpideEiology; Incidonce,
Geographic Distribution and Genetic Considerations. Charles Co
Thomas., Springfield^ Illinois. 1958.
CRP-2 KIKDB - Epidecdology Branc_
ms-ms.
Indlvid\xal Project Report
Calendar Year I958
PART A
11. Project Title: Multiple Sclerosis in Twins and Their Relatives
Principal Investigator: R. P. Mackay, University of Illinois
Other Investigator: N. Myrianthopoiilos
Cooperating Agencies: University of Illinois
Man Years: Total - 8/32
Professional - 2/32
oaier - 6/32
Project Description:
'i?o detesmine whether any hereditary factors are involved in the
causation of multiple sclerosis by studying the occurrence of the
disease among monozygotic and dizygotic twins and their relatives.
fi'ie first phase of the study is coining to an end hy Decemiber 31>
1.'58. The second phase, \diich will involve the reexamination of all
tvins, ^Till begin in I961
PART B
IfewAay, R. P. and Myrianthopoulos, N. C Multiple Sclerosis in Twins
and '."lielr Relatives . Preliminary Report on a Genetic and Clinical
Study. Trans. Am- Neurol. Assn., I95S.
Also accepted for publication by the A.M. 4. Arch. Neurol. Psychiat.
':MP-2 HIHDB -• Epidemiology Branch
FES-Wm
Individiial Project Report
Calendar Year 1958
FART A
12. Project Title: The Association of Blood GroTzps to Ajas'-otrophic
Lateral Sclerosis
Principal Investi^tor: N. Myrianthopoulos
Other Investigators: P. Schmidt, LBBP
Leonard T. Kurland
Coopejrating Units: Lahoratory of Blood and Blood Products, Division
of Biologic Standards
Man Years: To-fcal-V32
Professional - V32
Project Description:
A pilot study to determine if there exists any selection for a
specific blood type among patients with a motor neurone disease.
Present Status. 1!he medical facilities of the An^y, Navy, Air Forca
and Veterans AQministration, and many practicing neurologists are
participating in this project hy providing blood and saliva specimens
for analysis. A small nuriber of specimens from Guam has also been
received .
PART B. Hone
CKp-2 NHSiDB - Epidemiology Brsmcb.
TES'Wm
Individiial Project Report
Calenaar Year 1958
PABSf A
13 ■> Psxiject Title: ParklnsoniBia - A.taraxic Drugs Study
Principal Investigator: H. Mi^rianthopoiilos
Other Investigators: Leonard T. Ktirland
A. Kurland, Spring Grove State Hospital
Cooperating Unit: Spring Grove State Hospital
Man Years: Total - 24/32
Professional - 8/32
Other - 16/32
Project Description:
A study to detejmlne tJie occurrence of Parkinsonism among the
relatives of two groups of patients: tlaose who show Parkinsonian
synrptoms on high therapeutic dosages of certain phenothiazine
derivatives, and those who prove to "be resistant to the side effects
of these drugs. The two grorrps of patients, actually patients and
controls j, have been selected from the patient population of Spring
Grove State Hospital,
P'iHP B. None
(BP-2 KB5DB - Epidemiology Branch
IHS-HIE
Individual Project Report
Csilendar Year 1958
PAST A
ik. Project Title: A Storvey of ScMzoplirenics Among tSie Relatives
of Schizophrenic Patients
Principal Investigator: H. BSyrianthopoulos
Cooperating Unit: Spring Grove State Hospital
Man Years: Total - 18/32
Professional • 2/32
Other - 16/32
Project Description:
To determine the occurrence of schizophrenia araong the relatives of
patients who have already been selected for another project (13) and
to deteaanine the mode of inheritance of schizophrenia » This project
is a "by-product of px'ojeet Ho- 13 and has the advantage of overcoining
some of the biases involved in selecting an adequate sample.
PABT B. None
NBTOB - Epidemiolo^ Branch
ms-nm
Individ\ial Project Report
Calendar Year I958
PART A
15. Project Title: Some Epiaaniologic Features of Tay-Sachs Disease,,
Principal Investigator: K. Myrianthopoulos, NIMDB
Man Years: fotal - 2/32
Professional - 2/32
Project Description:
A study to determine the incidence and prevalence of Tay-Sachs
Disease among the Je\d.sh and non-Jewish popiilations of the United
States hy mortality statistics.
Present Statxis: Mortality statistics for the years 195^4-, 1955 and
1956 have already been collected. When data for 1957 become avail-
ahle, these will he added to the already existing ones and then analyzed.
PART Bo None
aRP-2 NIHDB - Epidemiologj'- Branch
PHS~NIH
Individual Project Report
Calendar Year 1958
PART A
16. Project Title: The ABO and Rh Blood Groups 'taong the Chaiaorros
of Guam
Principal Investigator: N. Myxianthopoulos
Other Investigator: Samuel J. L. Pieper
Man Years: Total - 2/32
Professional - 2/32
Project Description:
\ study to detexTnin^ the phenotypic aad genotypic frequencies of the
ABO and Rh groups among the Chamorros of Guam, with eiaphasis on
anthropologic and genetic implications.
Present Status: The project has been completed and a report has been
prepared for publication.
P\BT Bo None
cep-2 NIHDB - Epidemiology Branch
PHS-NIH
Individual Project Report
Calendar Year 1958
PART A
1?. Project Title: Skeletal Abnoymalitias With Motor Systam
Disease
Principal Investigator: L- T. Kurland
Other Investigator: N. ^Syrianthopoulos
Man Years: Total - l/32
Professional - 1/32
Project Description:
A detailed study of congenital skeletal defects in two families in
vo-hich amyotrophic lateral sclerosis is transmitted in a dominant
fashion, to determine whether an association, genetic or otherwise,
exists between these anomalies and motor neurone disease.
PART B
Kurland, L. T., and Myrianthopoulos, K. C.: Skeletal Abnormalities
With Motor System disease . Neurology 8:727-733, 1953.
CSlp-:? NIHDB - Epidemiology Branch
PHS-NIH
IniilLvldual Project Report
Calendar Year 1958
PART A
18. Project Title: Hxmtington's Chorea in Monozygotic Twins
Principal Investigator: N. ly^yrianthopoulos
Other Investigator: P. Rowley, NINEB
Man Years: Total - 2/32
Professional - 2/32
Project Description:
A comparative study of the onset, course and clinical findings of
Huntington's chorea in a pair of female monozygotic twins with
emphasis on some pathological findings and eugenic problems in the
family of the twins.
Present Status: The study has been completed and a report is being
prepared for publication.
PART B. None
CeP-2 NIKDB - Epidemiology Branch
PES-NIE
Individual Project Report
Calendar Year 1958
P<\BT A
19- Project Title: The Detection of the Hetero zygote in
Cerebroretinal Degeneration (Amatarotic Family
Idiocy)
Principal Investigator: N. Myrianthopoulos
Other Investigator: G. Brecher
Cooperating Unit: Clinical Center, NIH
Man Years: Total - 2/32
Professional 2/32
Project Description:
A study to investigate ihe possibility of detecting the heterozygous
carriers in the infantile and juvenile forms of cerehroretinal
degeneration, by changes in the peripheral blood
PART B. None
(M*-2 NINDB - Epidemiology Bi*anch
PHS-KIH
Individiial Project Report
Calendar Year 1958
P.'gCT A
20. Project Title: The Question of Penetrance in Peroneal
Muscular Atrophy
Principal Investigator: N. Myrianthopoulos
Man Years: Total - I/32
Professional - 1/32
Project Description:
A study of families with peroneal muscular atrophy to determine
whether the reduction in penetrance in this disease, as described
in the literature, can be substantiated after vigorous neurological
examination.
PART B. None
EatlK&ted €bligati«as for Wt 19^9
fetal: |l65,000
Beljaburaefflente: 28^000
AieDAL REK)JIT
Calsades- Ye«v- ISS"?
Bio»«tries Bresneh ^ Coliaborativa Hsseareh
Natioaal lostitQta of Ns^r^iogiesl
Disttasee and Bliadnaiss
KiBtionsl Inotitistn-'^ o£ H«alth
A» SCOPS OF PaOGBAM
The ioterests and lnvolvemen£ of the Biometrics Branch
during the calendar year 1958 may roughly be divided into four
major areas:
1. Collaborative Project of Cerebral Palsy and Other
Neurological and Sensory Disorders of Infancy and
Childhood*
2. Other sollaborative studies «
3. Statistical consultation and/or service given to in-
vestigators outside of NINDB on other projects «
4o Statlstieal consultation and/or service furnished tse
clinieal and basic research investigators at NINDB
ia the areas of neurology and blindness »
Aceosiplishznents achieved and probleais encountered during
the course of 1958, as well as proposed future objectives in each
of the above four areas ^ are reviewed below:
lo Collaborative Project of Cerebral Palsy and Other
Neurological and Sensory Disorders of Infancy and
Childhood.
In the intensive phase of the Collaborative Project » it is
anticipated that some 14 participating project programs will coa^
tribute enough pregnaneies over five year to account for a minlmuni
of 40^000 live births available for at least a six»year followup.
The purpose of this prospective approach is to relate factors in the
gravida (geaeties fasBily» medical historyp socio=eeonomic» prenatal „
etCo) to outcomft of pregnancyo In January 1958^ pretest forms cover-
ing tha various aspocte (prenatalg labor^ delivery^ aeonatalg ete-o)
were distribufced to each of the participating institutions » The pur<-
pos« of the pretest was to train local personael and to deteemiae
whether the types of data requested eeold feasibly and reliably be
seeuredo As the f®rms ware cotiipletedg they were seat by the
institutions te the Biometrics Brasch for editing aad critical
reviewc For «ach institution there were prepared periodiigally eval«=^
ationa of the qtrality of these complet&d forms and suggestions for
improvenaeat o These evaltsations were taken up with each institu-
tion separately^ If. should be mentioaed that in the above-mentioned
trial run only the forms were being pretested^ There was do attempt
to pretest case selestion or methodological procedures. From the
data received, tabulations pertaining to certain characteristics of
the obstetrical patient population of these iastitutions were compiled.
In addition,, statistical evaluations were made of data seeusred in
areas of special interest, such as an anlysis of the data received in
neuromuscular examinations of 166 babies at one of the collaborating
institutions, Diiring the course of the studyg it became evident that
the pretest forms would need considerable modification before they
were suitable for the full-scale investigation. As a result,
statisticians of the Branch worked in close cooperation with ob-
stetrical 5 pediatrics "and psychological consultants in the Project
Services Branchy Collaborative Division;, and with the staff of the
Bureau of Social Science Researchj, Washington!, D.C, in devising
forms that would secure tnore meaningful and reliable data in the ob-
stetrical » pediatric, and 3ocio»economic=genetie areas and that would
also be amenable to coding and tabulating procedures. Consideration
was also given to revising hospital methodology for the collection of
the data. In connection with the "^ranch's participation in giving
consultation on standardization sampling procedures for the Bayley
Tests Branch members designed an abbreviated version o* socio-
economic data sheets for use by psychologists during the 'standardiza^
tioa,'' This wou'd pifovide data oa the characteristics of those
included in the standardization and would allow a comparison of the
soclo»economie data of patients included for staadardiaatlon with
those who for various reasons refuse to participate.
It is planned to make the revised forms in all aspects avail*
able in January 1959 to all institutions in the project who are
sufficiently wall organized to be able to start the study at that
time. Furthermore, it is hoped that some information on population
characteristics and pregnancy outcome, evident at delivery, may be
secured by each institution from reviewing its past hospital records
so that declsiona pertaining to modification of sampling procedures
for each institution may be made by the Branch, The development of
data on expected incidence of maternal characteristics and fetal
pathology will be of great help in serving as a basis for modifica*
tion of the sampling design. The Biometrics Branch is prepaxad to
assist in the development of such data. In this eonnectiona it
should be stated that a source document and related punch card format
and codes have been devised for one of the collaborating hospitals.
It is anticipated that this would replacs their present adminlsirative
punch card and would serve both administrative and research purposes.
Furthermore 5, it would make it possible and relatively easy £or this
bospital to provide to the Biometrics Branch population and background
data needed for the Collaborative Project,
During the cotjrse of the year tha Branch Chief atteadsd
meetings of "-.he ■\dvisory Board of the Collaborative Projisci and
the Ad Hoc Review Board of whieh he is aa ex-officio meTObar, He
and other Eranftb staff members attended meetings of the Proj9c<.:
DireetorSs as well, as ronfereaces and trorksbops devoted to con-
siderations of the protocols dealing m.th cbe following aap&r:t:>
of the Project; (1) Sot:io»etORoro,ie''--geaecic5 (2) prenatal j,
(3) emofcioaal evaluation of gravida.^ (4) labor and delivery,.
(5) pediatric^ (6) aeurological, (7) pRychologiea.1. ^ ai3d (S) sta^
tisfciealo In addlfciong nmaerous meetings of ssialij working
subcoBTOittees were attended^ Pracfcicaliy every institution in
the Collai'Oraeive Project was visited at least once by Branch
staff -'^ either as raembers of Project Site Visit ComiaitteeSs or
as statisti'":al coasultants on procedure in aay of the above
aspes?.tSo The purpose of these visits was to obtain information
regarding the characteristics of the bospifcai populations;, routine
procedures for all hospital obstetrical patients « the way in which
the Collaborative Project patients fitted into the hospital rcutinei,
atfd other specific problems of concerrij such as selection of ob*-
stetrical patients for studyj selection of children for standardiKa'^
tlon of the Bayley psychological test., processing of patients ;.
completion and processing of study reeordSj and ofeher problems of
a statistieal nature with regard to the various aspects of the study.
Assistance in the formulation of st«dy design aad in earopliiig
eons i derations was given by the Branch to a study at the University
ef California, under contract with the NINDB^ of the Bayley Teet
with raferenee to its efficiency in detecting neurological daraage
at eh® eighth month of age.
Bxaneh mesabeics have worked closely with procedure analysts
of ths Project Services Branch» KIKDBj and of the Statistical
Procossing Braaehj HIE3 in the preparation of procedures to be used
in the collection of data eentrally^, storage o£ forms, and establish-
ing eecessary controls a© that up--«to»date information on number aad
type© ©f fon%s received, and patients processed may be available on
a curr®n.t basis.
Members of the Branch preseated a paper on 'Statistical Aspects
of the Collaborative Project,'' at a eonferenee on The Epidemiological
Approach to Problems of Pregnancy Wastage;, held at Arden House^
Harrimang Ne^ York, in March 195So Another paper^ dealing with the
deKeetability of differences in incidence rates of various neonatal
defects resulting from populations of gravidas with specific maternal
coraplicationss wai» prepared for presentation jointly with thjs
Assistant Director,, NINDBg at the annual aieetiag of the American
Public Health Assoeiation,- SC; Louiis^ Miosouri» in October 1958.-,
-k
A Eiultltude of statistlc&i prdbleuss reaaia to "be solved.
Asong these are: (l) The establisiBaeat of a suitable sajapliiig
procedtire adapted to the sitisatloas of each institution and to
the needs of the project; (2) the |npepara.tion of approjpriate
codes and isamials of fO£^s and procedures for each aspect of
the study J (3) the developaeat of adequate laethoddogical. tests
of reliability and validity of the data secured; (Some of these
tests Hill precede the inauguration of the stun^ ajid others wiH be
conducted concurrently vith it.) (is-) preliiainayy planning for data
She araach is involved in two studies^ in which it is offer-
ing consultation and/«r ser«i.ce, ^ii<di are indirectly related to
the intensive pfease of the Collaborative Project. In one of these,
coordinated by the Epidsaiiolo^ Branda, MMB, a collabomtive
prospective study of the relationship between Asian Flu during
pregnancy and the occurrence and course of neurological sequelae
in the offspring, the Biometries Branch will be involved as the
central statistical agency. Visits have been nsad© by Branch meiabers
to several of the collaborating centers in this stiu3y. Codes have
been set isp and procedures for proesasi3ag these fozms frcsa institu-
tions have been established. Consultation relating to the sasipling
of serological specijaens in the study has been given. The tabula-
tion and analysis of •Qiese data will become a anesponeibility of the
Branch « In the other study. Branch meBibers have provided con8\2lta»
tion in the foBsmlation of a studly design to the Dijpector of
Research and Statistics, Baltimore City Health Deja-rlanent, and his
associates, in conjuncrfcion with a proposed are search study,
"aaofeing and Preaaaturity." The purpose of this study is to detenaine
the incidence of prematiarity amcasg offspring of gravida ^o smoke and
among those who do not ssicifce, and to include within these groves
other covariables, such as work histoxy, education, blood grouping,
and personality characteristics.
Staff of the Branch have consulted with the Director,
Research and Statistics, Health Insurance Plan of New Yoxfc City
(EEP), and his associate, concealing the possibilities of coopera-
tion with SIKDBo Tbs HEP is currently conducting a prospective
pregnancy study based on ccaipleted report fosma received on services
rendered to aesibers of the Plan. The purpose of the investigation
is to stuSy pregnancy loss, congenital anoBalies, and morbidity in
early childhood as rslated to imtex^sal morbidity and other conditions
prior to and during the antenatal period, and for a two«year period
follosring birth. As a result of several conferences between the
Biaaetrics Bjanch and HXP staff EiKabers to det@smine how the HIP
study and the Coilab03Ps.tive Project could reinfcafce each other and
mutually take advanta^ Of both research pregrisms, methods of liaison
and areas of cootperation were established.
tfee sccoB^liBhiBeat* given above relat:e to the inteiosi'Pfc
phase ©f fche GollafeoratlTre Project, In view of the poasifeiXlt;;
that tha incidence of certein »eoiaat@;i deficitie is bq l.oa th«t
even 40,, 000 live births 'will not yield enough cases t© desonstr*tt
se<»£latie»l significance ^ an extensive phase has been postulated.
In this |>h&se a nwiaber of approachea are envijsioned «toerefef the
cases available for atudj in given categories of disease may he
increased, la those cojaBaunities in ^ieh collaborating institu^
tions are located, attenpts will be made to relate retroapecfcivel'j
during the course of the Collaborative Froject the occurrence ©f
neurologie^il disorder to prenatal « labor and delivery data
available fron hospital records, vital statistics records^ etc,
Hie inmig^ration of an extensive phase program in this Re%' fork
Git J fir«a has been undertaken by the Colua&ia Uuiversitf School
of Fublic H«<h and Adsainistrative l^dicinco The Branch Chief
has consulted ^th officers of that school in order to help co^
ordinate the respective efforts of the agencies concemedo
Additional studies, valuable to the condsacfe of the extensive
phaa®3 asaf h& ezeeuted by utlllzit^ copies of ptsneh cards of all
births and fetal deaths » piurehased by contract from cooperating
State mad local health departiaents in the cities concerned. Such s
8fe»^ ©f fetal wastage in Bew fork City is currently uaderway ©n &
Joint basis ^th the 'Hew York City Health Department, It is con-^
carfied ^th the tabulation and analysis of approsiiaately 380,000
punch cards for the period 1955''56. Xt is expected that a nuoiber
of valuable leads nay come to light ms & result of this investig@<»
tiOBo itete&^t® ^11 be made to undertake similar or related
studies during the coming year in other cities in uhich collaborating
inatitutions are located «
Ca) Collaborative Study of Etiology ©f Uveitis «
At the request of the Executivs Secretary- Treasurer ©f the
Aaieriean Aeades^ of Ophthala)@iogy and Otolazyngologye the Bioraetrle^t
Branch was called upon to set up a collaborative retrospecti'^e study
to evaluate possible etiological factors in the production of
Sraaulo9»teus and non^^granulomatous uveitis » After consultation
with meatBrea of the Acadesgr'a Coianitsae oa Field Investigation for
the Osa @f Magnostic Froeedures and Iherapy in irs'eitiSa a study
dasifn and study fonss ^«re devised by the Biots&trics Sranch <mA
approved by seeairgrs of the Goimittee. This study 'sd.ll secure data
froM IS- 20 eye institutions on fandLly history » patient's aedical
hiafeory and conditlonij asspoaure to rare diseases and InfecfcionssB mtS
laboratory findings with respect to blood teetSg feiopsieSa skin tmmm,
and skin sensitivity to varimss streptococcal agents for approxio^tely
3500 uveitis patients and 3500 controls over a fi'<ipe»year period, ffe«
il»@rlcan Aesdenip of %hthalfiiology and Otolaryngology has it&qu&stM
that this l»e a collaborative study involving the Biosi^trics Brunei m
tie^ statistical ageneyo It is planned that, if the study 1$
» 6
approved by £ha Field lavestigatioas C&miltte.e aad b^ She Advisory
Couoeilc, tha Blcmietriee Brsneb fd.Il render eoasultaeion to eaeh et
the collaborating ia3til:uti«ms %7ith regps^t to e:as9 iseleetion a£id
dafea eoll&etioa proe«dureSo eompleted data i^ill be forwarded to
the Bi€sai9tri@e Braaeh via the Meriean Aeademy of OphfchaI^>Iogy
affid Otolaryisgology for statistical proesasiagg tabulations and
aoalysid ,
<b) Cailaborative Stu^ of Evaltsatioa of
Non^-Stargieal Treatment of Strabi£
At th® request of the Chairmaa of the Coomittee oa Evaluafcioa
o£ thtt I^os^S^rgieal Treatmsnt of Strablstous of the Aiserleas Assadm^
of Ophehalnislosy and Otolaryngologyj, the Braaeh Chief reviewad with
several msabsrs of the Soimnittee a proposed study to ©vaiaate sueh
therapyo Tha aead for a clinical trial est up on a 'blind' basis
with adequate eontrols and with evaluation made on an unbiased basis
was streasedo It is plaansd to havs a Cemmifetee meeting in ths near
future ao that these facts might be brought to their attentioB and
so that the necessary groundwork for collaborations with the Bioeietrics
Branch as central statistical agencyj may be laid,
3o Statistical eoasultatioa and/or service given to
iavestii^atora outside of NIHDB on ether projects.
The folloviog represent the type of eosisssltation and/or service
rendered dtaring 1958 to outside investigators on other projects c
(a) Prsparatioa of age^specific mortality tabulations
on cerebrovascular accidents in the United States «
(b) Svaluatlon of study design of a retrospective study
©f etiology of cerebral palsy in Chicago, stapported
by aa NZH grant , and consultation given to help
B^ the £laii?s in the study design ,
(c) Consultation on study dasigng developaaent of adequate
tabulatioasa proeeduress and design and construction
©f codes given to the ©edical director of a research
foundations ia coxmeetlon with a retrospective study
of the etiology of mental retardation snwag children
ia Chicago o
(d) Censultatloa oa study design of a retrospective study
of etiology of cerebral palsy in several urbae areas
ia Minnesota o
Evaliaatioa of the study design aad merits of several projects
submitted for research grants to the Easter Seal Research Foundation
aad to the Office ©£ Vocational Rehabilitatioa has also beea randsred
«t the request of the Director of tha InstitutCo These applications
for research grants have beea coneeraed with The TU)le of Neonatal
Jasmdiee as a €&ns& of PreveoEable Physical and Meate .
■'StiKty of the Causes of Mossgollsm asd Other Congeal. tal Befect-t, '
'Correlation befcv^een Clinical aad Pathological Findings ia
Cerebral Palsy, ' aad 'Prepregaaacy Investigafiion of the Emoeicr.u^
Physicals; Eadocrinoiogiealg aad K«5tritiosal Faetors Involved in
Soageiaieal Hal formal: ioBSj, Prematssre Fetal Deaths^ aad Spon£aneous
Abortions « ''
Merabers of the Branch staff presently serve on the follow-
ing eommitteess thus beiag« in effeefe^ ehasnels of eoaaaunicatioa
whereby problesn areas in the aeurological field bseom© kaawa to
the Branch aad whereby the Branch's field of Interest aad activity
besoa© kaown to ofeher investigatox'sj
(a) Statistieal Advisory Consiiteee to 'A. Study of
CJse of Statistics osa Maternity and Newborn
lafant Case in KospitJ^ls. '
(b) Advisory Consnittee on Epidemiology and Biometry
(WINSB-Liaisott)
(e) Coniraittee on Komenelature aad Biometries of the
Ameyieaa Academy of Necrology.
(d) Ad Hoe Comnittee on Mental Retardation of KDiHo
(e) SIH Advisory Conanittee to Natioaai Health Survey
(NlKDB^Liaison)
(f) Panel for the Study Group on Guide Material for
Comparable Studies on Materrsal and Periaatal Events
Reported on Vital Records »
The Braneh Chief was designated by the Director, NINQBj, to
represent the lestltute at the 'National laistttute on the Role of
th@ Workshop ia Rshabilitatioffig" spoasored in April 1958, hy the
National RehaEE^ation Assoeiation,,
4» Statistieai consultation acd/or serviee furnished
to elinic and baaie resesreh iavegtigatox's at
RINDB ia the areas of n^rology and bli^daessc
Bislow are indieatsd the imits in NINBB that reeeivad atatistieal
aid itmi the Biometriea Braaeh ia IS 58 aad the problems iavolvsd:
(a) Laboratory of Neuroanaeomical Scienesso
Assisfcaicm® tn preparing data on the soelal behavior of £ree«
roxmissg rhestsss tEtonkeys for nzaehine ealeulations in order to obtain
correlations betiaeea the o€c«srr«nce of epeeifled soeial "aets,,''
(b) Medical Keurology Breach
Sfcatistieal analysis of the effects of eertain drag® am
jmacl® rsaeeion eo elecerical stiimiXio
(c) Swffgical Saurology Branch
Staeistieal aid ia pr©bleais involviag phyaleal dietrlbnticn
©f various tjrpes of cells in the pituitary glasid,
Stagisfcieal analysis of the effect of various types of hvaln
surgery oa chisapanEce body temperature » Coaswltafcioa givcaa eoseesa-
lag ways to laprove the study design o
(d) Ophthalmology Braaeh
Statistical help for a proposed correlatloa of physical
pararasters lavolved ia hmsan sight »
Statistical asialysis of data on the blood level of a eortaiis
drsiigo
Statistieai aid in analyaimg data oa the visual resposise of
tba h^saa esfe to light of different vave lengths o
Statistieai analysis of data on the effect of different
emsysa®® oa several types of tissue from rats' eyes at selected
ages*
(e) Epideaiiology Branch
Statistieai analysi© and interpretation of ineidsnce and
prevalenee rates for ParkingonissBj based on reswlts of a sjarv^y ecm'-
(tectsd ia Soshasterg Mismesotao Also iaseluded was a eomparia®a ©f
life axpsetsnsy -of case® of Parkinsoaima with that ®f the Uo So
popnlatioao
Statistieai eotEpMtation and analysis of ags-adjustsd d®ath
rates frssm varlomt asmrologieal diseases ia foreign cowntriesp tn
th« total msited States, and by State and region <>
Statistieai analysis In connection ^th an investigatioQ of
amltiple seleiresis in a e(»simsnity with a high ineiden@e rate.
Be PRQ^ES,.FllTljRE OBJECTHTHS
To date^ the energies of the Braneh have h®en direeted t©
the aspeets of the program mentioned above « With an Increaa® In
staff it is eKp®Eted that other aspects of pr®pesed Branch activi-
ties relating to developing a program fos statistieai data oa
prevalences ineid^mce^ and aortality due to nettrological and ssneorj
dieorderss idill be approacbsd. It is also hoped that a program
to design and implement studies in order to investigate the re^
lafcioaship of pre-valeneej, iaeideneej, aad raoa^tality of sueh dis-
orders due to vario^ss biological g geaetie,, aad eavlronmental
fae,tor§s, will be andertaken cooperatively vsieh the Institute's
Epidemiology Braneh« It is believed thatj, as the Branch grows
and as knowledge of availability of. iSs statistieal assistanee
beeomes naore widespreadg there will be iaereased tttilization of
its services.
The Bsaneh's budget for fiseai 1959 includes 20 positions
(12 professioaal'-statistical positiosts and 3 ftleifical"stenographic
positioas) , To daSej, 8 of the 12 professional positions aad 5 of
the 8 elerieal^aeenogiraphic positions have been filled. Efforts
at racruitment av& steadily being made. The employment office© of
th© >Aaaerican Peblic Health Association and of the American Statistieal
AssoeiatioR have been requested to lend their efforts in this diree-^
tion» LstS:ers have bean ?;ritten and int&svi&xm arranged whenever
there has been a possibility of seeuring a pramisins eandida£e„
Hoxvreverj, the great shortage of q«alifi©d biostatisticians has, to
date, made this a Snsstrating experience^ The provision of.addi»
tional spaee for the feo«sing of needed staff and files has become
an acute probie£ia» In Augtsst 1958, the Biciaetsics Branch was Eoved
off the KIH grounds to the Progress Buildings Bethesda, !5hiie this
mave eased t^aporarlly the need for additional space, it is believed
that the disadvantage of being off the reservation will act to deter
yossagcs' statistieians frsaj seeking smpioyment with the Branch. It
is expected that the proposed additional laove of the Branch to Silver
Springy Maryland, will only accentuate further these difficulties „
1ATIC»L JieSISUSE OF HEUROLOGICAL D3SBASK ASD BLMM^S
Esttoated Obllgafelops for Tt 1939
I Besearcih drazsts )
n Field lavestigatiojtjs aaad ) $l6, SS'^^OnXJ
Pilot Projects )
III Gpadjiate TroSsil&s (seaxxHiB U,O75»O0O
IV Special Sraineeship Program 1,500,000
V Sesearch Stellowshlps 536,000
71 Review aad Approval 551*000
lasalsi^Say fear, 1958
Extradural Progx'smiS B,x»as.Gh
Hatiosial Institute ot n«*vur©XegicsJl
Dleeases fend BliRcbaese
Rational Institutes ©f Seaitli
1 IffiSEAROS QRAISfS
1» Proggant developments
DwrlBg tfee year a ©ensiderabl® expansioR ©ecurred iia the amftt^r
cf researeb studies ©is neur©l©gleal and ee^.m<ry disorders, aad ia tli©
basic neurological esieaees f\;csdaasentel t«. sotmd ciltaieal inv®®Mga1&l«;'rfi.
fixis expassl©B was sade passiKL© by a marked isersase in the budgets
All i?esea3?(gh projects recea^aeEded fer approval durlBg tfee year h&ve X^e-en-
paido In addition, awards were ©ad® ©a several prsjects approved lagt
year, feuf unpaid^ then, because of laek &f fuEds.
It^ber®^ @f graata i - A® the year ended (i3ecajiber 1958) th«re were
&i'9 &®tiVe regeareh grants,, A year ag© at t&is tiaie there were 670
attive researeh grant® (a net Inerease ©f 2608^ occurred dxsring the
present year)o
BMM3get> distributions - Vhen the bud^t f@r research pirejeets is
broken dews int® bread pragrwa areas it is seen that 1^T^ was \iged for
support ©f individual researeh projects in neurologleal dlsKsrder® such
as epilepsy, multiple gcleresis, muscular dystroplsy, cerebrovaseular
disorders, degenerative diseases, neurologleal deficit® ©f the yeiasg,
and others o ffee previous year suck studies used 51^ of the budget »
Support of projeetB ©a sensory disorders consumed 26^ of the
researeh grsst funds this year; thi@ was the same as in the prevlexjs
yearc Abov^ two-thirds of these fussds were used for investigations of
vision and its disorders sueh as glaucoma cataraet, uveitis, and acci-
dents, the other ons-third was afesorbed in studies la hearing, ©peeeh
and ©cLuilibrium, isnd other sensory studieg as sasell, taste asd t®u«h
including pain. The balance ©f the budget was uged dxiring the year for
supporting field investigation studi«g@o
^gaadlag parograas ; - During the past year researeh into the
eerelsirovaseular' disorders e^anded rapidly, Kearly $800,000 went into
its support, as eoiapared wlSh $J*00,,000 last year and only a little iKrre
than $100,000 two years a^o fhese figures icelude the tw© ©©operative
studies ©B. aneurysm;® and antieoagulants that begsin la^t year and which,
this year, used $300, 000 0 KevertSseless, it is evident that even aside
from these eooperatlve studies a marked increase in infeerest in the
area @f eerebrevascxilar problesas has o«sc\irredo Undoubtedly the two
cooperative studies have stimulated omeh of this interest and are
directly related to the rapid esepanslon in researeh grant support flowing
into thig areso Ae'feivlty in this pregraia will remain hi^ is the future =
- s -
Ab acti-re interest in Eultiplfe selerosis aad oth«r daayelinating
diseases 4s ehsva by siiipport ©f resscarch projects at a level ©f $100,000
diuring the year, as ©oEpared with $Uc^,000 last jrear. Studies la Reuro-
muscular dieerder© Ineluding mtiscular dystjfophy lBcrea@ed to a level of
$800,000 this year as ccnspared irith $500,000 last year,
AnsDEg thiS disorders of vision, a marked inerease has occurred in
studies related to glausoma. During 1958 invsBtigatlons in this field
reeeived support of about $400,000; this was double -^le level of a ye&r
ago. Further rapid ©Kpaasion in this in^jortant area will undoubtedly
occur becaui© ©f the interest axoxised hy the field investigation study
and the research conference that are scheduled to be eupported next
year. A similar sharp increase is acted in studies related tc uveitis
and infectious eye diseases »
Efforts are tuader way to prcaaot® studies in speech disorders as
related to neurological deficits and mental retardation, fliis year sexr
8 projects started with $100,000 support. Rapid expaaeion in the im-
laediat© future is asatieipated,
2„ ReBeareb d^yelopB@nts_; Oontributiona from specific awarda
During 1958 reprints of h^ papers tfeat appeared ie Seientific
Joiaraal© were placed ©n file -by HIBDl Grantees ., Theg« were supported
by 296 research gsrauits. With 670 grants active at the start of the year
it is evident that nearly half of JJXKDB grantees published at least one
paper during ths yearo Coatributions from specific projects are listed
in the following paragraphs, bixt wittout any atteni^jt at covering all,
or even the nsBst iEporl-ant, discoveries »
Durinsg the year Dr, Heinrleh Waelsch of Coluaabia Itoiversity has
repoi-ted extensively on his studies on metabolism of protein and aaiBo
acids in nervous system and brala» fhis iiaportajEt work is fimdaiBental
to the understmsding of rormal, as well as ab:^rmal, activity of brain
and nsrve cells « Tiae relation of tryptophan to sejrotonin and other
CKS-sctive drugs exasjplifies this interest* It has beem Bbamx. that
gassaaa amino butyric a«id is formed by brain cells from glutamic acid
and that during convulsions of the animal the ability of brain cells to
proaste this eonversloh is lessened or atwliehedo Is this changed
metabolism a caiise or effect of the convulsions? Clearly much nasre
infonaatioa is needed as to the fuactioa of amines emd the role of
amiBO aeld metatoli^i in brain and nerve cell activity.
Dr. Eugene Kennedy of the U. of Chicago 2ms expanded his f\aida~
isiental work on th6 way the cells of the body produce cephalin, lecithin
and other phosphelipides important to nervous tissue eoi^oeltlon and
activity. He has worked out the pathirays by which these fundamental
•units are put together, and has reported on the enzymes which are needed
for t^is» Several ©tiier srsateejs continue the laborious atteaip^s at
sorfeiTig out aad idestifyiisg the various 'bizarT« lipid eoagpene&ts &t s^erre
tissue o Ilot«worthy in this ai«a ©f vtixk are Dr.. ^ordl Fol®li-Pi of
BodtTpA wljo has repeated oa s«fS7 llpoproteisB fro® th© eerrorta system, aad
Vr, H« Cax^r ©f 0* ef Illi!»i« wJje eontiaued g-tudies mk the eos^sltioT;.
of 8phia©D8ine and -^e various 8plii3agplipJ.dg»
Dir. Z.ar7abee @f ^ol^sus Hopkis^ coatyifiusd bis fuodaoee^al studless^
©f the metabsliaa and glyeolysis of cearve cells. He has oMalned evid&Eee
■§hat the souree ef eissrgy for serve ii^pulse in ga8sn.©tt eells involves
aome cen^sonent in additicn to aluccseo 2* tMa eesmeeti^n Dr^ Po 0*.
Sehaaiitt of MaSef » liafi x-e-es^liasiaed «hs thaory tha% the Setowan eells ef
the myelin eheath, far f rem being inert cells, are aetively iavelved Isi
sv^j^lytna the energy necessary for ien traaspor* and ©peratioa of tile
'@ediua ptxap in icsrve iaipulee iransod-ssloae
Daring a syiapogjism Bxx^^Tt&d by aaother Insti-fetifee, Dr. BeEay-lPovsi
ef Hartfayd, ais HBlOi grai!it«e, reported oa the iB^rtaace ©f n\itylti«sn to
neuropattelogy^ His s«vl(sw ef -^e neurologieal diserders y«a«ltinp; f^^csse
Biaple Kutritioaal deficiencies la oan aad antaals brought the fclJaewing
relatione to minds (a) Ifental d®terloratlon o«eur» from a deficiency
of nieotinle acid aad tryptophan (pellagra is huoia^). fhe stxiaetural
relation between serotosin, 'several ©f the Swllucogenic drug« and
tryptophan were recalled, (b) An extraue ceurologisal ay&dxoras result-
ing from peroiclcus anesaia, ean be preven'ted by therapy \rlth vitsffila B12o
(c) The epileptold convulsions reported as a result &f vitamin Wo
deficleney in Mce, srabbits and hisaaJR infants » (d) The epiaa3. eord
lesions and collapse of voluKtary nmscles in vitarain j&6 deflcieaeiee in
swine and E«fcbitSe (e) ffce polyseurltia in humans aad the iavolxjntary
rolling EEotion in rats deficient in vitamio Bl (tbiamine). (f) Speeiflf?
cells in the cerebellum of chicks fed diets lacking in vitamin 2 usdergc
necrosis; this is related t<3 the eacepfcalomalcie vhich can be prevented
In chicks by adeqiaate dietary vltaala E. (g) Sn rats a ehronle de-
f ieieaey e<f Tlt^slfi & £«salts in prcsounced lesions la the spinal cord ;
this is correlated with slowly develeping paralysis or muscular dysfcrcpK^"
in these aniwO.a. Its, rabbits fed tfce diet deficient la -vltaolA E the
Bussular dystrophy proeeeds es^losively to fatal texnieatlon in a few
weeks, (h) A deflsieccy of po^ja6»l\iBi Ift th« die^ of dcfls, x«t»«
rabbits aad buoaas results in total flaeid paralysis of ftrlated muscle,
similar to faoiUal perlodio par&lysis in humans, (1) A dietary de«-
ficiency of choline, the pareat substaiuje of aeetylcholine, and the
zaajor co^aponent of pbsspl^liplds aad fspblagiollpide/ results in a
SRj.3cle weakness and pas&lysis in rats, guinea pigs, and rabbits., (j) A
partia3. lack of vltaiaiffi A as well as other diet essentials;, lndividua3J.y;>
ireeults In abaciraal of f spring of vasfieus kinds; f«r ©Ksjaple, hydro-
cephalus can be produeed at vill by proper atteB^icn t-o '^ibe diet, of
pregnant rats, (k) A laak cf flopper in the diet of cattle or sheep
_ k -
produces very narked aeiiroiBUScular disorderso Dr. Semsy-Brova empisa-
sized tbat In spite of the clear esaiiq^lee of close relatioasMps^ tnxtrl-
tlonal scientists as a gffoxsp igaore neixsopatbolOGy and few neuKJlogical
iaveati£5ati©ns accord significant cpnaideyatiou to the nutritional com-
ponent of etiology of neixrologieal disorders.
t». Hactemsolm and Ms group at Colisobia Ue have succeeded is,
pjpeparing fat-soluble derivatives of acetylcholine* Thene compounds
act like acetylcholine in triggering ffl3ax>th muscle contractions and in
promotins synaptic trsmsmissioas of nerve inpxilsec However, since they
are fat-soluble they penetrate cells, aeuawaal aasons and myelin sbeatlis,
and are not 6ul34ect to inhibition by curare and other acetylcholine
inhibitors at tlie notor-end platei^ acd the nsuronmscular junction. The
possible and potential uses of such compounds may be very interestix^o
DTo Ws&ok Morrell at the Uo of Minaeeota has coatlxnifid his studies
on the epileptic lesion and the conditioned reflex are. Epilepsy ims
isdacted in imr^^ye by linplaating a disk of alizaina or by spraying a
specific area of the brain with methylchloride, A study vae thea made
on the ability of these animals t£» establish osw conditioned reflex
pathways, or to respoad to reflex stimulae established before epilepsy,
fhe epileptic aMmale vers sigsif ieaatly slowet' than Eonaels in both
aspects. SoTTever, vhen the eplleptogeaie lesion in the brai& v&b
surgically rasoved, the ease of establishing the conditioned reflex arc
was returned to norasal*- 'g^Be studies indicate the pathways of certain
legs of the refleE are throu^ specific brain areas « Xn addition this
study re-eii?>hasize8 that the spontaneous volleys of iaqjulses character-
istic of epilepsy originate from brain cells whose metabolism has been
altered in some way. RoKi^al of these cells by stirgery or other measas
frees the aaiiSBl from the burden of these loaverick cells, fhe nature
of the abisdrBaality in the specific cells is not k&own.
Drs. R. Mayers and y. Fry at the U. of lotra axe attess^ting to
remove the specific centers of abnonraal cells in the brains of h'saaans
suffering from epilepsy, chorea, parkinsonian aad other aovement die-
orders, fhey are doiiig this by Idlling the cells with focused beams
of ultrasound waves, fhese studies are in a preliminary stage with
reports of gsod success ia about ei^t patients. Dr. Spiegel of Tei^le
U. reports s^Keess in epileptic patients using implaafeed electrodes as
the techniciu« for killing the specific brain cells. Hueh of the success
of these efforts depend on the ability to locate the desired area of
the brain, £«ad several stereo-teetic devices have been developed for
this purpose.
A report on the vise of preserved human eye tissise for trasja-
plantation in surgical cases, has been given by Dr, J. Ho Kistg of George
Waohinglwn U* 5be conaea of eyes easa best be preserved for later use
by dehydiutlng them from pure glyceartne, sealing, and storing in vs^xso
at room temperature. Dr. Kiag fisids. These can be used even after two
-.5 -
years of stjsrage, Sise la over 50 patiesfes eIiow tJjat "^fes pz^servad eosaeas
aro as good as fresh eomeas- rased for siiailar ■feraasploiitatioa* Dj?. Kiag
also sspor&S'oa jnetbods f or pseserviac vitseons hiESir f or vst 4a special-
ized probleaasof s^tiasl fietaeismsnts and other «ses. S^aceessfvil use of
solera aad eoojuctiva is also • reported, Sssasplaw^tioa of - lejjs lias 80
far not beea euceessful. because of the. opacity that iiivarisTjl^ develops.
3, Major ipgobl^is eceotaitsged
So major pix>blei2S have been encountered during the year. Some coa-
eem voe eacpimtered illative to- a sl^ajge ^ rules goyeraiBg use of ?e- '
search gjraat f\md8 for forei©i travel. As of July 1, 1958 the new policy
on foreign travel required prior approval by the Advisory Gouacil, 5Ms
vas Dore restrictive thoa iSxe previoiis rule requiring approval oaly for
travel to interzmtional iaeetln(^. Tins Couwsil ^^nt on record favoring
relaj^atioit of the restrietioa ssather thoa the reverse, but agreed to live
\7ith- the. new ruling -for a tpiai' period,
k, Cbajp^ea os^ teproveEaggts in program
She year smr the liftiafl of the aaftitrasy f ivfe-year celling for
support of research graats. ^der Vae leadership of the Advisory Couaell,
12 gpaots were aade -Hlth a period of support resoEBended up to ei^t
years, SelectioB of resipieats of this loag tera aiapport vbs hosed on
the p3?od\ietivity potential of the ima as veH as tSie broodth and sigsiifi^
eaace of the project. ISnctoubtedly this type of si^port will confes^lbute
to stability and frcedcm of researclx la the broad area of interests in
basic aeiirological sclezuses, represerrted by >&h@se twelve grants.
Z^sring thd year a research gj^xcb ims ^ade to S^ssaehusetts Is^titute
of Te^ihBology for the total salary aad asicillasy si5>gort of a seniors
stabure Investigator, Dr. l&CuUcch. Piv* years -wess sreconsojended. Al-
tis^ug^ laade on tlie Mais of a research project and vsi&sr eonditloBS that
d&parfced froai cd rales^i nevertheless, this grant was widely interpreted
as a 'career ^vesti^tor'"' grasst by the consultants who reviewed the
application. As s^^h^ it siay serve as a prototype and precedBot*
5. ?rocgaiii objeetlyes fog 195>9
!3uryss the nest year ertrery efft>rt will be sEade to maintain the
proper balance between exrpport flowing into the various pxogroias aS
interest to SXSn3« Zosofar as possi'ble attst^s will he siade to pzoaiote
Ixxtsrest in certsisi areas that appear eosssvhat retarded; speelTieally,
one sv&h ar^a is the {Neurological aspects of speech developsaent in th^
youac, 2? ^he Prograia HLasjaiag CosKolttee of the Coumll is to carry
oist its In^rtaat role of mapping areas for special progjaaa developaeat
in the neurological and sensory dlsovd&T field, it is is^rtaiit that
reseasGh gz-aib funds be availsible la flexible sssoxsbAb, At least
$15*000,000 -iJiH be neeessazy duslns th© year for Research Grants.
6, ^j'oliattg of applife^ati&na
(arct including Field
1956
1958
1958
MOc
209
201
162
5?2
oasidered
AffiOURt
$3*613,515
$3,625,030
$2,764,T19
Jio.
1U9
119
111
3T9
Moveeiber
$2,02U,39I
(55?^)
$1,929,963
C53?£)
$1,5?2,V4^
fotal
$10,o63,26i*
^5,526,800
(55^)
T» Sfcafg asgleeaaents
DTe Sdwin Hove (since Jfareh 13, 1958)
/
1958 AKS«AL BSPORT
of the
l^tloeal Institute of neurological
Diseases end BUsSzidss
13:. FHXJ} IBVSSTZGATXaR ASD PILOT mMSCS (StASfJS
1. BftfflreaB dgreloiiBEeats
!^« purpose of tlsis progreaa is to l>3X)adeQ €!ie research graot-ia-
eld base aM to facilitate the AsTelopss^nt, wfeere seed is iMicatsd, of
epidSEiological studies, aatioaal surveys, cooperative acd collaborati'tts
studies t^aat call for a Qiulti»institutional aaS, of ten a oulti&lscipliisary
approacis, as well as pilot projects established to work out leads, i,®,,
t3se most proaisizis spproada for furtJier research. la maaay rsspscts it is
a progre® of applied r^searclb, ia tfeat it sodeavora to enlist teie aid of
a certaiis segsssnt of tb© Batioa's scientific ussmpower i» an effort to
ccsobine forces, utilize the information at &aM, es& obtain soss^ aasvers
relatirely soon, instead of depej^ing on tfes gS33©rally elow course of
events. 5^i8 prograai got uxderway in January 1957, assd now consists of
6k projects ia tlie eaoxint of $**-, 329,196.
T^is program is one vhtdh has been established 8]^cifically to
ws&t urgent research needs. In order to assure coin^tsnt, unbiased,
objective review of applications, two prellMaary review ec!«Eaitt«es hs.r@
b€«n establisljed. T^ Field Im^stlgations Cossaitt*®, so broadly cob-
stitaited t%at a particular special-^ or area is usually repr^seatsd by
only one ssmber, reviews 'all applications except t^ose ii&^olved in. t&sa
Collaborati'^ Stu^ of Cerebral Palsy aM Saurological Deficits of Xafascy
aad Cliilfl&odd« Applications for this latter study ere reviewed by l^,s
-ad Iffioc Ccsiaittde on Cerebral Palsy 4 SSsrou^ t&is Hii^ehanissx it is possl^
Sle to bris^ to bear on tl© C.P, study, a coaceatratioa of esipert a^cLee
necessitatod by Wis breadtl^, size aM intricacy of tSiis p8rticulas>
researcfe effort ,
Mtlbott^ s Tsas^ter of single isstitution ss'ojects of a pilot siature
sre fizuascad through tkis program, 92 per cent of -^se funds is «sg:pen^d
ia the Buj^rt of cooperative aM collaborative studies i^volriiag a
araaber of institutions. To provide a means of differentiation between
nulti-inetitutioaal ^^ojects larolvimg an eK^oditure of lastltuta dlrset
opsrations fuMs and latraisural i^reon^iisl ttcm tl^ose eupr^rted ei^tirely
frca esctrs^sral j^iu^, tim ters "collaborative" '%as been adopts^ for t^
fo£ffier aM the t&sm "cooperati'is'ie" for V&s latter. 'Sm tesm "eooperati^%*'
ie, titerefore, utiliz&d to describe a sitslti-institutional projaet in wMc^
a BuaeS^er of outside isastitutions are eooj^rating witSi ea<^ oVmv, witSst m>
couatftTport Institute actirity as& sisEportefi wholly by ertrcsaral ftoSs.
em to
M^ TMi^m .mwi mMMmi-
During 1958, tke Collaborative SboA'";- c4 iienibTol Palsy aud Sfsuro-
locicctl Deficits of .I-afoiicy ajad Chil^ood ho-s oai.cst reacfesd optiinuB
developaent. This study iicm iavolves ih central isasti'fctitioaaa, vith oae
laoye to "be ecLded in orfier tlaat "fciie stu&j cover a total of itO,CX)0 isiJfeuats
cs .x'S'-uired for veQldity. It is cow being fiusaced io the assotqat of
$j;^07j99^i-« ^is Is a lo3g teaaa KHilti-inatitutioaal laaaitidiecipliaaey
Btu(3y froa whida little significant data is espsoted until follow-up is
relatively complete in I967 or I96S. .
The Cooperative Stu^ of Aneurysias and Acate Subarecimoia Hesiorrlia «,
involving 20 iastitutioas aud fiiaasiced la the amount of $200,000, reached
full dev^lopEent in 1958? Kie saxrpose of tljis study is to amass a "bo^- of
baseline data on tiie laedxeal versus surgical treatment of- apeui^rsiBs,
utiliziEig a nuaber of optional fonas of ti^eatasent xiader coEditions governed
by a study protocol.
!Sse gecansdieal Collaborative J^idssaiological Study of Selected
KeujTological Deficits Involving South Carolina, Hova Scotia and Japea^ will
b© eoapleted in 1959* Bata from this study are not yet available.
Use Cooperative Antie9agalant Tlierajy Study involving six institu-
tions at a cost of $5^'^ 000 per annum has one laor© year to run before
eoi^ilete data will be available. At that tlase it xaay be indicated that
a broader approach, testing a larger number of drugs, would be prcmisiag.
Suriag the year, tfor&ing with the Chjponlc Disease Dirision of the
Bureau of State Services, planning has been e<s©pleted for a Collaborative
Glaucoma Detection Evaluation St-^dy to start early in 1959. Ssis study,
vhieh has as its purpose the developa^at of techniques and proeeiSures
leading to improvesrasnt in preHreatioa and detection, srill ijEKrolve 5 research
institutions, and cost epproisi'aately $150,000 to $175,000 per year for
five years.
In fedditioa, the Institute's Biometries Brancla, -fe-orkiisg vit3*
officials of the ^jjsriean Aea^^i^ of O^-b&alffiology asA Otolarysagology and
research opltijaljaologists, has developed an acceptable protocol for a
uveitis study. Siis study to he eatitled "She Collaborative Study of
Etiolo^ of Uveitis", -arill imolVQ 20 institutions, cost about $2^0,000
per year aM. ^t xinder^ay in the fall of 1959.
'S&Q National Adnriso^y Ifeurological Diseases ■ sM BlisdaesB Couscil
has made tentative plans to sponsor an asisual series of conferences stsr-t^
iag in I96I, on Variovis aspects of glauccaaa resear<^. It is believed
•Siat liiese eonfereaees and the Collaborative Glaiiccasa Betsetioa Sralua-
tion Study will provide Goasidsrable i^^pst^s to rese^'ch in the field of
vision.
Piass have- beea ccs^letad -to hold a natioaal working eonfereaee on
the conservation of heariag to be jointly spoasored by tS*e CliilSrens
Bus^au, -iije Claronic Disease Mvisioa of the B^ m^ i^is Isstitute,
during the last veek in May 1959. "Hse purpose of tMs eonferftace is -fee
deliJMsata tli© ■various facets of the probleai, map out -^e most prcsaisiag
attack and stimulate i^seeych efforts in a field that is sadly lacMng
in activity. A conservative estimate of the nuniber of persons presently
feandicapped "by iaipaired hearing is 10;5 or 17 million. Xn industrial
coHisuBities, saaple surveys place tlse figure as hi^ as 20^.
2. Cpatrlbutions result ia^ frcai particular ttfcragi£s '^ ■
Siace t^ais progrsBi got uaierwoy in 1957/ aJ3d siace almost 100^
of the erpeaded funds iax^ifolve multi^institutional studies^ the sbortsst
of wliicji ere tbre© yesrs, no research contribution is yet ready for
eimouskcacent. It is knoun that several studies such &k the cooperatire
ojoeuryaa study, the coo^arative anticoagulant stuSy, the natioaei survey
of hearing ia children and ot^irs are resulting in data which se^ at
this point to he highly sigaiflcant. There is no douht that this pro-"
greja \jill serve to open up asjqy nev research leads as veil as contribute
definite advances of marked sigaifieance. Specific citatio^i, however^
snist he delaysd at least one sore year.
J. Major grobl^s encountered
T3ie Field In^'eetigations program has grown fro® 51 projects in tfee
saiouat of $1,961, fel5 ia 1957 to 61^ projects reiuiriag $i^, 329,196 in I958.
(See attached table entitled "Field laarestigations Krogram, Fiscal Years
1958 CBd 1959") Concurrently, other extramural programs have similarly
«jGpcnd«^ necessitating additional staff* Baccuse of -i^is pressure it
hCG hesn decided to ea^loy a full^tiae stc^'f pe3^s6n to take jjss^di&te
charge of this progrsffii iastead of relying on tS^e part-time efforts of
the C^ief, Ejctraasiral frogs-affls Branch. A ftill-tii»e staff icdivitShial,
serving cb Executive Secs^etory of the FieM tEjRresiisations CoaBKLttee,
will be able to devote laore tla^ to graatees, keep the progrem ussder
closer surveillaaee and thus have a laore intisaate knowledge of probli^s
as they occur, and also be of greater aid to tb^ Cc®aittee in ccrrying
out its plans.
^» g^rosrcaa ob^eetiyes I'or 1^3§
Progreas objectives for 1959 8«<© (l) to get the collaborative
glauccma PAid uveitis studies established on a souM basis; (2) to develop
a Isrger research prograsi in i^K& field of hearing; Ci) to get uMsrway a
cooperative brain tuator chiQswthera]^ project, aiad (4) to stli»ilat@ in
foreign eouBtries projects id^ch because of tlieir imi<j.ue aspects \j111
ffiaterlally contribute to research progress in the U. S.
■^iOldo'xq, m^
/©
5» Vohxm pf appilleatioas
Ifeetlng Ko. Amoujit R6.. -^ Amount i
Jferci 1958 9 jS9f7l9 5 55.6 ai7,5i6 66.0
^fene 1958 li* 1,028,3^ 6 h2,9 27^>0lj.5 26.7
Ho^embar 1958 22 1,1^55,155 1^"* 6j.6 l,00j,311 68»9
II ■ 1 1 I »■ I I I ' II mil ». nil. II niiiiii»i uMii I ini I n Him—within HIM I i m m ly i n m «ni «ii 1 1> >n i Ti-i«ri.i»n.fiiij.«j h j,iin«j.»a
TOTAL I1.5 2,8lj,220 25 55-6 1,^9^,092 53.1
6» Stsjff asslgaments
Ilr. Segey, Acfciag Esscutiv® Sdcyetary, Field iQTestigations eas&
sd hoe Cerebral Palsy Cosanittaes,
«w/> -ff' ■..•^•'^'- • W'.;iHMy> •|.1>L»i^«Ml«>>*«■;<b•*t\r.«'n^M-<^^
Site
(Fiscal year ba^is)
//:
1956
L DXSOBEOSBS
"ifoo toPisgl^ "~~fWW
1939 «
i, of $
51 $1,961,415 100 6h $4,329,196 100
tufological Defieits of lafanej
k CMldliood •- fotal
aUab. Study of eoPo]
f«'bjfal "Ifaseuiar Disorders - festal
jop, St.udy of Aaguryam)
jopo Study of Aatieoagulants)
Llobo Epifis^, Siirvsy Seleeted
feurologieal Dlsoytiers
Borders of Aging
Llepsy
Ltiple Sclerosis & Other
Oemylisiating t)i®easfe8
arlng & Glance
feinopatiiy
Eroleiafeal Fibroplasia
sitls. Keratitis & 0%>er
EnflajEsatory ^ Pasaeitie Diseases
15 1,295,515 66 „1 25 3,639,151 8U„l
(8) (1,101,506) (56„2) (16) C3,W,99i^)C7Bo7)
ainistrati'^m ©ffarat
-7)
6)
375,^5
(253,033)
(56,71^8)
19.1
Ciao9)
(2.9)
25
(6)
318,810 T«^-
(188,514) (kj^y
(53,964) flea)
3
81,183
4„l
2
55,241
1.3
-
„—
.-
1
23,621
0,5
2
22,607
1«2
1
19,60?
0,5
-
-„-
*».—
1
18,501
c.
3
111,992
5.7
3
100,552
a.3
-
—
—
1
19,103
0.4
1
6,597
0,3
a
59,370
l,k
1
43,056
2.2
2
54,935
1.3
»
—
-»
1
20,355
0.5
i
10,000
0,5
1
15,000
0»8
Awarded t© d&te
1958 .animal B«port
of the
Katiosal lB8tit«te of Neiaroiogiejal Diseases and Blindneas
III. Graduate frainiag Sr^xts
■^ • ,^^-?,^<^Q^ Mgagology and gedlatrlg Mgurology
Gradaat® trainisig gmnt aid is offered in Clinical Neurology and Pediatm..:
Neurology to facilitate tb.B establiehsjent jmd develojasent of postdoctorsl
traixiiEG progx«ms ia eacfe field, f&e priizsary purpose of these programs is to
aid 3j3 the trailing of postdoctorsl clinicttl personnel for careers as tescfeer-
investigators in t&e field for ■v&ick "ihe gs?©at is given. In essence^ tfee
grant- iE.-aid training pKJgraa of t&e HXHDB i;aa prinjas^ily established to
facilitate thm research effort in ttee ne^arologic and sensory disease fields
tfero^i^ t&e prodiffictioa of career investi^tors .
fraiaisg g^saats in GliEieai Hexi3?ologi<' provide stipend swppott in
^ole or in part for traieees j^osen Iby th& grantee institution. S&e l^ovevah^jx-
1956 Soancil Mopted a policy, effective Jxlj 1, 1957* ilsitiag t&e stipend
^feids €Oifild "b^ paid © ts^inee* in •wfeole or in part froia a training grant to $3;,60C
^i*-,2O0, aad ^h-^O:^, for l&e first, seeoad, and tfeird y@arsj plus ;?350 T^r depend;
fels is a j^simun iiMt that n^y not t^ exceeded wit&out prior spproval .
TsBine© stiij^snd f^aads sa.'^ not provided on t^iiniag greats In Pediatric Neurology j,
sine® as -wili "b* iadieated later, trMaees In '^is field satisfy residency
s^gui^reu^nts ia Pedletyies prio? to efiterlng into Fediatsric Efe^sirology trainlBg
saad -^erefos^ or© eligible to apply indl'^ieiiaily for Special f raineesfeip
suj^rfe . Special fS^ineaafelp support is f telt to "b€ sore appjfopriate since
t&@ ffiwsrde are at a fei^er level -l&an laad^sx' s trsiiaing gx^tsit sad in line
witli t&e s8or© advaae^id stage of Pediatric I^arology traiaing.
'^bl® I IjsIov^ outlines l^e fis^sl details of grant smyport in
Clinical H®»ol6^ ^id Pediatric Hig^arologjf diiring Calendar Year I958.
Tsible I
CliMcal
Pediatric
Total
Amount
Ho. itesiint
Wo.
AiHoxmt
Ik,
Aative &^ts-»lg/3l/5T $1,559,255
PSPogjraaia A\mr3ed laitiel Support 52,^
Bew Pejdod of Cjssid'tsjen.t 750,^6
Dleappz^yved for eosatimiatioa ©i^pjrfc —
Teraical G8?ssscta 99,733
Coosasltted ceaxbixiuations 7X4,332
Active Grast®— 12/31/^ I^599.,@5
33
|^^^035
g
$1,5^,^0
m
3
23,7^
1
76,^9
k
22*
5
h
—
—
750j^6
gfe
J
—
..
99,733
ss
i«),008
2
75^,3^
31
55
63,T^
3
1,^3,^53
^
0
^3,733
i
1 6^,163
X
CllMeal HetiS^log^ ?apz%@entjS es^e of ^bie az^ae of lasitial
p^ogmadsag OBsSiasiSi. IJtyward the eM of ealerwlar ^ay 1957, tSfee rate of gpcwr&
of this program rapidly Secyeased. As cea "be seen fjpom fible 1, a total of
55 active psogras® vers receiviijs s^port oa BeeeEaser 31, 1957- BisafiKg
ealesidar ^rear 1953, tki^ee cev pTOgramd vere avtr^deS tx^ziiiig @rs^t aid "but
at ^3® saste tiiae five applleatioms foar coatiscuatioB s^^^^rt ve?e disapproved.
Sa?ea of the lattsa* teKsdaateS <m JiflSft 30, 1958, wi12i tie yesult tiaat at t^
ead of ^e ealeaSar }'<^ir 1950, the maSasis of gra&t mg^jwrted prograo^ resnedas
at 55 > ^I^iese pro^rax^ oas^ st^poz'tM at 1&@ lev«l of approadsutely $1,600,000
or at B3X average of $29,091 ^? ssogrsm. H&e largest grast is $59,009 asd
involves 15 ts-aij^ss^ "^^ ss^illest is $12j(X%), for \diid» as y@t co troissee©
h&ve ^esn srecxuit®^*
T^le n
Aeoitejie 'fear
7/1/5T - 6/30/58
7/1/58 - 6/30/59
"fear of trai^ijjg
AH
^ear of traisie^
1
2
3
1
2
3
eiiaical ^iimslo^
Sto. of Si?ais4§«®
88
Ih
3fc
196
105
83
6i
2^9
Average Stiii^esas
$3,^
,$3,753
$if,oi6
$3,522
$3,^2
$3,6T8
$1J,112
„i3aM
?9€iatrie Hiaurology
So. of l^e^tsees
2
2
3
7
€
2
g
la
Avegaa^ Stijpas^
SH5CZAL THAIlSKifllFS
/*
A® casa bs B&ma tTcm Sable II a'bovaj, 196 tnaiaees wers ia tral.aiag
for s career in Cliiaieal JTeux-ology diiyiag -f^e period of trainiag ensiiag J?>jse 3*5 3
1958. At present 2^9 3Jidivi<Saa3.3 are is training ead 61 are esrgnect^d to
coarplete t5?alnijag ia Jisane 1959- It is px'Ojected tliat eventually 80 to SO
iadividfemls will regalarly cojHplete trsining each year as specialista in
Clinical He^syology.
Pefiiatric ileuarology trainijog is as yet in its initial stages of
clev®lo|aB©nt . ^Is eonditioa is due to tfee asEiyked de»3Ptfe of persoasnal ia tSie
field capable of carryiag out traiaiaag progpan», ead to the feet tisat only
recently feas a demand for such hi^ly trained persoaael evidenced itself.
One of tbe importaat factors coatiriTMiting to this denjssnd is tbia recently
eotabllslsed Coliobosativa Study of Cerebral Palsy sponsored by tfeis Institute.
During Calendar ye^r, 1.93% one rmv pgog^^m x^as establielied bringing i^e
total active progratns to -^ree. ^@8e programs are being B&psorii&d. at the
level of $63j,T68 or an average of ^21 » 256 per progmm. As is indicated in
Sable 11, there are at present 10 IMlvidaala in training for a career in
Pediatric Keorologjr.
2. Contribation remiltlag froa x»gticulax srj&t&b
It is as yet too early to evaXv&te t^e contribation of the varioms
training programs. Eadi institution varies in its approach. Since t^e
product is tfee timined investigator j, only tijae iTill reveal the caliber of
a program as measured by individaxal successes, and failures in 1^© investigative
field-
3- l^^or j^roblema encouatgred.
JJp ymtil -H&e present tii^, t&ere has been a i^rked dearUa of
Clinical Nemrologists In academic positions. Sowever, as more and mor@
specialists in ^Is field complete training and steadily fill available
positions in t^e madleal ss&ools across the ooimtry, and private pra.ctice
opportjmitles develop, l&e problem of attracting iadiviAsals to investigative
carwors will beeora© Inereasin^y difficsalt. flie answer to tfee probltaa is
aailtifaceted and prinffiUPlly consists of providing adequate wapport of Clinical
n^u^logist c^jr'e«r Iscveetigators ei&d keeping t&e training in "^sis specialty
in Imlanc® vit^ national needs.
In t&Q field of Fediatrie N«arol©gy, it is obvioas tfeat -ttiere is
a EjaK&ed deartli of fei^^y trained personnel available to condsict txuialng
pro^E'aaiB. l&ea all available persannsl h&v® been recr^it^ to the training
effort, a lag period vlll ens^e lantil t^e programs can propagate -^emselves.
^^s, tfe© aeute lack of Pediatric IJaarologists will contiime for oany years.
k. Chaama ioad IniproveBjignts ssade.
Woxr t&at th'^ training prograaa in Sllnieal Wewrclosy feas developed
to a point Tf^r^s it esia. be seen \^lels progr^rs are succeeding in training
nearologiste and -v/lfelcfe, for one reason or ^aotSaer, are failing, eontiimstlon
support l8 being danled iSm Wim&emBBtvd. and additional support a\mrd@d to
ptroven pvo^cers. In tvo Instances, tkls Ms involved tfee avrarding of long
terminal grsmts. TtmSf to make long terminal grants tinnecessary, review of
eontinuatloa applications nov t^es pMce f^r enou/^ in advance so tlwt
deeieioa is kno'aa at least ons year before the tenninatioc of tfe« ciirrent
couBiitaie^t period- lifEder this system tfce only teanainaX grists awarded axe
for the pisrpose of fiaasEJcing the progreaji to tl^ end of tba aeademic y«sar.
Bj as atteEtpt to establish soite geser^ guidelines as to the
trairdcg likely to prodtss® aa investigator i© ttes field of i^diatrlc Wsusolo^r^
asi ^ feoc Cojsaitte® was convened on May 20, 195S, to consider the problem asd
offer eoacrete reeoiBEeijd&tioas. ffee Gomaiittee eonsieted of representatives from
the fields of Pediatrics, Pediatric Slem-ology, aM Clinical Haiaroloesr. It i^as
conclMed that as prerequieit® traiatag, ose year of internship azui tvo ysars
of Pediatric resideusy would be essential. Trainees meeting these cpalifications
are eligible for Special SlraiaeesMp asjards.
In rega?d to ttee prograsa, itself j, it vas the ecm&^sisiss that the
period siaould be a minSMm. of three yeai's^ eonsigtiag of Mmlt fetarelogy,
flliaical Bsdiatris Ne^^^ology aad sxperieace ia the Neurologic B^ic Seieases^
It vss noted that ia the ease of special t^est^ further traiaiisg ea^riejace
wonsld he desirable to aake it possible for a trainee- to develop regeasrcfe pro-
ficieeey ia his special area of interest, fl^ae reeannssndatioJEiJs were adopted
by the J'sase 1958 Council a^ a gaide. In light of tlis above recoanaeMaticsiB^
training program grant® In this field do aot provide traliiee stipends, asjd tha
10 individualg saow ia trainisg a3?e ^J. supported on Special ISraiaeegMps,
Prior to caleisdar year 1958? three graduate traiaiag grants ^lere
awarded ia fei^os'iS'gesy. I& Wi effort to elarif^ the maeser iji tiMch training
ia this field could be best g\:^orted hy this as^ltut^^, as ad hoe Qajsiitte®
was egppoiffltsd to consider laie matter, fMa CosaBittee jaet oiTFebnary 22, 1958^
end r®cc8sniei^ed that stipport of traini^ dairisg t^ residesicy period through
th@ atrara of program grasits did not represent a gesaral seed at tke present
tiiB^. It MOM the. consensus that 'vh&r^es t^re is sot a recognised need for bb.
iaereased anaber of private practitios^rs ia this field, there is a dearth of
trained investi^tors end thafe gi^pport by Re$ea?el5 Fellovship ^d by Special
fS-eiiaeesMp awards d'^irijsg aisd after tJis resideecy period wotald be iS^ laost
effective metfeod to aid leurosisrgery.
/^
Tt-Q^%m
setxFi
It "Isss beeoaje appB-rasst t'&at iihe evoXntlon of the CliBicsl Heurolog-y
grant-in-aid trainiiig yrogrwi -will consist of cuitivatioB of prodxactire progr-ccsSj
the aiding of those irith real potential and tlie cessation of ©appoi-*-c to
programs -whldh} for one reason or another, lisve failed to develop after a
reasonable trial period. Furt&er, It is recognized that a longer period tb.aa
tfee usual t&ree year training program is required to trada an investigator
in Clinical Bsurology. Accordingly, various Progj-aEi Directors are 'beginniHg
to think In terms ojp enlarging tfee eeope and capacity of the prograa in
order to be aljle to offer training for a period of from fo\ar to six years.
Indivifittais csa "be supported on Special TraineesMp awards daring iihe period
■beyond three years. It le expected that a core of t&e nnst pri^ductive programs
will develop along these lines.
It ie estimated in regard to Pediatric Neurology training tanat
optiimns groTrtis Trill prolsaibly "be reacihed ■vftmn 15 to 20 progrsaasJisve "been
establifilhed. Daring the next calendar year, it is projected t&st fiv© ntpsf
progreusBiflll evolve. As was mentioned previously, slow groirfch jresialts froa
tJse HsirlEed dearth of adecpiately trained individuals to establieSi progs^ms.
Wou tisat definite training gaide lines feave Ijeen estsblisijed, tfeis prolialjly
represents tfee greatest difficulty la t&ls facet of t&e training progrsm.
6. VolwB» of AppiicEtlona (Clinical Hearology emd Jediatrle
[—- ^■— — -— ~-~"— — — '
Reqaeated
Approval
fo Approvea
Councils
AnRxarfe'
Ho.
Affioant ^
Eo.
, AmoiintJ Ho . J
Ifercfe 1958
$1*78,301
16
l5195A3Jv
T
hQ,e$
113.8?'.
Jtoe 1958
^222,332
10
$150,806
8
68.0^
eo.o^
Hoveober 1958
■:?ao,5i3
8
$iU6,609
T
70.65&
&!.^
TotAl
$909,16?
3^
^>k$5M9
22
5^.5^
sh.rf.
f. St4tff Aaslgnment
aavy®nce A. Far"b»r, M. ©., Esetmtiv® Seeratary,
Heurology GraSimte fraisilng Grssit Coaomittae
19^Q Anmal Beport
of tfee
Hatlosiai Institute of Ueu3X>iogicsil Diseases aad Blimdaess
III . Gxiadsaate Traislsg Gie^^ts
ffee ps'iiBa^ pss-pose of th© training prograses in this area is the traiaiBg
of postdoetoysl lassie ecieace personsel for careers as teacfeer-iavestigators .
fjrograajs ar@ b®iiig estslilisfeed ia t|j^ "baaic departsaents sucfa as latfeologyj
fiDStoaiyp pteyaiology, ^arsaacology, acd biocfcieaiatry. ^eept for patSiology moai,
trainees will prol^bly be fte.D. 'a. It ie hoped tfeat ti^iaing pjrogs^aas iri.ll be
broadly enoag^i oriented so sa to produce a!at«re> protoetiv© investigstors .
TitB fields iacludesd is -Qiis category are Hearopatliology, KcuroanatoEiys
Neoxo^yBioiogy, SestieojfeBXBSiisology^ and Heia2«odiensi8t.Ty.
Table III
Frogwuas
ActiY® A-TO2ded Active Ia©3?©aae
Grants Initial ContlE. Conanitted Grsaats lOiJiring
12/51/57 SteppcMTfc grants Ooatia. 12^31/58 dy_l^
{«») C8) Cl) C3l (12) (8)
H«»sppa^ology $ 96,C?2i $151,1^9 $ 2?,©93 $ 59.536 ^31,7Uo $l4i,719
C3) C3I (3)
Kesssraamtomy — 823706 »«- --« 82,T06 82^70€
Cil CD Cil (k) " (3)
Nmso^yaiolofisr 2J*«tT9 lOk.QkB =«« J|0^503 lM»,55l 119i.?72
m CD Cil (2) (1)
118,650 29,^6 =»« 305 56J* 6D,HfO ai»U9Q
Cl) C3) Cil C*^} (3)
l^msodkmLnti^ 17.^ 13^736 -^-° 11^|85 ISO^ja 132,l*CU
(T) (16| Cl| (6) (25) (18)
I. ggograaalag AeeoaipIldaHMaits
fyloy t© eftleodsur year 1958, t&e»« were foia^' tsminiBg progsttms 1®
Re^iisopat^ology^ nos« in Hssasoematoaiy oM ose @ad& in H^gaxopisysiolo^^ H@t£7o«
T^isvmsolosff and Bmroe^estlst^, for » totod of seven, flis px^gram r<epresecitB
«& w^ of sa^or progswQliag @q;i^Asls during ealeadar y@ar 1^8. Tfee typ@ of
p^gr«m sTolTlsg is mia ^a^le 8ei«3iicc a£«a Is broad 'bftsedj, designed to zaot oeQI?/
sl-loxf ax5 inaLivid-jsal to attain technicises!; gkI Jmowledge in a narrow area of
Ms OV13 particular iiD.tei-est, but. also to siske J\ia avara of the work teiiig
cerrie^ cat ia each speetrvim of Ms field of eadeavor sxA broaden his horlsos-
ffcis i.« some-what different tralcing thaxi that eustosja^-ily raceived by a i.vmMrm
B©rviag as a reseansfe sssistaBt or fellov on a researela projee.t.
Ei^t 5ESW progi'aza gi-awts were awarded In lejsrop&tfeology Isrii^giag
the tot-al to twelve, Siesa prograKt^ ©re supported at the level of $237??^ ;»
8Si average of $19,812 per program.. It is th® "b<mt of our consultaats tfcat
the mogt desirable prerequisite trsdning in ttoee years of residency in
geaer&l patfeolcgy. ffeus, most in^vi duals are eligible for Special Trais^g'^sfeip
support aad accordiagly vhea greats are awarded ia tMs field, tiiey provide
at mo®t os« trsinee stlpeM for the occasional traiaee -who for one reasoj^ or
aaother i§ not eligible for. sueh aa award. Ci^sequently, the average traicdisg
grant in lietsropat^logy is somevtiat smaller in amount 6xts to the absence of
larga suats for trainee stipestds.
fferee nsv pro^aaas were egtaS)ligbed is SteuroasmtooQr, ttosse in
IfsMTopSsysiology, om: ia feuropharaBacology, aad three i.n Seurochonistxy, briBg^-
isg th^ total programs in these fosar f i'^Ms to 13. fhey are snapported at tl»
level of $^38*718? aa average of ^33?_7^8 V^^ program- Sie basic scieiMe prograat
graat^ ia thes© f ieM^ 8or« saaewhat M-^smrf dise to tfe© ased for fus^ to
provide specialized equip9»»i aad STi£pplies as well as trainee Etipe»d^.
Since tte majority of the basic seieace traiaisg prograns wier^
cstablisijfid d\jriEg the latter half of caleMar year 1958> tfe® aiaaber of train* sg
is as yet quite low. i^roadn>at@ly 20 l^lTlduals are in traisl&g for caa^ers
iE ^ssropatisology. It is e^ttaated t&at a^ro%ls&&t^ly 20 or more are in tralEiag
in the re»Rl.alBg 13 basie gclesce prograaiSe Mthomg^ tha mimber of traiaises is
atlll relatively ^sall^ this progrm has lRer6a;ial by 3^ ^^r cent is ntanber of
gra^g aM by 3^0 psr cent is. tbe amount of fmids involved d^urisg 1958'
2. CoatributioE^ rgaaltlEig froa pagticsalar awardg
St Is aa y©t too easrly ia tfee basie seiessee traiBisg progrsa to
eval'K&t® it« eoatributlo^. K&ip)®ver, siiy;» tfe@ prodiaat of this ar«a of
trailing will b@ a ba^ie «cte»,tlst, tiie private praetlee estieeH»»t do®s iKst
exist. Zt is believes that alnost all individuals traiaed will sntrnf into
ftsil°°time imvastigati'^'e csis^em,
^* !^^^ pyg'^^.^jg eaec^mtered Ist pgpgraaaiag
Major pfobl^as^ es^o^mtsrad in pr«3^amiag in these basic sciesee
arsas 1@ the Biarjted dearth of highly traix^ per@«:m%€l avalleble to carry o%t
postdoctoral traliilsig. W^r^ag 8lsu>st all gr^uate isstltutloes provide traln^^
i%sg ia basic seiesiee aareas for th^ doctorate^ mily a vary tmt im the past hmvt-
beea imteTested or have deveeloped postdoctoral prograsss-. At present all are^^
poteiQtially enable of estabXlghisg traisisg pro^^^ are being eceouragsd
to do so.
Secniiil^i^t of traioees to estabilsbtd t^r'a^lsig prograans is
aaotfegy uajor problesa ©seota^tes^ed by the vario^ ^g^graia Dtreetors, fM®
problem ha® as'isssi partially from ttee lack of Jmowled^ that pOiSt^octoral traisiag
opportunities exist, bat primarily because of tbg dearth of Pb.D<= j^ipgoaseel to
uKdsrtaks advasDsed, highiy«>epecializ®d trai^l%g. fl^ou^ aanouseeaients i& th^
literatm-'e sad disty^'ibistioii of 'i>T(yd:rvi'sNi&, t»:e se-sraitaiest -prdbi^n la
gariidssally Iseijsg eoKcra4»i'^.. Favorable r*su.lts are very great in sorae
cases, teus illaxsti'^ting a deiinite Seasand fcr saoa training. 'Mcvever,
the primary proxies; atiii exists . It is expected ttet %\b predoctoxal
tralciEg scs.pi>orted. by -tfe* .^Ivlsiou of Geaeral Medieal Sciences will
p^^ilaiiy sojA'e t&e sit^isatiosj.
k. @£aogea aEd Isiiprovem^gDts ig prograias
Witfe the atelft of pi^Dgravaing emffcasls to tfee basic Bci®ns#
ar«a, it beeaase evidsat t&st t&« Nassroiogy Onsufeats TralciEsg Grant Ccnsmittee
^icSi consisted largely of slialcal ae^sirologists ■wo^aia teavis to teave tlbie
l^sEeflt of advice fron Indlvifeals w:Lth apeciai basic scleace coaspeteaee
fHierefosMij, <asBtri»g 195S* & Keuroaaatoraist ssad a Bfimroj^emlst were added
to t^e Comaittee, aad a Nenr3]^ygioXogist has 'been nomliiated asd is
isxpected to h^ appoiatcd in W.p, near fsat^ire.
5- Program Objaetivee
It le estiuatfed t&a.t. 1b order to aie^jt aatiojsal Keeds tfee
eatabllebmeat of appztoxiaiateiy 20 trsiiaicg ■progr^ajsB in esc& of tSse five
iMiBle seleace aress will b* K^ceBssa":-'. ffeis woi4ld rcsmlt in 25 to 30
ifidividsaels ssompietlsig tral&lag in @i»<^ field every year. Baring t&X'
n«xt e&leadar year, it is cacpscted t&svt four or five new progrsaaa will 1)6
tatabiisked Ib etaefa field. It is (pslte apparent tM&t since this program
1b as yst in its iaifelai stages j its Impaot upoa the research effort will
be slow is d«veloplag. Oadoubtsdly tjfeere will be a coasiderabie lag in
growth ^jatll t&e eetablisSied prograsic. pros&ee enoegfe trained investigators
to sssectiaiiy propagate tfeensBelves. I>ariag t&e next calendar year, two
aiffis will b© hl^ly sigaif leant. First, prograaing will be very aetive
If t^e tsmt^T of programs indicated &.bove a3<e to be realized. Second,
exist««t progrsms will b^ developed end aided in all possible ways
QomiSiln
iSI©ijS€st®d
App:f0vai
^ Appsov&l
Ajttouat
No.
Afflcunt
Ko.
Aincunt
Nc
March ]9:?8
J?sne 1958
Hovffifflber 1956
$2l6,i8?
T
?
9
^227, 3S3
5
6
8
55.6?^
5?. 5^
6T.9S^
11 M
88.95i
total
$aii8,283
23
a.'„r..v.
$511,229
19
60-1^
82.6^
leavsiBase A. Fs8%®?, M. ©.,, 8xe®stive S^sa5?<etaryj,
;, »d Atmual Report
nattcjaal laotltute of ii«i^^roli3gi€aa, Bis®a»©8 aoi Bllnto«e»
*-d''^jS%© I'rsijsia^ Grauats
-■' feo^^_ «ceomgl£shm@ats
In tli# pus'Butt of the go«l o? fievsio^tng aM malntaiaiufi
postdoetosai ts^tolag fg«^«Bi8 to t»ain eaap©ey t^Ksher-ia^teti-
£^targ la oj^tbAlaologyj 195S 8*v aa tncf«a»© of pyo^«BS fe«8
35 to 380 T3i©s® srog^^eas ftovM© % total of 2$6 tsminees wiiai
aprroxSmately 87 iadi-^Miisls |f©? yeag> finishing g®sid«ttcy
tyalsiinc ^o «^^® pot,@tttial teachey-intrestigptorso Ki® tte^i@
jaw sg©0?«B® ia4tiftt®a ia I958 loejre aM@£ to th© eliaieal
'^thiOadilogy ti«.lBliag iro^«a besaus© of th(§S.r Jwtggi pot<ssjtlAl
for timialag persom®! »limtat®a toiwi*€ treeeftreb easng^fs. Tb^«
wai. a EmsU aa&sunt of ^<jrfeb of tb© eltaieal tsainlBg fTOgy«K
iuMaig; th© yiaypo Thlst@@n a^iicatioug for a y@qv.®st«si ^92^039
w©r© ffe€@ivea, aM of thee© 9 werfe ap3?rcivea f®? a total of
$175,ij62. Wlthia feh® same peytofi of time ficasseiaS. guppcrt of
thr®« ^-©sraiBi® was ■ ai@CQCtlnu©4o
a. e»tgtbi:tions gesultlog tstsa^ yaaftieular aiwgias
Bu® to th@ short, pearlo^ of time tb€ c#ithalaQlogy tsaintne
prograaa has betn In ap^ratioa. It Is not yet possible to singl®
out ®p®cifle ^o^Httss iA>icb ar® outstajoiing la coafiist®atly
pyoSueijng s^en of high eslibrst as t^chejp-iavgetlgatorSo No
pyogjaa has yet ru« for a loog «nou^ period to ©stabllsh a
tiMition of tralntctg tsacbsr-lnvestigataps aM thug attract
ap^i©antas pieiamrily iixt©i?est©a in sueh career© «M train theas to
Uim llait of its potentialities o
3" Major ggpobljaas ©a^ouat<tg^
Two major ^ohlmm havc^ hmeu ©B.eourit®rs8lo (1) Th# slow
®voltitloai of progyaiB® of tsainisa^ fyoB thos® trsditi<»aly
orgEUiiztd to .d©v®lof eKtri^reaeiifg lato those with suffleiiat
flexibility to i^wrid® ejEp€rlene®8 , especially In th# basi@
sci@nefeSj nsceseasy fc®" txainlDfi t®aeher«>in'restigatoriSo (2| %«
g.#eoea froblaa is ixshereat ia th® fi«ld of ofhtbalmology » that of
y^eruitiag Indiriiualg isato acadenic ca^mmre.. 'Vhm lee^eulog of
6hie ®®caoa problem ts«ou.sb th« passage of tiB*s is psMietoi as
fiswag reseapeh tsaiaM indiviauala h^ecm® Proggaai Directorg aM
iaH-u^cc® tsata®gs in tb^lr prograjtt&o
As of ths May Ccsaaitt«« sievtlDg^ ]C^« J«f>aae
&as«ufciv« S®es?«ta3fy of tLe OpiithalmoloGy Gioduat® Tapalntnc
Cremt CctaHltt«to Ills appointA^nt acbi@v«8 au adteinl^'tintiY®
organizatloQ «&dch ps-ovid^s conetaat c?^idaae«i of and attsatloo
to th« d«tail« of Conndtt«« bu«in«ss.
In th<g monUi of Septmbm? a asBall eonf@rea@« via« h«M
of pxt3f«s®ioi3al perecsmtl lnt«re8t«A In th« subj«@t of olauecaiao
FartleipeuitG v^xm Dr. BfBleyy ])r« B«ektr; aod Bro SIia.ff@ro A
geccmraartfinticai >Ae fos^uas4«d to Couoeil that a e«f>l«s of fiv®
or six yearly v^AiLng eoofsreoesfi oa th« 8ubj«et of glauecsa
b« h«34 becinnin^ th® aead«odLc y^ar I960-6I0 It ■ms Invlslouied tlut
th«y b® laterxutiooal in ehajpact@y, Inclvifte both clialclans and
basic scl«ntiste» acd that invitatioua b«i iseu@d to a anon number
of partieipoats wMch probably would not uiaab«r ctvsr 3O0 Tb©
Nov®Bb(ir CouneiJL authoriiied the hoMiug of sueh a s®rlffi®«
A T^roc^^om Dirsctoars Con.f©y®ace tias b«il4 in th® moath of
January,. Thl« ya@ att.«ad®i by agproxlfflat@ly 10©- invit««So Tfe« aa.y
uafi sptnt ia dArifyiug th® goals of tb@ ti^lning graat piro'graffi aoS
diaoiasiiag bofUi SBSthoSology and eoatfmt of ellzilcal ai^ basie
•ci«ae« a«p«cts of tjaining tiachtf-iiiViistlgataffSo
Porfey-tbf^g youag, pot«atially produetiv® tnv®sti<^toiPS! in
ophthalmology w«r® gganted fiaoneial help to attendine the l8th
Inttsnatiooal Con^@0« of DphtbalatolOQy h@ld at Bruss#lfl!j Btid^sium.,
Th« piijfpos@ of this was to provide «x|»eri«nce and etiaxulatiou of th@S€
youn^ investiffitors at a cidtieal rtage in th«i^ Hevelcfpni^nt as
Cliangss iu tb@ p«r»onn«l of the txainiijg grant Cciiattitt«e
occurring durirjg I958 are as followBs (1) appoJntatsnt of DsTo G«crg®
LssaSoy^ Assoeiat® I>iaji of th® Biologieal Seitsaets, IFiii-ffigrslty of
ChicagOj to the CoBwit«e fear a four year t«2a, (2) appotataaent of
STo B«raa£igl Beei&gs' to th« ehaisRaonchip succeeding Dr. Al.son Bralsy^
and {3} T&tixmimt of Dr. Braley f^tm the Qaamittee as of
31, 19580
At its Nov®BB(ber neetiaG the lldtioml Advieory N«urologieal
Diaeasss and Blindness Council adjusted its ceilings on trcijie®
8tip@ad« f^cB $3^30O« $k3X> and |4oOO plus $3^ psr dopendent for
«ach of three ysare to ^h^OO, $5000 ajad |550O plus $500 per
d«p@ndent for «ach of th« thr<^ y@ara eonetituting the rsgular
3!«»ideacy ptslodo
In October at th© r«sque«t or th® Dirsctor.^, Dr. Braley
sp@nt 10 days viisiting institutions In Eiigland^ Scandinavia^ aoS
on th@ continent vhich train ophthaljoolosiets and hav@ faciliti@s
for ophthalffic^ie resoarcbo Wail% thm trip utas made with thft
prinaxy goal of evaluating ppportuaitias for the trainto^ of
special Tr&ia««ieij luforBtatioa of valu« to th« eauSuct of th@ oi^ihtbal-
ittology tgnining ©rant ps=OG3raa> «l»o wa« catUered.
Progyaa Objeetiyea^ for 1^5g
Th« pH]&ii3?y goal for thl® yiar wHl b® to ictjcsoti© the training
of grsatftr 0mb«rs of t«a«her»lnv«etlsators by Inesr^atsing th» jteaw^nt'"
age of tra.ine«8 twva, present programs who e«l@et this typ@ of ^sas^mro
This idll b« achieved by dlseeainatinG the goals of the progrean to
tyain®«is through the cooRperatlou of py^s^nt Tro&nm Dir®etors» eSt®
visits la ectinsction td-th coatlnuation aoS su5j^«mtntal appllcatioasj,
aod 41is««tlne attention to the organization and eontsnt of tiralning
prograois vhi«h appear euecessfUl in pfodueiiis men orl«ntat«di towaM
ear@«yfs as t»aeh@r»lnv«sti^tors father than as entrepreneurs. Thee
abov@ goal in no way rul@8 out aMInc proGreoos which appes^ likely
to txaen out highly traln^ Q@n of th« type d«e-lr©io A 8«eondary goal
of th@ ecntsg y«ar is to e^.th«r general idaait ate to th« types of
training programs and areas of spscial cn^phaBlii incorporated in thos®
vhleh nay have ip«ei&l elgnifloanc® in Ssvslaping '&^aeh@r'» Investlcatars
Staff _a8gigaafeat
suMMAiry OF counceLp actions
Ca^tbalaology Aaolleattoaas
Itfcb
Aato Reqo /totto Agpo Moo Reoo WOo Atm^
.«w or Revl»«^ ^7777 ^30000 j— *— -Ji&.
ootlnuatlcos 20304 20304 1 1
ajrpleB«nt« £$600 12043 2 2
atol l93aai |ot3i^7 "T" ^T^
i Amto Appo 66 ^ NOo Aiqpo 300
$17280 2 1
10800 2 1
P 1_ 0
5 T
9w or R€vis®i
aottnufttiottiS
Jtal
^ AKt.., ApfSo
$75031
^2826
4320
11^79
21
sw or Revtsad
?ntimia,tioas
)tal
)» Aatc Appo
$ 0
36320
29S59
$65979
9^
^ Koo Appo 40
$ 0 0
30320 1
25659 3
IS1579 "5"
^ NOc Appo 69
(Mm TOTAL ;^^39~ ~™ $1754152 ' 13~~ ~
'-^. .. fj tloo Appo 6^
1958 Annual B«sport
of the
national Institute of Neuroloslcal Dlseeuies and £Iindaeo«
3XEo Graduate Txuiniog Graat8
9„ OtolAyyngolOjgy
1. Program Deyftlopaente
This y^r 19^8 saw an lnc?«ase in number of jirogamais traa
6 to 180 While this Is not ths grovth s«>oJ«ct<^, It still is a
sizeable inc?«ag®a As of Secember 31# 1957 the 6 programs wer*
financed in th« anount of $101,976 and the new proe^ajas establisheii
increase this total to IS^!*©,?!©. Present number of traiiiees total
116 of vheai 25 ar« in th« third yaar, 9 ia the fourth, and 1 in
thsB fifth c At th# present rat© of flow of trainees ^ b«tvs«in 20
and 30 potential tfgacher-investiaatcTS will be available each
y«aro Far %hm next few yesss'® this figur® can be e:q>ectfld to vary
ccffitsiderable as aev prograais vithout an @vem flov purtieipat© in
training. Also, for th@ $est few y^ars many residency pro^'ams
in ototUxyogGlofiy \mv® filled mostly with for^i^nerso As
Aseriean citizens a|!ply in Sjctcreasing numbiers fos^igner's applim-'^
tiaos ar@ usually ff^jectefi by Frograoi ©Ir^ctors even though th®
total rtisidente on a service total fever individuals.
So Ccotributions resulting frcn partimilay a^sa^s
It will be a tsatter of scxse years before outstanding
progreme can be identified. Stable pro^^sms with a eoitinuity
of truining may now appear to be outstanding only becaus<fe there
is not a sufficient maBb^r of canpetitive prosraas ftar valid
cctaparists^ 6
3" Major prdblans eneounterad
There is an Insufficiency of adeqiiately trained Ijadividuals
to take over positions as- Prograja Dir^ctoys and it is joredicted
that this situation will ccsatiau® for at least 3 to 5 years. The
economic lur« of life as en entrepreneur is well known in oto-
larynco3.ogy as the need for such speei^iets is extensile » As
yet there is apparent wide spread Meuoderstaoding of the goal's
of the Otolaryngology Tiraining Grajst Frogrem. Soaie of this
otisunderstanding is aorm apparent than real. Gradtially, however,
the point is being driven b«3n@ that th@ purpose of these gzrnits
is not to train private practionere.
^^ Changes and impraviBnents in proGyam
The establlahiaent of dinieal Audiology Prograas ha® been
apprcachM ulth cauticm. This has been necessitated because
naai*4iiedically atti non>researeh orientated predoctoral prograas
in speech d^artmgnte are aiixious to e:3qpau3d into the postdoctoral
training field without change in orientatioo. Due to ground wo^
7<-?v«»»-, sr^; n©*i«^ o:-.li06i. :,^iu-iM V^^ Ca«si»te« {>ctvb«r i f-^r
. f't.'jp ;m.r tes'sa.. Br. Glfti ^^^eviar tuf 2Ylac«toE. n^« b^fc; lii^'l''«®d
..c aeeer.t C.:-j»9Bdvte® 5«sB>i»r«l:lpj, TM-^i Dr-» Soles of th* 'i/al.-.-aifsivv
..vf Ki:.o-.««:>t.-A>, l:r<. 5oey oi' tne Ifel (r«r si t.y ot CJallfon^t;? «i'-^
i-f'(.)Si«r p«r©i3im@l of this type 1« of prto© ii^parS^AXse® «:-4 '-'.s.; ■
r.ie?^S,oji t.« t© be a'^'o.il
■:*■ afxtr Jul:' i<, l-^'v v-ili be ir'«--^'-,irrf* to «£aats>l«-.« -^ rcft.'jf y^j-r
At i%» BeptOBi.-ef a®«tl8a,3 th« C'-JBml*t«# •. ■- ©A '».-. . ■.V.4 ,.
2*<tt«ri@d ft.ftsy "u:n@ fosrasi-t ■of the St 5 I'^ovis c-nnf^T^n®® livs
tf'js. t-'re /tJTteiee F.^csso Fi-uittla,'s fo>r ■';'-i6 toy C'o::>fsx«E.<:ffi Ir-
;aiii*Jy-- >?ja a. trip to Is'^laaiS oaS Sca^idlaSvie. ;,
l?l'^^''S-iiki '««tiv'e!*__£iOT- 10 ;>
i'^it.ely ;i; 5r-.vatii is «avis!tmi@a, ?hisi sS.?®® aat Ir.sl-ud® tb@ pre, ■?:
S^ t^o elt:'si«&X a^j&iologj' progra.-JU8o I'o a.ce«js5)lish this t-.t t:-€
|^4fe#®iit St«a>,u8 of a'/ails.bl® Frotifftixii Dl.r@stC6rSj it win b® aa^sty? :
nay b« i|^p«ad«d upcm to bmkp In aehl«vlng this goal. Outstanding
trainees twom gira&t supportM progs'aas vUl b« waeouret^®)} to tak«
further training &s Special Traiogcfio Doly a vcxy f«v appllcatloos
hav« ecne fIfCBa otologists for Sp@cl&I Train@@ahipa and this timlA
ne^AB th« hij^y traiA«d ImtlvHiual which ean be sxptct®! to bs
d«rv-«lop«& aft a recmlt of five c^ six yga?e of tralnlngo
60 Staff asslgaatent
Dr. J^roM
Nsireh
New Off- Revlffisd
Contlmiations
^ Anrto App« 26
SUMMARY OP CamCtL ACTIONS
Qtolagyiagology A:^plieaticaaig
Anto Rfeq<.
$300302
0
l^83'32
$77367
0
|B539f
NOo R«qo Noo Aj^^
^ No.
8
1
0
9
Appo
56
4
1
0
J\m«i
Contlmi^itlons
^ Aato Appo Mk
Noveab^r
1 991^
0
I809S
$a6i6
0
$ te660
0
■f5ao6o
0
0
0
3
0
2
^ No, Apfpo jO
2
0
0
2
^ No» App» 2
2
0
1
3
New or Revised
CcaittayatioQ©
Supplaaents
0
0
0
Total
f^ Aast. Appo 0
0
0
GRAND TOTAL
5& Atato A|Jpo 28
■■ l^tBiD ■
^X37^57
16'
,.i.^« ^^.,50
"■■"■■B
1958 Aanual Beport
of the
NatioQGl In8tit^Ite of Neurological Die«ae^ az^ BliMneefi
IIZp Graduate Training Gstmts
lo SottBory Fbyglology
1. Pgoggaia aeeeagllebmgnti)
Barly in the divslosiBeut of tbd Nsuroj^ysiolQey TTalulng
Grant Frograa it bccaiaa appas'^at tljat it vae not attraetiog the
intar^st of certain individuals vhoe® main areas of endeavor ve:r«
thm ssnseso In order to interest theee in training, tiae Senscay
Physiolc^y Training Grant ^ograia vas institutod during the latt«r
half of 1958. Thie is a po8tdc«toral pSKjgram design^ to train
basic eeiantiets for highly specialize irtsearch internets « The
area of the spseial senses needs cultivating if significant research
studies are to he expected. While the oain goal of tha Sensory
Phyaiology Program is the training of basic science personnel, it
is not th® plan of this training program to exclude M.Do's ftrcm the
progromo
nine potential training institutions have been contacted to
ascertain their interest© tojward maJdug grant applicaticaas^ Thes®
nine include Interests in audition, vision, and olf^tlon. At
j^esent, four aprplleatiaue for training prograffls in this area in the
aanount of ^^9,^75 are being readied for review by the Mirch Council*
The University of Michigan has applied for a ©rant to train both in
auditicai and vtsionj Florida State University to train in dfaetlonj
University of Colorado to train in olfaction; and the Childrens
Hospital Society of Los Angeles to train in audition. Applicaticnji
will be reviewed by either the Otolaryiagology or Ophtbaliaolosy
CcQimittee or in ecoe eases both, before Council eonsideratione
2o Contributiong resulting ffcm particular awards
Hot applicable
3, Ma;}ar problens ^leouHtered
It has been the policy to avoid ai»y real or apparent conflict
with the developnent of Neiirophyeiology Tsaining Grants. Where
overlapping interests of potential Program Directors oecu?, there
has been hasltaney csa. the part of sonie men to apply for grants.
iVp Changes and JB^^arovgaenta in proggeaa
Not apocO^lcable
Pro^dia obj^<?.1;ly:t« for 19^9
A call of 10 -ETOgxaxoB by the etid of 1959 ha.8 b«eaa e®to Tli@e«
psfogyaasfi to cov«r the areas of audition^ visiooj and olfaction
(including tast«)«
6« Staff aagigiiaBent
Dto Jerosa®
of tha
national Xastltuie of Ilmirolo^coX Di8«M8fis and Blindaese
IV SPECIAL ^PRAINSESHII»S
1, Pyograat aceoB^llabaaBnts
Subst«iitial sdvancts b&v@ b@en mad® during 19>B in. tb<ii
developussit of this program vblch foms th® cap^ston^ of tb@
Institute's total trainints program ty providing the advaacsd^
highly specialized muSL diversified training indispsnsahl® to
the preparation of & coaip®t«mt invasti suitor.
A total of $905,750 vms at/supded to 125 trainees, an iacr®ftst
of appEXjacijnately 50;j ovtr 19^7 <> Thes# trainees ar® located in 50
institutioas in the Oaaltigd St&teo, Canada.^ Europe aisd Sowfch America,
undiir th« direct g^danc© of 79 cotstm^ding investigators in th@
neurosensory fields
On® h«2ndr®d of th&a® \rmr% first ai^rds to thost vho had
not r©cf!iv@d sup^rt previously, and 25 "wex* for th^ eontlnuation
of trainlEe already su^orted for 1 or 2 yearso
^«se awards are "btias «s«d by investigators at all stages
durin;:; their canterBo *Jhs postdoctoral @xp«ri®nc® of ajjAioants
for Special Traineeshipe this year raagtd from 3 to 21 years ^ with
an evtfrag® of 7<.5 years. Although, as always, th© uaajority of the
ap^lsantc wer® satdically trainied, lncr@aa@d e©n®ral auargnees of
th« availability of tMe award to basie scigntlsts is reflected in
the 10 awards aad® this year to per eon® holding th@ IlioD» d@gr®«c
Anatomists, biochemists, ^larsBacologistB, aund psychologists
have h^&a. th@ first to apfly for and r©ceiv® support for advancing
their knowledg® and sitills in the neurosensory asptcts of their .
disciplines o
la the fsc® of an acute need for seuropathologtats, aeuro^
eh«ial8ts, aad thos@ trained for research in the ne^arolosical
deficits of the yoinsg^ the Special Kraineeship program contributed
materially to training in tl^se ^p areas s I6 awards in neuropathology,
7 la neurocheaistry and I8 in pediatric neurology were tade in 1958,
which doubled the sup^2ft given in 19->7 in these areas «
mch of tMs lacnsas® r®fl«cts tht tacfaawiicu of th€ laotitMte's
frsiaisg Orsat frog^^m la tfats^ ansas i&lch ppovid®s a,]^aie«atSo
A coi::^?#j®asiv® report* on th(i dc^vwljjpiii^nt of th« Special
ftMai<t0hip pp&iSMto, frou its inespfeioo to Jtia© 30^ 1956, sad an
analysis of ite pj^sciidujfSs ©sd aetivlties during this Initisl pme®
xme paresex^d this y^jf s^rvljos *o p^o'ld^ <tet®il®d ittfonaatioa for
our advi8®rs &nd eojosultants on this p^asg of th® ITationsl Iaatit«t#
of !lem''oloc^c&2. Bis^as^s asd Blirtdnfi^^s ©ctrsBit*^ isn^srsa, ai^ f@r
us® la duidliig f\itu3!^ ppo^'Sn dtv^J/.jpagnto
2o ContyibMrtioaa resultlae i^ou i^irtJLeii;dar a-tiaMa
Iter® th^a 60^^ of tbt Sps^jlfsl f r^3a@«e sta^^rl^Jl to th© ^®s®at
tirae ©;pi stlH in traialac statmi, sad thg rsmaindsr (;onclttd«<l trainlag
a© loag®r thsn a ygar agOc For taio x^asoa, inf oms6,t:l on has saot yet
b#«ii aftth®y®fi syst^matlcsU^ fx«j. thii ©poup, &s to their subsequent
©ctivitieo and aeeomplisbuisatc.. now®v#r^ urn do know that a. large
SK-oportioia of thos® vho h&v® coc|jl.#t®d t^tiialng havt bt®ia selected to
fiUL €iGad@Ld.€ postS; sM tlmt r»st of th#s« syr@ uud^rtskiag liid€p@ad#sjt
3=. i^or-^Qhl&as @Rcount#i^<'
^«> cajor problg^'^j he.« beta ®nuount-#s«d this ym^, both
relatiaa to th# 2^vi#v «md f.j^o'val of a^ille&tio£Sc
Thm first pr'Oleia t-^sultsd ft-om shifting the Sp^ocial TrsltmrntM^
a|i©lieations fron' 6h@ llatiorjal Institutt of H<iu.roloci®&.l Biseffi,s©s aasd
Bliadatos fx-Mja^-'sMp Revieu Bcami to th« 3 fraiaiag Co!'JLiitt«@s for
rtvl®^.^ lJba». ^"3P consultante \i«pe drairn f3P9in th# s^oj' scitntists
of til® Iaatl^-«^®'s intWBnuewl a-^aff, jni!®tiase could b® &-ifx®iig©d as th#
volujr* of ^vpUcstions r«quir«d:, It ho« h%mn fouad thst restriction
of th# rfr-^'^f of as5iUeatioa0 to the 3 taaeetings held yearly by @acb.
#p-i»rt on th© Special Emlas^ship ^o^ma from its iacgption
.o Jmm 30^ 1958| loC« Hfesr&aaa, 9/15/58.
tKttl»ins consoittes x«sult0 la uadu» ddngr lA acting on a coa«id$z«bl@
nuBtb^r of r^ueste. Xt tmeaem lticv«ulngly a$par«nt tbat ouch dslaars
would soon 6«riou8ly ImpsAx the osttfuloess of the -gseogpemao Accordingly,
f<^ thm cbttirnan of each txaini&e coianiitttes to
3 iBeBib«2°s to smx-ve as an ad hoc Interim Special Sralneashlp
Coii8i!ltt@«j, tiblch voiald considftr afplicatioos b«rtv«a2 thit rvgular full
coBBsitt^s mMetlngs^ as the trolums of a]p|illcatlox^ »sccifisitat«da In
addition to aacj^ditizig astion oa a|irglieations^ th« laterlm Conmitt®®
cottXd ajjao sm^m m& iis^ortaxrt function 1a coordijoating program policlse^
eus th@y d@Tialt^ in th@ ar«as of special, i&t®r«Bt x^pres^attd, axsd la
proiaotine; ssatual uM®ratanding "brtv^sxi tbm %hr<m Graduate frainii^
©cNnsdttemso fhio ffoc^ur®^ stiH x^fsi>a^m6. as an ascperiioint, has vamt
with aotm resistance fs^m cur consultants, pertlcularjly those on th@
(^thalmology ma& Otolaxyngology Coimaitte@8» fhls reolstaace st^xm
largtly faresm th« disproportionately sia&ll amib^s' of afg^aicatiaas tn
th@8@ ar^£ as coo^iaj^d with thoaie In Va% nevtrological fi«ld, ^li
m^m^ixmAt i» ImlnQ contlnutd^ eincm it is ballavtd that this ■ppocm&ur®
will p*ov» to i3® to the (advaatag® of th« reviev |ffoc(g88 as a wliol©o
Tbm eacoxid fxvhlsm aros% heeauA® of thtg incs^^slng nmAmr of
apstlieatioss to go to Europsem ixastitutioas for trsdninso A &e^&t
deel of heeitancy ym^ f«lt' by our consultants In r^coQsnsading afprovssl
for txalnlng at institutiona vbmxe facilities and conditions of
txwdnii^ wei^ largely unknovn to th@sio Xhis problgin v»s ingt by
azTSo^Lng for the chairsaeB of the 3 training comndtte^s (Drs^ Bordl«y,
Braley and Sahs) and th@ Chief -of thg EstranBiral Programs, DTo Seger,
to visit a considerable nuaibsr of European inetitutiona where training
ia aost frequently r«^ue8t@do Tim observations of this croup are heing
loade Bvail&bl® to th© comaiitt#«c, snd should result in more ©fftctive
revl©u of these apgpUcationSo An add^d divideisi of these visits is
an incrsgassd underw^ndlng of our progran by the sponsorc who "wera
vislt#d,
h<, phsingts aod_lEg>yo-ym3igHte in fgfigcaa
Thm only major change in th® prograja this ymir has bgen en
incr^sa in th# aaount of fm^B eaxTtcarked for -^is ppogram, ftoia
$1^000,000 to 01,500,000, T,^ch ms de«ni@d mecassery in ord®r to
mmt as®ds stlmlatied by the taqganaion of Institwt® srosraraa ia
IsaslQ sci^ice and hearing and eptttch floXdSo In addition, this
iuer@es« hsiJi flOIoviid us to uteet th« needs of an increasing ntmbtr
of tminoee desiring renewal of awxtba to cou^lete a pro(£raQ of
training on i^alch tli@y bav« cOroady cmliarked.
Othmr chignges lieiv@ b^tm aimll ones, as requix>«d in eda^lng
our pirocedux«8 to tha major changes oads lAst ysar, i»9<. enlarging
Xb» foeograin to includu ha&ic scientists, exui. shifting of tlMS
rwspoxuiibillty for apj^cation r«-rt.8w to the frslning CoiaaitteeSo
5- Program objectives for 1999
fMs ^ogjrsm viXl continue to bt ex^nded as necessary and
as fund availability allows to provide the highly trained investi^tors
aeedsdo A continuous survey of former trainees viU be initiatfgd^
Questionnaires vill b@ s^nt to each individual during the sscond,
fifth, and t^ath year following conclusion of traineesbip support »
Analysis at regular intervals of information derived from th®
questiowjaires -uill eearve as the basis for evaluation of the progress
"by the Special Traioeeship program in reaching its objectiveso
6° YoluBi® of afpUeationa
AjpHcatians revi®u»d: 1T5 $1,371,500
Applications approvtds 125 905,750
Averiigt avard « ^l,2k7
7° staff assifflaneatB
llrso Hartaan « Training AnaJyat, in iimnediate charge of the
program, vith the advice of s
©To Parber « Executive Secretary, neurology graining Grant
Coiamittee for apf^cants in the neurological fltldo
DTc Jext>io© » E3£««utiv@ Secretary, Otolffir/ngolosy and Ophthalmologsr
frainlng Grant Comtnittees for applicants in th@
otolooical imd opJathalBaloGical field.
i4
1958 AanuAl Report
of the
Nation&I Institute of MfsttrolofifLcal taeeae*© and BUadn««B
V EESBABCIi £%LLCXf^QLFS
1. BfOgy^ffi a,ccoigpilsbiaBat8
This srogg^ffli of ftwaxds, dtsi^asd to supparfe th* r«8€arch trsiMng
of candidates qualifiM for Investigativ® caree?-©, continued at apsroxiffls.t®ly
the 6&a»e lev@l la 1958 &s io 1957 «
One hoadr^d ninety on© awards for a total of $^^3,65** wer« mde
ia 1958, SXii^tJy iaor@ thac half of the funds weat to 32 I'redoctos^l
Reeearcfa H^llovs^ vhoa^ tredsaing \flf»« directed towasrd the Bs,D. ^^.fi^.m,
msA 36 Bastdoetoral BeBefir<± PeHovs for r®s#ajfch training su:B?ort
duricis th© yeays iaanediately foUowing ngcslpt of th€- JiioD- t* K.B,
degsree^ Th® 3f^miii±)ag funds -were distributsd on «& inatitutioaa-l l«3i®,
and yer® used tc sm^oirt 9^ studentB for psrt-tim^ trainisSG^ ssnd 24
nsedlGSi «tud®nt6 electiag to Interruj^ misdical coi::xBec to secure a 3?ear
of .speci*lized lxs,6la sci@nc® traloiac" t^e iiidlvldu&ls %o >■© Bu|^srt@d
are BeXected by tli«e iastitution» As the IiMstitttt«*s coatributioia tc
the est&blishiaejat of the Fof^ic^ Ffellovahip pffoepcam, a mmXl portion
of the fellowaMp funds vbs. nwde Available to th® Katioaal Institat©®
of l]e<h a-Qd. us«d to swggort '} European scientists selected ia th#ir
r@s=5«8ctive cotMKferies to take a yfear of tr^ains ia the ?S2lt«a Statte,
2o CoRtribtttlons resultlni^ from particular s^pjagjis
Detailed inforcsitioa about the activities and contributions
of research feUowg subsequent to coa^etion of their sufisort is
«»av®ilabl« to «© ®t th© pcees^nt tiin®,,
Th® !_ffl,jor pfobI@i22S <eiwous3,tered in adainisteriaj th<i Research
F€llo'.r8hip proi^fSift stejffiaed this jnsar, as ia %hm past, froa; tlse fact
that ciiiitrcl of th® progsaa operation and responsibility for it has
' '^en placsii alLMj^t ©xsloaiviely outside the JJatlocal. Inatltut® of
. «5iiiroloi;y.s«y, Bleiea8@s aM Blindiiess. The result iBts latsfc of lustitut®
jarticiptttion In £md lntiB>»t@ knowledge of the Bese«rch Fellowships
sas^yte€ by its fuads ha» led to difficulty Is adapting the program
to ise^t Institute pragras a®ed*» la additio®, tjtier^ v«a frequeatly
^^'otjactied d«lsi,y in acting oa apgiiic&tioas^
i^
It is koped tS%&t cert&Ua cMrages Initiated as of July 1, I958
nay serve as an l&itial step la isqproviiig l^is UB>3»slrable situation.
**■* Cteaa^es and_ isEproyegents ia progrea
A. As of Jtily 1, 1958 fluids availalsle for tSais prograac vere
increased fra» $525,000 bo $5j6,00O. ^Is increase is soerauSiat large?
then tise figures indicate, since t&e total anount \jill be available for
Fredoctoral and Postdoctoral Fellowships; fellovsbips in tSse post*
so^dKcobre, part-tl^iie and foreign categories are being supported by
the Division of General Msdieal Sciences recently created.
B. Since July 1> 1953f Researck FuUovsbips kave undergone a
coqdetely revised procedure for review and award, Kodclled ia a
general way on tit&e method of Research Grant review. Applications
are received by "ttae Research Fellows&ip Review Brands, and assigsied
to tlbte appropriate discipline panel (ccogposed of intraxaural sciesttlsts)
for evaluation, and to tSie appropriate Institute for support .^
The recoa^iendations of the Review Panels, and the priorities
assigned the applications are made available to the Institute, which
tlten advises the Research Fellowship Review Branch which are to be
paid. At the present time, the three National Institute of neuro-
logical Diseases and Blindness Training CoMnittdes (ifsurology,
Ophthalaology and Otolaryngology), are acting as our consultants on
Research Fellowships as veil as Special Traineeships .
All procedures relating to %e duplication of applicatioses
and supporting credentials, preparation of applications for review,
preparation of routine letters aM es3icua3>raace lists is the responsibility
of the Research Fellowship ^view Branch. Snexisbraace lists ar<e signsd
by the Chief, Extrainiral Progress Branch and tSie Director of tte
Institute. letters of award sad disapproval are signed by the C&isf,
Extraaural Programs Branch.
T^ above procedure is still frankly ea^erissental and
subject to further modification as the needs arise.
^^
^J^^»d^ &0xam laav^QMe Itx direct conmuQlcatioja b«tv«m3i th@
Fellovd a»a tMs Institute bag o<scisrr®4» as & x^sult of tii@
iii,;pmtw© om tls® l#tt#rs of mmrAc It t» taeYitsbl® that this
iJiM l®«d to ii!^£>ov®Bsgat In our c«14tiaQs with the Fallows sujferted,
>ny th® JJatiozal iMtittttt of Meurolo^cal ms««ases aad Blindjats®,
ttjAd l2ic£«%3tid. iinterst^mdiiag ofi oour part of th@ kind of people aad
th@ sa,tur« of tb# tr*itiij\j tht lastitut© is supportiag,
^■'' fiy^^am Q 1;. j_f cti y-fes f or^ ^9'y9
KiTlae- 'fe® coiaiS8('i yeskr^ the gaias uiads duyiii{^ l^-, -. ia
laetltsstt parti eipatioa it> tfcie progrSBa ^11 b© sseploitsd ' '
■aeofar afi. pc"6®ivl@„ It le realized that a,^lissi.ility tc« th®
iiiStitut©'s5 a^ftti®! i.ilssion should be intejfjr^tsd io it® broad^rt
mum, aal timt tbe im.j©rity of th® aijards la tfa© prectoctorsl are®
:>dii ?^ fi^am for tsmiolEMJ that i@ sot strictly eat^gcrl.cale Howsv^r,
■is effort sAU >b# csdjg iia th® ^etdoctoiml fitld to «B^o,y a «®rlaia
,a«j:ili«r of til.® R*6«iiix-eh feUm/sMps to trala those i-ho js®@d special
EkiUi?' for rsssareh i» th© a«tiiros©n@e2^' dii
6. S!i^Sj££-.^^iS^i€^:.
-4K*li eat loss atJardM?
Rs^ilar ft-gdoetoral assd
P&atdoctoml aes@arcb Pellovshigs 63 ,/•. , , . -^
Postsoffcoiaort FeUoysMpg. 24 86.^2?0
fotal 1^ ■'■ :'-,\' 'k
■■. jlt^f^t>a-L^!jqe&tB
'■ ■ ' progysa, -iri^th'tii© &dvi5# of?
!&»=, FayL#r - Biecutiv© Seeretasy, NauroiosA f}ETSiQi.ag QrmA
CoEssitt«@ for fefplicsffite ia the laeuyslogical fJ^I-.^-
Iir. Jmr&sim = Eac«cutit«6 S«cy©t®ry, Otolsr^iigr.lc 2?/ aad Offctlmic»vic.,
fff^ril2(i_'. Gnifflt Coriultt®eg for 18,5^1 caatis la tii#
ctoiogissl and o^thalaiologisal ft©ld<,
• AHHUAL REPCSer • • •
Calendar Year I958
Eictramural Programs Branch
Batlonal Institute of Seiuroloslcal
Diseases and Blindness
Kational Institutes of lealt£i
VI Vmtm AfiD AFEROfTAL OF GSiABSS
1. PR0JEC3? DESCRIEEI<m
3Mb aotivity provides for the suppost of the activities
of the Rational Advisoiy Council on neurological Ittseases and
Blindness, the advisory training committees in neurolc^gr,
c^^ialmology and otologyji the advisory conimittees revievdLng
applications fear field investigations and pilot projects, and
the professional and clerical staff engaged in -Mxe processing,
analyzing and management of the grants and avards. Included vithin
this activity are funds for the Institute's proportionate share of
costs for services performed centrally l^ the Division of Besearch
Grants.
Tbe progress and problems in the isanageoient of the extra-
sEural programs are discussed in individual progress reports for
I Research Grants; II Field Investigations and Pilot Projects;
III Graduate training Grants; IV Traineeships; and V BesearCh
Fellowships.
Sixth Maual Beport of Clinical Isi¥@stigatioES of the
national Issstitute of leurological Diseases amd Blincimess
19S8
The Cliisical Myector's Report
The Sixth Asmual Eepos't of the Cliaical Savestiga-
tioBS Uiiit of the Hatioaal lastitute of neurological Dis-
eases and Blindness iacludes 94 projects » aad 73 publica-
tions eithex> published or is press « So^e of such publi-
cations obviously reflect results obtained from previous
f eases „ Six-hundred and six patients were admitted, for
a total of 19 8 278 patient daySo This is aa increase of
total patients of 81 ^ but a decrease ia patient days of
&ppro3Eiffiately 2„0CM)„ This reflects,, in part, the tuasor
program utilizing radioactive isotopes,, in which patients
were admitted but for 48 hours <>
Two in'restigators were lost through death during
the past year: Dr., Jo Godwin Greenfield died of a heart
attack on March 2 c, 1058 „ duriag his third stay at the
national Institute of neurological Diseases aad Blind-
ness ^ aad i5r. Francis Enomoto aet an accidental death.,
also durii^ the first part of the yearo "She Institute
also lost two of its senior investigators: Dr» Senneth
Mali accepted a post as Associate Professor of Anesthesi-
ology at Diske University v in charge of research, and BTo
Jose del Castillo, Chief of the Section of Clinical neuro-
physiology, will undertake his new position as Associate
Professor of neuropharmacology at the University of Puerto
Rico, Clinical Investigations gained one senior investi-
gator in DTo Herbert Lansdell^ who has taken over the
Section of Psychology.
As silvs&ws, the Unit has benefited by visiting sci-
entists and guest workers ffom Abroad = Frcm England were
two senior investigators -- • Dr„ Greenfield ^ &nd now Sr.
Tansiey frc^i the Institute of Ophtfealffiology in London;
troffi Japan,, l^o Eyoji fasa^i and B^o Snossbto; fro® India,
Dro Lele; from Austria,, another senior scientist DTc Dodt,;
froffi Sweden,, a senior scientist I^» Widen,; from Australia,
Dr„ Strang; frois Canada, Br,> Wherrett; frosa Spain, Dr„
Miqwei; from France » te., Geriaj and from Mexico ^ te,
Orti55„ nrso TaESley„ LbIb, Dodt, and Tasaki were or art!
attached to the ispasch of Oplitfeal®ol©gf ; Dars^ Orti® and
liqeels to the ^anch ©f l®ur ©surgery; DrSo Wid^ts, Q&Tim,
and Knosoto,, to th© Brajscfe of Electroencephalography mxd
leu^'ophysiologf »j assd Brs„ Greeisfield aad Wheryett^ to •:
-2-
in 4h@' tsrue naiee,, aa.d th&f ar© ^itfe the Institute for a
period of six months to two jears,, aad will b© returning
to their "xsativ# countries „ The specific sreseax-ch under-
taken t>f these investigators and their contribution© to
the research projects m&y be found in the Branch Reports
included herein.,
Specifically; the Sections of Electroefficephalo-
graphf and Clinical Neurophysiology » report the follow-
ing projects: Under the combined Branch of Electroence-
phalography and Clinical K®uroph|rslologf :
Dr. Snoisoto» before his untimely deaths was work--
ing with Br„ Ajmone-larsaaa on the epileptic activation.
of unitary elements of the cat*s cerebral cortex » and
their relationship to the ESS discharge => In this parti-
cular stud|f"5 epileptic foci were produced experisaentallff
on the suprasylvian gfrsis of the cat bf means of local
application of different convulsive drugs. The develop-
isent of the slow SS^ discharges was monitored from a
routine surface electrode and upon their appearance a
survey was then undertaleea on the various units within
the different layers of the nearby cortese by aeeaas of
tungsten ©icroelectrodes . The investigators report
several thousand observations on such units analyzed in
29 ©sper iaients carried out on cat. They noted that
such unit behavior occurring in coincidence with an SEG
discharge was characterised by a paroisysaial appearaac®
of high frequency bursts, plus a marked tendency towards
synchroniisatioa of different units. This property s how-
ever, was not absolute in that a given unit may be char-
acterized by rhythmical high frequency firing in "rest-
ing" condition 5 in which case it tends to become inac-
tivated in coincidence with the EEG discharge. They feel
that 5 in the latter such case^ the unit recorded is very
likely injured » I^„ EnoKoto and Dr„ Ajmone-Marsan con-
cluded j, from these observations, that probably the number
of units activatedjin a certain instant j their firing
pattern^ their location, and teaiporal - inter-relationship
are at least closely related to» if not even responsible
for^ the final shapes amplitude, and polarity of the
slow BEG event.
Still using the tungsten microelectrode y Ite>o AJmoae-
Marsan and J}Tc Widen have recently emfoarfeed upon a study
of the relationship of the cortical unitary eleaients to
slow surface responses evoked in th© visual cortesj after
sti@»ilation of the lateral geniculate nucleus » This prob-
1&& has just been initiated and as yet these investiga-
tors do not report definitive results.
BTc Gordon Loag,, ia the sail® latoos-atos-f ^ has
finished a eompl®te study om the modif icatioi^ of sensory
raechanisBia Ijf subcortical stnactiares,, is which h© has at-
tenpted to ©lucidate the @f fact® which the braia stea
reticular fosnatiosig the sioa-specific thalamic s^sten,
as&d othef subcortical .structures (basal s^as3glia» thalassic
associative aucleis rhijaeacephalic foraatioos) liay have
upon cortically evoked potentials from peripheral sensory
stiffiulationo DTo Long reports his ex^riMentm on some §5
eats. The subcortical structures were localised stereo-
taxicalljf and histologically controlled « Varying para*-
ffieters of conditioning stisiuli were used, 2^„ Long found
that in the unaaesthetiaed preparation fee could »ore
easilfr modify somatic and visual potentials « and that
this effect was nore prolonged when the conditioning
stiMuli were applied in the reticular formation of the
brain st^a. In decareasing order, the sajoe '^as found to
be true of non~specifie thalanic s^t^i^ the aa^irgdala^
the putaffleas the globus palliduss. and the lateral half
of the head of the caudate nucleus. Stimulation of the
pul'^inar-lateralis posterior coeplesc of the thalamm pro-
duced s«»ie iiodification of -risual response onl|^, fhe
changes recorded in the evoked potentials i^ere more learked
at the cortical 'level than they were at the specific
thalamic relay nuclei, ^us, I^e Long confirmed pre-
vious impressions that high frequency stimulation of tte
reticular f©:rmatioa will depress the aaplitude of evoked
s<matic and visual responses ^ and that this depression
was ^ore consistent and of longer duration than the
soaato-sensory syst^i. In addition ^ however « it was
shown that lower frequencies of stimulation to th® reti-
cular formation and its projections will augment the a^'^^-
plitude of the evoked visual and s<matic responses. Both
sBodifications of the sensory potentials were abolished
or diminished by barbiturate anesthesia. Dr. Long sug-
gests an analogy between the reciprocal effect of
tation and depression of afferent conduction £, and the
facilitation aad inhibition of motor responses by the
reticular f ©station.
I^. Abraham and Br. Richards, in Dr. Aji
larsan's laboratory,, after noting certain chai'acteristics
in the SEG of patients receiving steroids^ ui'idertook a
study of the EEG changes induced with photic stimulation
in patients treated with ACm and adrenal c©rticoids„
Sighty such patients were reviewed; 9 of these 80 showed
an unusually marked response to photic stifflulation. Thi?3
plac^ the occurrence of such responses in the group
study at 11.2 percent; however j if on© takes a populatiora
of non-selected subjects , such an instance is only 1»3
percent, ^ese investigators concluded that ACTH and
adrenal corticolds contribute to the lowering of the con-
vulsive threshold,, probably acting at the brain stem level,
upoa probably already abi&orsal steoctures „ and the ab-
iiosmal reactioa to iatesmittemt photic stiEUiatlosi iB^
ia facts »• iffl»Bif©statioH of such a coss^Mlsi^'® teMeaciTc.
They also iradicat© tfeat suck a cois¥ulsi'«'® tendmncf w&j
be iffi fact dependent upon th® disease stat®, la that a
«iaJo$>itf of tfeeir patieats liad elthes' lupus ©rytfeeMato-
sus or Ifuplsatic l©ulKeiiia» eitlaer of which saf Jta^e^ at
sxam time during their course , central aenrous sfstasi
lesioi&So
Ifeo AJffioae-MarsaE,, Dr., Abraham and Dr. Vaa Burea
ha'y© continued theig- studies of depth electrographi' ±n
seisure patients » asd have reported their first findings
ia a temporal lobe epilepsy monograph by C„ C. Tht^aso
As Qoted in the 1957 report ^ these investigators found
the ssetrazol activatios of extreme iMportaace is such
ind^^elling electrodes, and fouad many cases in which
epileptic discharges aay he present at a cortical level
but would not be recorded with the routine SEG. On the
other hands a certain nuasber of discharges will^ instead,
be recorded fro» the routine ESG^ ^xkd their aj»plitude ap-
pears to be directly proportional to that of the cortical
discharges. But' another group demonstrated amplitude
which is definitely not related to that of the original
discharge, and aay vary from &s high as 58 to 1« to as
low as 2 to lo Such amplitude and ejctent of area
covered by the discharge,, however;, were not the only-
factors responsible for the presence or absence of spread
to the scalp J and spikes do not appear to project more
easily or more constantly thaa the paroxy^raaal waves, A
complete analyses of this report may be seen in the re-
ference listed above,
Br. AjKone-larsaa and Sr» Baldwin are continuing
their observations on t^^poral lobe epilepsy and they
have reported the electrocorticograpfeic findings ia a
large series » again in the leonograph by C„ C„ Th<^&s„ dusk-
ing the past year., fhe procedure and ultiaaate coisrse of
this project follows the lines reported in the 1957 an-
nual report o
Ifefo AJmone-Marsan is also still continuing, with
the cooperation of Dr» Abrahams the atlas of seiaur© pat-
terns accompanying Metrssol activation,, and the correla-
tion between such moveaests and behavior with siffiultaneoas
electroeacephalograns c Th& publication of this atlas has
been undertaken by the Journal of ffiG and Clinical Neuro-
physiology o For the latter Journal » DTo AJsone-liarsan
and Iteo Eenry have also undertaken a bibliography cover-
ing the last ten years ^ froas 1948 to 1958.
In addition to this produetii'e research, the Uait
has carried oa a large ser¥ice fuactioa ia which a total
of 1^502 electroencephalogaraiss were coaipletedo Mskuy of
these T^ere froie other institutes - thus, the Matioaal
Cancer Institute accounted for 306; 23 eIectroco2'tico~
graphic studies were performed on neurosurgical patients
ia the operating roosBj and extensive SEG studies were
carried out on aanj patients with indwelling cortical
electrodes „
The Section of Heuroph^slologf^ has been closely
aligned vith the Branch of Biophysics of the Basic Unit
CHIHDB)j with Dr. lenneth Cole. One of Dr, del Castillo's
largest projects was in the study of excitation in Medul-
la ted nerve s in that the results previously obtained in
the giant aisons cannot toe applied indiscriainateiy to th®
medullated fibers of vertebrates or man,, 1!he extremely
small surface area of the nodes of Eanvier malees, in j&my
wayss myelinic fibers particularly appropriate for ©s-
ploration of certain aspects ©f nerve excitation » ^hich
could not be resolved when the «hole axon is studied.
Counteracting this^ however, is the difficulty of iso-
lated single nodes of Eanvier, The study was a combina-
tion of an iaproved technique to perform voltage claap
esperiaients in the membranes of the nodes of Eanvier^
and to ccaibine this with the electronic resistance multi-
plier aiethod of Frankenhauser . Taese investigators fouad
©riginal difficulties due to the high longitudinal impe-
dence of the inner nodes through which the controlling
ionic currents are injected. These were overcooe eventu-
ally by the resistance multiplier method, which was also
adapted in these esiperisents to siniaize the external
lealss of the controlling current projected into the in-
terior of the clanped node. Dr. del Castillo and Mr.
Moore found that depolarization of a mammalian nodal sec-
tor asie conforms to pattern similar to those found, and
thoroughly analyzed ,, by previous investigators in inverte-
brate Material. They isade an important incidental obser-
vation» however, that when a sedullated nerve fiber is
sectioned J the cut end of the syelin tube tends to close
in such a way that the leak of aiioplasaa is minimal and a
high electrical resistance is maintained. ThuSj one has
a basis for providing an artificial single node of Eanvier.
'DTo del Castillo ai&d Moore have reported in a paper the
technical methods utilized ia this procedure, entitled
"An Electronic Electrode", 'Rhlch is to be presented at
the 1959 National Convention of Institute of Radio Engin-
eers. Iia the future B these investigators wish to study
the nechanisai hf ^ich certain organic cations ^ such as
hydrazinium ions may replace sodium ions in the excitable
aechanissi of the
BTo del Castillo's other project pertaiias to the
nechanism' of traiissaitter liberation at presynaptic aeyre
endings J, in whicli intracellular capillary aiicroelectrodes
i»ill be used to record potential changes at the endplate
Biesbrane of nerve to sroscle^ or of s|maptic potentials,
lonophoretic methods will also be utilised to apply sub-
stances to the localised spots of the nerve ending o In
their initial studies j, these Investigators found that the
depolarization of the post-synaptic nembrane produced hj
externally applied acetyl-choline is laarkedly influenced
by the pH of the extracellular solution. This project
will« in essence;, place at intracellular level s<»^ of
the findings reported by Br^ Irwin in the Section of
Keuropharsnaco logy .
In the Section of neurological Disorders ;, a ne^^
investigation as to the iiedical treatment of seizures
has been undertaken s, which is dependent upon the findings
of Brody and his colleagues in the National Heart Insti-
tute j, of new monoamine oxidase inhibitors, lonoasd.ne 03zt-
dase is the primary enzyme necessary for the break-down
of 5-hydroxy-trypta»ine to the 5-hydroxy-indoles, the
isost important of which is 5-hydroxy-indoleacetic acid.
The formation of ,the sulphate ester of this group in the
urine has already been associated with a neurological
disorder characterized by cerebellar syraptasiotologyc
dexmitis, and ai^atal retardation^ under the name of Jep-
son's disease. Brody and his colleagues have found that
the utilization of monoamine oxidase inhibitors in ani-
mals markedly reduces the epileptogenic threshold o Thus
a double blind procedure has heen instituted in which pa-
tients with centrencephalic seizures ^ having as jsany as
50 or more attacks per day^ have been adi&itted, and a
double blind procedure initiated ^ using the new monoamine
oxidase inhibitor JB-516o As this is a double blind pro-
cedures the results irill not be known for approximately
six months' time. At the present ti^e^ ten patients have
entered into this project,, This project is being carried
out by HTc Bushnell Sisith and BTo Darwin Prockop,
It has been noted that cases of orthrostatic hypo-
tension have been noted to have many neurological disor-
ders » in particular loss of sweating;, loss of external
sphincter controls, impotence^ mental dulling » and^ in
some cases 9 a Parkinsonism-like syndrome , with or without
ciliary atrophy o Three such patients have now been studied,
and one such patient has coige to post mort^. To date no
thorough anatomical study has ever been accoas^plished on
a patient dying from orthrostatic hypotension » The im-
portance of this single case., hence., is not to be under-
estimated o It was the decision that serial sections
~7«.
should b© accomplished tlarougla the hypothalswus j syiapa-
th@ti® ganglia 2, the iater^ssdiate cell colusms, asid the
cranial nerve nuclei of III, Tj ?!!» IX » and X^ as -well
as the basal ganglia^ anterior horn cells s and corteXo
This necessitates literallf thousands of sections ^ and
the strict correlation of anatomy and pathologjr. It is
anticipated that the thorough studjr of this one post
nortrai case 'will need, in tine, Invest igati'^e us® of a
neuroanatosiist for at least six sonthSo This is being
undertaken s at the present time, by !>rs» Brager and Shf^
and to date important findings have alreadj been found
in intesaediolateral cell co limns » the ventral cell co-
lusmSs in Clarke's coluiin, the dorsal nucleiss of the
vagus, the ventricular graf^ and in the inferior olives...
Degenerative changes in the cerebellum were also found
with sanf torpedos. There were marked degenerative
changes in the substantia nigra and in the mesencephalic
nucleus of the trigeminal ^ as well as in the larger cells
of the corpus striatun and the pirraaidal cells of the
cortex »
Siailar to this is the study of a new syndroae re>
cently described with rapid central nervous sfrsten dete-
rioration ^ central blindness ^ nyoclonuSf, and death in
approximate!!' three to lour nonthSo Here again » a long-
terra anatoffiical and pathological correlative stud^ is be-
ing undertaken 5, with serial sections » This cas® will he
studied extensively hf Drs» Drager and Bushnell Ssiitho
As in past jears,, mamj of the patients admitted to
the I^anch of Medical Keurologj are suffering from dis-
eases of th© motor uffiito Hecent advances in isotopic pro-
cedures and muscle pathology have changed radically this
prograffi froai the past f@ar„ In combination with the As-
sociation of Research in Nervous and Mental Diseases „
this Institute undertook » during the past fesx^ a review
of the effects of aetabolic and endocrine abnormalities
upon diseases of striated susclec This was an over-all
survef :, ^plof'ing chemical studies of Muscle biopsies.,
c<»ibined with various metabolic tests with gonadotrophin,,
corticoidS;, ketosteroidSj TSHj, etc. Of particular in-
terest were two disorders: Faailial Periodic Paralysis
and so-called McArdle's glfcogen disease of ssiuscle. In
the foraer disorder ^ aldosterone levels were deteraiined
bf the double isotope derivative isathodSo Intracellular
cations on Muscle r^ioved both before and during attacks
were also studied «, as was pathology before and during at-
tacks c And finally;, giicroelectrode recordings of single
ffluscl© fibers in vivo before and during attacks, Potassiias^-'
turn-over® ■were also studied in this disease. The pathoiog;^
of tills disorder was quite striking ia that large ac-
cuiiulatioi&s of fluid appeared isitracellularly lis ap-
proxJjaatelir oae-third of the liters o Chemical deter-
siaations showed that^ in spite of this accuMulatioa
of fluid J, the catioffiic coaceatration of the cell re-
maimed appr©si»atelf within no:raal limits „ This was
coi^fismed bf microelect^ode recordings ^ which showed a
resting potential of 71 o 2 =,v 11 oS^ which is what Might
be anticipated if iatracelTular potassium were at nor-
Bial levels o Studies on aldosterone on these particu-
lar patients revealed that there was no increase pre-
ceding the attack, as previously reported by Conn.
There was also a decrease in potassium in the urine
preceding the attack,, which indirectly confirsi^ the
latter observation done on double isotope derivative
methods,, in that if there had been aldosterone eiecre-
tion there should have been a potassium diuresis ,,
Twenty-three cases of infantile neuromuscular
disorders associated with hfpotonia were also studied
in reference to pathology » electrosyographyj, and clin-
ical course. Froai thiSy five different types of dis-
orders were found in the disease state ^ which h-t-ve
been recently grouped into but one disorder. These
findings have been reported by Drso Greenfield ^ Corn-
and Shy J, in the Deceaiber issue of Brain.
The recent findings that BMA is probably inert
in non-proliferating cells is now leading to th® util-
ization of tritiujss labelled thymidine o Ihis will be a
powerful tool in the study of regeneration and growth
of ffiuscle, and this^ combined with electron microscopy,
will be undertaken by the Section of Biophysics «
i^« Haase has^, in addition ^ undertalsen a long-
range study of the pathological findings of intramus-
cular »otor and sensory nerve endings in nomal and in
neuroiMiseular disease states ^ using the Coers technique
of intravital nethylene blue staining. He has confirmed
axonal regeneration in neurogenic diseases ^ but the
other abnormalities described by Coers and Wolfe hav®»
as yet not been verified.
The Section of Biophysics has completed its in-
vestigations on th® localization of cerebral neoplasia
by collinating techniques;, utilizing various isotopes.
Over 200 such patients now have been studied ^ with a
confirmed accuracy of 86 „ 2 percent « The final techniqiues
and instrumentation utilized in this study ^ as well as
the statistical evaluation^ have been reported in nono-
graph form by So & So Livingstone o This Monograph was
also utilized at the International Conference for
Peaceful Use of the Atom, Similar p^'ocedures have aow
been initiated hj th® lastittste foi* the Johns Hopkias
Uaivesfsitfj, the latioaal Ma^f ^©dical Center ^ Oak Ridge
Hatioaal Laboratories, and mow at Los Alai&oSe
The studies of microelectarode srecordiag is siagle
g»2scle fibers has heem utilised ±n familial periodic
paralysis and mys^theaia gs'a^is. Due to the scarcity of
the first disorder ,, this «as dose by cut-dowa nethod. la
the Myasthenic patients ^ coatiauiag attempts are made to
record siagle nuscl® fibers through the iatact epidermis.
The Bak Unity 6aia Amplifier has been utilized as optimal
with a constant current seat back into the grid of the
cathode follo'wero "This latter allo'g^ constant sasipling
of the condition of the probing electrode » To date, ©n~
dcmysius and perimysial connective tissue has been the
chief stufflbling-block, in that the electrodes intermit-
tently plug or break o Of the literally hundreds of re-
cordings i^hich have been attempted to date^ only 5 suc-
cessful intracellular penetrations have been ^ade through
the intact epidermis. The continuity of this project
will depend upon the ability to overcome the difficult
techniques listed above.
In the Clinical Director's Beport each year, an
s^e^pt has been made to select areas of outstanding con-
tribution o This year the studies conducted in the lab-
oratory of clinically applied pharaacology , under Dr„
Eichard Irwin s, has accomplished much which '^ill ehow con-
siderable insight as to the interrelationship of blood
and tissue cholineste^ase systeiis^ their substrata, other
enzyme systems working upon such substrata $ and basic
fundamental knowledge as to the differentiation between
depolarizing and competitive blocks, as i^ell as insight
as to where in the muscle fiber the blockading compound
has its maximal effect, Thus^, Dr^ Ir^in and his col-
leagues have demonstrated that competitive blocking com-
pounds j, such as d-tubocurarin@ and depolarizing blocking
compounds such as deca^ethonium^ may be differentiated
in their action lyf inhibition or excitation of muscle
cholin©steras©j thus^ th® competitive block of d-tubocur-
arime is reduced or prevented by inhibition of muscle
cholinesterase » On the other hamdj, the block of depolar-
izing drugs is prolonged by the jiiihibition of plasaa cho-
linesterase or muscle cholinesteraseo In the case of d@-
camethoniumj, this cannot be due to destruction by cholin-
esteraseo per 8®g as decamethonium has no ester group &nd
hence could not be destroyed hj cholinesterase. Succinyl<-
choline, on the other hand, has an ester groups and thus
could be destroyed by cholinesteras© . It is of interest ^
-10-
however J that the prolougatioH of the blockade by iishi-
bition of plasiEa choliaesterase is identical to the two
substances p thus showiag that this iuhibitioa proloiaga-
tioffi is not of necessity due to destructioHjj or the de-
polarizing compound. Thus one can assusg©, I believe cor-
rectly«, as Dr. Irwin and, his colleagues have assumed j,
that Hiuscle cholinesterase has but a minor role in re-
lation to the total block „ If this substances however,,
is not aietafeolized by plasisa cholinesterase o then inhi-
bition of ffiuscle cholinesterase has a marked effect on
the blocking activity, and the non-depolarizing sub-
stances upon such inhibition of muscle choline demon-
strate a decrease in their blocking power ^ whereas the
depolarizing substances demonstrate an increase in their
blocking power.
Dr, Irwin and his group have continued their stu-
dies on the action of directly stimulated innervated and
denervated iimscie. In this they have been aided by a
device 3 created by Mr, Wells, of an optical-isotonic le-
ver systeas,, recorded through a cathode ray oscilloscope „
With this mechanisms they have been able to deawanstrate
that the block is not due to increased beuscI© compliance,
as added compliance in series does not give contractile
responses similar to those obtained ^ith succinylcholiiie
or decaraethonium. If this isotonic system is observed
closely « one may see there is less shortening of the fi-
ber and reduced velocity of shortening ^ again showing
that this is not an increased coiapliance of the stuscle
fiber o The isotonic-optical system allows this^ in fact
that it reduces the elastic coisponeat of lausclec This
system J, however, does demonstrate a prolonged latency
froas the onset of the stimulus to the time of contraction
after administration of depolarizing compotmds. These
investigators feel there is a spatial distribution o£
the depolarising blockade over the muscle membrane j, in-
dicating either saultiple end plates upon the lauscle meia-
braae, or a teisporal spread from a single ffiesbrane, i^e,
one end plate. These investigators point out that siuscle
cholinesterase is low in quantity and is not uniform in
various species and/or organs, and hence has a species
and organ specificity. It is thus depeadeat upon the sub-
strate and enzyme activity. Thus, muscle cholinesterase
studied as to substrate specificity and well-known inhi-
bitors would give considerable information as to the chesss-
ical interchange between the substrate and the ©azyiaec
The cholinesterase of muscle hosaogenates « in which
the blood was reasoved so the plasma cholinesterase was
not present, was studied. Such homogeaates hydrolyzed
acetylcholine more rapidly than benzoylcboline , or butryl-
choline. An excess of the substrate » however, would inhibit
"•11-
such hydrolyzes, the optimal level being 5 x 10""^ c The
optimim level of coaceatration for substrates other than
acet^rlchollne are higher. Thus,, muscle choliaesterase
is highly specific o HoweYer, sisace feeusoyl- and butryl-
cholJjs® are hydrolyzed at measurable rates, sisaall amouats
ol noa-sp©cific eraz^e saust also be present . It is of _
interest that neostigsiin© depolarises the uembraEie at 10"**,
whereas pyridostigiaiae (laestiaon) will note This becosaes
of double iaterest in that both drugs are highly useful
in the treatiaeat of myasthenia gravis, Galanthaasine ^
which has been isolated tTom an alkaloid in the United
Soviet Socialist Republic, and utilized in the treatreent
of myasthenia gravis j was also studied by these investi-
gators o Galanthamine is a phenanthrene derivative and
not a carbamine ester „ Dr. Irwin and his group found a
50 percent inhibition at 6 s 10""® „ The value for the in-
hibition of plasiaa cholinesterase was the sajae„ Neostig-
mine and physostigmine inhibit at lower concentrations as
far as cholinesterase in the aiuscle is concerned; but in
vitro inhibit more rapidly than with galanthamine.
Finally these investigators are studying the pos-
sibility of choline esters other than acetylcholine oc-
curring as natural constituents of biological systesas;
the object being to determine to what ©stent the choline
esters are found in such biological systems and related
compounds p and how they depolarize tissue membrane. Se-
condly, to relate the depolarizing properties of these
compounds to their stimulation or blocking activity of
synapses J, and finally to study the metabolism of these
compounds h^ tissue enzymes. To study this, the travel-
ling fluid electrode technique is used to measure depo-
larization of the isolated frog sartorius muscles,, and
ffiicroelectrodes will be utilised to determine the rest-
ing membrane potentials., presumably through the Bak
Unity Gain Cathode Follower. These investigators have
found; in high concentrations ^ i.e. 10"^ molar , that
butrylcholines benzoylcholine, and imidazoleacrylcholinej
all resemble acetylcholine ia their depolarising prop-
erties, Methacholine, however, does not depolarise muscle
membrane. These investigators have also found the plasma
from myasthenic patients have bees observed to metabolize
imidazoleacrylcholine at the same rate as plasma from
non-myasthenic patients. And finally ^ these investiga-
tors are attempting to find to what extent depolarization
of the muscle membrane may effect the efflux of enzyses
from inside the muscle fiber, in particular aldolase.
This latter project is projected into the coming year.
The Section of Neuroradiology suffered in having
its chief investigator, I>r, Giovanni Di Chiro, undergo
surgery for a major illness. In spite of this setback.
-12-
however 9 Dr« Di Chiro ^as able,, upon his return to dwty ,
ia addition to his heavy service resposisibilitieSj to
carry outs In combination with Dr„ Martia Eubln of
Georgetown Uaiversitj', a research project which culmia-
ated in a paper concerning the sietal chelates as pos-
sible contrast nedia for myelography, 'Kaese chelating
compounds were tested against coamonlj used iodinated
contrast snedia. Different concentrations of the various
chelating compounds were tested in order to determine
the concentration for opti»al opacity. Once such opacttf
was detersained in vitro, it was tested ia vivo on dogs
and rabbits. Chelating agents used are listed in Br.
Di Chiro's report, with primary interest on lead ethyl-
enedis^inetetraacetic acid. This substance was adminis-
tered at the dose level of 10 milligraras per kilOj and
appeared in the urine to the extent of 85-89 percent of
the injected dose within two days. Of that retained in
the aniiial^ i.e. 10-15 percent,, 50 perceat was found in
the liver and sossie 20 percent in the bone marrow „ This
demonstrated that^ despite the laxge amount of excretioii.
the anount retained is not to be discounted. The ex-
perlsents in vivo show that studies of good diagnostic
quality siay be obtained as fax as X-ray contrast and
detail are concerned, with radiopaque ssetal chelates.
However, the acute toxicity of the asetal chelates in
ayelography,, as well as in nost of the other X-ray ex-
aminations carried out, proved to be too high,^ Accord-
ingly f. Drs. Di Chiro and Rubin are going on to undertake
studies ia other metal chelates with high atomic nm&hew ,
in hope that ia this screening one agent of local toxi-
city would be found which was so low as to sugg^ it
could be used in clinical isyeiographyo
The Section of Keur ©chemistry continued its ef-
forts in the ®aJor fields listed in the 1957 report. Dr.
Horvath continued his studies ia the distribution of
actin and tropomyosin in noraaal sjad diseased auscle, his
coiRparative biochemistry studies of sjsooth muscle and
striated suscle, and alterations of aetosayosin tensile
strength and muscle proteins in neuroMiScuIar diseases,
Dr„ Tower and his colleagues have continued their
studies on the aietabolis® of r-^^iaobutyric acid in
neural tissue , with the aid of »r„ McEhann and J^To Wherrett
Studies on the relation of pyridosine to certain seisure
states, in particular in those cases known as pyridoxiae
dependency continued. Dr. Tower Continued his elabor-
ate studies on aaino acid metabolisi® in nor^ial aad epi-
leptogenic cerebral cortex in vitro., aad in electrolfte
energy asetabolism in normal aad epileptogenic cerebral
cortex. The unit as a whole continued its clinical
evaluation of aaiao acids aad related compounds jja con-
trol of seiasures in man.
-13-
Dr, Curtis contiaued In the realise, predomiaaatly „
of surface-chemistry,, aad la other pb.s'sico-cfeeaical ise-
thods la deteraiiniffig constituents of hussaa spinal fluid „
ocular fluid, etc„
Dr„ Tower's studies specifically aow revolve
around C^'^ amd H^S la&elled cosapouads. Two-deoxyglu-
cose was utilized as a coapetitor for glucose utiliaa-
tioffi, by inhibiting the hesokinase step primarily due
to depletion of available ATP required for this step.
Dr. Tower found it was possible to overcome the 2-deo36F-
glucose block in glucose utilisatioja by adding either
ATP or glucose-6-phosphate to the slices in anaerobic
conditions o Wo effect of these additions,, however ^ was
obtained in aerobic netabolisBi,, presumably due to their
failure to penetrate the slices « Dr, Tower felt that
2-deoi£yglucose inhibition did not result in any acti-
vation of the glucose-6-phosphate dehydrogenase or iu
any oxidative shunt pathway » These findings were cheeked,
by incubating the control and inhibited slices with
glucose-1-C-^^ and glucoso-6-C^^ phosphate,, determining
the utilization of C^^2 ^^^ C^'*-lactic acid production.
Since the ratios of the C^^ lactate froia the C~6 cosapared
to C-1 saasples were 1„0 in both cases, whereas C-6/C-1
would be less than 1,0 if the shunt pathway were uti-
lized, this would indicate that this inhibition was
not due to an oxidative shtint pathway. This was in-
directly confirmed by the finding of low level brain
TPN by other investigators in that TPK is the necessary
coenzyme for the shunt pathway, Dr„ Tower found also
that 2-deoxyglucose inhibition not only resulted in
marked decrease in glycolysis, but also in oxidative
metabolism o Thus^ with glucose-U-C^'* ., less C^^Ogs less
labelling of the free aaiao acid pool,, and less C^'^
lactic-acid were all obtained. From these studies with
(jM-iabelled glucose 3 the distribution of glucose
utilized by norasal slices to various intermediary steps
could be estliaatedj thus glycolysis to lactate, 70 per-
cent; asBino acids aieasured by glutamatej 22 percent;
respiratory C02» 7 percent, and other interuiediaries .
such as lipid and protein, 1 percent. Sf one calculates
the oasygen uptake as SSjiM, /g/fer o , it is clear that if
30 percent of the latter is accouated for as aasino acid
and respiratory COa^ this almost exactly balances the
oxygen uptake, asstming 6 moles of the latter per mole
of glucose oxidized „ This is consistent with studies of
other laboratories s, and this laboratory ^ that non-glucos®
substrates » such as amino acids,, normally support ox:id,a"~
tive asetabolisas by th® brain and they are repleted subse-
quently hj part of the glucose utilised. Studies with
"14-
2-d©osf glucose clearl^^ desoastrate , accordiag to Bi't
Tower 5 that glucose is ziecessary to ma&e repletion of
aon-giucos© iaterjaediates possible j aad enevgf produc-
tion rapidly falls In its afesemce, aad that this is
not only by depletioa of ATP and creatiae phosphate,
but also by deleterious effects ou glutaaiic acid «iad
electrolytes in the inhibited slices. As isa his 1957
report, Dr, To^er poiats out that such isihibited slices
fail to extrude excess sodiuis aad recoaeentrate potas-
sium in noneal manner „ This is similar also to defects
seen ia slices "which have been removed from epilepto-
genic patients,, and Dr. Tower has also found this in
cortical slices fro® cats with seizures enduced h-^ 3-
iaethyl-3-ethylglutariaides and by Methionine sulfoxi-
iftinea Dtilizing the Cot love apparatus, Dr. Tower and
his colleagues fiad that the spelling of norsal and
epileptogenic slices dt^ring incubation is confined
to the chloride space, and that calculations of electro-
lyte concentration per litre of non-chloride space
water at the end of slice incubation demonstrates again
a loss of potassium and a gain of sodium «
DVo Tower has continued his studies on incubating
slices of cat cerebral cortes with L-glutaiaic acid
labelled with C-*-^; L-glutaiaine labelled with C^^;
^-aiainobutyric acid labelled with C^^; L-aspartic acid
labelled with C^-^; D-L-asraaragine labelled with 2.B-C^^;
D-glucose labelled with C^^; Sodiua Pyruvate-S-C^"^ ,, and
2-pyrrolidinone-2-C^'*. Using these compounds,, Dr, Tower
was able to determine the order of labelling ia amino
acids » and was able to show this had considerable sig-
nificance since the aspartic acid could priiae the Krebs
cycle by providing both oxalacetate aad acetyl-CoeaJsyme
A (from pyruvate) in the absence of the latter froai gly-
colysis. Dr. Tower concludes that these studies indi-
cate how active the components of the glutaaiate-aspartate
aaino acid group are in metabolic participation in the
Krebs cycle, and feels that the release of COo laeasured
by C-*^"^ liberated during these experiments confirmed this
conclusion.
Ia the second part of his experinsentj Dr. Tower
analyzed the liberation and forBnatioa of glutamic acid,
glutaaine, 5?^~ajffiiaobutyrie acid and free asmonia aietabo-
lism in incubated slices frosa non-cortical areas of the
cat brain; these were the sub-cortical white siatter^
the thalamus, the caudate nucleus, a^d the cerebellar
cortex. He found the levels and aetabolic behavior in
all gray 5 i,e, neuronal areas, were similar to that pre-
viously observed in the cortex, but that the white ssatter
©3Ehifeited extremely low levels ajsd. little cSsauge o» 1.%-
cubation for glutaasic aad j,--aMiaobut.yric acids » wMle the
white »atter glutamiae was mot greatly different frost
the cerebral cortex. In studies oa the levels of these
substances o ,t>r„ Tower felt, using the calculatioos of
Elliott aad Hellers that at least 85 percent of corti-
cal glutamic and ^^--a^iaobutyric acid content ^sls associ-
ated with neuroaSa is?hile oaly about 5 perceat of the
glutaaine appesired to foe aeuroQal ia location =. The cere-
bral cortex was fractioaated by the Brody aad Bain tech-
nique j, and Dr, Tower found the majority of glutamic aad
I'-asinobutyric acids ^ere associated with fraction R3 or
"the aiitocbondrial fractions whereas glutaaiine was distri--
buted aisffiost equally between that fi'actioa and the cons'-
bined Rj > Eg fractions which contained cell debris , axoB
fragments , nuclei, etc. Ko content of any of the three
asiino acids was found ia the laicrosomal fraction. The
finding of these substances in the mitochondrial fraction
is 'compatible with their close association with the Kretos
cycl©c
Dr, Tower's studies also indicated that the in-
hibition of glutaaine synthesis by isethiorblne sulfoxi-
jiine is primarily an interference with aamonia moiety .,
possibly by the isiine group of the toxic compound j and
that by adding only a«moniuBi chloride such a block could
not be overcome unless adequate ajssounts of glutaiaic acid
are available to aisidate to glutaasiaeo Studies with
sisiiilar epileptic agents., such as Megiiaide showed that
the glutamic acid metaboiisiii was blocked to include y-
a^inobutyric acid,, in that the latter co^spouad was sig-
nificantly lower than normal. The saise was true when in-
activators of pyridoxai phosphate were used. If raaloaate,
however s was used, the aiaount of glutamic acid and y-simlno-
butyric acid in the slices rose to double the aorsial
values. The action of Bsaloaat© is to inhibit succinic de-
hydrogenase. Since this was accompanied by reduction of
oxygen uptake, it was previously not clear why such slices
did not also show succinate accxuiulation. Bt^ Tower's
data suggest that in the whole cell preparation it is
glutamate and i^-aminobutyrate rather than succinate which
accumulates and requires a study of the relationships
among these three components of Erebs cycle. Dr, Tower
plans to continue these interesting experits^^nts , using
the laicroanalytical siethod of Dr, O,. H„ Lowry.
Clinical evaluation of various awiao acids and re-
lated compounds in the control of seizures in vivo is asaji
has been continued by Dr, Tower aad Dr, McJOiann., and the
Branch of Slectroeacephalogr aphy . Patients on |^-amiao-
butyric acid have continued to do wel3^ ia Dr, Tower's es-
tiisaation., one patient beisg seisure-free after three
three mositlas oa tlae coi»poimd„ compared to multiple
daily seizures previously , to stoppiag the compomsd, the
seizures returned and have again been abolished by start-
ing ^''-aiffiinobutyric acid. Several other patients are get-
ting jsore benefit froia y-ajainobutyric acid than from 1-
asparagine. Gamaa-aiaiffiobutyric acid has been given in-
travenously to levels of 4 asMAg. body weight j with no
untoward effects, in dogs. However ^ when 1/200 of this
dose is adaiiaistered to maa^ there is iamcdiate agita-
tion, flushing, hyperpnea,, and a drop in diastolic blood
pressure. Recovery occurred within 5-10 jsinutes. Dr.
Tower rightly points out, despite the reports by Elliott
that such occurrences can be ignored ^ it would seem that
this potentially is a dangerous drug given intravenous lyo
Another case of pyridoxine dependency has been worked up
by Dr. McShann and Dr, Tower, These patients were also
studied by the Kry^ton®^ technique for laeasuring cerebral
Bietabolism developed by Sokoloff . The original case of
Hunt was restudied, and the patient now 7 years old is
still dependent, regularly developing seizures withia 72
hours of omission of her regular daily dos® of 10 sag, of
pyridoxine. Typical EEG abnormalities could be abolished
ia 30-60 seconds by intravenous pyridoasine-HCl (15 aig.)^
During a typical period of depletion, cerebral aaetafoolism
was iseasured by. SIrypton®® technique, and the decreased
oxygen consumption during the depleted state in this case
was similar to the situation reported by Sokoloff for hy-
poglycemia subjects. Thus 5 the interpretation tentatively
put upon the data obtained in this case is that during py-
ridoi^ine depletion a deficiency of the substrate for cere-
bral oxidative metabolisis exists which is promptly cor-
rected by pyridoxine administration. Since pyridoxine de-
ficiency affects ;r-as®inobutyric acid metabolism priBsarilyj
and since that compound appears to be a significant sub-
strate j, Dr. McKhana and Dr. Tower rationalise that this
case may actually represent an example of y-amiaobutyric
acid deficiency^ with a consequent reduction in oxidative
metabolissic Drs, Mclhann and Tower have continued their
studies of the metabolism of i^-aisinobutyric acid in neural
tissue by using the fluoriasetric methods as described in
th© 1957 report. They appear to have demonstrated that
the shunt pathway j i^e. glutamate to ^-aiainotoutyrate ap-
pears to be active and iaportant in cerebral oxidative
iffietabolism, and is significantly involved ia certain dys-
functions of the brain, such as seizures. They plan to
undertake further studies to see how such a pathway may
exert a regulatory control on oxidative metabolism and
hence on energy production in teras of normal function and
of seizure states.
-17-
Br. Curtis is contiauing his studies on physico»
chemical atethodology in aa attempt to obtain quantita-
tive data frora fluids ^hich contain estremel^ small a-
mounts of organic metabolites. He is working particu-
larly oa the surface tension of urine » and in particu-
lar optical raeasureffleats. hj polarised light aad its re-
flection off of surfaces utilising the elliptical po-
larization as an indication of the thickness of the sur-
face interface. The apparatus has been built in coEsbina-
tion with the Naval Besearch Laboratories and explora-
tion of this approach is now being orientated towards
the use of photomultipliers^ so that the square function
m&f be utilized, and saonochromatic light. Parallel with
this he is continuing his studies of adsorption on solid
surfaces 9 such ss coluam resins «, in foams and interfaces
in larine and water-iEmiscible liquids « Br. Curtis has
now found that there is so i^uch gross interference in
the acetylcholine-boron-flavonaol reaction to biological
materials as to make this procedure unsuccessful, in the
determination of microchemical amounts of acetylcholine.
His studies on guinea pig serum asparaginase, detailed
in 1957 J have now been completed ^ except for some elec=
trophoretic and ultracentrifuge data now in progress »
He finds that the purified easyrae preparation contains
two macromolecular contaminants which have defied at-
tempts at separation by electrophoresis or ultracentri-
fugal means; that enzyme can be quantitatively adsorbed
on a modified cellulose and in carbon dioacide foam,, and
purification by these means is currently being attempted «
Dr. Eorvath is continuing his work on proteins of
mwscle in normal and diseased states, and has calculated
total solids 9 total protein ^ non-protein solids (i.e,
fat), non-col lagenous proteins g collagen » water-soluble
proteins J, myosin ^ alkali-soluble proteins j, non-protein
nitrogen, electrolytes , and tissue water., He finds dif-
ferences in the normal and dystrophic muscle analyses
are reflected by connective tissue and fat, and by an
increase in sodium and chloride in dystrophic specimens.
He finds there is a relative increase of myosin and de-
crease in alkali-soluble proteins in most cases. These
changes seem to be independent of the remaining muscle
mass. The water-soluble proteins appear to be increased
relative to other proteins in most dystrophic samples and
an inverse relationship is indicated between the remain-
ing muscle mass and the percentage of water-soluble pro-
teins in the muscle on the other hand. He concludes that
samples of dystrophic muscle sot only contain less muscle
and more connective tissue and fat than normal muscle,
but that the protein composition of the remaining muscle
is different from the normal »
-18
Iq the studf^ of actin and tropomyosin in normal
and diseased siuscle^ Dr. Elatzo and Br. Horvath are turn-
ing to iBuauno logical properties of functionally important
muscle proteins. They find that rabbits immunized against
serum tropomyosin A, clam tropomyosin A, mammalian myosin ^
and antisera to human and cat myosin precipitate clam
tropomyosin A<, :^o such cross-reaction was found between
antisera to chick tropomyosin B on the one hand and clam
tropoBQrosin A or mammalian myosins on the other. Using
antibodies to myosin conjugated with fluorescein, myosin
in sections of normal human muscle was clearly and dis-
tinctly demonstrated under the fluorescent microscope «
Preliminary sections of dystrophic muscle similarly treated
showed myosin in residual islands of muscle and a sugges-
tion that in areas of active degeneration myos in-reactive
material was present in macrophages. Thus the immunologi-
cal findings are consistent with the present concepts of
the myosin molecule consisting of subunits - tropomyosin A,
3 and actin, the latter can be prepared in a higher state
of purity than myosin itself ^ so that it is more suitable
for investigational purposes. Since these proteins are
also iso-antigeniC; the immunological response of the or-
ganism may be important in conditions where destruction
of muscle could permit these proteins to escape from the
usual confines of the muscle and enter the circulation of
the body. Dr. Horvath is continuing the same studies in
muscle protein and electrolytes in dystrophic aic® ob-
tained from Bar Harbor.
Hs'o Korengold and Dr„ Hampp have concluded their
studies^ which were an attempt to confirm the findings
of spirochetes in the cerebral spinal fluids, with patients
suffering from multiple sclerosis. Identical material to
that vised by Ichelson was utilized, and a trip was made
to Dr. Ichelson 's laboratory to be certain that there w@r@
no differences. Twenty-two patients were studied in the
outpatient area^ at which time spinal fluid was removed.
Ho positive cultures were obtained^ and it was felt de-
sirable to terminate the project, after this number of
studies 0
The Branch of Ophthalmology haSj, over the past
year J, continued in its investigations directed towards
further imderstanding of the metabolism and growth of
the lens of the eye in relation to cataract; basic studies
and clinical studies in the function of retinal elements;
studies directed towards further knowledge of the forim-
tion and outflow of the aqueous of the eye and its rela-
tion to glaucoma 9 and studies of primary tumors of the
eye, and infections of the uveal system. Although orien-
tated in such given areas j, a multi-disciplined approach
is used so that widely dispersed laboratories may be en-
gaged on different aspects of a given problem. This co-
ordinated research is possible $ largely j, through the able
direction of the Branch Chief.
-19-
Specifically^ basic studies of the retiaaa from a
unicellular approach have coiitis&ued by Br. Fuortes, Dr.
Gourasj -aod Ur, Tasaki, ia an attempt to study the fea-
tures of the activity of the visual aerve cells ia the
eye J, as well as the more general problem of the trans-
ducer action of sense organs j whereby external energy is
transferred into a change capable of stimulating nerve
cells. Dr. Fuortes has fotmd the frequency of impulses
discharged in response to light stimulation is approxi-
mately a linear function of the logarithm of light in-
tensity in the single cell. The frequency of discharge
of the sane cells in response to depolarising electric
current is^ however, a simple linear function of current
intensity o It would appear, therefore j, that the logarithmic
transformation which is typical of light perception may
be exerted in this case by the photochemical processes
inherent in the perceptual cells » Sf light of supraliminal
intensity is utilized there is a sustained depolarization
upon which may be seen superimposed impulses. If a sub-
liminal intensity is used in the natural stimulus; only
a sustained depolarization is recorded. It is apparent ^
therefore, that light evokes the firing of the nerve
cells by depolariszation of the membrane. Dr. Fuortes 's
analysis of the interaction between light and electrical
currents in a single cell of the limulus indicates that
the depolarization evoked by light is the result of change
of conductance of the nerve cell's membrane, and that iss-
pedence measur^nents show directly that a change of the
membrane conductance occurs during illumination. In con-
trast, no conductance change occurs during electrical
stimulation. It is of interest that it has been reported
in other laboratories that the eyes (of certain fish at
least), respond with a depolarising change to lights of
one wavelength and a hyperpoiariziag change to other wave-
lengths. Dr. Tasaki plans to work in this particular area.
Dr. Gouras, before leaving to Join Ru^hton, at Cambridge,
brought to a conclusion the work on relations between
slow electrical waves and impulse activity produced by
illumination ia araphibiaa retina. His results suggest
that both the ganglion cells and receptor cells produce
electrical potentials during illumination and both con-
tribute to the electroretinograffio During this observa-
tion j, Dr. Gouras also described a phenomenon ccmparatole
to "cortical spreading depression'^ occurring in the e:s-
cised amphibian retina. This process is spontaneously re-
versible «, and recovery occurs in S-15 minutes.
Just recently Dr, Katharine Tans ley » from the In-
stitute of Ophthalmology J has joined the Ophthalmology
Branch as Visiting Scientists for a year. She plans to
continue EBG work in pure-cone maasmalian retinae. In her
studf she will use, nes-rlf as raossibl^, 3. BtoBcchromati'C
-20-
light source o Both flickering and single flash stiaiuli
will be used to stud^ the responses and dark adaptation
curves o This study is importasat la that one of the great
difficulties in studying the human ERG is the separation
of the photopic Ccon©) response from the scotopic (rod)
response o Many ssember® of the squirrel family j, on the
iU&erican Continent , possess pure double-layered cone re-
tinae. It is hoped that further study of the ©lectrore-
tinograM in these anismls ^ill lead to further understand-
ing in the ERG in Esan.
Studies in man have continued with Dr, Copenhaver
and DTo Gun&el, in which they coaibine electroretinography
with adaptometry; the latter is a dark adaptation plot to
determine the paramacular retinal area on the GoldMann
adaptometero In the past the EHGj, in the hands of Dr„
Bornnsheinj I>r» Dodt, and others^ has yielded information
of significant value in the diagnosis of laany retinal
color-defective subjects j coaaprising 8 percent of the
laale population. The defects responsible for the typical
color abnormalities are demonstrated to be retinal in lo-
cation rather than in the optic pathways or the cerebral
cortex „ These findings are of some importance in that
Le Gros Clark postulated that color reception was done
at geniculate level. The electroretiaographic method al-
lows the determination of the type of defect and to some
extent the degree of deficiencyo Thus the peaJs absorp-
tion due to the red-sensitive pigaseat erythrolabeg which
was found by Rushton^ was found to be absent in the re-
tina of this type of color defective. The sensitivity
loss in deuteranopes agrees well with the green-sensi-
tive pigment present in normals and also in deuteranopes ^
and hence suggests an interruption of the electrical im-
pulses from the green-sensitive cones at a retinal level
rather than a loss of pigment.
Further ERG studies were undertaken by these in-
vestigators with Dr. Bodt, a Visiting Scientist to the
Ophthalmology Branch, These studies were directed to-
wards spectral sensitivity curves on deeply pigmented and
albinotic human eyes. These investigators found that the
relative spectral sensitivities for wavelengths longer
than SSSeii were high in albinos and low in negroes, while
the dark Caucasians and subjects with "blond" fundi showed
intermediate sensitivities. The maximum sensitivity in
the albinos occurred at 61(^ as compared with a peak
sensitivity of 558m|j!. for Caucasians and negroes. These
investigators felt the difference in spectral sensitivity
in th® albino and negro is due to the reflection of light
by blood in the former. By traas-scleral illumination ,
these investigators fom&d that only selective absorption
of light in the tissue coats of the eye was due to blood.
It was also determined that the blood voltime in the
-21-
sclera and choroid cajinot be ascertained with this method.
Hence, this woTk. demonstrates the isiposTtaat effect which
the density of the pigmei^t epithelium has on the electro-
retinal spectral sensitivity.
Other studies on such pigments were carried on hf
the Section of Cytology and listopathology, of th© Oph-
thalmology Branchy by DTo Wolf, Or» Aronson^ and Mr,
Caravaggio» Tissue cultures of choroid pigment epithel-
ial cells and ciliary body pigment epithelial cells ^@re
raised in the Paul Chamber in the presence of staining
concentrations of Acridine Orange, and the fluorescent
image observed at regular intervals through the lifetime
of the cultures. Although Acridine Orange is toxic in
tissue culture, at concentrations of 1:100,000, it per-
mits growth at concentrations of 1:1^000,000; it has a
photodynamic effect in stained cultures, and makes them
more susceptible to light injury than unstained controls »
Healthy cells ^ill fluoresce green predominantly in the
nucleus and nucleolus. With continuing illumination, the
cells become brighter, and red granules appear in the
cytoplasm. At this stage the light injury is still re-
versible. However, if illumination is continued, the
nucleolus and the entire cytoplasm acquire nonspecific
fluorescence and at this stage the cell is irreversibly
injured. Thus the study of staining of living cells may
provide important infoxmation about the chemical state
of the compoaeats of living cells. For esasaple, the
metachromatic granules in irreversibly injured cells
probably are not ribonucleic acid, because the cells ob-
served do not contain granular aggregations of ribonu-
cleic acid large enough to produce the image observed.
These are some of the major studies concerning the retina
and the choroid.
The Branch has, in addition, continued its studies
on experimental cataracts, and growth of lens tissues.
Dr. von Sallsann has continued his studies on diet and
drug induced experimental cataract by directly applying
Mimosine, or to initiate Mimosine cataracts by studying
the effects of high pyridoxine and niacin levels upon
the toxicity of Mimosine. Such lens were subjected to
electron microscopic examination as well as to histo-
chemical examination; the first by Dr. Waako, and the
latter by Dr. Suhlman. Tryptophan deficiency cataracts
were also used in this study, and the eyes were studied
biomicroscopically^ as well as histologically ^ for six,
eight, and fourteen weeks after the five-day-old animals
were put on the diet. The Mimosine cataract has a unique
histologic picture in that, initially;, there is selective
4fimi^^ of t&@ «-^lis of tli€ g-eimiBative zoia# is th© ©a;.
*?t?i^es>f, asdprol if ©ration ©f tli©®® e#lls Is a ci.ffc.i.«i©-
^mp&wto Thm coffibiaatio® of sweh l®s® ehaasg®® wi*Si eos-
Jimcti^alft eonseal and aaterior uveal eliajag^s, ®ugg@st®«l
t!u@ local us© of th© conpoimd ia th® towm of fr@cs.ii#st i&-
©tillatioa of 1/2 pere©iit solutioa im th@ eoffijimeti^al
sae> low©v©rs, sueh t3P©at«©iit coaipietelf falls to pr©-
du&m nmj of th© f^u.rfae© el&&isg©e or signs of lesm daaag®
stm tli©f ai°@ g;@@Ei i% lilsosia©»f©d anii^ls. Hi© cstugnieai
gt^uetuif© of Missosts® is ibucIi liM© tliat of th® vitasiis
pfyidosis© Cs©© Ei5fo To'wigr's studies) $, a©d t© a Imsm @x-
t®st to that of siaeiBo thmxmiowm^ to ©xaaia© tli© po@-
sjlfeilit^ that ia this ©as© eataract fosmatioit is €omMm.tm4
ifith B& aetivitaMia ©ff©ct of th® toxie c^eponsd, h.igli
i#ir#l® of iritaffiins w@r© adainistei-ed in a© ©ffos-t to pre-
t©et th© aniaals froai such ©ff©cta of l!i3io6iii©o How#¥#¥..
sueh t3P@ata©at does mot ie au,^ way alter th© oculaif
®f@t^ic Miiiosia© ®ff©€t8., llects^os ssicroseopie ©xa^ias.-
tioffi of such cataracts bf Dip. Waafeo will show aeoaspl^M-
&vm d©v©lop»@at of ©udoplasmic r©ticuluffi ia th© ©qu&toriai
e©lls« aad dist©atioii of cfstie spac© b©tw©©a th© mm^'^
WwRmm of the f^tieuluMc fh©?© Im a dispersion of th#
SUA granules and ae^uffiulatioas of abaorsal^ fia© graa^la.r
»at@riftl is th© c@ll aucleus, fhu® th© abaoraalitf is-
plicate® Ijoth th® aueleus aad cjrtoplasiio Ttm sp@eificitf
of such chaBg©s to leas e©ll structure, howmwer, ±m t© Im
doubted ia view of the f iadlssgs ia isusel© disea®@ hj
S^So Waalio aad Shf^, %m disorders of sBuscle, Sr, lahlm^m':^
iav@®tigatioas ©f ©azpaes, particularly dehydrogeaas® ,
haT® aot led to ©oaclusiv© results is these Miisoeiae eat.-
araetSo la tryptophaa deficieaei^ cataract,, th© #€|tiatpri.a.l
%oa© reaaias unaffected ;» and th© structure of this ar^a.
is sow pr©s®nr®d, while th@ lens cortex and Immm mnel^ui
are destroyed == Here th© first change® ar© seen in thm
p@ri-ffiucl©ar sgon® abound th© anterior pole ©f thm Im&m
nucleus o fliere is a progresslv© d©coas>ositi©e of tilsmrm
which spreads later to th® surface of th© lens alosag t&#
sutures „ While th© ®pithellui& domm not uadergo muvM
chaages as described above tnitiallf ^ ia advanced sta«@a
it proliferate® to for® iBultilaf @r®d plaques or knotes.
These changes resemble those see^ in galactose and
alloxan cataract. Th® D"is<Hier towm of tryptophan mm
fed t© such animals on a tryptophan deficiency dieto
Clinical ©xaalnation did not werm&l any difference® in
th® utilissation of th© ©-isoiser fey itself or when f@d
siMRslt&aeously with th© L foifs, ll@ctroffi microscopic
studies of the epithelius, th© capsule and th© fib®r@ «f
the leass and on the epithelium of th© ciliary bodisgs aaa
optic nerve,, were mad® by t^o Wasko and Or^ voisi Sailisaim,,
i.s an atteisapt to iavestigate tke s:^orTO.al cfearacteristics
of lens tissues, as sees xu tfee electroa nsicroscope; to
iavest'i.gate ttee ultrastructure of the leas ©oitfeeiiiffls
aad tfev* leas fibers after cataractogejixc agents fead toe«?a
acteiiffiistered; aud to sttid^ the iftorpteologf of tfe€ ciliary
epitoelitffi T^xth the ©lectroE stdcroscope^ In additions to
the 105? studies reported last year, tte cortical lafsrs
of leB,s fibers were studied ia rat, rabbit,, mosikmj aad
calf. sSucto fibers represented elongated, priseiaticailf
shaped cells , outliis,©d bf deuse messbraass aad separated
from ©ach otiier by small iat@rc©llular spaces „ Tb© au-
ciei, tb© fflitochottdrias, t&e eadoplassiic retlculuis, the
Golgi comples:, and low densxty el<sac*®B£s in tb© aormial
leas irer© all described hj these investigators. Ixperi-
Mental cataracts were induced fef 1500 rad X-rays, aad t.li« .
l^UB studied in the electron microscope. Structural
changes is the mitochondria were aoted and th© isucieo-
plasM appeared in dense masses inside a lighter siatrix.
In the cytoplasm » profiles of endoplasisic reticuims ap-
peared la3fg©r than aorssal and there was a considerafel**
increase its RHA granules „ In Mfleran cataracts a grmx
quaBtity of low density filaasents i2i the cytoplasm of tJ^f
1&&S ©pitiasliuifi u'as acted „ as v/eil as a deposit ioa of a
desss© asaorphous substance In areas beneath the ©pitteliu;®- ,
The findings in the Miisosine cataracts were described
abov«„ Investigations on the ciliarf bod^ has beeis in-
itiated, and &B'f&t no definitive findings have ba<ea re-
ported by the investigators,
Dr„ von Sall»aun has^ in addition,, described tr^e
submieroscopic structure of the lens tissue by phase cosi-
trast microscopy i, in tissue culture of lens epithelitm,
Cultures in the Paul Chamber have been successfullf maiB"
tained for periods up to one usonthj while cul tares om
roller tubes have been asiaintalned up to two months „ Bag:
isiediuffia with horse seruss has been used as the culture imdAum
These investigators s feel in their preliifeiisarF stages that
there is some evidence to indicate that elongated Sorwm
of cell® are asore constantly produced in cultures €mu-
tailing chick, embryo extract ^ while cells grown without
this ^edia grow in sheets saor© analogous to the in vl^o
condition. The^p feel that the use of. phase mtcroBcopy
and tissue culture techniques enable the direct obse-^rTa-
tioB of' th^ effects of cataractogenic agents on livings
cells, aed that artifacts of the histological method art?
eliminated and the cell responses that escape detection
axe recorded b^ tiase-lapse cinematography for further Btndv
Investigation of the enzfEaatic systesis pr®sei5!t la
th© lens, cornea, and aqueous humor, has beeis under tafe«?si
by DTc Kuhlsisan and Dr, Resnik, with particular reference
to lactic dehydrogenase in the corneal epithelium,, is
glucose 0j©tabolism of th© cornea, using tracer exgerim^nt-
aad similar studies upon the lens a®d cornea after ac
tratios of 1000 E of X-rays „ Three speciest were iav^
gated;: the rat, the rabbit and the cat. Th^ rabbit -
-24
epitlseliim had the highest geaeral level of ©naimsatic
activity o Two eas^mes of citric acid cycle,, aamel^
malic assd isocitric dehydrogeaase were found, in all
three groups to be present at levels equivalent to
those present in cellular areas of the brain and retina.
While all species also contained glucose-6-phosphat©
dehydrogenase at a level equivalent to brain and retj-na,
aldolase and hexokinase were lower » The rabbit corneal
epithelium was unusual in that it had a verf high lactic
dehydrogenase activity ^ being 20 ti^es higher than that
in the cat to rat. These investigators found that the
whole cornea oxidizes glucose at a rate of approximately
one-hail that of liver or diaphragss^ and the presence of
a direct oxidative shunt in corneal Bsetabolisia is con-
firmed o In additions they found that the cornea is able
to oxidize lactate « and laasr do so even in the presence of
glucose o The removal of the epithelium fro» the cornea
reduces this ability to oxidise glucose by a factor of
80 to 97 percent 5 "Whereas lactate oxidation is reduced
only 27 percent o After irradiation,, although there laay
be morphological changes^ there was no alteration in the
lens content of hexokinase or glutathione reductase.
Preliminary investigations of such enzymes in the aqueous
husior are now started,
1^0 Resnik has continued his studies concerning^
the primary proteins of the lens. In his additional
studies 5 he- finds that the value for the sedimentation
coefficient of alpha crystallin is 17,0 x 10-13^ Thlm
value is slightly lower than that reported previously j.
but is based upon additional datac He feels the isiole-
cular weight of alpha crystallin is now 900^000 to
950 9 000 o In collaboration with Dr, Wanko^ isolated pre-
parations of this protein, and low density elements seen
in sectioned lens fibers «, were also carried outo At the
present time these investigators cannot state whether the
elongated structures in preparations of the low density
elements are alpha crystallin. These studies do indicate.
however 9 that such low density structures are proteins
and that the soluble lens proteins alpha,, beta^ and gasmz,
crystallin are present in these structures. Thus again
a coordinated prograas is in force in the understanding
of th© nonsal amd abnoraal lens^ in relation to growths
degeneration » and cataract forssation.,
Studies in nonsal and abnomal control of intra-
ocular pressure are also continuing, Dr, von Sallmann
and Miss Grimes report on the anatoisy of the posterior
ciliary nerves in cat and monkey » as&d the preparations
froas these species have in comsion the fact that nerves
close to the globe are generally mixed nerves, and they
contain fifth and third cr^ial nerve fibers. Isolated
-25-
"long" ciliary nerves may be found in the cat, however,,
which are not observed within the orbit , asid which do not
fuse with any of the post-ganglionic branches of the cil-
iary ganglia. In the monkey, the ciliary ganglion re-
ceives three or four coasmunicating nerves fTom the fifth
cranial nerve, lerves arising ia the ciliary ganglion
going directly to the eye without joining branches of the
fifth cranial nerve have not been observed in either spe-
cies j, but the segregation of fibers within mi^ed nerve©
and the subsequent branching » Might , iei the opinion of
these investigators J give rise to a few nerves which enter
the scleral coat and which are purely parasyaipathetic «
Covering the long ciliary arteries are nerves which sup-
posedly contain only fifth cranial nerve and sysspathetlc
fibers o These imrestigators feelj, however , that these
are mixed nerves which also carry post-ganglionic para-
sympathetic fibers.
A study of afferent electric impulses induced by
intraocular changes has been undertaken to see if such
impulses may terminate in diencephalic centers as origin-
ally postulated, toy voa Sallmanns et al. This study is
being undertaken by Dr. Lele in aniiaals in which external
ocular ©ussles and connective tissue and the ciliary
nerves are all diseeted from the globe j and connective
tissue excised. Pressure within the globe is maintained
by 22 guag® hypodermic needles j, connected by short lengths
of salia© columns to a pressure transducer. The nervous
activity was examined by placing recording electrodes on
each of the dissected nerves , and recorded from one chan-
nel of a dual-beaiB cathode-ray-oscillograph „ The trans-
ducer, in turns is recorded into the second channel of
the sasie scope. Dr. Lele finds that branches of the
ophthalmic division of the fifth cranial nerve did not
show any spontaneous electrical activity j, but in every
instances afferent impulses were evoked when the cornea
or conjunctiva were mechanically stimulated o Neither
spontaneous nor evoked activity, however ^ were recorded
fro© the short ciliary nerve originating from the ciliary
nanglioid and entering the globe. Responses obtained from
ciliary nerves of ©ixed origin were essentially similar
to those of the long ciliary nerves. All the long and
mixed ciliary nerves tested showed a response to increased
Intraocular pressure. In each instance there is a sharp
Imt transient increase in the frequency of the impulses ,,
lasting as long as the increased ocular pressure rises. Ihe
imximum frequency was proportional to the rate and the
height of the rise of increased ocular pressure. In ap-
proximately 60 percent of the preparation j, such activity
was sustained while the pressure was sustained. Every eye
tested showed this type of sustained response in one or
^ore of the serves „
B^„ Macri,, Dr. ^oa Sallisaara, aiid Miss Griaies Isa^®.
coffitiau©d their studies on th© effects of drugs oa iiitra-
oculaj* pifessure„ ©«■» Macs' i repoyts tiaat muscle felaxaats
such as decamethojsium (see Dro Irwisa's studies) ^ and suc-
cins'icfeoliis® probafolf have their effects oa spasEss of ©3k-
traoctilaf ssuscles (duriag 1957). To furthesp clariff this
asi atteiapt was isade to record intraocular pressure upon
stisralatioffi of the third nerve intracraBialiy » a®d after
rigor mortis o Th© procedures were essentially those de-
scribed in the 1957 reports These investigators find
that spasm of the eistraocular striate muscles induced bf
third nerve stimulation aad/or rigor ssortis produced
changes in th© elasticity of th@ eye similar to those ofe-
taiaed after the administration of thes© two drugs o This
effect could be abolished toy resection of the extraocular
striate muscles or enucleation. These investigators feel
that since three different methods inducing muscle spasm
produced similar changes in th© elasticity of th© e^e,
it appeared very uniikelir that factors other than lauscles
could be iis'g'olvedo If the ©fe was also placed in a cham-
ber filled with saline ., aisd th® pressure of the chaasber
raised to various levels and th@ elasticitf determined „
the effects on such elasticity of the eye^ under these
conditions 9 'were almost identical to those obtained by
muscle spasn. Utilizing the saetSaod reported last year
for the determination of aqueous outflows, devised in their
laboratory 9 these investigators find that now there is a
second biphasic outflow pattern which is not proportional
to th© internal ocular pressure throughout the pressure
range examined „ The biphasic curve was characterised by
a veTf fast outflow at lower intraocular pressures., which
then isiflected at pressure levels between 35 and 50 am.,
of mercury e Such an outflow pattern could b© induced by
parenteral administratioB of Diaao^o
Since it has been reported that the pressui'e in the
veins to which the aqueous humor flows is essentially con-
stant and independent o£ internal ocular pressures, then
th© outflow pressure should be th© difference of pressure
values between two ends of the chsmnels, i^e^ the intra-
ocular ^id th© venous. These investigators felt it was,
therefore 9 iasportaat to study the venous pressures oa th©
surface of the ©y®» Three ipeias,, th© anterior ciliary »
the long posterior ciliary , and the voltes ^can be casiau-
lated,, A cast material was injected iato the anterior
chamber -im^eT continuous pressure until »aay of the epis-
cleral vegisels were seen to be filled o The material was
allowed t© harden, and the tissue wa© digested away.
Thus the aqueous outflow channels were demonstrated in
their course from th® trabecular area to the Circle of
Hoviu®. Pressure readings in the su&terior ciliary and
-27-
voTtex veiasj and those of intraocular pressure , ap-
peared almost identical under resting coaditioaso How-
ever, if the intraocular pressure was either raised or
lowered, the venous pressure fello Thus they f©lt that
thej?- had shown that changes in internal ocular pressure
can alter venous pressure, Thej surasarize this by four
points:
1) The feiphasic outflow patterns become more
pronounced when the outflow pressure is calculated as
internal ocular pressure against venous pressure o
2} Acetylcholine » Arterenolj, histamine, sympa-
thetic and parasympathetic nerve stimulation all pro-
duced changes in the venoiis pressure which paralleled
the changes in the internal ocular pressure c
3) Trauma of the eye induced identical eleva-
tions of internal ocular pressure and venous pressure,
4) Diasiox lowers both the internal ocular pres-
sure and venous pressure.
Thus, for example 5 Diamox may have a double action in
that it may reduce the aqueous inflow, but it laay also
reduce the venous pressure in the eyoo Such studies
are important in the further understanding of glaucoissao
Glaucoma studies at clinical level are being con-
tinued hf Dr, Paton and D7c von Sallmann» in an attempt
to determine the most valuable diagnostic tests and the
prognosis and adequacy of glaucoma therapy. With pre-
sent day techniques J measuriament of intraocular tension ^
visual fields i, and the individual response to test situa-
tions^ a diagnosis of glaucoma is often uncertain. This
study is concerned with information gained from tonography
in borderline cases of glaucoma. It is also concerned with
distinct subdivisions of glaucoma noted as "low tension'%
"hypersecretion",, "pigmentary narrow angle";, or "inflame
matory" torsm of glaucoma. Patients are accepted to this
study by admission for a minimum of several days in order
that an extensive glaucoma workup may be performed under
rather constant environmental conditions, and at all
times of the day or night. Tests include tonometry with
day cuj-ve determinations,, applanation tonometry, measure-
ment of depth of anterior chamber , biomicroscopy p gonios-
copy^ visual fields ^ and photography of the optic discs.
At the present times cases of borderline glaucoma have
not been followed for sufficient time to judge the value
of th® data obtained. In addition it is apparent that
more normal control subjects must be studied. These then
are the primary studies concerned with increased intra-
ocular pressure.
»28~
A Study of the isflaMsator^ disorders of tfe© eye
is coatinuiag is two ©epa^'ate axeas: The first one toy
Dr„ K&utm&m. om toxoplasmosis a^d its tkerapy, and the
second one by Dro O'ltourk© aigd his colleagues in deter-
Hiiaiag the effects of endociriae glands upon esascerba-
tions of iaflai^iatory disorders of the uveal t^actc, In
the fis-st study l^r. laufiia^ is siaisEtainiiig straisis of
the organism in the chick embryo « and frois the chorioal-
lantoic S£^ibran@; as reported in the 1957 report o These
were then kept alive in tissue culture roller tubes o Hie
tisie of attachisent of the micro organism to the cells
could be deteranined by washii^ out the inoculum at de-
sired intervals 8 The organisafflt were allowed to multiply,,
tout th© culture could be fised and stained before cell
destruction had occurred c The effect of Darapriss on
tosoplasBiosis in vitro was invest igated„ Testing of pa-
tient siaterial was continued as in previous years. It
was found that slow growing organisiss were much more re-
sistant to Daraprim than the rapidly growing organ issis.
These studies also showed that appreciable time is re-
quired for the organism to be in contact with the cell
before an invasion of the cell takes place:, ^^^ that
chronic infection of the tissue culture can be produced
with slow growing strains o £n such chronic states c the
organism and culture sees to be in symbiosis «, and the
damage to th^ culture is not apparent o When the organisms
are incubated with serum containing a high titer of dje
test antibodies p these organisms were killed hj this
seruKs suggesting that the dye test antibodies smy^, in
fact J be toxoplasmocidal.
As noted ±m. the 1957 report ^ Dr„ O'Rourkes in an
attempt to ©xplaira the multiple reroissions and ejtascer-
bations of uveal infections^ has turned his energies to
endocrine studies,, in particular thyroid hormone turn-
over f, and he has now coispleted radiothyroxin® turnover
studies in 30 uveitis patients » aod B norsial controls o
His data suggest that in the main uveitis patiects show
retarded rate of utilization of circulating .thyx'o id
horiffione as compared to the normal. The sajor (difference
lies in the daily rate of degradation of I^^^ thyroxine.
Treatment with thyroid hormone hasj, in a few .watients,
in DTo O'Eourke's opinion 9 resulted in correction of these
afenoraial results 0
Finally^, Dr„ van Alphen is studyii^ immunological
relations in ocular tissues ^ in an attempt to determine
the possible antigenicity of lens capsule, and to produce
cataracts immunologically by immuniaation with lens cap-
sule and lens proteins; also to see if the various tissues
-29»
of the @y© are is»,ynol©gically related. DTo Yaa Alphea
finds that th@ sera of gisisiea pigs iismisaized witlx cap-
sule aad guiaea pigs' lens proteias or with calf lens
capsules aad calf leas proteins, show cross-reactions
with cos^eal epithelitm asid vitreous, but do not react
with doaor Mood, -iris, -retiaa, or -aqueous, . As&ti-
calf vitreous sera show strong cross-react ios^ ■with calf
blood aiad noae with other calf aaitigeias. Aati-calf cor-
aeal sera react with coriseal ©pitheliua oal^j aad not
with calf blood or ocular calf aatigeaSo Is none of the
animals irasiunised with lens capsules and lezas proteins
did cataracts appear, although repeated paraceateses
were carried out aiad some leases were traumatized. These
studies then represeait the collected research directed
towards infection aad iiaiEunization in ocular disease.
Dr. O'Eourke is also coatinuiag his studies on
the detection of ocular tuaiors by isotope tracer methods ,
using radiophosphorus , aad traiss-scleral counting doae
as a surgical procedure. E® finds evidence in four pa-
tients aost recently studied, the trans-scleral counting
results were correctly positive, although results of the
trans-coajunctival method were negative or equivocal.
The former aethbd seesis to be the present one of choice,
as might be anticipated by the low energy range of the
beta particles of radioactive phosphorus. Thus it would
appear that surgical procedure aiust still accompany a
diagnostic method of determination of intraocular neo-
plasia.
The Ophthalmology Braiash has initiated a ae^v pro-
ject this s©ar, studying th® basic factors in refraction
ancmalies. This is an effort to complete a statistical
analysis of the interrelations of the five optical ele-
ment ® in th© hissaa eye^ aad to test in part a theory which
assus©s the tension in the choroid by reducing the pres-
sure oa the sclera as a factor in determining the size of
the globe. It is clinically known that wherever the
choroid is absent, th© sclera becoaies ectatic. A quan-
titative conf irmatioB could foe obtained by measuring th©
pressure in the subarachnoid space, comparing this to
the intraocular pressure. The subscleral pressure in 12
eyes appeared to be lower than the intraocular pressure.
The differences asouat to 2 to 6 sm. of Bercury. Para-
sympathetic stimulatiosi of the ciliary ganglion leads to
decreased pressure, and sympathetic stiaulatioa of the
cervical sympathetic leads frequently (but not always) to
iacres^ed pressure. Several iavestigators have considered
th© choroid as too fragile to stand pull aad pressure. Dr.
van Alphen, however, h:s^ trephining scleral windows ia the
posterior pole, asay make the choroid bulge out and cause
-3®~
it to 3fetract on. paras^patlk^tic stlmulatioKo If a
lurge s©l®jpal wissdow is cut a tfaere is a large Mraia,-
tloia of tfee choroid ;, Mt evea wSaem o^e^stretclied tfee
choroid i© abl© to ©tajsd 90 ism of isstraocuLar pressure
Sr, G»mk©i. ia coatiisKiBg lis progara® of desigu
amd COBS true tioa of optical iast2'iim©ats ,j asad haSp dwriag
t&e past fear, ia corr©latioffi witfe Dr, CopeEha-s^er ^ad
oW.ewSg ©stalblished r©tisal profiles usiag new color
filters and smaller test sj?otSc. Dark adaptation curves
wer© oMa.iis®d with tli« iK)dif i«d iasti"imeat which have
b©©ffl fouBd to fo® quit© satisfactory c T&is aad othes"
tentative data ^ith otiier ©y© disieases indicate the po-
tential lasefislE^ss of such testing proceduir©© for a
^arietf of disease ©ntities,
Br, lau£i»ai&.« JDr. ^as& Alpiieii, a^id Dr. lyon Ballmaisnc
ha'¥© reported upon th© Ssigtol^ iE&t^restiag f iadiags ia
til® vitr@oiiS im primary faiiiXial amyloidosis,, Th@ mamcl®
of such cases were also examined in t!i© Medical Msurolog'f
Branch s and tter© is no doubt this almost pat&ogaoaossic
appearaac© is th® vitreous cosafinsjs the diagnosis of wi-
sysp©ct@d Bss^lotdoBls , In non® of the fi^if© cases seeiK
hf th© Clinical Director has ou.® presented with th© clas-
sical f ladings of am^loido Biopsy of gism, m&mclm,, skia^
new® and vitreous paracentesis, howeirex',, have coa-
firis©d the accisracy of this fiadiag^ aisid indicates that
a do®® ©^KaiBlnatioffi of th® iritreows is iEportant if this
di®®as© is to be coiasideredo
DTc Bruc© Cohan has undertaken a study in intra-
ocular angiography,, using radiopaqu© dyes after replace-
ment by such djem of the aqueous huMor, He utilise®
laiainai^raphic tachj^iqueB^ using a 0,3 m&^ focal spot
tube« Th& anterior ciliarf r@lM of the cat's ©f® is
also cannulated,, and radiographs axe ta^en during hand
iiBjection of radiopaque dyes into the 'S'enous systaa^ with
assd without paraorMtal tissues. 1^©- stud^ has resulted
ia the successful demonstration of th© Xr^'a^r anatcmf' of
the intrstocular vmimum s^stepjs in the essentiallf intact
cat e^® which will. allo« & sor© detailed study of the
aaatoagf of the iatrascleral ■venous plexus ;, and th© d^»-
n:?i!®i©s of intraocular vasculatur<i »
Finally 8 KTo Wanko ie continuiBg with his oteer-
Tatlon® of ffioraal and atonorsial striated muscle,,,, as ®^~
assim^d hj electron microscopy o Four norsel specimens
hair© been examined s, four cases of mi^otonic djrstrophy^
and one case of Werdnig-Hof f isan ' s disease have been studied c
In th© m|?otonic dystrophies,, there appears to be an in-
crease Jjs th® EWA ^aisules, sand a rareficatioa of th©
electron ssicroscop® , coafirmiag the ^Isidiisgs in the light
siicroscop©o la Werdjaig-Hof f ®aa ' s disease, peculiar
shaped mitochot&dria ha-^e bees observed. The total series
of all such cases is too ^m.11 to make definite coaclti-
sioBS at this time.
The Branch of Surgical Neurology reports iateasiv®
investigation of 119 patieats with cerebral seizures, th©
fiajoritf b^iag temporal in location. The pathology «, the
physiology Cat the operating roosa) ., the autonomic concom-
itance of temporal lobe epilepsy p the language character-
istics, and the psychological abnonsalities are described
in detail. Study of micro^lectrode techniques in tissue
culture in neurological and muscle elements have continued ^
as ^ell as studies is cortical neurones, and the effects
of hallucinogenic £^ents upon higher primates after re-
moval of specific areas of brain « The anatomical effects
of temporal lobectomy have continued, and a new stereo-
tascic device has been developed which will be directed to
the treatment of involuntary movements. In combination
with the Cancer Institute, studies in hypophysectomy have
continued. Further develoi«ents of the underlying fac-
tors in cerebral palsy have been reported, and attempts
to correlate this with the embryology of the central ner-
vous system. Tissue culture studies have continued ,, as
well as the effects of hypothermia upon the central ner-
vous syste® and -cerebral edema , The pathological charac-
teristics of a rapidly degenerating disease found in Mew
Guinea are described. New anesthetic agents, and their
effect upon cerebral circulationj, have been studied o Spe-
cifically,, the following investigators have reported their
projects as follows:
Dr, Baldwin reports 119 cases of cerebral seizures,
the majority of which are afflicted with temporal lobe ab-
normalities. From his sost recent studies he feels tl^at
a cryptic angioma is a significant cause of this form of
seizure ; and that this vascular abnormality is found more
commonly in the mesial temporal structures close to the
Junction of the circulation of th© middle cerebral and
anterior choroidal artery, Dr, Baldwin suggests that the
peri-insular tissue^ through its epileptogenic character-
istics « may initiate a perceptual process in the opposite
intact temporal lobe, I^, Baldwin reports a series of
patients admitted as probable temporal lobe seizures, which.
after further study » appeared to arise from the cingu-
late area. These patients had a clinical seizure pattern
characterized hf epigastric aurae, altered affect , altered
awareness, posturing and adversive movements, as well as
autonomic changes,. Dr, Baldwin is studying these cases
in conjunction with a similar series at the Mayo Clinic
under Dr, David Daly, and he hopes that this series will
provide a means for differentiating those seizures arising
from the cingulate gyrus. Dr„ Baldwin has continued his
-32-"
of the motor phenomeiaa of the temporal lobe seizures;
he !£ow feels that there are certain raoveiEaent patterns of
the hsmds and upper extremity which are characteristic
of epileptic activity in one or both temporal lobes. Ee
feels that such movesieats have a lateraliziag sigaifi-
caace, and that they occur oa the side opposite to the
most active temporal lobe. The aiovemeiits of the head
and neck in a taiaporal lobe seizure are usually such
that there is a turning to one or the other side; this
Btovement is a slow postural movement, and is thus dif-
ferent from the adversive novement, which is pathogno-
monic of the suppleajeatary motor area, and the chin
points downward, Se notes that diiring epileptic auto°
Bsatissij, th© fine digital movements are lost^ and the
hand is used apparently en bloc. He has now photographed
1,721 such phem
Dr. ¥an Buren has continued his studies on the
series of patients with temporal lobe seiauresj by
use of polygraphia nieasures&ents of autonosiic concosiitants
of such temporal lobe seizures. He has noted a hyper-
tension , tachycardia, respiratory apnea, a fall in skin
resistance and skin temperature ^ as well as swallowing
moveaients and inhibition of gastric motility. There is
no strict correlation of such autonomic activity to the
electrographic tracings. Perceptual aberrations do not
always coincide with clinically recognized seizures.
They saay occur without other stigmata of temporal lobe
seizures. The perceptual disorders of space and color
perception are most frequent. Dr. Baldwin feels such
perceptual aberrations are never separate from differences
in affect. In fact, in disturbances of the temporal lobe
by epileptic processes, the most frequent combination
is that of fear and perceptual aberration. Dr„ Baldwin
feels the physical basis of fear may be one of the most
significant sources of the clinical characteristics in
temporal lobe seizures. This has prompted him to turn
his research in the direction of searching for catechol
amines or other adrenaline- like substances, which may
increase in amount as a result of mesial temporal dis-
charge. Dr. Baldwin feels j, in addition, that during a
seizure, the patient does not have the usual appreciation
of body image. The "memory difficulty" which has been
noted so frequently in patients with temporal lobe seiz-
ures has also been studied. It is the impression of these
investigators this difficulty is not so much in memory
as in relating space and time. In the laboratory, Dr.
B ildwin has continued his seizure project utilizing peni-
cillin-induced seizures. Penicillin lesions within one
or both temporal lobes will usually project first to the
eoa'tex of tfe^g hemispheres iia th© parasagittal area os
tlb.© side opposite to the involved temporal lobe. As
sucb a seiKiire discfea-rg© spreads across tfee cortex,, it
is preceded by discerjaifele Tascialar cfeassge, a«d if Mas-,
sii^e may b© followed by sever© aad occasiousallf critical
@de!Ba<,
la the operating roo«s S'*- Baldwin, has coatiaued
iis th© electrical stimulatiom of liisffiaB aad higher pififfiiate
temporal lobes o la the tsumaa operatiixg roosi tlie in-
terest has been focused on the so-called psychical re-
sponses o The majority of such respoases haire coas© froai.
depth stiisulatioiii, but leay also be fouad from surface
stimulation o Approximately 200 positive responses feav©
been obtained from the chimpanzee cortex,, which were
Hiotor in nature j, and DTo Baldwin feels there siay be a
centralateral inhibitory motor areao The study of abla-
tion preparations continues j> and chiMpanaees have been
studied now lup to four year® after bilateral temporal
lobect^syo Dr., Baldwin finds the aniaial now adjusting
more socially s and remains More placid than his contem-
poraries » After four years,; in th© cas© of the bilateral
frontal lobectomy j, however , a similar animal does not
regain his place in the social hierarchy and his indi-
vidual and social habits remain abnorsal, Dr, Baldwin
notes the leesia'l teroporal lesions affect cossaunicatioa
in the chimpanzee for approximately four weeks after
their creation. He also notes that hallucinogenic sub-
stances which are contained in th© Mexican Mushroom do
not affect the chimpanzees whose temporal lobes have
been removed, yet such substances affect the normal
chimpanzee as to teak© hi® tame, relatively unaware of
his surroundings., and soaewhat ataxic » Dr.. Van Buren,,
and DTo Paul Yakovlev froffi Hariirard, have been studying
the anatomical pathology following temporal lobectoisyc
With an anterior teaporal lesion » nuclear degeneratioa
appeared in the inferior and lateral portion of the pul-
viaar<) and the posterior portion of the medial genicu-
late body 9 and the lateral part of th® lateral geaiciilate
bodyo In posterior temporal lesion the degeneration ap-
peared in the iBiddl© and posterior part of the pulvixiar.
the anterior portion of the medial geniculate body, asd
the aiedial part of the lateral geniculate bodjo Tfeey
note also a loss of cells in the posterior part of th©
nucleus ssedialis dorsaliSo These investigators feel
that the stria terminalis in man appears to arise from
the cortical and medial accessory basal nuclei of the
amygdala since it reasains intact when the lateral por-
tions of the amygdala are destroyed by surgery. Th®
an.terl.or coaaiaissure , bowewea^., was nearly SBtirely d®-
geaerated from such lateral iesion of the ajEygdala^ ang-
gestiag that the retaiffl©<l aaedial portions of the amyg-
dala affld region of the uncus received r^Tj little pro-
jection trom the aaterior ccwisissure. A desceadiag patb-
viSLj frcsai the amygdala to the brainstem is the lateral
part of the cerebral peduacl® is also fomido This could
foe followed as low as the lower poos=,
Br„ Yan Burea reports in souse detail his experi-
ence in hfpophysectom^ of graded nature in aiano Thir-
teen cases formed the basis of this studi'. Serial sec-
tions of the sella ^©re obtained insofar, as the size of
the retained pituitary fragiaentj, and differential cell
counts were made in this fragment c Such findings were
correlated in each case with the patient's clinical
course and the response of the thyroid and adrenal func-
tion,, and the level of gonadatropfeins j, and the presence
or &hB@mce of diabetes insipidus. The amount of pitui-
tary r©ffiaining after surgery varied froa 0„3 cubic im..
to 160 cubic wmo Iffifflediately after such surgery there
was profound depression of thyroid and adrenal acti'S'itys
and the gonadatrophin levels fell to negligible figures.
It was furing this time that remission of a primary tumor
might occurs, and this was seen in approscimately 50 p©a»~
cent of the cases , The most striking feature noted by
Dr, Van Buren was that there was no correlation between
the amount of pituitary tissue left and the amount of
bypopitMitarlsUp or tuaior reisissioa present in the pa-
tient a In a patient having 160 ,am. j, the thyroid and
adrenal function returned to normal , but th® patient'®
16 month post-operative gonadatrophin levels res&ained
near negligible figures. Thus, there was strong sugges-
tion that depression of individual trophic pituitary lso:r-
mones are not the same for all trophic hormones, fh©
only feature coiamon to all cases was surgical section of
the pituitary stalk and this saay indeed be the 'essential
feature 9 according to Dr„ ¥an Buren » The aaiount of dia-
betes insipidus present could also not be correlated in
any way with the amount of pituitary tissue resaiaiago
An initial rise of cholesterol was noted in 5 cases that
were seen to fall to normal in on© to three Month® fol-
lowing surgery c The initial rise nor the ultimate fall
again did not correlate with th® amount of pituitary tis-
sue left. Histologically tla©re appeared to be a lack or
decrease of specific granules of the chroaophile ceils ^
presumably to ®®@t the increased demand of pituitary hor-
aoaeSo These assm cases also provided Wo Tan Biiren witl^
valuable post mortem material for study of the visual
system.
In tfee visual Sfstem studies j which were a con^
tisxiation of the 1957, 38Cc) studies 5 Dr, Van Burea
studied humaa and primate retiaa; the effects of lesiozjs
of the optic pathway© upon the retina; the effects of
lesions of optic pathways and the lateral geniculate
body; and th® visual field defects following temporal
lobectomy o Dr. Van Buren and Dr. Baldwin reported the^^-
fiadisgs partially ia the 1957 report 5, and a paper no^
has been pubiishedj, in Brain, 1968 „ That portion of
the study having to do with the visual field followii^u.
temporal lobe defects teiminated with this report »
Sight additional retinal studies aai,d four lateral gen a
culate studies are still in progress „
Dr, ¥asi Burec. has now tested his new ster@ot;a---. 1
instrument on cadaver material at HIHo Obtaining of
such ssaterial has been difficult,, and the first such
cadaver was undertaken in liay^ 1958, Since this tis^
only five other cadavers have been availableo Dr,, Yau
Buren feels 5 however, on the whole the results have
been encouraging in that they show the principle of the
arcuate electrode carrier is a souad one under practical
operating circumstances „ a^-d that the apparatus is me-
chaMcally accurate o Simultaneous ventricular and cis-
ternal punctures were necessary for good paeu®iOgraphf iu
such cadavers o Dro Van Buren is continuing in the pre-
paration of a brain atlas for the utilization with this
stereotaxic instrtiiaent. The utilization of this stereo-
taxic instrument will be exceedingly important in the
futfure approach to minute lesions in the treataieat of
involuntary isoveffi©nts» Measureneats of such involuntary
isovemeats again is difficult and the surgical unit is at-
tempting the preparation of acceleriineters as a mea&m of
sisple graphic recording of such movesentSe
la 1957 Dr, Van l^raa reported that his findings
suggested that patients complaining of pain which appeared
Bore functional than organic in origin., had unusaally
unstable autonomic responses. He has continued thes@
esaffiinations during the present year in an attempt to
correlate the degree of th© autonomic responsiveness
with other features of the patient's clinical picture,
Ke feels J however 9 that his results have been practically.,
in this case., of no value. Autonomic responses to ap-
parently the same pain stimulus varied from examination
to examination p on the whole tending to decrease as the
patient became more used to the exa^iiser and the testing
situation 0 He f©©!®^ stt thim timm, th&t autoffiosiic ,t©«-
cording do@® siot s®®^ a pr©bafel# l®ad for aeca^at® ai^a.-
muremmsitm of paia i® aa obJ^eti¥® faslnioig,
BTo Li laas eoffltiawed laiB @tMdi®s of iiatrac^lluiaj'
recordings iu th® c©referal cort©^ aad iss tissM® cialt^ared
ffi#y¥® assd siBiffi€le c«ll®5 as ®«I1 as stMi@s of n^iarotraiis-
!gi3@ioj& in hypothermic o Dr, Li lias foimd fi^e s@paxat@
tfp@s ©f iistraeollular potsatials recorded trcm th® cow--
tex^ t^@ first tfp@ l>eiag & st®adf potsatial «mr®epoii.-
Btvtt to aff@r®nt stij^ulatioffi mid/or local application of
sts>fclyiiii@o 1# feels that tl&es® potentials orifgisiat#,
p^obablf trcm glia elesests. Secondly^ larg® slow po-
tentials oi'igiaates, prohahlj from glia ml«msiRtm. S®-
condlfp large slow potentials , proh&hlf origisatii%^&c%
deiidfites; tliiird^ &Sk&ll potentials pr@mmm.hlf origisat-
isg fron sfimptic s'egions; fourth;^ brief 0pik@«s witli aa
iffif legion im tke risisig pl&aise^ presimalblf rmcor^md from
ceil bodies; a,ffid fiaalliTs simpl® brief spikes froiB asEoas,,
■Kie missiatiasre potemtlalSg arisiisg from pr^siaw&blj sfii»
aptic regioffiSft ®bow a marked si^dlaritf t© that F#e®riil.#4
py«vioi»sl3f hf Fait affid lats., is Ssglasde in fflesas^oig^jBesila'r
JusictioigSo le f^rtfeer feelii t&at tlie meebaitisiii of e^.cli
sfmaptie traiisiiissio^ im tlse eemtral ^er^o^is @^tig%
lie^ce indicates tlie l%pos°ta%ce of dendrites i£i t.te pro-
d«a€tio!& of electrical actiwiti^s of t&@ cereteai c©rt®s
liaf b© o'irer-empbasisedo
Ee has coffitiffiyed t© ^ssiu'® t^e actlTitf of m@rwm
«®l]ts ta tfe® ffiotor cortesc «itSi ®icropip@tt@s w&ii® ^lem--
trie stisastlatioa was applied to ^arioiss ^^ibeoftical
structMy@s and peripheral ^essorf aer'sre®,. Thm cmllm wls.icfe
are iiatiaaateif related to liotor famctioa, a®d «&icli &a¥®
desceadis^ axo3%@ to i»edulla^f pfrasid were idejs&tified &;?'
tlieir reisponsiee to astidr^c sti^nlatioig., smd tb^os®
cells is tis© iiotor cortex wkicfe do sot fea-ire dm^^mmdlmg
SM>ms were identified as istters%i3icial cells. Dr. Li
verified him pre'vious report to tlie effect tlaat thm mu-^
cl@us ires&tralis lateralis of tl&e tt&alaigys activates th^
cells ^itb desceadi^ axos^ but suppress^ the actiiritf
of tbe isitensiancial cells ;, asgd lie suggests that thlm
th&lsMtQ nucleus m^w h&-wm sose control o¥@r tbe mttor bjc-
tivitf of tlft® es&perineiital aniiealo Se also foui&d that
wliat hm identifies m inters^mcial cells in tb<i motor
Gorteii; could be iaf lueaced bf a &®^Borf wollmj from the
peripfeeral cells o BuGh. a seasorf vollef, how&wers was
also capable of exeitii^g a isotor seisrona i^ tte G®rmhit%m.
Th® stwdf deaoastratedj t© ®r„ Li's satisfaction, that
tfee refractorf periods of th® pfraasidal f ib»s^ varied
froK loS to 2.5 sillisecondJS, and tisat th® coadsactioss
^elocitf was 3 aeters to @5 a@t®rs per secossd.
-3I~
The sfttc&roaoias activitf of umrv^ c®ll® in tfee
cdretoral co7t«:s w&m thm subject of a further studf hf
DTo Xiii^ is whicli he foui&d o»If a irerf f®w !£«s°^€) c@Ils
ill a sphere of 1 an. In th® cerebral QowtmK ^ould dis-
charge precisely at the sasie iaistast; secondlf^ that a
sfachronoiss irollef eiroked discharges of swerve cells wit!&
tiae discrepancies varyisg froai 2 isillisecoads to 20
aillisecoaids; third, that the applicatio£& of strrchsiisi®
activates about 85 percent;, but siot all of th@ nerve
cells; and fourth, that there is a tiae relationship
between neuroiaal activity in the "aroused" corteSc
Since it is generalli^ pictured Csee the reports of te„
AJaone-Marsan, @t al>, that neurons in the epilepto-
genie cortex tend to fire in linisonj, the present studf
suggests that this is a generalisiation with a certain
degree of truths fros the strychnine espuriaents but that
in the normal cortex in an alert subject cells are fir-
ing randemliTo
Xn his studies in hypothermia » Hr, Li has con--
eluded with publications on the effect of cooling on
the neuroiBuscular trassnission in the rat. Iliis studf
indicated that there was a critical bodf temperature
in MaiSBals below which the transmission of iiiipulses a-
cross the neuroipsscular Junction could not occur , and
if the bodf teffip@ratur<@ would be further lowered to
4®C, transmission is^ coepletelf blocked „ Sinilar studies
on conduction of impulses in cranial and peripheral
nerves were carried out by Z^„ Li and Dr. Orti%, in whicli
siaall segisents of the optic nerve and sciatic nerve w@re
subjected to -ISO^C for 30 seconds. The animals were
then kept alive for 1 day ~ 4 lionths, and inpulse con-
duction was tested at various intervals o lliis studf is
in its initial stage, and is designed to see if extr^gielf
low temperatures locally applied to tis^u® ham &bIatioi»..
experiments by sui^ieal procedureiSo
Finally j, in coordination with Or^ Baldwin's pro-
jects listed above, Dr. Li and DTo Ortis have been @ti%df-
ing the effects of the hallucinogenic activity of the
SSexicsm nushroos in both cats and mo&k®wm. Multiple
electrodes were used which were capable ®f injecting
s^nut® quantities of the testing chessiical agents ^ and
inserted into various depth structures of the brain. E@~
cording of the electrical activity and responses to stim-
ulation fro» th^^e structures and fro» the cortical sur-
face were made. At the present tii^ such studies are
still inadequate for conclusive stat^ients to be amd^o
lis. the Section of Clinical neuropathology » Dr,,
Elatso reports him initial imv<»^tigatioms in pinocytosis
of lab@li®d pr©t@iffis i» tissu® cultur®„ fliis con-
sists of i&'ls@lliiig proteins ^it^ a £Iuor@se@i&t c<sm-
poaeat aisd f@«disg cultures with tli@s@ labsll^d pro-
teins. H&KS diff@rdiic0s ^tw@@@ iMividtial €®11 tfp@!S
could b@ demonstrated in this a@p€Hct as cellular pro-
tein giotabolisifi m&f b@ studied hf alt^fiisg pE^ tempera-
ture» asd ch4^ical substi°at®o l®wl>om kitt®^ aiad rat
c«reb«Ilu!i %@r« grown in ^itro^ Cat s^ruM albumin ajad
rabbit serum globulin were labelled with fluorescein
isotbioc^anateo The cultures were "starved" for a
period of tiiree hours ^ and coi^equentlf fed witb the
labelled substance. The preliminary' findings indi-
cated that it was possible to denonstrate protein up»
take bf living cells gro^is in vitro „ A sigi&if leant
difference in leetabolism of proteins bf variosas cellu-
lar elffiients was observed. Cultures washed for a brief
period of tise after feeding showed abundant labelled
proteins in the aiaerophages and onlf a few fluorescent
droplets in the glial elements. Cultures washed for
several hours in balanced salt revealed abundant grmmn
fluorescent droplets in glial cells, whereas the nm-
crophages showed iscMStlf autofluorescence of various
lipid substances, Ihis studf is also important in re-
lation to collii&ation techniques for detection of brain
tunors with radioactive serusie albusin labelled with
l^^^. This would s@e% to indicate that the uptake of
such substance was not onlf due to breale in the blood«
brain barrier, but to actual ingestion of the labelled
protein bf the tuiior cell.
©To llatzoj, Ikto Sorvathj, fmd Dr„ S^imrtf, are coss-
tinuiffig their studies of the localis^ation of mfoslia. i@
hus^an striated ansscle bf fluorescent antibod^^ using
the Coons' fluorescent antibody technique « fhef find^
in normal nuscles the specific stain for Myosin was
observed in the A band, and that the I and 1 band ap-
peared unstained with the Z band showing an occasional
non-specific autofluorescence o In studies of the various
patholf^ical processes in husan muscle , there was a
striliisg persistence of antigenic reactivit|r of li^osiis ±m
fibers with far ad'^anced degeneration. Regenerating
fibers observed in cases of miscle injurj and polfn^o-
sitis showed similar features to those siosscle f iberis
grown fros chick es»brf^o« Am occasional macrophage aliso
contained green-fluorescent inclusions in their cjto-
plas»o This observation m&f be of isaportance for in-
terpretation of possible mechsuiis^ of hfpersensitivitf
due to release of ^ssuscle proteins. Attempts at the pre-
sent are now being ^ad® to induce allergic sfopathf i.m
laboratorir animals <.
Br,. Slatso aiad .^... , .,.,„,^du3@fe have eow completed
their fiudifflgs ±n Koru disease ^ tfo® ffi&iia patfeological
findings beiug a widespread neuronal degeeeratioia;
mf^ltu degeaerat ioHt affect iag predoffiinantly cortico-
spiaal aad spiao-cerebellar tracts; iixt®iase aad wide-
spread astroglial aad Microglial proliferation; peri-
vascular ciaffings with Moaoaucl@a^ ele»©s:&ts; aad the
presence of peculiar plaque- lile bodies ira half t&e
cases studied o to masij wa^s this disease resembles
t&at described tef Jakob-Creiatafieldo Together with Dr„
Ortiz-Gal-faUg aB.d Dr, Laskowskij Dr. Klatzo reports
some stuxdias oe rsgeaeratioa ia the central aervous
systems, after tfee application of cold,, and then injec-
tion iiitra-cysternally with prednisolone. The prog:ress
of the regeneration will be followed hj photic stiasu-
latioa recordings froia various parts of the central
nervous system „
DTo W„ Eo Engelj Dr, Li, aad Dr, Klatsso report
on the feistocfeemical and electrophysiological observa-
tions of muscle fibers grown in vitro » The iEuscl®
tissue is obtained frcm 14-day--old chick embryo or new-
born rats and studies in the RHA (ribonucleic acid) coa-
tent was demonstrated with gallocyanln and Toluidine
blue methods. The first appearance and localizatios^ of
myosin in myofibrills has been followed with specific
fluorescent antibody „ Data on the electrical activity
has been obtained fross cultures several weeks oldo This
study has demonstrated that spontaneoiis activity ©ay
occur in such fibers. In earlier days of culture ^ spoB-
taaeous pulsation of asuscl® fibers may also be seen, D.r„
Bngel is testing specific blocking agents to such tissis©
culture after administering electronic stimuli to the
muscle fibers 0
DTc Miquel and Dr„ Horvath,; and Dr^ Klatsog ar©
utilizing a new quantitative aethod for estimating pre-
cipitin reaction J, by applications of antigen-antibody
»i3Etures to chr«Matographic paper » By using fluoresces^ t
aatibody instead of seruM ia the tests p th© ratic be-
tween the amount of antibody to antigen ia th® precipitate
may be quantitatively measured o This method has been
applied to the precipitin reaction between antigens of
contractile muscI® proteins and their respective anti-
bodies. The quantitative data obtained by this method
is in agreeissent with the mach more complicated and cum-
bersois© K;jehldai nitrogen deteraiaationSo Th® sensitivity
of the isethod was estimated to be as low as 1 gans&a of
nitrogen »
„4©"
Dro Laskowski aad BVo Klatzo are coat iauixig
their studies on the relatloaship befwees edeiaaj, blood-
brain-barrier aad tissue elemeats iss experimeatal braiE
imjurf o Sodium fluorescein was us@d for this studf of
th@ blood-brain-barrier „ These fiudings have aow heen
published In the Journal Neuropathology aad Experimental
neurology 3 is i»hicfe the development of edema was ob-
served within 6 hours ±n the white matter umderlyiag
the site of cold application » This edema ©.xhibited strong
PAS-positive staining of astrocftes aad less iatease PAS
staining of interstitial spaces o The histocheisical
analysis of PAS positive staiaimg in the edematous whit®
matter suggested a glycoproteia nature of the substances
involved o Electrophoretic studies performed at the
tim© of maximal intensity?- of the ©desia and foreals-dowa
of blood-brain-barrier indicated an appreciable increase
in total proteins with a striking elevation of alfousBins
in the area of edema » This fluorescence in the super-
ficial laf®rs of the cortex persisted one month alter
injury j, and was associated with presence of s»all as-
trocytes ^ while that of the deeper layers » io@, white
matter,, disappeared in this period of time„
In the Section of Developaaental Neurology ^ Dr„
I>ekaban is continuing his studies concerning the site,,
type, and extent of lesions involving the central nervous
systeia in cerebral palsy, and allied conditions. Dur-
ing 1958 Dro Dekaban reports 56 patients studied In
great detail as inpatients ^ and 28 as outpatient® „ An-
alysis of his results shows that in 62 percent of the
cases the site of the lesion was d@teria.ined; in 27 per-
cent such an abnomaality was of diffuse character,, aad
in 11 percent the localization was not possible. In
29 percent of such cases the lesion was compatible wit,li
a destructive process j, in 12 percent with a congenital
malformation , aad in 21 percent it was of diffuse charac-
ters and in the remainder of 38 the lesion could not hm
estimated with confidence. An etiological diagnosis
was nade in 43 percent of all patients by elimination
of antibodies 5 lipid contents la cerebrospinal fluid ,
special retinal stxidies, and estimation of amino acids
in wriae,, pbesaylalarsiise ia blood ,, aad a geaetic assef .
This brings to a total 141 patients that Dr. Dekaban feas
studied since the toegimsing of this project,
Dro Dekaban's extensive survey of 4,480 products
of pregnancy at the lational Navy Medical Center aad
Walter Eeed Hospital ^ between March 1, 1956, and Marcfe I,,
1957 J, is contisiuiffigo Over 8© percent of such products
have now been evaluated., and the final statistics of
-41-
In^UTf to the central ner-vovM sfstea ia sucls a large
group of pat lea ta tBk.®n at raadofe is of ©xtrem© iM--
portaisce in. ®st teat lag tfee incidence ia caisse of cere-
tea.l palsy „
®r« Bekafoass has also coatiKued his study of tfe©
pathological lesioas in patieats eoisiag: to post ®ortem
who ha-r® eeati-al u®twqub eysteis lesioas oecorrisig d^r-
iiQg presaatal,, ixitraaatal aad ©arly poBtisatal iif©o
T%'©2ity bralas h&ve isow fe®®B oMaiaedj which are teiiag
processed and. studied « Analysis of tfee pathological
ficdiBg® repeal tfc,at ia 9 c&ildren tb© atoormalitf of
the central aer^'ous system i?ajs of preraatal origin; i:a
fiT© it was coapatifol© witJs birtfe issj^iryp amd ia oj&lf
four was it tiae result of iatracraiaial lffif©ctio2!o In
cae cerebral K©oplas» m'as preseat., aad so ceatral ner-
Toms sfstem atonoraalitf was detected,
Bt. Dekatoau asd Dr. Baird are cojatiiauiag tfe®ir
studies of tMe products of diabetic BEOtlaerSj In wfeicM
th^j u,ot@ tfeat th® total for all wastage of pregasaiacies
in diabetic aotfeers was 43 o 4 percent as coapared to 1?.6
percent ±n the Boxaal control o Of tiae starviviug off-
spriag born to tfej© diabetic motli®s°S9 6o7 percent slj.o'w©d
congSEital abixorMalities or various neurological areas;
tills compares with onlj 0„48 p®rc©Bt of sttjaorsial
eliildsr#n in t!i® xstora-diafoetic control group. Tte© tallies
of t&s;ir findings m&f b® seen ia their detailed pre-
ssBitatioKio
Br. Bekabaa feas also coatimued Mis i®ea®ur®ffiei&ts
of ®xtea*aal aad iateraal orbital distance ia isal©s aad
fenal®® fro® birtfe to adulthood 5 as3,d lias aow accomplished
tlies© measureroeiits 03a approxiffiateij 60© raortial cliildremo
This material is teisg ciirresitly validated ajisd sutese-
quesatlf will fee s^fejected to statistical ans.lYB±B,
His sttidy of the Bonsal deirelopffierat of the laotise
tos-aia is coatinuiBg ajid has bow resulted ia as atlas of
tfee norm&l mouse teaixs, Caxeful disisectiOKi of the brain
aad teaiastem in 11 age horiaoas have tosea pesfoymedo
Productioa of gialfonaatiouis by meaas of .s-rsdiatiora has
also he®n attempted „ Appros:iMat®l|' 10 percesat of the
litters of x~a*adiat®d mice have aajor ataormalltias;
about 25 percent misior ataoamalitieSs a.nd the rem&lnlng
ai'© fr@e of detectable pathology ,
-42-
Dr. Lsfflsdells, ia t&® Section of Cliaical Psf-
choiogys feas uadertakea studies to the effect of "fear-
provokiiag" stimuli on visual discriaiaatiosj ia primates ^
aad aE iffiteissi'?® stud^ on t&® psjcteo logical evaluation
of temporal lobe disease. H® and Mrs. Weissbach &nd
Miss BleviaSj Isave reported a teadeacy for patieats
witto left temporal lobe reao'^al to be "poor cosaunica-
tors". Since Dr. Lasisdell has recently joiaed tliia
Clinical la^'estigati'^'e UEiitj, smcis of feis prograsi is
projected into the future „
As lioted by DTo Baldwin, the Clinical Unit has
been injured by the loss cf Dr. Kenneth Hall, who has
joined the staff of IDiulse University as Associate Pro-
fessor in Anesthesiology s, in charge of research o Be-
fore leading , Dr. Hall terssinated his fluothane studies ^
showiBg that fluothaae has proves to be a potent nos-
combustibleg aoB-tosic anaesthetic agent 9 and that Flu-
ether was found to b© non-combustible ajsd a relati'v'ely
stable agent hj various chemico-physicai criteria.
The physiological effects of the latter drug in acute
dog esiperiiiients generally paralleled those of fluothaae ^
the latter being the more potesat of the two. Dr. Hall
s«id DTo Norris have reported their findings in dog in
Anesthesiology., in May„ 1958 3 and Septembers 1958. Dr.
Hall continued also his work ±n hypothersiia in neuro-
anesthesiology g and in the use of succinyl choline in
the awake craniotomy. This latter project has been
developed to the degree of proficiency that it repre-
sents an isiportaat adjunct to the surgery of epilepsy »
and renders more successful the electrocssrtical studies
performed in human patients.
Finally 5 Dr, Pritchard aad Dr., Edgar have been
studying the effects of hypertonic urea solution in re--
ducing intracranial pressure in patients undergoing
surgery with suspected brain tumors and with ward pa-
tients with clinical evidence of increased intracranial
pressure. They find that the brain voluae and iatra-
craaial pressure may be reduced by the application of
urea. Thus far no serious side effects have been noted s
but not enough cases have been reported for full docu~
It at ion at this time.
Sn closings Clinical lavestigation® Unit also
again acknowledges its debt to Miss Huiburt and her
staff J particularly through a period of difficult tran-
sition during which each of the major branches trans-
ferred the majority of its admissions into given areas.
Much of the data reported herein is done with th©
cooperation of the Cli®ical X-ray Department j, Clinical
-43-
Pathology, asid tli® Iisstrtment Section of the Ceatral
Services, Fiaallf » we would like to acksowiedge the
cooperation and aid of tia@ Atos^ic Energy Cosiaissiong
Central latelligemce Agency » National Na^-al Medical
Center., School of Aviation Medicine ^ and Walter Eeed
Army Hospital ^ as well as other Institutes of the Na-
tional Institutes of Health , with whom many of the
projects were undertaken.
At the request of the Director,;, national In-
stitutes of Healths the projected program of the
Clinical Investigations Unit to the year 1970 has
been suhnittedo The suggestions of the Board of Sci-
entific Councilors have been incorporated into this report,
Calendar Year 1958
SEBVICES GIVEN BY THE CLINICMp XIWESTXG^XONS UNIT
OF THE M^IONAL INSTITUTE Of NEUROLOGICAL
DISS^ES AND BLINDNESS
Services given by the Clinical Investigation® Unit to
the Clinical Center of the National Institutes of Health are
as followss
One thousand four hundred and eighty- five (1,485) conssults
were rendered in either the in-patient ©r out-patient areas of
other Institutes,. Of these, one thousand and thirty- three (I9O33}
were Ophthalmology, ninety- six (96) Neurosurgery, and three hundred
and fifty- six (356) Medical Neurology. Electroencephalogrsphic
Laboratory carried ©ut one thousand five hundred and two (1,502)
exaninationaa Of thase, five hundred and forty-eix (546) were
patients referred by other Institutes o These were distributed
as follows;
NCI 0.00.000.. 306
NHI O , O 0 O O O O « O 76
NLaMD 00.0 CO 00 00 81
NMI coo.oonoc 56
NIMH =,a<>..ooc,» 2?
Twenty- three electrecorticograans w«sr« performed on ffl«lsure
patiente in central surgeryc Indsfelllng electrode® w@re in^lanted
on thirteen subject* <,
These figures show that the consultive services of the
Institute have now levelled off with ceo?sl®te activation of all
beds of the Clinical Center s&d reflect altaost the exact figures
given in the previous year»
S®rvic(g@ by th® Neurosurgical Unit ware continued in
which major intracranial or intraspinal operations were done on
patients who w@r@ adsaitted to other Institutes^ The collaborative
project with the National Cancer Institute ©n hypophysectomy has
continued and will be detailed in the report by Dro Van Buren^
Eighty- seven (87) radioactive scan techniques were done
on patients as a consultive service during the last year^
The Neur^athology Laboratory processed one hundr@d and
twenty- four (124]) surgical specimens » eighty- five (85) ©f these
b@lng muscle biopsies and eleven (IX) of the latter were from
outsidei sourceao One hundred and aiusty-one (191) autopsy
cAaes vers processed sad these included thirteen (13) Kuru
cases and sevsn (7) cages from the Belgisa Congo c
Collaborative work with the Physics Division of the
Atomic Energy Comaisaion has contiauedg as well as with the
Central Intelligence Agency, National Kaval Medical Center,
Walter Reed Army Msdical Center, sad Johns Hespkinffl Unlversityc
Calendar Year 1958
SERVICES GIVEN BY THE CLINICi^ INVESTIGATIONS UNIT
OF THE MTIONM. INSTITUTE OF NEUKOU)GICAL
DISEiUSES AND BLINDNESS
Services given by the Clinical Invest igafc Ions Unit to
the Clinical Center of the National Institutes of Health sr©
as follows^
One thousand four hundred and eighty- five (1,485) consults
were rendered in either the in-patient or out-patient areas of
other Institutes o Of these, one thousand and thirty- thrse (1,033}
were Ophthalmol ©gy^ ninety-sis (96) Neurosurgery, and three hundred
and fifty~siK (356) Medical Neurology^ Electroencephalographlc
Laboratory carried out one thousand five hundred and two (1,502)
esaminationso Of th@s@, five hundred and f@rty°°six (546) were
patients referred by other Institutes o These were distributed
as followss
NCI oooocooooc 306
NHI C O O . O 0 O O O C 76
NIAMD occooo=o.o 81
NMI co«co»o.,o 56
NIMH =oo»oo„ooo 27
Twenty- three electrocortic^gnons were performed on seizure
patients in central surgery » Indwelling electrodes w@re iniplanted
on thirteen subjects c
These figures sho^ that the consul tive ssrvlcee of the
Institute have nmi levelled off with coa^l@te activation of all
beds of the Clinical Center and reflect alsnost the exact figures
given in the previous yearo
Services by the Neurosurgical Unit were continued in
which major intracranial or intraspinal operations were done on
patients who wer@ admitted to other Institutes c The collaborative
project with th© National Cancer Institute ©n hypophysectomy has
continued and will be detailed in th© r^ort by Bro Van Bureao
£ighty°3@ven (87) radioactive scan techniques were d^sne
on patients as a consul tlve service during the last yearo
Th@ Neure!f»athology Laboratory processed one hundred and
twenty-four (124) surgical specimens, eighty-five (85) of these
being muscle biopsies and eleven (II) of the latter were frsia
outaide aouixeSc One hundred eiad tiiaety~one (191) autopsy
c*3es were processed and t-heae iacluded thirt«ea (13) Kuru
esses find »ev@ii (7) cases iteet the Belgium Congo «
Collaborative work with the Phytic® Division of th«
Atomic Estergy Conaaisslon has continued, «8 w®ll &■» with the
C^Btrsl Infeellisenee iigency, National Naval Medical Ceater,
Halter Reed Amy Medical Center, and Johaa Hopkins University c
SUMHARI
As in the previous years^ the Braneb a@tivi^ has consisted of
rontina d3.agnostie ses'riie© (for the entire Clinical Cent^:') and rBseaitsb.,.
the latter chiefly latilizing part of the patient population of NINUBj, but
also of other Institatesp as well as animal material for expexliaenfcationo
Since the last report tip to the date tiiis repc^rt is being pr©icH.red
(November 30» 195^) a total of 1^502 eleotroeneepnal®gi-apbie examiiiations
has been carried outj, patients from the various Institutes being dis*-!"!--
buted as follows s
NCI o . o o 0 o . o o o c 306
Nffl: o „ c o o o o e o c o 76
MMD o o o o o . o o o o o 81
MI o o o . o . o o o o o ^6
imm „ o = . o . o . o = c 2?
NINDB o o o o o o . o o o o _956
Total 1^502
In collaboration -with t He Branch of Neurologieal Surgery --23- -e-leetrei^
soiiiicograpnlQ studies were perfo-rsued on flcsasions of cortical expesttre
during the siargical treatment of apileptie patienteo In a few epileptie
subjects in which eleati^odes were chronically implanted -within subeortioai.
structures for diagnostic loealization purp^seSj, extensive EE3 studies
were carried aut under dif fersnt ecndi-tieKSa
As in the pastf, a considerable miisber oS BEG examinations were pes -
foiled as part of research projects ©utsid© of our Branch and as a requts^st.
adjunct to rsaear-ch projects of Institutes other than NINDBj and this
service has taken up a relatively l^'ge portion of the total aetivity of
tlie Braneho Fortunately the active and ps'ofielent help ©f some of the atafi
roembergi and of Dro Ko Abrahastt in par-ticularg as well as skillful teehnieiant
and secretaries has made this collaborative service pe-ssible and^ it is
hoped^ of some practical usefulness «
From tte Braneh of Eleetroeneephalegrapl^ a total of eleven rsseareh
projects are in prcsgrsss or ha^e been compjleted vdfein 1958 » Of thesS;,
eight are xram. the Section of EEQj, five are new {kC-s, SOg 6Cj, 7Cj, 8C t
three are eontiauations of loiog range pix>jects pre-elously outlined in tlm.
1956 andf<^or 1957 reports o Three are frora the Seetion of Clinieal Neur©i=
pt^siGlogy ,j
Projeets IC^ 2G and 3C are all related to clinico=-electrographiG
problems in the field of the epilepsies and their description has already
been given in detail in the 1957 reporto
Some data pertaining to projects 2C and 3G (790=1957 and 81t 0=1957 «
arespectively) have appeared in printed foMa in three papers during 1958
calendar yeas-o
Project IC (continuation of 78C-1957) is progressing very satis ^
factorilje The investigators aire now processing and preparing the wealth
of material accumulated in the last 30 months in form stiitable for print-
ingo Arrangements have already been irade with the editor for the piiblica"
tion of a monograph - Atlas illustrating in detail the multiform patterns
of the epileptic fit as well as their electrographic correlates in a large
series of cases »
Among the new projects of the EEG Sections, one is considered canpleted
6Cj some of the data of two other projects are ready for publication or are
actually in press kO and 7G and tie remaining two 5C, 8C are still under
way and will be carried out through the coming calendar yearo
Project 6C was carried out and ccmpleted by one of the research
associat-es and the final paper has been accepted by the Jo of Neuroplysiolo
It deals with the "control" exerted by a number of subcortical structures
upon the thalamic and, chiefly^ cortical potentials which can be evoked by
peripheral i^rve stimulation ami which are considered as one of the elee=-
trographic manifestations of the arrival of centripetal seasery messageso
This project had the honor of being officially commended by "Uie Chairman
of the Editorial Committee of the NINDBo
Project ho is an experimental approach to the problem of epilep^
and specifically deals with the investigation on the nature of those EEG
discharges considered as the characteristic and typical eleetrsgraphie
signs of epileptic lesions » The first part of this project vjas ccmplet-edj
the results presented at a National meeting and a paper is now in press ^
The study of other facets of the same general problem is now in progress «
Project 5C was initiated recently and no results are yet available
for this ps^sentationo It deals with the general problem of the relation=
ship between relatively slow EEG changes in the cat's visual cortex and
the behavior of local unitary elements o Most of th@ technical details
involved in this experiment have been solved and ths first results appear
quite promising and of interesto It is hoped that some definite answers
to the problem tinder issrestigation can be obtained within the first half
of the coming calendar yearo
: a eliadcal 5,n-78stigationo It teas been e-osxpS-eieid
-„ies are stiD:.! collected, before thrj papsr is ready foe
liblic^itjxtJio The pro Jest* deals icith the ■urmrmal (.and abnormal) EEG- :re'=-
:.u3tion wM.cVi can he elicited In certain patients on steroid treaianent wheij
submitted to intermittent xjfeotlc stiBi'al.atioac. In view of tte 6bser¥ation
that on!J.y patients mtk ces-tain -t^^jpss of systemic dJ.sord@rs present tlil.s
photic aeti^'etdOB;^ a synergistis meebaM.sai was tentatiTely suggested b©--
tween the existing CNS patfeol®gr asd. Uie s teres id effeeto The high in--
cddenca of seizijres in tlie ssjm group of pat-ienta sonfir?!® the relatiosisixl
betx^-eea conviilsi-ws tendenag- ard t-te observed typa of photic aetivatioE
andj indireetly^T eautions against the (ab)use of steroid therapy in thcs©
G£ses in ^^hich such an activation is presento
feet 80 does not arealijc deserve suck a qualification and it is
mentioned in this report only for the salc@ of offering a ccsnpleta picture
of the aetltd'ty of the Bransho Noti^dthstaiiding the non-researeh charaet«r
of this f'pro^eet", howeverj, it is felt tliat the little time spent in its
actuation is far from being useless o /t the present rat® of seientifl@
publications th© a"5ailability of coroplate and speeiallzed bibMagrsgiphiis
raferenees becomes a3isiost indispensable andj nnforfeunatelyj, the task can
only be successfully undertaken by people ha¥i,ng g,?eat familiarity mth
the fleldo
From the Seetion of JleTiropi'^siology thjree projects were undertaken
and partial results are now reac^ for publieationo
Project 90 deals with tha investigation of excitation and conduction
of the nervous impulse in layelinated fibers andj, in particular^ with e%=
pcriaents on maimaalian nodes of Ranviero Prelisiinary ©bserirations suggest
a similarity of patterns of ionic currents in marnEialian and irwertebrat-e
nervous tissueo Son® of the results chiefly relating to teehni&al details '
are to be presented in early 1959 at the Natio.oal. contention of the Insti'^
tute of Radio Engineers-,
Project lOG is a study of the meehaniisms of synaptic transmission.,
Easides a few interesting obserrations on the effects of the pH of the
extraeellular astdiuB on ths artificial depolariaatioa of -the post^syimptie:
aiejabrancp Bost of tbs work is still coBcent.rated on teehni©al=imethodological
a^jpects of ths problem o
A farther project of the Saetion of Neurophysiolog;^ is only briefly
mentioned here because it appears in fall in ths report, of the Laboratory
of Biophysics of NIKDBo It was carried out in oollaboraticn. at til©
I'ferl.ne Biological Laboratory at Woods Hole and deals with •toe int©3^reta~
tion of r^rve funetion in terins of the fast transports of ions across the
membrane of the squid giant axoiio Prelimiisary results have been obtained
and a paper (Jo del Castillo and Jo Wo Moore s "On increasing the velocity
of a nerve ir^ulse") has been submitted for publication to the Jo Physiolo
TsTO papc of" Electros ncephalographj related to researc-
projects otitl:',; rcl oovacletacl in p.t^vious ariKiial reports have
appeared ±n tj^eir final ic 73ar3
lo Ralston. Bs Lo s The msctianisK of traiasition of intei'ictal spild.Bg foci
into ictal seizure dischargeso EEG Clin<, 'Ne^^rop!:5'•siolo WS^si 22.* 217-£.
2o Ajiiions Marsap.j CoS fecruiting response in cortical aM s"ubcortical
stnict-ur'ss 0 Archo i'balo Biolo 1958s 96! l<=l6o
A considerable contribution to the routine activity sxid research pro-^
ductivity of the Bi'anch was prcyrided by tb^ n-usasrous scientists who selecix-;:
the Branch Itsell for either traixiiisg or active cooperation in original
irRrestigatiiJB worko Visiting scientists (2)j, clinical associates (2)5
research associates (2) and guest -^sorkers (1), have taken active part in
several research projects and in tte diagnostic serviceo Some vieK! alreai^!"
td.th the Branch in the last caleMar year^ ^shile others plan to contintie
their work thro'Kgh 19$9o Their enthusiasm and eagerness are only isatched
hj their high sense of adaptability to the precarioijs space situation whichj,
as alreac^" mentioned in the 19^7 report^ represents thg only facet in the
BraiHih physical organisation pjhieh could stand sosie iiapx-ovemento
It is with great sorro-Kr that -we -sd-sh to mention the untimely ani srsdden
death of Doctor To Fo Enomoto "Kho was one of our most promising and efficient
research associates©
In closing this report, tl:^ Ch3.ef of the Branch wishes to esqpress his
sincere appreciation to the Clinical Director for his help^ guidance and
constructive cooperationo
ItetiJisaal lasfclfcut© of Neur©logics;l
Diseases and BllBd&@S3
Cliaical' Eeseerch
Electroencephalography Branch
Serial Nutssbers of Projects:
NINDB-lCc), HIKDB"2(c), WIHBB-=3Cc), MMJB'=4(c),
NINDB°5<c), HXKI5S"6(c), KII®B-7Cc), Kim)B-=8(c),
NINDB°9(c)p aad NINDB-10(e)o
Estiaiated ObllRatloas for FY .1959
Totals $96,500
- Direct! $84,000
aeifflbursments $12^500
ORP-2
Serial Noo KINDB'^l(C)
lo Electroenespnalography
and Glino Neuropbysiol
2o EEG
3o Bethesdag MaxylaM
ho Continuation of 80C, 19?6
78c, 1957
PHS<=NIH
Indiiridual Projeet Bspoyfe
Ca3.endar Tear 1958
Part Ao
Project Title; Analytical sliaidy of focal cerebral seizuras
Principal Inyestigator s Cosiiao Ajmone Marsan
Other Investigators; ICristof Abraham
Cooperating Unita; None
Man Years (calendar year 1958)?
~ Totals olO
Professionals olO
Others dO
Project Description:
Objectives s Outlined in the title? described in previous
reports and in the Methodo
Me tn<?ds_employB d ; Described in det^dl in the 1957 report
"(780} « Briefly it consiste of a special photographie tech-
nique by which one obtains a series of closely spaeed siiagl
fraroea synchranised vath the EEQ tracingo This provides a
continuous and pesroanent recording of all the electrographi
clinical events taking plae® throughout the developsnent of a.;.
epileptic seizures and^ chiefly^ it permits a very analytical
study of all the details >7hieh would likely escape tte sifflpl-*;
visual observatioHo It furtherraore enables or^ to closely
coirelate aotor^EEG phencsnena thtis obtaining resxalts of tm---
questionable physiopathogenetic interesto
No fx)
0HP"2(a)
Major findings; Over 150 seizures of different tgrpes ha^B been
elicit8d and recorded -Kith tha i^xf methods These are routinelj
used in the weekly conference in the discussion of the patient "s
caseo About 60 cases j z^presenting the most interesting, i3.n«s-aal
or demonstrative examples are now being selected for display and
analysis vjith the respective portions of the EEG recordo It is
of interest to note obvious discirepancies betwsen clinical and
eleetrographie behavior* changes in the former showing no cowelates
with changes in the latter or vice versso On the other haMj cer-
tain motor phenomena appear more often acccmpanied by EEG modifica-
tions than others^ etco
Significance to the program of the Institutes This project is part
of a vast research program related to diagnosticj, etiopathogenetic
and therapeutical aspects of focal epilepsy and of temporal lobe
epilepsy in particular^ which is one of the main projects carried
out by the Branch of Neurosurgeryo
Proposed course of the project; Continue tlK routine collection of
seizures for discussion of each patient's casso Goacplete the
selection and analysis of the demonstrative examples and publica-
tion of an Atlas of such analyses to illustrate clinical^electro^
grapliie correlates as well as the details and the variability of
patterns in foeal cerebral seizures o The Atlas should be ready in
early 1959.
0RP»2
Serial NOo NIKDB-l(C)
lo Electroencepnalbgraphy
and Clino Weurophyaiolo;
2o EEG
3o Bethesda^ Maryland
ko Continuation of 80C, 19"
78c, 1907
PHS^NIH
Individual Project Heport
Calendar Year 1958
Part Ae
Projjsct Title? Analytical study of focal cerebral seizures c
Principal Inrestigator s Cosimo Ajmone Marsan
Other Investigators; Kristof Abrahaia
Cooperating Units; None
Man Years (calersdar year 1908);
Total! olO
Professionals olO
Other: „10
Projeet Description;
Objectives s Outliied in the titlej described in previous
reports and in the Methodo
MethcKls employe ds Itescribed in detail in the 1907 report
T78C)o Brie:^^ it consists of a special photogsaphie te«sh"
liique by which one obtains a series of closely spaced single
fraines synchronized ra.th tte EEG traeingo This pi^jvides a
continuous and pesraanent recording of all the electrographie-
clinical events taking place throughout the development of an
epileptic seizure, and^ chiefly ^^ it peraiits a very analytical
study of all the details which would likely escape tl^ simpl©
visual observationo It furthermoi^ enables one to closely
correlate motor^^EEG phencmiena thus obtaining results of tm<=
questionable physiopathogenetic interesto
Part B included Yes /7 N© /^
0RP«2(a)
Major flrjdings; Over 150 se3.zuares of different types have been
elicited and recorded xd-th tte i^^ inethodo These az'e routinely
used in the weekly conference in the discttssion of the patient "s
caseo Abotit 60 cases, representing the moat interesting^ umxsiaal
or demonstrative examples are now being selected for display aM
analysis with the respective portions of the ESG reeordo It is
of interest to note obvioiis discrepancies between clinical and
electrographie behavior j, changes in the former showing no eorrelatt;
xd-th changes in the latter or -eice versao On the fether handj, cer-
tain motor phenomena appear more often accorapanied by EEG modifiea^
tions than others^ etCo
SignificaiEe to the progran of the Institute? This projeet is part
of a vast research prograas related to diagnostic, etiapathogenetie
and tHerapeutical aspects of focal, epilepsy and of temporal lobe
epilepsy in particular^ which is one of the main projects carried
out by the Branch of Neurosurgery o
Pz'oposed course of the project; Continue the zxjutine collectiorx of
seizures for discussion of each patient's caseo Complete the
selection and analysis of the demonstrative examples and publica=
tion of an Atlas of such analyses to illustrate clinical=elei3tro=
graphic correlates as well as the details and the vazdability of
patterns in focal cerebral seizures o The Atlas should be ready in
early 19$9o
ORP-2
Serial Noo NIMPB^-aCC)
lo Eleetrcf>enceptialography "
and Cliiio Neurophysiology
2o EEa
3o Betiiesda, Maryland
ho Continuation of 8lC^ 19^6 j
79C, 1957.
PHS-NIH
Individual Project Report
Calendar Year 19$8
Part Ao
Projapt Titles Deptii electrography in epileptic patients
Principal Investigators Gosimo ."jmoiB Marsan
Other Investigator; Krisfcof Abraham and John Van Burea
Cooperating Units t Kora
Han Years (calerriar year 1958)!
Total: oio
Professionals olO
Others olo
Objectives i See Project 790^ 1957
Methods employed; (See Project 8lC<.1956s 79C°1957)o Sine®
the last report only three new cases were studied in whiefe
electrodes were implanted in the depth of both teitpsral lobes
and also on the cortical (or dural) surfaes of temporal and
frontal lobes of patients affected ra.th various iypes of api-
lepsyo The electrodes were kept in place for about 10 day^
during xfhich time daily EEG tracings in various conditions
could be obtaixiedo
Major firKJings; The findings described in projects 81C-1956
aiKi 79C--1957 have noi^ appeared in published fonsi (see Part B
of this project) o The data from the more recently collected
cases are still in the process of being elaborated^ parti-
cularly in regard to the relationship between deep and cortical
elect rographic changeso Of special interest is the stu«^ of
the electrographic modifications, as recordable by means of
implanted electrodes, during metrasol~induced (or spontaneous)
seizures s The latter have been ai^l3r2ed and will be presented
togetner mtb the large series of seizure siaidies described
in the previous projecto
Part B included Yes /x7 N® /T
0HP-2(a) Saspial N@o NINDB-SCd
Sifflfictance to the prograsi of tae Institutes Saise as in previoiis
prbject>^
^'roposect coDirse of the projeot; Patients suitable for this study
have to be carefTSl7~iil!ectecrin view of trse possible risks in^^'
volved ill the techniqae aM for this reason their naaiber has been
necessarily liraitedo Collection of furtaer cases is contemplated
and the proposed course of this pro^Ject remains fundamentally tin-
changedo
Serial Noo NINDB 2(0
PtS-NIH
Individ'oal Piroject Report
Calendar Year 195^
Part 3; Honors, Awards,, am Publications
Publications otiier tnan abstracts from this projects
lo AbrahaKg K and Ajsnon© Marsan, Co : Patterns of cortical
discharges and their relation to routine soalp eleetre-
encepiialogrsplijo EEGo Clino Neuroph3rEiola, 195B« 10 s
2o Ajmone Marsanj, Co and Van Buren^, Jo ; Epileptiform Actiyi^r
in Cortical and Subcortical Structures in the Temporal irf>be
of F-an - in TEMPORAL LOBE EPILEPSIp C. Co Thomas, Spring-
fields Illo, 1958g 78=1080
Honors and /wards relating to this project,
NONE
05P-2
Serial MOo KIlTOB^lCe)
lo Eleetroenceplialography
and Glino Nextrophysiologj-
2o EEG
3o Befchesdaj Tlaiyland
ijo Continuation of 79C-1956s
8UC=19^7
PHS==KIH
Individual Project Report
Calendar Year 1958
Part Ao
Project Titles Elactrocorticographic studias in temporal lobe
epilepsy and in focal cerebral seiztareso
Principal Investigator; Cosimo Ajmone Marsan
Other Inyesti gators : Maitland Baldwin
Cooperating Units? None
Han Years (calendar year 19$^^) s
Totals olO
Professional : olO
Others olO
Project Descriptions See BkC - 19^7
Part B included Yes ./k7 No
Serial Noc JONDE-MC).
PHS-NIH
Individual Fro;)eet Report
Calendar" Year 19$^
Part Bj Honors, Awards j, and Publications
Publications other than abstracts from this pro^jects
Ajmoi^ l-Iarsang Co ard Baldwin^ M«s Elsctrocorticogs^phy •
in TEMPORAL LOBE EPILSPSy, G. C, Thomas, Springfieldp 111
1958; 368=395
Honors and Awards relating to this project:
None
0HF»2
Serial Noo JlJIB22sM5l— ..— —
lo EleetroenciptSography
and Glirio Naxirophysiology
2o EEG
3o Be the s da if Marylarsi
lio New
PHS^NIK
Individual Project Report
Calendar Year 1958
Part Ao
Project Title; Epileptic aGtt.vation of taaitary elements of the
cat csarebral cortex and their relationship with
EEG discharges o
Prlncix?al Investigators Takayiiki F« Enomoto
Other Ing-eatigatorss Cosimo Ajiaoaise Marsan =• Paul Gerin
Cooperating Units s Nons
Man Years (calendar year 1958);
Total; 1
Professional; 1
Othej?; 0
Project Description;
Objectives; To inwestigata the intiiaate nature of the EEG epilep-
ti?om paraxysmal patterns commonly referred to as "sharp waves'*
or "spikes"*
Methods employed; Epileptic foci were produced exj»rirnantally on
the gyms suprasylvian of cat by means of local applications of
different eonvulsant drugs (strjchninsj, penicillin^ ciirareg etCo).
The development of ths relatively slow EEG discharges was monitored
with a roatine surface electrode and^ upon their appearance, a
systesnatie survey of the behavior of the various units within the
different laysrs of the nearby cortex was carried out by means of
Tungsten laicrcieleetrodes made according to Hubsl's descri-ptiono
A similar method was applied in a few experiments in wlii^ paroos-
ysmal discharges were elicited following intravenous administration
of different drugs o
Seversl thousands of units were recorded and analyzed in 29 ex-
pejriraents carried out on cats©
Part B included Yes /^ No /~7
0RP-2(a) Serial Noo NINDB°l4(C)
Major ftrriirigat Observations were made on a) tte gsnsral pattern
of unitary activity i, b) the general and detailed relationship be-
tween tmit activity and slow EEG paroxysital discharges^ c) topQ>=
gs-aphieal distribution of the various activated unitary elements ,
d) ana34'j|,des araong the effects of the various corriralsant drugSj,
and e) in-fceraetion between unitary elements o
It waa foundy among other things, that the two most characteristd©
features of unit behavior in coincidence wLth an EEG discharge are
the paroxyOTial appearance of high frequenc?y bursts and a marked
tendency towards synchronization of a very large number of dif^
ferent units o Tnis hypersynchroiy is not absolute beeause when a
given element is characterized by rhsrthmical, high frequency firing
in "resting" eonditionSfi the oommon pattern in coincidence wilii.
the EEG discharge is an arrest eittier temporary or permanentg of
the firing itself o This characteristie behavior was analysed and
dissussedo
Unit activation may take place in correspondence with any phase
of the EEG events however, for a given unit the tiriB course of itis
firing and the pattern of relationship with the whole of tte sIoh
event or a given phase of it» tend to remain quite canstsnto From
these and other observations it is concluded that the mmhsT of
units activated at a ceirtain instantj their firing pattemg
location and tanporal interrelationship are closely related to = i
not responsible for = the final shape g amplitude and polarity ©f t
slow EEG evento
Signifieanc© to the program of ths Institute; A better undarstand --
ing of~the~3isinjei~of tliose EEG changes, whfdi are considered
almost pathognomonic in hustan epilepsjj, is greatly naededo The
signifisanc® of the abov®«mentioned data^, obtained experlraenfcally^,
but closely pertinent to such a problem is evident if one consides-.
that on© of the main dinical projects at NINDB is the study ^^f
foeal epilepsyo
Proposed course of the projects Part of this project is eoaripletecli
the results presented at the Juik meeting of the Americaii EEG
Society and a paper sulaaitted and aec^pted fos? publieationo Furtfc.:
studies on the saa^ line sltq now under way with tbe puipose of
a) compariBg local ("original") discharges vrith distant ("projeGte::.
ones in regard to unitary behaviori b) investigating the unit
aetivi^ during fully developed seiaures (the above stu^ was
limited to the inter=ictal discharges) | c) esteM the analysis @f
cortical unitary patterns following systemj.c adm3.nistration of soh-
vulsant drugs p
Serial Noo NI!©B=.li(G)
PHS^NIH
Individual Projeet Report
Calendar Yeaif 195B
Part Bs Honors J Awards, and Publications
Publications other than abstracts from this projects
EnoiaotOj, To Fo and Ajmois Marsan, CoS Epileptic activation
single cortical neurons and their relationship with EEG dis-
charges o EEG Clino Neuropl^siolo 1959s in press.
Honors and Awards relating to this pixsjeets
None
OEP=
Serial Noc NIMDB<=='g(G)
lo Electro eneepnalographv
and dirso Netiroph^siology
2o EEQ
3o Bethesda^ Maryland
llo New
PIS<=NIH
Indi'Tidual Project Report
Galeniar Year 1958
Part Ac
Project Title; Urdt amlysis of the responses elicitable
in tlia visual cortsx
Principal Inyestlgator; Cosinsj Ajmone Marsan and
Lenimrt ;^den
Other Iiwestigatora t None
Cooperating Units; Nom
Man Years (calendar year 195B)s
Totals olO
Professioiml? olO
Othey; olO
Frojeet Description;
Ob jectlvea ; As implied in the title^ the pujpose of this
research is to analyze tte various components of the ccwiplex
potential evoked in tl^ -^sual cartes (following stimulatioa
of the lateral geniculate nracleua) >d.th particular regard t©
the activation of tl:^ ccsptical unitary elemente and their
relationship with the slow surface raspoasso
Methods eBiployed; Acute experiments in cats either X2eja<=
butalized or only curarized after a brief period of psntothal
anesthesia for the craniotois^o Subcortical stmctures
localized stereotaxically for stiimilationo Recording from
go lateralis with silver siacroelesti'ssda and w3.th tungsten
raicroeleetrodso Systematic suricey with the latter through
depth of cortex and un<ferlying white matter o
Part B included Yes /~7 No /^
0RP-2(a) ferial No, HINDB^$(C)
Majjor fiixiings; This series of experimeiibs was started only
recently and at the raoment of the present report the various
data have not yet been elabcra-fcedo From the first experiments
howeverj, there appears to be a wealth of interesting findings o
Significance to the program of the Institute; The study of the
Behavior of unitary elements in various cortieal areas has been
carried out quite extensively by a number of other imyestigatorso
In this project the emphasis is placed on the relationship be-^
tween slow cortical event(s) and single cell activityo Its sig^-
nificance rests on the information one can obtain thereby for a
better knowledge of the intimate essence of the EEG phenomei^io
Proposed course of the project; Carry out tbds recently started
pro j act o
0RP=2
Serial No. NIKI!B=6(C)
Electroe ncephalograp^
and Clino Neiiroplijrsiology
2o EEG
3o Bettesds.^ Maryland
^m- liin Uo New'
Part Ao Calendar Year 1958
Projeet Title; The modification of sensory mechanians by sub^
cortical structures o
Principal Investigators Ro Gordon Long
Other Iiwestigatora s None
Cooperating Units; Nojkj
Man Years (calendar year 1958)
Total?" 1
Prof es sional : 1
Other: - 0
Project Descriptjon?
Objectives s To further el^icidate the effects which the brain stem
reticular formation^, tte non=speeific thalamic system and other
subeortieal structures (basal ganglia-, thalamie associative nuclei j>
rhinencephalic fonnatioias) may exert on peripte rally evgked sensos^-
potentials at thalaraiss and cortical level So
Methods employed? Experiments carried out on 50 cats induced with
etherj curarized and maiiitained on artificial respiration (some
stwdies als® performsd on nembutalized animals )o Bipolar stiBiula-*
tion and bipolar and monopolar reeording teehnique was usedo Sub^-
eortieal structures for recording and stimulation were located
stereotaxieally and histologieally controlledo Frequer«y, for tim
test stiuiuli (to contralateral peripheral nerves) at Ooj/se®
(rarely at ^--lO/see)., Conditioning stimuli deli vended at ttes©
approximate frsquenciess Oo5/see| 5=10/se@s 250/seieo Cortical
reeording from primary reeeiving areas (mostly somatosensory and
visual) by means of Tektronix amplifiers and IXtraont CROo
Part B included Yes /x/^ No /~7
0RP-2(a) Serial No, KINDB°^(C_)
Major findings; In the unanesttietized preparations the modifi-
cation of soinatic and visual potentials was obtained most easily
and the effect was most prolonged when the conditioning stijttxili
were applied to the reticular formation of the brainstenio Gon^
ditiordng stiiraili to the non-specific thalamic ^stem^ amygdala^
putamenj globus pallidas and lateral aspect of the head of the
caudate nucleus were, in this order^ deereasingly effective in
raodif^ng sensory impulses; stimulation of the pulvinar=late rails
posterior <x>iaplex produced some modification of visual responses
oalyo The changes recorded in the evoked respoxises were more
marked at the cortical level than at the level of the specific
thalamic relay nuclelo In genaral the modifications were of equal
degree and duration in the primary and secondary cortieal sensosy
areas o
The obaeryation made by previous investigators j, that high<-fr8queney
stiraulatJ.on of the reticular formation will depi'ess the amplitude
of evoked somatic and visual responses^ has been confizmedj this
depression of amplitude was obtained more consistently and was of
longer duration in the somatosensory systemo In addition^ it has
been shown in this study that lower frequencies of stimulation
to Hie reticular formation and its projection will augment the
amplitude of evoked visual and somatic respor^seso Toe phenomenon
of "rebouTid" increase in amplitude of evoked potentials after an
initial depression has been obsai-ved and described- These modifi^
cations at sensory potentials were abolished or markedly diminished
by barbiturate anesthesia <»
Oes'tain mechanisms by which these modifieaticns may be produced
are raentioiBd and discussedo It is suggested tl:iat the reeiprQca3.
effect of augstientation and depression of afferent conduction is
analogous in certain respects to the facilitation and inhibition
of motor responses by tte reticular systsng and that the augment'
tation of afferent signals may represent a meehanism which permits
limited focusing of awareisss or attentiono
Significance to the program of the Institute; To quote from the
comments of the CteiKnan oi tTie NINDB Editorial Csmmitteoj "there
are veiy few subjects of investigatioa which could have sueh im-
portant implieatioas, not only t® neurology and neurosux^egy but
to the psychological and episteaologieal branch of phylosophy
as well"-
Propesed eoigse of the projetsts This projeet is complet/sd.,
fxhdiitigs arid conclusions war® written up and a paper submitted
and accepted for publication in the Jo Keuropbysiolo
Serial No^ NINDB=>6(C)
PnS-NIH
Individual Frojest Report
Calendar lear 1958
Part B ; HonorSj Awards, aLti Publicatiom
pTjblicatioEs othsr than abstracts from this projects
Longg Ro Gordoni The modifica-aon of sersory meehanisjns I
subcortical stnicttireso Jo Nerrophysiolj 1959j) in presso
Honors and Awards relating to th:U' projects
None
ORP-2
Serial Noo KLWB-'liG)
lo Eleetroencephalographj
and Cldno Neurophysiolos""
2o EEQ
3o Betliesda.fl Maryland
PHS^NIH li^ Ke-^
Individual Project- Report
Part Ao Calendar Year
Project Titles EEG changes induced ;d.th photie stimulation in
patients treated with ACTH and adrenal corticoidso
PrLnsipaJ. Investigator; Kristof Abraham
Other Iiryestlgatora; Nelson Go iiichards
Cooperating Units; Nors
Man Years (calendar year 1958)
~°fotals nlO
Professional; dO
Other: olO
Project Itescription;
oyeaU^es: To describe the type of EBG changes obtained -with
photic stiiHulation in patients imdergoing steroid treatiiien:^, re='
view their clinical diagnoses and investigate possible pl^io=
pathogenetic mechanisnso
Methods employed; Tha record file of the last fcmr years of the
EEG Branch was reviewado Ont of 80 patients who had been on
steroid treatsnent at the approximate time of their EEGs, 9 were
found who sho^^d an tnmsuaily marked response to photie stiMila-
tion» All of their charts were reviewed and their seTspal EEQs
including follow=^ps, were re>--analysed in detailo The clinical
diagnosis of the 9 patients was; lupus eiythsmatosus (li)^
lymphatie leukemia (3)^ rheumatoid arthritis (1) and prs»gressig-e
ossificans myositis (!)<, Six of these patients developed seisures
during corticoid or ACTII ssedicationo
Part B included les fj No /S7
0-flP°2(a) Serial Koo mipB:^7(G)
Major fJEdiagss Sight of these patients showed a "recrxrltirsg"
type of response and one presented a "t^TpersyriGhronoTis" response
to intennittent photic stimulationo Although the occurrence of
such responses in the group studied is relatively low (llo2^-)s
orja must consider the fast that in a population of Ron^selectedj
non^epileptic sijtojects it has an incidence of only lo35So It is
concluded that AOTH and adrenal corticoids contribiite to S.owjjidxjg
the convulsive thr-eshold, acting at -the braiji stsra levels upon
probably aixead^ abnomisil ststictures and th© abnormal reaction fe>
inteimttent photic stiissu-lation is, in factjj a manifestatdon of
such a convulsive tendency »
Significance to the program of tbs Institute; In vie^ of the fac-t
that the occurs^nGe of seizures in those patien-ts i^a shoTo- an ab-
normal photic response is significejitlj higher thasi anong those
who di-d not present any particiilar activation to th.e sasie stirsila-
tion, practical considerations of prognostic value ms^ be inferredo
The abnormal photic response could actually be an early sign of
impending coiwulsiv© disoi^er Td.th manifest clinical tits and^
therefore^ caution should be exerted in such cases toward the s^on----
tinuation of a given steroid treataiento
Proposed course of the projects The first data have been collects':;
in one paper almost ready for publicationo Further cases and esori'--
trol studies are now gathered to better determine the role of 'bhe
systeiaj.c disease or/and of the corticosteroids in the pathogenasig
of the abnormal photic respsnse and seizures o
ORP-2
Serial Moo_ _ _ NIKDBJC G)
lo Electroenciphalograi)ny"
and CliBo Neuropi^ysiology
Co EEG
3o Bettesd0.j, Marfjlarid
ho New
PHS-NIH
Individual Project Report
Caleadar Year
Part Ao
Frojeet Title; Bibliography on Eleetroeneephalography aiid
GliJiical Neurspl^siology, 19li8=.195?8o
Principal Irtvestigategg ; Cosimo /jmone Maraan aixi Charles Henry-
Other Inyeatigators i Nona
Gooperatisg Units; Institute of lAvingj, Hartford, Connecticut.
Man Years (calendar year 1958)
Total? oOl
Professional.! oOl
Othsr: ol8
Project Descriptions This is a minor projeet which tte two iraras-
"tigators (both in the editorial board of th© Journal of Electye-
encephalograpl^'- and Clinical Neurophysiology) have been asked ta
undertake as a continuation of a similar work done by M«AoBo
Brazier in I9U80 The projeet title is self^explanatorjo Most of
the work is routine secretarial and is peirformedi, as nrneh an j?os-
siblej xsithout interfering with active clinical and espenlmenta;:.
research projectSo In view of the trsBiend.otis niiraber of publica-
tions appearing in the last 10 years in the various ssieatifie
journals, both Awierican and foreign^, there is a need for a com-
plete list of pertinent papers in tnis field, properly divided
into subgroups and iMexed in a practical and rational wayo The
organization of a eumulative 10-year indes of the EEG Jotjrnal is
als© part of this project o
Part B included Yes /7 No /^
ORP-2
Serial NOo _ Jj|E) j°9(C.)__
and Clia. Neuropliysiol
Pi!S-H2K 2o Clinieal Netarophysiol-
ladividual Project Report 3o Sethesdaj, Marylaad
Calendar Year 195S 4o Mew
Pagt Ao
Project Title; Escitatioa in medullatsd aes^'S'eo
Pgincipal Investigator; Jose del Castillo, Mo D„
Oth@g Invest igatogs! J, Wo Moore;, PhoDc
Cooperating Oiiitg; Laboratory of Biophysies ^ NINfjI;
Man Years (Calendar year 1958);
'S^tai: „5o
Professional; ^50
Other; ,50
Pro jest Description;
Qfojectives: The study of the basic laechanisiss of
nerire excitation and ccndi^ction of impulses has
been isainly carried out in amyelinic fibers ©f
invertebrates 9 which are estreaiely suitable »
because of their large sis©, for the application
of intracellular recording techniques. The
results obtained in giant asons cannot, ho^^everj
be applied isdiscriiBiaately to the. medulla ted
fibers of vertebrates and of sano The first
objective of this project is, therefore, to repeat
in vertebrate nerve some of the esperisiental w>rh
done in giant invertebrate fibers and obtain
enough quantitative information to allow the coai-
putation of the equations governing the behavior
of vertebrate es;citable isesibrane* On the other
hand, the e^tresely ssaall surface area of th®
laembraa© exposed at the nodes of Baavisr asakes
the myelinic fibers particularly appropriate for
the exploration of certain aspects of the es=>
citation process which cannot be resolved ^lien
dealing with larger areas such as those offered
by non<=3a©dul la ted asoas, muscle fibers « 0e= neuron
bodies.,
Fart ® included Yes .^_. jj@
Methods ■:3€: The experimeats on jaedxallated
fibers ■ T ear-ricsd out to begin withj in isolate
motor as;;wiiS g* t&e frog CRq pipieas) and toad
CBoffiarimss) asd ©veattsally'TiiESiffialian Ciaou^e)
fibers o T^e basic preparation ^11 consist of a
single node of Handier separated from the adjacent
nodes hy seals which create high external resistances c
lltie potentials generated across the nodal membrane
tmder study ^11 be amplified and recorded hj
means of a circuit provided with a negative
capacity inp^t stage. One of the adjacent nodes
will be used as a recording probe into the inside
of the central node v^iile the other ifill ser'^e to
inject the electric currents needed to stimulates
polarise and "clamp" its membrane.
Major findings; Siace October 1S57 taatil th© ead
of Aprils iSSSTj fwll tirae ms devoted to the
development of an improved technique to perforiia
"voltage clasap" experiments in the membrane of
nodes of Banvier of vertebrate medullated nerve
fibres. Isolated motor axons of the frog iE,plp±ens)
and J in some experiments » raamaaalian nerve fibres
have been used.
The iffiiaediate objective of this m»rk was to
combine the electronic resistance multiplier
method of Frankenhaeuser vith the special
instriMentation for voltage clasip technique
developed by Dr. So S. Cole and his collaborators
at the Biophysics Laboratory j, HIMDSo Many
difficulties were at first efficountered due to th®
esctreisely high longitudinal isipedance of the
internode through ^^ich the controlling currents
are injected. Eventually « these difficulties
were overcosae and a aethod for the study of the
permeability changes and ionic currents uaderlyiag
excitation processes in vertebrate nerve is now
available. The resistance multiplier method was
also adapted in these ezperisaent® to minimize th©
external leak of the controlling current injected
into the interior of the clamped node.
Preliminary experiments performed, for the first
time, in maMsalian nodes of Banvier gave results
that while pure qualitative in nature are of
great interest from th® viei»EK>int of the cosa-
parativ© physiology of ©scitationp as they have
shown that th® ionic currents elicited by controllec:
0EF'-'2 Cb) Serial No„ NINDB^;9XC)_
depolas'isatioa of the smismaliaa nodal meiabraa®
eonforis to patterns ©imilas' to those founds, aad
thoroughly aaalyseds, by previonas investigator's
ia invertebrate materislo Their results c&n now
tee applied witli coafid©a©e to ^aMsalian nernroiss
tisBueo Oa© of th© original objectives of this
research project aay b© considered as acoaplishedc
Ae ia^ildental observation ^ ^erortSa asationingg aade
doriag t!i©s® essperiaents is the fact that when s
aedtsllated fibre is sectiojaed tbe cut ©nd of the
Bsyelia tub® teads to close ia such a way that
th® leak of asoplasae im laiaiiEal and a high
electrical resistaace is sjaiatainedo Thig
observation is interesting as it aaight explain
why th© resting potential of neurons in slices of
nervous tissue is higher than one would expect on
the basis of the short leagth of the out nerve fibres,
It might also have sobs© technical significance as
it provides the basis for a "single node of Hanvier"
preparation which might have useful applications ,
A paper dealing with technical details (Jo del
Castillo §s J, w, Moor©p "An Electronic Electrode") p
will be presented in the Technical Prograia of
the 1959 National Convention of th© Institute of
Radio Engineers o
Significanee^to the program of the Institute; The
Isasic mechani^as of nervous activity, both ia th®
CKS and the periphery, is the excitation process
in which nerve ispulses are generated o As
pointed out aboveg practically all th© infonsatioa
we possess on those process©® derives at th© present
moaent fro® studies ia organisias widely different
fro® th© huiaano A reinvestigation of these
ffiechanisms in vertebrates and, eventually^ is
majaaaliaa nerve fibers is considered to be of
iaapsrtanc© for a ssor© eosaplete understanding of
th© physiology of the hvmsLU nervous systej®, Sioth
health and disease « FwrtheriEorep such knowledge
is necessary for th® elucidation of the aechaniiisB
of action of several types of drugs » aainly of
local anesthetics 9 whose selective blocking action
on the eascitation process of nerve and muscle
membrane is still in need of clarification
0RF=2 Ce) Serial Noo Nirn)B<^9(C)
Proposed eoiars© of tb® psfoject; Apart from
@is.perlm®nts designed to obtain quantitative
iafog-mation on the behavior of the nodal mesabran':
in conditions of coatfolled Membrane polas-isatio:
a number of subjects can be investigated with
the technique developed o One of the most
interesting being the study of the mechanism
hy ^ich certain organic cations , such as the
hydrasiniuia ions m&y replace Ha ions in the
excitable mechanise of the membrane o
Serial No, _ NIMDB-^IO(C)
2„ Clinical Neuyophfsio
3„ BetSiesdas Kept land
4o New
PHS=NIH
Individual Project Report
Calendar Year 19511
Part Ac
Project Title; Studf of Meehanissas of Transmit ■i::,v
Liberation at Presynaptie Ner^e
Endings o
Principal Invegtigator ; Jose del Castillo ^ McD-
Other lavegtigators; None
Cooperating Units; None
Man Years (Calendar year 1958):
Total: ■ o^O
Professional? o50
Other: o50
Projeet Description:
Objectives: Tbe studies of the aeehnnissa® of
synaptic transmissions) both at the Merve-aiuscl©
junction and at th»5 bk> toneurons of the spinal
cordj. have so far not thrown nsueh light on the
ph^siologf of the presynaptic nerve endings ^ i,€
on the liberation of the chemical transmitters.
This is due both to th(» small miz® of th&mm
nerve terminals and to the fact that^ although
a number of druga acting on th« post=sfnaptie
membrane are available^ very few agents capable
of influencing the activity of the presynaptic
nerve endings have so far been discovered o Aa
attempt will be made,; therefore j, to find sub-
stances influencing the liberation of chemical
transmitter, and to develop new methods for th©
investigation of the meshanism of action of
those so far kno\mo
Methods employed ; The frog (Re pipiena) ner«/e
muscle "junction Ts the synapsis to be us®d
because of its ©nsy access ^ large size of the
Part B included Yes j^r No ^^H'
OE.P-S Ca) Ssidal Noo NINDB--IjO(G)
post-SFaaptie cell asad tis© possibility of
direct mieroseopie observatioa of the Eotor
aerve ©sadiagSo Intraeellular capillary Esies=©=
electrodes will lb© lised to r©eord potential
ehanges at the ©nd=>plst© msmbifaaeo Mew optical
aethods will be employed to obtaia a better
view of the presynaptic nerve endings and to
position external recording sii^roelectrodes in
its iia^adiate vicinity » lonophoretic methods
i^ill also be employed to apply substances to
localised spots of the nerve endings o
Major findings; Experiments have been performed
XncS-fefeFToTsftady some of the ionic factors that
influence the liberation of chemical transmitter
and its combination with the post°-synaptic
receptors o In the course of this investigation
it was discovered that the depolarization of th©
post-^synaptic merabrane produced by externally
applied acetyl'^choline is markedly influenced
by the pH of the extracellular solu^lono This
phenomena may help to elucidate the nature. of
the acetyl'-choline receptor reaction,.
Significance to the program of the Institutes
the physiology and pharmacology of the pre-
syji^J^-,n@rye endings are still in its very
beginningj'' 'however 9 its importance ..for our
understanding of nervous function doe's not
need to be emphasized ^ as well as the interest
of its pharmacological and therapeutic
implications. Although we possess today a
number of pharmacological agents ^ich produce
post°synaptic blocking on potentiating actions
(ranging from d-tubocurarine to prostigmine)
the potentialities of presynaptic influences
on nervous Junctions have not been explored
so faro
^oposed course of the project; a) Setting up
th© necessary experimental apparatus for the
electrical study of nerve«»muscle junction,,
b) Exploration of new methods for direct
microscopic observation of presynaptic motor
endings^ c} Future course of research to be
determined in view of the technical improve<='
ments achieved.
Annual Raport
Medical Neurology Branch
National Institute o£ Neurological
Diae&ses and Blindness D«ceoaber, 1958
SUMMARY
The Branch of Medical Neurology admitted during the period
covered by this report 215 patients for 5,974 total patient days.,
Thus each patient stay tfas 27.8 dayso One hundred and fiftysist
out-patients were seeno
The Branch of Medical Neurology reports specifically on
the following projects;
In the Section of Neurological Disorders, a nev investiga--
tlon as to the medical treatment of seizures has been undertak^a,
which is dependent upon the findings of Brody and his colleagues
in the National Heart Institute, o£ n«» monoanine oxidase it^ibitoxm,
Monosi&ine oxidase is the primary ensyme necessary for the break"
down of 5-hydroxy-tryptaaine to the 5-hydrosy- indoles, the most
ii^ortant of irfiich is S-hydrosy-indol-acetic acido The fonasfcioa
of the sulphate ester of this group in the urine has already been
associated tfith a neurological disorder characterised by cerebellar
symptomatology, dermitls, and mental retardation, under the name
of Jepson's dieeaseo Brody and his colleagues have found that
the utilisation of monoamine ojcidase inhibitors in animals markedly
reduces the epileptogenic thresholds Thus a double blind procedure
has been instituted in ifhich patients vith centrenc^halic seizures,
having as tasay as 50 or more attacks per day, have been admitted,
and a double blind procedure initiated, using the new monoamine
oxidase inhibitor JB^Sldo As this is a double blind procedure,
the results will not be kno^m for approximately six BKJnths" timeo
At the present time, ten patients have entersd Into this project o
This project is being carried out by Dra Bushnell Smith and
Dr« Darwin Prockop,
It has been noted that cases of orthrostatic hypotension
have been noted to have many neurological disorders. In particular
loss of sweating, loss of external sphincter controls, isf»otence,
mental dulling, and, in soae cases, a Parkinsonism" like syndr«ne,
with or without ciliary atrophy. Three such patients have new
been studied, and one such patient has come to poet-mortsno To
date no thorough anatomical study has ever been accoa^»li@hed on
a patient dyiag from orthrostatic hypotensionc The itsportance of
this single case, hence, is not Co be underestimat^do It was the
decision that serial sections should be ecc@i;^lished through the
hypothalamus, syo^athetic ganglia, th© intermediate cell columns,
and the cranial nerve nuclei of III, V, VII, IX, and X, as well
as the basal ganglia, anterior horn cells, and certexo This
necessitates literally thousands of sections, and the strict
correlation of anatoaiy and pathology » It is anticipated that
the thorough study of this one post-morten case will need, in
time, investigative use of a neuroanatomist for at least six
months o this is being undertaken, at the present time, by
Drso Brager and Shy, and to date in^jortant findings have already
been found in intennediolateral cell coluasns, the ventral cell
columns, in Clarke's column, the dorsal nucleus of the vagus,
the ventricular gray^ and in the inferior olives^ Degenerative
changes in the cerebellum were also found with many torpedoso
There were marked degenerative changes in the substantia nigra
and in the mesenc^halic nucleus of the trigeminal, as well as
in the larger cells of the corpus striatum and the pyramidal
cells of the cortexo
Similar to this is the study of a new syndrome recently
described with rapid central nervous systan deterioration, central
blindness, nyclonus, and death in approximately three to four
monthSo Here again, a long-term anatomical and pathological
correlative study is being undertaken, with serial sections c This
case will be studied extensively by DrSo Drager and Bushnell Smiths
As in past years, many of the patients admitted to the
Branch of Medical Neurology are suffering from diseases of the
motor unite Recent advances in isotopic procedures and muscle
pathology have changed radically this progran from the past year.
In combination with the Association of Research in Nervous snd
Mental Diseases, this Institute undertook, during the past year,
a review of the effects of metabolic and endocrine abnormalities
upon diseases of striated muscleo This was an over°all survey,
eiqploying chemical studies of muscle biopsies, combined with
various metabolic tests with particular reference to hormonal
levels of aldosterone, gonadotrophin, corticoids, katosteroidsg
TSH, ctco Of particular interest were two disorders; Familial
Periodic Paralysis and so-called McArdls's glycogen disease of
muscleo In the former disorder, aldosterone levels were determined
by the double isotope derivative methods ^ Intracellular cations
on muscle r«sioved both before and during attacks were also studied,
as was pathology before and during attackSo And finally, micro*
electrode recordings cf single muscle fibers in vivo before and
during attacks <> Potassium^^ turnovers were also studied in this
disease a The pathology of this disorder '&m» quite striking in
that large accumulations of fluid appeared intracellularly in
approximately one~ third of Che fibers <> Gh«nical detexminatloas
showed that in spite of this accumulation of fluid, that the
cationlc concentration of the cell remained {^proximately within
normal limits t> This was confirmed by microelectrode recordings;
which showed resting potential of 71=2 + 11 =3, which is ^at might
be anticipated If intracellular potassium were at normal levels <>
- 3
Studies oa aldosteroae on thsse particular patients revealed
that there was no increase preceding the attsck, ao previoualy
reported by Conoo There was mlso a decrease in potaseiiim in
the urine precediag the attack, which indirectly confirra® the
latter observation done on double Isotop® derivative methods;
in that If there had been aldosterone excretion there should
have been a potassium diuresis o
Twenty- three cases of infantile neuromuscular disorders
associated with hypotonia were also studied in reference to
pathology^ electromyography, and clinical courseo Frc«a this,
five different types of disorders were found in the disease
state, which have been recently grouped into but one disorder »
These findings have been reported by DrSo Greenfield, Coraman,
and Shy, in the December issue of Braiuo
Th® recent findings that DM is probably inert in n0n<=
proliferatit^ cells is now leading t@> the utilization of tritium
labelled thymidineo This will be a powerful tool in the study
of regeneration and growth of muscle, and this, combined with
electroraaicroscopy, will be undertaken by the Section of Biophysics »
Dr^, Haase has, in addition, undertaken a long-range study
of the pathological findings of intraiBuscul&r motor and sensory
nerve endings in normal and in neuromuscular disease states,
using the Goers technique of intravital methylene blue staining o
He has confirmed asonal regeneration in neurogenic diseases, but
the other abnormaliti«8 described by Coers and Wolfe has, as yet,
not been verified.
The Section on Biophysics has cec^leted its investigations
on the localization of cerebral neoplasia by collimating techniques,
utilizing various isotopes. Over 200 such patients now have been
studied, with a confirmed accuracy of 86 » 2 percent. The final
techniques and instrumentation utilised in this study, as well
as the statistical evaluation, have been reported in monograph
form by Eo & So Livingstoneo This monograph was also utilised
at the International Conference for Peaceful Use of the Aton,
Similar procedures have now bean initiated by the Institute for
Johns Hopkins University, the National Naval Medical Ciaater, Oak
Ridge National Laboratories, and now at Los Alamos c
The studies of microelactrode recording in single muscle
fibers have been utilised in familial periodic paralysis and
B^asthenia gravis ^ Due to the scarcity of the first disorder,
this was don@ by ciut-dowu methodo In the myasthenic patients,
continuing attempts are made to record single muscle fibers through
the intact epidarmiSo The Bak Unity Gain ^nplifier has been
utilised as optimal ^ith a constant current sent back into the
grid of the cathode follo«rero This latter allows constant
- 4
sling of the c©ndieion of th« probing ©Isctrodco To dat©,
endomyslum and perimyslsl connectiv© eissu® ha® b®en feh® chi@f
otumbling-block, in that th« electrodes intenaittenely plug
or brcako Of the literally hundreds of recording© which h«v«
been attea^ted t© date, only five successful intracellular
penetrations have been mad® through the intact epidermis o The
continuity of this project will d^end upon the ability to over-
the difficult techniques listed above o
In the Clinical Director's Report each year, an attecft
has been ntade to select areas of outstanding contributiono This
year the studies conducted in the laboratory of clinically implied
pharmacology, under Dr« Eichard Irwin, have accog^Iished much
which will show considerable insight as to the interrelationship
of blood and tieeue cholinesterase systems, their substrata,
other enzyisse systane working upon such substrata, and basic
fundamental knowledge as to the differentiation between dsf»ol aris-
ing and cffiapetitive blocks, as well a@ insight as to where in
the stuscle fiber the blockading compound has its taaximal effects
Thus, Drc Irwin and his colleagues have deoionstrated that coaspmtitivm
blocking coB5>©uad8, auch as d-tubocurarine and depolarizing block-
ing cota{>ounds such as decasaethoniua, loay be differentiated in
their action by inhibition or excitation of muscle cholinessteraseg
thus the c«aipetitive block of d- tubocursrine is reduced or prevented
by inhibition of rauacle cholinesterase o On the other hand, £h«
block ©f depolarizing drugs is prolonged by the inhibition of
plamta cholinesterase or muscle cholinesterase o In the case of
decamethcnima, this cannot be due to destruction by cholinesterase,
per se, as decamethonium has no ester group and hence could not
be destroyed by cholinesterase <. Succinylcholine, on the other
hand, has an ester group, and thus could be destroyed by
cholinesteraseo It is of interest, h^ever, that the prolongation
o£ the blockade by inhibition of plasma cholinesterase is identical
to the two substances, thus showing that this inhibition prolonga-
tion is not of Qecessity due to destruction^ ^r the d(^olarisins
conpounda Thus one can assume, I believe correctly, as DTo Irwin
and his colleagues hmm assumed, that sbuscI® cholinesterase has
but a oiiaor role in relation to the total block. If this substance,
however, is not metabolised by plasaa choliQest^rase, then
inhibition of muscle cholinesterase has a marked efface on the
blocking activity, and the non-depolarising substances upon such
inhibition of muscle choline demonstrate a decrease in their
blocking power, whereas the depolarising Siibstances d«Bonstsat®
an increase in their blocking power.
Dro Irwin and his group have continued their etudiee en
the action of directly stiasulated innervated and dcaervated asusclec
In this they have been aided by a device, created by Hr^ Well®,
of an op tical« isotonic lever systcsii, recorded through a cathode
ray oscllloscopsc With thie mechanitsn, they have been able to
- 5 -
demonstrate that the block is not due to increased muscle
coa^llance, as added cos^liance in series dees not give
contractile responsea similar to those obtained with succinyl^
choline or dec^aethoniumo If this isotonic systen is observed
closely, one osay see there is less shortening of the fiber and
reduced velocity of shortening, «gain showing thstt this is not
an increased cos^liance o£ the muscle fibero The isotonic^
optical systan allows this, in fact, that it reduces the elastic
coffiponest of imiscleo This system, ho^sver, does denonstrate a
prolonged latency from the onset of the stimulus to the time o£
contraction after administration of depolarising c«3q>oundSo
These investigators feel there is a spatial distribution of the
depolarising blockade over the muscle membrane, indicating either
multiple end plates upon the muscle msabrana, or « temporal
spread from a single membrane, loeo one end plate. These
investigators point out that muscle cholinest erase is lov in
quantity and is not uniform in various species and/ or organs, and
hence has a species and organ specificity., It is thus di^endent
upon the substrate and enzyme actlvltyo Thus, muscle cholinesterase
studied as to substrate specificity and weli°kno«n Inhibitors
would give considerable Information as to the chemical intern-
change between the substrate and the ensiymeo
The cholincsterase of muscle heaaogenates, in which the
blood was rsE&oved so the plasma cholinesterase was not present,
was studiedo Such homogenates hydrolyzed acetylcholine more
rapidly than benzoylcholine, or butrylcholineo An excess of the
substrate, however, would inhibit such hydrolyses, the optimal
level being 5 s. 10°^o The optdLmum level of concentration for
substrates other than acetylcholine are hlghero Thus, muscle
cholinestigrase is highly speclflCp However, since benzoyl^ and
butrylchollne are hydrolysed at measurable rates, small anounts
of non-specific ensyme must also be present o It is of int@r@st
that neostigmine d^olarises the m^abrane at 10° 3^ whereas
pyridostigmine (tsestinon) will not a This becomes of double intsrest
in that both drugs are highly useful in the treatment of myasthenia
gravis o Galanthamlne, «diich has been isolated from an alkaloid
in the United Soviet Socialist S^ublic, and utilised in the
treatment of myasthenia gravis, was also studied by these
Investigators o Galenthamine is a phenanthrene derivative and not
a carbamine ester o Dr^ Irwin and his group found a 50 percent
inhibition at 8 x 10°^ o The value for the inhibition of plasma
cholinesterase was the samso Neostigmine and physostigralne
inhibit at lower concentrations as far as cholinesterase in the
muscle is concerned, but in vitro inhibit raore rabidly than with
galanthaalneo
Finally, these investigators are studying the possibility
of choline esters other than acetylcholine occurring as natural
constituents of biological syetemsj the object being to determine
6 -
to %3hat extent th« choline esters are found in such biological
systems and related compounds, end how they depolarise tissue
menbraaeo Secondly, to relate the depolarizing properties of
these confounds to their stimulation or blocking activity of
syn^ses, and finally to study the metabolism of these compounds
by tissue enaymeSo To study this, the travelling fluid electrode
technique is used to measure dqpolarisation of the isolated frog
sertorius muscles, and microelectrodes will be utilized to
determine the resting membrane potentials, presumably through
the Bak Unity Gain Cathode Fol lower o These investigators have
found, in high concentrations, i.e^ 10"3 molar, that butrylcholine,
b^isoyl choline, aad imidazoleacrylcholine, all resenfcle acetyl-
choline In their depolarizing propertieso Methacholine, however,
does not depolarize muscle membrane <> These investigators have
also found the plasma from myasthenic patients have been observed
to metabolize imidazoleacrylcholine at the same rate as plasma
from non-n^asthenic patients., And finally, these investigators
are attonpting to find to what extent d^olarisation of ths
miscle msB&rane may effect the efflu:^ of enzymes from inside th@
muscle fiber, in particular aldolase^ This latter project is
projected into the coming year.
The Section of Neuroradiology suffered in having its
chief investigator, Dr, Giovanni Di Chiro, undergo surgery for
a major illness« In spite of this setback, however, Dro DiChiro
was able^ upon his return to duty, in addition to his heavy service
responsibilities, to carry out in ccMsbination with Dro Martin Rubin^
of Georgetown University, a research project which culminated in
a paper concerning the metal chelates as possible contrast media
for myelography o These chelating cospounds were tested against
coBBBonly used iodinated contrast oediac Different concentrations
of the various chelating coo^ounds were tested in order to
determine the concentration for optimal opacityo Once such
opacity was detexminad in vitro, it was tested in vivo on dogs
and rabbitSo Chelating agents used are listed in Dro Di Chiro' s
report, with primary interest on lead ethylenediatainetetraacaeic
acido This substance was administered at the dose level of 10
nilligrms per kilo, and appeared in the urine to the extent of
85°d9 percent of the injected dose within two daySo Of that
retained in the animal, i»eo 10-15 percent, 50 percent was found
in the liver and some 20 percent in the bone marrowo This densn^^
atratad that, despite the large amount of excretion, the amount
retained is not to be discounted. The experiments in vivo show
that studies of good diagnostic quality saay be obtained as far
as s-ray contrast and detail are concerned, with radiopaque metal
chelates o However, the acute toxicity of the metal chelates in
B^elogr^hy, as well as in laost of the other x-ray examinations
carried out, proved to be too high. Accordingly, OrSo DiChiro and
Rubin are going on to undertake studies in other metal chelates
with high atomic number, in hope that in this screening one agent
.„ 7
of local tostcity would ba fsuad which was mo Imi slm fe© ^ugg®st
it could b® used in clinical myelography c
Th® Section of Neuroch®mist3ry continued it® efforts in
th® major fi«sld» listed in th® 1957 r^orfco Ur,. Horvsth continued
hla fltudisK in th® distribution of actin and tropomyosin in
normal and diseased muigicles hi® coiBparstiv© biochemisstjcy »r,udi©®
of smooch muscle and striated mascles and alteration* of acto-
myosin tensil® strength and muscle prot^inss in neuromusculSE'
dl»eaiii«So
Drc> Towgr aad his colleagues have continued their studies
on the metsboltsm of ) - aminobutyric acid in neural tiasu^j, with
the aid of Dr^ McIChann and Dr^ Wherrettc Studies on the relatl<Dn
of pyrldoa:ine to certain seisure states, in particular in th@ise
cases known ss pyridoKinii dependency, contlnuedc Dr^ Tow@r
continued his elaborate studies on amino acid metabolism in normal
and epileptogenic cerebral cortex la vitro, and in electrolyte
energy metabolism in normal and epileptogenic cerebral corteXa
The unit as m whole continued its clinical evaluation of Amino
acids and related c^sanpounds in control of selsures in mana
Drc- Curtis continued in the realm, pred<»iinantly, of surface^
chemistry, and in other physico-cheaical methods in determining
constituents of human spinal fluid, ocular fluid, etCc
Dr, Tower^s studies specifically now revolve around C^^ and
j|l5 labelled cempoundSo Tvo^deoxyglucose was utilised as a
competitor for glucose utilisation, by inhibiting the he^okinase
st^ primarily due t@ depletion of avail^le AFP required for this
step. Dro T@wer found it was possible to overcome the 2'°deoxyglucose
block in gluc®®e utilization by adding either AIP or gluc©8®"6-
phosphate to the slices in anaerobic conditions,. No effect of
these additions, however, was obtained in aerobic metabolism,
presumably due to their failure to penetrate the slices, Dro Tower
felt that 2»deo3Eygluc@iie inhibition did not result in any actlva~
tloa ©f the glucose- 6-*phosphate dehydrogeni^se or in any osldaeive
shunt pathisayo These findings were checked by incubating the
control and inhibited slices with gluc@se-l-C^^ and gluc©8e-6=C^^
phosphates determining the utilisation of C^^Oa «nd C -lactic
acid production. Since the ratios of the C^^ lactate from thm
Q-6 ccstspared to C»l samples vere loO In b®th cases , whereas
C-e/C-l weuid be less than IcO if the shunt pathway were utilised^
this would Indicate that this inhibition was not due t@ an
osldattve shunt pathway o This was indirectly confirmed by the
ftadlEjg ®f l&w level brain TPN by other investigators in that TPN
i® th® necessary co^izyme for the shunt pathway o Dro Tower i&nnd
also that 2»deoxygluco8e inhibition not only resiuited in m^rk<@d
decrease in glycolysis, but also in oxidative metaboliamo thusj
with glucose-U-C^^, less C^^02, les® labelling of the free amin©
acid pool, and less C lactic-acid were all obcained. From
these studies with C^^-labelled glucose, the distribution of
glucose utilized by normal slices to various intermediary steps
could be estimated, thus glycolysis to lactates 70%; amino
acids measured by glutamate, 22%; respiratory CO21 7%, and other
intermediaries, such as lipid and protein, l%c If one
calculates the oxygen uptake as 85^o/g/hr,, it is clear that
if 30% of the latter is accounted for as amino acid and
respiratory CO2 that this almost exactly balances the oxygen
uptake, assuming 6 moles of the latter per mole of glucose
oxidisedo This is consistent with studies of other laboratories
and this laboratory, that non^glucose substrates, such as amino
acids, normally support oxidative metabolism by the brain and
they are repleted subsequently by part of the glucose utilized a
Studies with 2-deoxygluco8e clearly demonstrate, according to
Drc Tower, that glucose is necessary to make repletion of non-
glucose intermediates possible, and energy production rapidly
falls in its absence, and that this is not only by depletion of
AIP and creatine phosphate but also by deleterious effects on
glutamic acid and electrolytes in the inhibited sliceSc As in
his 1957 report, Dro Tower points out that such inhibited slices
fail to extrude eKcess sodium and reconcentrate potassium in
nonnal manner o This Is similar also to defects seen in slices
which have been renoved from epileptogenic patients, and Dr^ Tower
has also found this in cortical slices from cats with seizures
enduced by S-methyl-e^ethylglutarimide, and by methionine
sulfoximineo Utilizing the Cotlove apparatus, Dr, Tower and his
colleagues find that the swelling of normal and epileptogenic
slices during incubation is confined to the chloride space, and
that calculations of electrolyte concentration per litre of non-
chloride space water at the end of slice incubation demonstrates
again a loss of potassium and a gain of sodivmio
Sr, tower haa continued his studies on incubating slice®
of cat cerebral cortex with L-glutamlc acid labelled with C^^»
L-glutamln@ labelled with C^^; jt^-aminobutyric acid labelled with
C^^; L-aspartic acid labelled with C^ ; D-L-asparagine labelled
with 2,3-C^^j D-gluc©se labelled with C^^; sodium pyruvate- 3-C^^;
and 2->pyrrolldinone-2"'C^^c Using these compounds, Bxc Tower was
able to determine the order of labelling in amino acids, and was
able to sho^ this had considerable significance since the aspartic
acid could prime the Krebs cycle by providing both oxalacetate and
acetyl" Coenzyme A (from pyruvate) in the absence of the latter
from glycolysis o Dro Tower concludes that these studies indicate
how active the co&^onents of the glutsmate-aspartatc amino acid
group are in metabolic participation la the Rrebs cycle, and
feels that the release of CO2 measured by C^^ liberated during
these eKperiments confirmed this conclusiono
In the second part of his experiment, Dr<, Tower analyzed
- 9
the liberation and forcsiatlon of glutamic acid, glutamine,
^-aminobutyric acid and free ^uomoaia metabolism in incubated
slices from non-cortical areas of the cat brainy these were
the subcortical white matter, the thalamus, the caudate
nucleus, and the cerebellar cortexo He found the levels and
metabolic behavior in all gray, ioe<, neuronal areas, were
similar to that previously observed in the cortex, but that
the white matter exhibited extr^nely low levels and little
change on incubation for glutamic and^^^'-isainobutyric acids «
while the white matter glutamine was not greatly different from
the cerebral cortex° In studies on the levels of these substances,
Dro Tower felt, using the calculations of Elliott and Heller,
that at least 85 percent of cortical glutamic and>^-aoinobutyric
acid content was associated with neurons, while only about five
percent of the glutamine speared to be neuronal in locationc
The cerebral cortex was fractionated by the Brody and Bain
technique, and Dr^ Tower found the majority of glutamic and
X-aminobutyric acids were associated with fraction R3 or the
mitochondrial fraction, whereas glutamine was distributed almost
equally between that fraction and the combined Ex -1- &£ ^^actions
which contained cell debris, axon fragments, nuclei, ctc» No
content of any of the three amino acids was found in the micro-
somal fraction. The finding of these substances in the mito-
chondrial fraction is compatible with their close association
with the Krebs cycles
Dro Tower^s studies also indicated that the inhibition
of glutamine synthesis by methionine sulfoximine is primarily an
interference with axoaonia moiety, possibly by the imine group of
the toxic coQ¥>ound, and that by adding only anaoonlum chloride
such a block could not be overcome unless adequate amounts of
glutamic acid are available to amidate to glutamine^ Studies
with similar epileptic agents, such as Megiraidc showed that the
glutamic acid metabolism was blocked to include i^-amlnobutyric
acid, in that the latter coi^ouad was significantly lower than
normal c The s^ne was true when inactivators of pyridoxal phosphate
were usedo If malonate, however, was used, the anount of glutamic
acid and F-sminobutyric acid in the slices rose to double the
normal values o The action of malonate la to inhibit succinic
dehydrogenase c Since this was accocapanled by reduction of oxygen
uptake, it was previously not clear why such slices did not also
show succinate accumulation.. Dro Tcwer"® data suggest that in
the whole cell preparation it is glutamate end j^°aminobutyrate
rather than succinate which accumulates and requires a study of
the relationships ^oong these three coa^enents of Krebs cycle.
Dr. TcDEffer plans to continue these interesting experiments, using
the microanalytical method of Dro 0. H. Lowry,
Clinical evaluation of various amino acids and related
compounds in the control of seizures in vivo in man has been
10
continued by Dr, Tower and Dr. McKhfinn, and the Branch of
Electroenc^halographyo Patients onj'^-aminobutyrlc acid have
continued to do 'well in Dr„ Tower's estimation, one patient
being seizure^free after three month® on the cosipound, coaipared
to multiple daily eeisures previously.. On stopping the coEapound,
the seizures returned and have again been abolished by starting
j^^fianinobutyric acidc Several other patients are getting more
benefit from ^fioastinobutyric acid than from l-asparagine<, Gmssaa-
aminobutyric acid has been given intravenously to levels of
4s]H/kg. body weight » with no untoward effects, in dogSo However,
when 1/200 of this dose is administered to man, there is
immediate agitation, flushing, hyperpnea, and a drop in diastolic
blood pressursc £Lecovery occurred within 5-10 minutes » B?c Tower
rightly points out, despite the r^orte by Elliott that such
occurrences can be ignored, it would seem that this potentially
a dangerous drug given intravenously <> Another case of pyrldoxine
dependency has been worked up by Dr^ HcKhann and Dro Tower <.
These patients were also studied by the Krypton^^ technique for
measuring cerebral metabolism developed by Sokoloff o The original
case of Hunt was restudied, and the patient, now seven years old,
is still dependent, regularly developing seizures within 72 hours
of omission of her regular dally dose of 10 mgo of pyridoxin® o
Typical EEG abnormalities could be abolished in 30-60 seconds by
intravenous pyrldoxine-HCl (15 mgo)o During a typical period of
depletion, cerebral metabolism was measured by Krypton°^ technique,
and the decreased oxygen consumption during the depleted state in
this case was similar to the situation reported by Sokoloff for
hypoglycemia subjects,, Thus, the interpretation tentatively put
upon the data obtained in this case is that during pyridoxins
depletion a deficiency of the substrate for cerebral oxidative
metabolism exists which is prsn^tly corrected by pyrldoxine
admlnistrationc Since pyridoxins deficiency affects ^-aainobutyrlc
acid metabolism primarily, and since that c«Qq>cund appears to be
a significant substrate, Dr, HcKhann and Or^ Tower rationalise
that this case may actually represent an e^eample of -aminobutyrlc
acid deficiency^ with a consequent reduction in oxidative metaboliemo
Drso McKliann and Tower have continued their studies of the metabolism
of ^-•aminobutyric acid in neural tissue by using the Fluorimetric
method, as described in the 1957 report ,> They appear to have
daaonstrated that the shunt pathway, ioOc glutamate to ^-«nlno-
butyrate appears to be active and In^ortant in cerebral oxidative
metabolism^ and is significantly Involved In certain dysfunctions
of the brain, such as seizures » They plan to undert^e further
studies to see hcfw such a pathway raay @s@rt a regulatory control
on oxidative metabolism and hence on energy production in terms
of normal function and of seizure states «
Dro Curtis is continuing his studies on physlco>ch^nical
methodology In an atteQ^>t to obtain quantitative data from fluids
which contain extremely sisall aaaounts of organic metabolites » He
11
is working particularly on the surface tension of urine, and in
particular optical m®a®ur®nent8 by polarized light and its
reflection off of surfaces utilizing the elliptical polarisation
as an indication of the thickness of the surface interfaceo The
apparatus has been built in combination with the Naval Research
Laboratories and e:q>loration of this approach is now being
orientated towards the use of photomultipliers, so that the square
function may be utilised, and monochromatic lighto Parallel with
this he is continuing his studies of adsorption on solid surfaces,
such as column resins, in foams and interfaces in urine and water-
iisniscible liquids^ Dro Curtis has now found that there is so
much gross interference in the acethylcholine^boron-flavonaol
reaction to biological materials as to make this procedure
unsuccessful, in the determination of microchemical mounts of
acetylcholine. His studies on guinea pig serum asparaginase
detailed in 1957, have now been completed, except for some electro-
phoretic and ultracentrifuge data now in progress o He finds that
the purified ensyme preparation contains two macromolecular con>
taminants which have defied attccq>t8 at separation by electro-
phoresis or ultrecentrifugal means; that enzyme can be quantitative-
ly adsorbed on a modified cellulose and in carbon dioxide fosm,
and purification by these means is currently being atteaptedo
Dro Horvath is continuing his work on proteins of muscle
in normal and diseased states, and has calculated total solids,
total protein, ndn^proteln solids (ioeo fat), non-collagenous
proteins, collagen, water-soluble proteins, myosin, alkali'^ soluble
proteins, non^proteln nitrogen, electrolytes, and tissue water.
He finds differences in the normal and dystrophic muscle analyses
are reflected by connective tissue and fat, and by an increase in
sodium and chloride in dystrophic specimens o He finds there is
a relative increase of myosin and decrease in alkali- soluble
proteins in most cases <, These changes seen to be independent of
the remaining muscle masSo The water-soluble proteins ^pear to
be increased relative to other proteins in most dystrophic sanities
and an inverse relationship is indicated between the reaaining
muscle mass and the percentage of water-soluble proteins in the
muscle on the other hand. He concludes that amsplma of dystrophic
muscle not only contain less muscle and more connective tissue
and fat than normal muscle, but that the protein coaqpositien of
the remaining muscle is different frcm the normal.
In the study of actin and tropomyosin in normal and diseased
muscle, Dr<, Klatso and Dr. Horvath are turning to immunological
properties of functionally important muscle proteins. They find
that rabbits isamunized against serum tropomyosin A, clam tropo-
myosin A, mammalian myosin, and antiscra to human and cat myosin
precipitate clam tropomyosin A. No such cross-seaction was found
between antisera to chick tropomyosin B on the one hand and clam
tropomyosin A or maraoalian myosins on the other. Using antibodies
12
t0 myosin conjugated with fluorescein, myomiti. in ««c£ions of'
normal huntan isuscl® was clearly and distiucCty d@n®astraced
over th® fluorescent leicroscopeo Preliminary sections of
dystrophic muscle similarly trested sh@w@d myosin in residual
islands of icuscle and a suggestion that in areas of iictive
degeneration myosin^reactive materia.! vats present in tRacro^
phages o Thus the iaasunological findings jire consistent uith
the present concepts of the myosin molecule consiistisg of
subunits - Tropomyosin A, B and Actln, the latter can be
pr^ared in a high state of purity than myostin itself, so that
it is; more suitable for investigational purposes c Since these
proteins are also i so- antigenic, the iacounologlcai response of
the organism may be inqjortaixt in conditions where destruction
of muscle could permit these proteins e© escape from the usual
confines of the muscle and enter the circulation of the bodyc
Dro Horvath is continuing th<g same studies In muscle protein in
electrolytes in dystrophic mice obtained from Bar Harbor o
Flnallya Drc Korengold and Br^ Haopp have concluded their
studiesj, «;hlch were an attetq>t to conflnn the findings of spiro-
chetes in the cerebral spinal fluids with patients sufferiog
from multiple sclerosiSc Identical material to that used by
Ichelson was utilized, and a trip was made to Drc Ichelson"©
laboratory to be certain that there were no differences c Twenty^
two patients were studied in the out°patlent area, at fd;lch time
•plnal fluid was removedc No positive cultures were obtained,
and It was felt desirable t@ terminate the project after this
auB^er of studies^ This concludes the major findings of the
Branch of Medical Neurology.,
Katioasl Institute o£ Neurological
Diseases and Blindness
Clinical Research
Medical Neurology Branch
Serial Numbers of Projects:
NIKDB-li<c), NIM)B-12(c), NIKDB-13(c), NINSB-14(c) »
NIl®B-15<c), Wim)B-16(c), K1HDB-17CC), and
NX»DB-18(c)=
Estiaated Cbligatioae for FY 1959
Totals $6918 000
Directs $227 » 500
Seiiaburement s $462, 500
lo Medical Ksurology Branch
Z« Section on Neurological
3« BttlMsda^ Haryland
Uo New Project
PHS - NIH
Individual Project Report
Calendar Tear I958
P^^ Ao
Project TitlQs Th© Use of a Monoamina Oxidase Inhibitor (JBc=5l6)
as an Anticonvulsant Medication for C©ntr©nc©phalie
Seisureso
Principal Investigators! Darwin Prockop
Bushnell Smith
Other Investigators s Andrew Engsl
Mark Lane
Cooperating Units s Kristof Abraham 0 Electroencephalography
Branch
John Oatesp Hational Heart Instituts
Man Years (calendar year 1958): Patient Days (calendar
Totals 1 - year 1958)? 40
Professionals 1
Others 0
Project Descriptions
Objectives;
Chen et al<,o have demonstratsd in aniinals tliat reserpin©
lowers th® threshold for ©lectrically induced convulsions possibly
by lowering th© amount of brain serotinin (5 hydro:^ryptaja3.n©)
and norepinephrine while Prockop ®t aloo showed that laonoaBiino
oxidase inhibitors 0 which increased the level of serotininp
pr«\'ented the tonic extensor phase of ©lectrically induced seizures
and generally raised the convulsive threshold to ©l©ctro shock asid
pentylen© tetrasolo
The objective of th© present study is to evaluat® the
effectiveness of JB=.5l6 (fphenyl isoprofiyll-hydrazin©) as an
anticonvulsant in persons with centrencephalic seizures o
tfeitliods Eerployed-i,
The centrsnoaphalie seisui'® with its syinBietrical electro
encephalo graphic abnoriaality (3 per second wave and spike) as
well as its clinical frequancy O'f manj per day is the type most
su.i.tabls to evaliiation on a shoi'fc tana basis o It aay possibly
be the one to ba most affected t^r JB=516 bacauso of the higher
concentrations of serotiirin in th® sxipposed sites of oi'igin of
th© seiaures (Brain stesij, Hypothalaiaiis „ Thalaioic Projections),
Patients with c©nti'enc®phali.c seizures will be maintained on the
anticonvulssmt oiedieation prescribed for them before admission
(which in thsse selected cases will not bs completely controlling
th© seistjx^s) to prevent wide swings in the number of seis\ires
that occur o An squal nvuabsr of patients will bs placed on placete
and on JB=5l6o The cas@s to be given JB=.5l6 will bs determined
arbitrarily hy the pharmacist who will have no direct contact with
the patientSf. To determin© if the JB=5l6 produces any inhibition,,
dsten&ination of 5 Hydrojtytryptamine in the lu'ina of all patisnta
at the end of one week wi,ll be mad©.; Electroencephalographie studies
will be auad® at regular intervals,.^ These will serve as a basis for
compilation of effectiveness of medications^ Clinical records will b©
kept of seiawr«>So
jla;ior Find3.n^ss It isf too early for any results at this tiiM©,>
Significanoe to Netirological. Research g If the ibhibition by the monoaadno
oxidase inhibitor (JB=5I6) of the ojcidation reabtion of 5 hj'droaybrytamiKK-
ta 5 hydroj^indoleactic acid also results in marked inhibition of seiKures
of the centr©ncepha3.ic type,, new areas of investigation will be opened
as to the nature and possible etiolo^ of ©entrencephalic ssizuresc.
Proposed Corn's® a Tan patients previously studied and known to liafe
centrsncephalic seizures,, according to th© classification of Penfield,,
will bs admitted to th® hospital for a period of 6=7 weeks = A basalina
period of one to two weeks will bs used to evaluate th© intensity and
nuEiber of seisures,. Elactroencephalograms „ level of 5 hydroxytryptamln©
in th® urine and coordinated studies with Drso Van Buren and Mirsky will,
b© carri®dc During the remaining five weeks ;, the patient will receiv©
either JE^5l6 or placsbo in addition to th© prescribed anticonvulsant
msidicationr. Electixtsmcephalograms will be don© at the same hour of th©
sarae day each week by the same technician.. Clinical course will b©
closely followed c
A comparison of all data and correlation with the drug that tixs
patient took will bs inade after the last patient has been studisdc
Part B included
2o S®eei©sa ®a EeuTOlogieai
4 a H^r Fsr@j®efe
€al@a4ar Year iSSS
Page Ao
Pff®Jeee Ti£l©s Sliaic«l Patitalcgical Q®zmlatlv& S£a%
@f Che Meswwa© Syss^ ia ©re&©r<©se®&ie
Oe&eff Iw©s£iga£©rs Go Mie®a Shy, M* So
fetal s I
As a eliaieal siaiey ©ssfessseasie fejss@g®asi©a sr©f®ss
£® a e®adi£i@a clsas®c£affis®d fey a sigMfieaae fall in fel©©d
effee£ poster® »
th® e®adlei©a t^as £ir©e desesifeed by Btamusy ©t alo
ia 192So SiKC® £feae tisi® appE©stea£©iy MO g«^»@s£s h®iy®
ap^®fflr®4 iia da® litesa^urep dealisg p?teeipally ®ig& the
hma. c€msimt®d ©a, sad it is seafc®d tfeas messsolegie sigas •
a?e fouad ia at Imet. ®a&»fou?&S& ®£ £lse
Serial No.
-2-
It has been assumed that the site of the lesions are either
in an autonoBiic center or in an efferent pathway o or that it is
generalized in the central nervous system^ efferent pathway ,> or
nei*ve endings » This assumption does not explain the manifestations
of extra=pyramidal and motor neuron systems g which are often associated
with orthrostatic hypotensiono
The literature fails to reveal any post mortem studies of
the central nervous system of patients with this disorder in which
a clinical pathological correlation was made» The present investiga-
tion concerns the post mortem findings in the nervous system of a
patient who had this disorder and was thoroughly studied from the
clinical standpoint »
PATIENT MATERIAL;
One patient 0 a 5^ year old white male^ was admitted to the
Clinical Center in January g 1957 with a 6=year history of episodes
of dizziness 0 loss of libido,, nocttiria changes in coordination f,
speech changes r, and a resting tremor of both upper extremities and
of the jawo Anhidrosis had also developedo Orthrostatic hypotension
had been present for at least six years o
The patient was completely studied in the hospital for a total
of 135 dayso Terminally j, he developed fever of central origin and
expired o The central nervous system was obtained at autopsy »
In addition 0 one other patient is being studied in the
Natioiuil Institute of Neurological Diseases and Blindness with a
similar neurological ^mdrome assosiated with orthrostati© hypotension
Other patients with this clinical syndrome have been examined in other
Institutes 3
MEIHODS a^ff^LOYEDs
The brain 0 spinal coaxlc and autonomic ganglia were removed
from the boc^ within a few hours after death and fixed in formalin c
The brain was cut and microscopic sections were prepared in the
usual mannero The sections were stained or impregnated with technics
that demonstrate neurons ^ glia and lipids o To adeqviately study this uniqu©
opportunity serial sections through the hypothalamus » the III5 V^ VII ^
IX „ X cranial nerves = dorsal root ganglia ^ and intermediate cell columns
of the cord is necessary o This necessitates the careful study of literally
thousands of sections o
Serial Moo „___________
MJOR FINDINGS;
In the spinal cordj pathological ctsangss were fotmd in the
Intermediolateral cell coliaan and^ to a lesser extent o in th©
ventral cell colusms and in Clarke's colusoio I^a^linization was
observed in th® fasieulus gracilis o This was aarted in th© cervical
segments of the cordo Gliosis uas also present o Lipids, t^hich stained
with Sudan III 0 was present in manj cells of the spinal cord but was
particularly prominate in th© ventral horn cellso
In the jaseduUao degenerative changes were found in many cells o
The dorsal nucleus of the vagus demonstrated a marked falling out
of the cells o Gliosis was also pi*esent around the ventricular gray
and in the Inferior oliveso As 5Ln the corda considerable lipid was
found in the motor neuj^onso
The cerebellum showed a marked decrease in the number of the
Purkinji© cellso Many torpedos were present „ as well as increased
gliosis „ and considerable lipid,, in the cells of the dentate nucleus o
Some diffuse degenerative changes were found in th© pons and tegmentum c
Marked degenerative changes were observed in the substantia
nigrao Considerable^ extracellular pigment was present. Diffuse
degenerative changes are present in the motor and reticular neurons o
Gliosis was found in the central aqueductal grayo Lipid was present
in the motor neurons aad in the cells of the mesencephalic nucleus
of the trigeminal nerve « Changes may be present in the Edinger=Westphal
nucleus but these are questionable ^ and more sections are being prepared
for study o
Diffuse degenerative changes have been observed in the larger
cells of th® corpus istriatum and the p^amidal cells of the cortexg
however„ th© study of these areas of th© brain have not been completed-
SIGMIFICAMCE TO MEDROIDGIGAL_.RESEARGHg
To study and report the degenerative changes found in the central
nervous system of a patient with orthrostatic hypotension and th© associated
neurological syndrome « So clinical pathological study of this syndrom®
has been madeo
PROPOSED COURSES
To study additional patients with this disorder and^ if possible^
obtain additional material for pathological stu<tyo
Part B included No
HIMJE"13Cc|
1 , Med icalTleuro logy Braacli
2, Sectioa on Neurological
Disorders Service
3, Betfeesda, Maryland
4„ CoEtiauatioa of l?IHDB-23Cc)
PHS-HIH
ladividual Project Eeport
Calendar Year 1958
Fart A,
Han Years:
Pattest Days
Total:
,20
Prof ess iosial:
,20
Other:
,20
Project Title: Tfe© Histopattiological aad Claemical
Investigations of Meuroauscular Disorders
Principal Itovestigators: G, Milton Sfey and
Theodor Wanko
762
Project Description:
Qtejectives: TIae objectives determine (1) variables is
MFTSTSe growth of muscle; (2) endocrinologic and
iietafeolic correlations of disorders of mwscle associ-
ated with aietabolic or cationic disturbances; (S) a
study of the value of muscle pathology in determining
the etiology of the "Floppy Infant"; (4) electron mi-
croscopic study of ffiuscle in the normal and diseased
states 0
Methods Esaployed; Becent advances in isotopic procedures
and amscle pathology have claaged radically bs prograisffied
during the past year. The laajority of accomplishments to
date are fouad is nuasbers (2) and (3), A study of 166
cases of myopathic disorders associated with endocrine
aMormalities were undertaken during the past y©ar„ A
list of these is as follows:
Part B included Yes
"•3-
' 1^'0„
NIHDB"I^«">
TtjTotoKtc Myopathy
4
Sscoplithalssic Ophthalsiopl@gia
7
Hypothyroid
5
Familial Periodic Paralf'sis
5
GajpstosTp-laleabeyg)
36
Central Core Disease
Glycogeo-Storag© CTfpe SIo)
IcArdle's Disease
Late Spontaneous Ufop&thj 79
B^ial Acidosis
H^Tpoparatfefroid
Addisoii'^s Disease
Cushimg's Disease
Amyloid Disease
Salt Lcsisg Sepbritis
Paroxysmal Ijoglofeiauria
Weraer's Sy;adroKe
Total
166
-.3-
Serial Mo, Nli©B»13Cc)
This is a© overall survey j ©niplofiiag clieaical studies
of muscle biopsies $ cossbiaed witfe various metatoolic
tests with particular refes-ezic© to feoraoaai levels of
aldosteroffi© J, goaadotropia p eortieoids, ketosteroids j,
TSH, ete, A swsmaxj of tla® fiadiags ia these disorders
has b®®ffi presented at tli® ®eetiag ot the Association
for Research ia Mervous asad Meatal Diseases „ Of partic-
ular interest is the isstensive study ia two patients
with Familial Periodic Paralysis = In thiSj, aldosterone
levels were detenaiaed by the double isotope derivative
methods. Intracellular eatioas on lauscle ressoved both
before aad during attacks were also studied » as was the
pathology of muscle before aad durisig attacks ^ aad aicro-
electrode recordings of si^le asuscle fibers in vivo be-
fore and duriag attacks, ^^ turn-overs were also
studied iss this disorder.
Twenty- three cases of isifaatile aeuroasuscular dis-
ease associated with hypotonia were also studied ia re-
ference to the pathology, electromyography ^ and cliEical
course 0 Frosi this^ five different types of disorders
were fouBd in a disease state which has beea recently
grouped iato -but ©ae disorder.
The recesit fiadiEg that DM is probably inert isa
aoa-proliferatiag cells has led to the utilisation of
tritiun CH^) labelled thymidine as a powerful tool for
the study of regeaeratioa of muscle „ The electrosi-
Eicroscopy studies during the past year have heeu liaited.
by Dr, Waoko to normal human JBuscle obtained at biopsy „
Major Findings; In Familial Periodic Paralysis it was
found that there was a large increase of intracellular
fluid, the esiact composition of which is still entirely
unknown. The cat ionic concentration » however j, demon-
strates that potassium eoncentrat ioa is not increased
in this disorder. To maintain suclTa concentration in
the face of increased intracellular fluid means that po-
tassium must enter the cells from the extracellular
fluid; the paralysis, however, cannot be due to increased
potassium in that the cosceatration is the same asj or
less than 9 in the nos^al cell. This has bsen confirmed
by micro® lectrode recordings showing the resting potea-
tial to fall with 71 ± 11 ®v„ Our present conception
is that the paralysis in this disorder is due to the
siechanical distension of the cell raeaibraiie aad not due
to high eat ionic values.. Studies on aldosteroae » usisig
double isotope derivative aethodSj have deaoEStrated tl^aft
Serial Mo„ KiKDB=l3<c)
aldosterone is not a factor in this disorder, as postu-
lated by GonQo Tliis is confiroied by the fact that there
is not a potassium diuresis before the attacls. The other
studies of emdocriae abaorasalities have beea suBraiariaed
ia the Tables preseeted to the Assoc iatioa for Ees®arch
ia Merwons asd Mental Diseases o
Fi-^e distinct entities have beeji implicated is th®
"Floppy lafaat" by BrSo Greenfield ^ Conman^ aad Shy,
These are Werdaig-Hof f raann ' s disease j Coageaital Mus-
cular Dystrophy s Ceatrai Core Disease « Beaiga Coageaital
Myotonia s aad Arthrogryposis. This phase of the study
has been completed aad the paper «ill appear ia the De-
cember issue of Braia. The electroa microscopic f ladings
ia aormal adult huiaaa muscle have beea suamarized by
Dr. WaakOs aad appear ia the December Proceediags of the
ABMMD.
It is of iaterest to sote that the oaset of late
prosysmal Myopathy ia the male is associated with aeo-
plasia ia 90 perceat of the cases ^ but ia oaly 10 per-
ceat of the females. This is based oa a study of 79
caisesj, isicludiag 33 Males aad 46 females.
Proposed Course of the Project: The growth of muscle
will be oae of the primary iaterests of this study in
the coiaiag year, using tritium CH^) labelled thymidiae
for studies of DHA with autoradiography a OTo Waako will
coatiaue the electroa Microscopic studies of diseased
siuscle,
Sigaif ieaac® to Neurological Research: The correlatioa
of aaatoiffiical ; chemical' aad metalKslic abaon&alitles as-
sociated with disorders of striated muscle as well as
further iasight iato the growth of smscl®:, gives a
powerful correlative uaderstaadiag of SEUscle ia disease
aad ±sk the aorm.
Serial Mo, MI85BB«l3Ce)
Individual Project Eeport
Calemdar Year, 1958
Part B: Honors , Awards 5 aad Piafelicatioas
PiitolicatioBs other thaa abstracts from this project:
1. Slayg GoM. So®© Metabolic aad EisdocriBO-
logic Aspects of Disordei's of Striated
Muscle o ProceediEgs of the Association
for Researcli ia Her-^ous and M©atal Ms-
for Beci^ber,, 1958 ,
2„ Gresafieldj JoGc » Coramaaj, To aiad Shyj,
GoMc The Progffiostie Value of tb® Miscl®
Biopsy iss tlie Floppf Iisfaato To fee pub-
lished iss Brain 3 Decensber, 1958 =
Serial Ho. MIHDB'.14(c)
1. Medical Neurology Branch
2. Neurological Disorders
3. Bethesda, Maryland
4 . New
PES NIQ
Individual Project Report
Calendar Year 1958
Part A.
Project Title; Pathological Study of Intramuscular
Motor and Sensory Nerve Endings in
the Normal and in Neurtnnuscular
Diseases,
Principal Investigator: G. R. Haase» M.D.
Other Investigator: None
Cooperating Units:
Man Years Patient Days: 25
Total : 1
Professional : . 5
Other: .5
Project Description:
Objective; The study is concerned with an investi-
gation into the histological structure of the motor
and sensory nerve endings in normal muscle and with
changes occurring in these structures in various
diseases affecting the neuroonascular system. Various
investigators ; ioe.» Schwenn (Deutsches Archiv fur
Klinisch Medicin« 70:193^ 1901); Schiefferdecker
(Deutsches Zeitschrift fuer Hervenheilkunde , 25:1);
and Caap (JAMA^ 48; 1230 j, 1907) have reported changes
in the muscle and specifically in the muscle spindle
occurring in cases of Parkinson's disease, Coers
(Acta Meurologica ©t Psychiatrics Belgicft,, 55; 741 »
1955) reported changes occurring in the terminal
fibers and in the motor endplate in a number of
different diseases ^ including myotonia of either
the congenital or dystrophic variety ^ in "amyotonia",
is polymyositis and in various neurogenic lesions.
The objectives of the present study are to determine
whether these reports can be verified or refuted and
whether any additional changes can be discerned by
histological means.
Metltod BaipXoyed; The methods cossist In the us© of
intravital istjection of ffiethyXene blue at the time
of biopsy as indicated by Coers^ and the use of the
acetyl cholinest erase stain as modified by Coers
and finally in the employment of silver staii^, in
particular the modification indicated by Winkelmann <
Patient Material: The material is usually obtained
in the course of routine biopsies j, e:n:cepting for
such variations as are imposed by the methylene
blue method. In all these pathological specimens,
routine staining methods are also employed by the
Section of Neuropathology. Control material has
been obtained frc»a a number of cases dying of
diseases not related to the neurxMnuscular system.
Finally f material has been obtained from various
laboratory animals.
The patient material utilized includes pre-
dominantly those patients admitted with neuromuscu-
lar disease in the course of studies concerning the
general pathology in various disorders. Several
patients with Parkinson's disease were admitted for
the primary objective of obtaining muscle specimens a
Major Finding; "Rie study is still too much in its
beginning to permit definite conclusions. Some of
the difficulties in the interpretation are due to
the great variabilities of these structures in the
normal The occurrence of axonal regeneration in
neurogenic diseases has been verified.) The other
abnormalities which have been described have not
been verified so far.
Part B included. / / Yes / / No
i„ Medical Neurology Branch
2„ Section on l^euro logical
Disorders Service
3 c, Betlaesdaj Mdo
4o Coatiauatlon of M21IDB-21Cc
PHS-NIH
Iadi¥ldt2ial Project Eeport
Calesdar Year 1958
Part A »
""^oject Title: The Physiology aad Pharjsacologj ina
Myast&eziia Gratis
Principal Issvestigator: Go Miltoa Shy
Other Investigators: William Matthews
msn Years: Patient Days: 560
Total: ao
Professional,; olO
Other: dO
Project Description:
Objectives: To continue studies of intracellular rest-
ing potentials in ayasthenia gravis to determine the
estent of depdlarization or hypopolarization of the
Methods Employed: Consist of intraf iber recordings with
microelectrodes in the single muscle fibers of afflicted
muscle in myasthenic m&a^ After much trial and error.,
the Bak Unity Gain Amplifier has been utilissed as opti^^l
with a constant current sent back into the grid of the
cathode follower o This latter allows constant sampling
of the condition of the probing electrode a
Major Findings; But a few amscle fibers have been sue-
cessfully, penetrated to date in myasthenic Man, As an-
ticipated in the 1957 Beport, this has becoase a difficult
and time-consuming procedure » Besults of present records
would indicate that in igyasthenic isan we have been success-^
ful in penetrating about only ten fibers.
Course of the Project ; The course of th® project will
resolve around the overc^sing of the difficulties listed
above. Many hundreds of recordings must be made to de--
teraiine a statistically significant resting potentials
At the present tiise this has not been feasible. Studies
will continues however » ©v^r the nert yearc
Nlb^BB-
T2iis project may fee abolished if it is found t&at nk&
results are uffirelialsle or th© teefeaiques so difficult
as to make aaf coxiclusiojis difficult to interpret o
Fart B iacluded Mo
Serial HOc jmiJ)B^l6(c)
1» Hedical Kettrology Branch
2o Section on Neurological
Disosr^ers Servless
3« Bethesda^ Maryland
'4-0 Kew Project
PHS ^ NIH
Individual Projact Report
Calendar Year 1958
Part A<
Project Titles A Study of Progressive ParoncfcQninatous Degeneration
of the Central Wenrous System »
Principal Investigators? Bushnell Siaith, M» Do and
Glenn Dragerg Mo Do
Other Investigators!
Cooperating IMitss
Man Tears (calendar y^^r 19$8) s Fatlsnt Days (calendar
Totals 1 year 1958)g 92
Professionals 1
Others 0
Project Descriptions
Progressive parenchyiaatoiis degensration of the central
nervous system refers to a group of patients described Tby Hfsidsnhs.:
n&joTo Leigh S: Baggs Pallia and Spillaneg Jones and Kevin? as well.
as Foley and Dem^-Brown in which they (age rang© ^0-55 years) she-
progressive involveiiffignt of the central nervous system with Miauros
persoiiality changes c J^oclonuSo ataxia and finally a eomatos© stat
The onset is insidious and th® clinical course is a progressive
deterioration of th® f\anctions of th© nervous systeaio
A 51-year old white female was admitted to the National
Institute of Heuroiogical Diseases and Blindness after 2f saonths
of such an illness o %oclonu3(, cortical blindness and stupor
were noted on admissiono Dsjiring th© 92 day hospital stay,-, th©
^tisnt continued to have niyoolonus and becaffl® progressively
more decerebrate with opisthotonic postttring until death„ which
occurred approximately si:c months after th® ons®t of this conditio-:
Post mortem ©jcamination t^as performedo
Studias of th® braiii are bsing «ndeii;akan (a) to establish
a possible ©tioiogy for this part.iciilar illness ^ (b) to establish
tha anatomical structures of the central nervous system that have
bsen pathologica3Jly affeetad in this illness o (c) to eorrelata
these findings with th© cliiiioal data and th© findings from a brain
biopsy obtained dijring tha tS-jird month of the disease „ and (d) to
correlate th® findings (clinical and pathological) with p8rti:!?,ent
cases in the literaturso
Methods Employed g
Th© brain and th© spinal cord wer© removed approxiEatelj
eight hours after death and fixed in formaline The brain was cut and
examined for gross pathology after ten days of fisationo
The spinal cordp brain steiSo cereballum and cerebnani will be
sectioned and stained with th® routine techniques for dsiaonstrating
Korphological changes in th© nervous systeEo In addition,, special
stains will be employed to demonstrate inclusion bodies and lipids
of the neurons*
Major Findings;
Gross ©xamination of the brain has shown the entir© cerebral
cortex to be affected.; In addition the caudate (and possibly other
t^sal gang3.ia)o the cerebelluaio the pons and the spinal eord all show
changes o
i
Significance to Reurolof^cal Re search g
This is a relatively rareo progressive degenerative disease
of the central nervous system^ The etiology is quit© obscur® and
the pathological process is controversial o Th® clinical cours© of this
patient from beginning to end has been well docurosntsdo k cortical
biopsy was obtained during the third aonths of the disease o The
pathological finding of the biopsy whan compared with th® autopsy
laaterial obtained six Eonths after th© onset of th® condition shotsld
reveal inforaaation concerning th© progress of th© disorder^ Special
histological studies may be helpful in establishing the etiology o
It is also of importance to correlate tJie clinical picture with the
pathological findings o
Prpposad Cpursa of Pro.leats
To make a clinical pathological correlativ© stwc^ of th© r
system of & case of progressive parenchimatous degeneration of th.e
oenta-al nervoas system^ This will entail a detailed study of th®
nervous system employing th© ustoal ixmtina stains and special technioxx^:
for demonstrating inclusion bodies and lipid content of neurons ^
Part B included No
I,, Mmdicsl i!teurol©gy
2o Neurological Di&oxd^x^
Service
3, BethgsdSj Maryland
4o WINDB=-24(c)
PHS-NIH
Ittdividual Project R^®r£
Calendar Year 1958
Prrajfisct Tiel®g Spirochetes and vlxal antigeas and their
relation t© th@ spinal fluid and bl©6d o£
multiple sclerotic patisntSo
Principal Investigators iteryin Co K@reng©ld
Other Investigators; Edward Has^pg Ao Sabin and Egbert Hu<Bbn@r
Han Years; Patient Days^ 22
Totals o20
Pr® fsesienal ; o 10
Oth®rg olO
Project Descriptions
Oblecfciyess To att®Hf>t t© isolate spirochetes in the spinal
fluid o£ multiple sclerotic patientso Samples of spinal fluid
and blood were obtained from 22 well^screened multiple sclerosis
patients and referred to Oro Huebner°9 laboratory for special
viral studies^ Saoples of blood were obtainsd from 22 ^®ll<°8crees@d
multiple sclerosis patients and sent to Dro Sabin" s laboratory
for special viral detenoinations against th@ antig@n obtained by
Drc Sabin frosa the USSE.
Methods EBg>l®yedg The !a®th®d @f B®»® lehslsoag a@ described
in th@ Proceedings of the Society for Eseperimental Biology and
Medicine, Mays 1957, will be ussd to attsaapt t© Isolate spir@ch®£®?s
fr^a the spinal fluid of multiple scl@r©tic patientSo A special
laboratory procedure and chssical media has been described by
Miss Ichelaon and an attempt to duplicate this procedure will hm
followsdo 15 cco ©£ spinal fluid will be obtained from 30 patientss
of wigll'^d®cum@nt@d multiple sclerosis (based on the criteria of
other imitiple scl<groais projects) o This fluid will be transferred
to Bro H^^p"8 laboratory lAere special procedures will be under°
taken to att@E^>t to isolate any possible spirocheteSo Each patient" a
spinal fluid will b@ 8tudi@d under the Ichelson medium as w<@ll as
under various other standard spirochetal mediums currently in
use for other purposes c
Patient Mafegrials Pafeieets w«re obfeaiagd freaa th«
previously w®l'i"®cr©eaed patianes with multipl® ®cl®£©si®r
HaJQg Ftadings; Twenty- twe patients with tsultiple sclerossis
w®r® admitted to the Clinical Center Out=Patient Dsiparcm®nt f^oE
special spinal fluid studies r. Sanqslas ©f spinal fluid wgrg ssnft
t© Dtc. HaDi>p''8 laboratory on all pati®ntSr. Th© specific tgchniquftg
as described and us@d by Miss Rose Ich@lg@n w@r@ dupllcaCeds and
in no instances, wer@ any spirochetes founds It was felt aft@r
22 negativig d@t@rmination£, that £urth@r patient studies would
not b$ n@c@ssaryo Th@ project was, eher$for@, concludedo
Significaace to Program ef Institutes The results of this
study indicate Chat no spirochetes can be iioplicated in th@
etiology of multiple eclerosiSo These studies have now be@n
CQs^letedo
Part B includsd Y®» [__| K© [x_J
Serial lo„ NXKDB-I8<c)
1= Medical Mexirology Branch
2, Sectioa oa Biophysical
Applications
3= Bethesda, Marylaad
4. Old Ko. irilDB~20Cc))
PHS-NIH
Individual Project Report
Calendar fear 1958
Part A.
Project Title: lavestigatioms in Localization of
Cerebral Neoplasia by Isotopic De-
tection
Principal Investigator: Go Milton Shy
Other Investigators: Robert Bradley and William Matthews
Cooperating Units: Oak Ridge National Laboratory
Oak Ridge J, Tennessee
Man Years: Patient Days: 483
Total: ^50
Professional: '50
Other: [50
Project Description:
Objectives: In the 1057 Annual Report this Project was
listed as an attempt to localize cerebral neoplasia^ Its
value has since been detenaiaed., and the prissary objec-
tive now has been to determine what percentage of accur-
acy this technique offers «
Methods Employed : This has been a systematic study of
collisaation upon highly sensitive large sodium iodide
crystals which are juxtaopposed at 180° from a moving
.electronic scanner » Since the last reports a coincidence
circuit has been added to the gasma spectrometer „
Patient Material: Over 200 patients now have been scaaaed,
^hese patients' are froE HIHDBj the national Cancer Insti-
tute, the national Navy Medical Center ^ Walter Reed Hos-
pital, Mount Alto Veterans Hospitals, and referring neuro-
surgeons in the surrounding area.
Major Findings : In these 200-odd cases an accuracy con-
firmed of 87 percent was found - 14 of these patients had
normal contrast studies.
-2- Serial Ifo. NIM)B-18Cc)
Proposed Course of the Project: Now tisat the useful-
aess of this procedure has been determined ^ this ap-
paratus ^ iffi its es&tirety, will be transferred to the
Central Diagnostic X~ray Department, who will now ue-
dertake this as one of their service functions « The
Unit also is superirising the coastruction of ast iden-
tical apparatus at Johas Hopki&s Uaiversitya
SigsificaBce to Heuro logical Research; This procedure,,
in desnonstrating its usefulness,, has relieved one of the
major hazards in the treatment of intracranial tumors ^
namely ; contrast studies with the hazard to patient life
and the necessity of being followed by iiraiediate opera-
tion <>
Part B included Yes
Serife MXN0B~l8Cs])
PHS-HIH
ladividtaal Project. Bsport
Calendar Year 1958
Part B: HoaorSj Awards ^ aad PublicatiODS
Publications other thass abstracts from this project:
Shy, G.M, , Bradley, R„Bo aad Matthews, WoB,:
E:sterisal Collimation Detection of Intracranial
Neoplasia with Unstable Nuclides o Published by
Eo 8s So Livingstone Ltd„ „ Edinburgh.. Scotland,,
1958 „
Hafcioi^al Institute? of Heurologicsl
Diseases and Blindnses*
Clinlc&l Research
liedlcal Neurology Branch
ScctlOQ on Neuroradiology
Serial Jiun&er of Projects
KIKBB°19(c)o
Eatimated ObliRafciona for FY 1959
Tofcal; $20 » 500
Directs $19,900
Reinibur6«a®nts $500
2o Neuroradiology
3= BethssdcB Maryls
4 o New
PHS ° NIH
Individual -Project Report
Calendar Year 1958
Part A.
Project Titles Metai Chelates as Possible Contrast Media
for Myelography
Principal Investigators s Giovanni Di Chiro, M. Do and
Martin Rubin, Ph.D.
Other Investigators J None
Cooperating Units s Diagnostic X-Ray Departments CoCo and
Georgetown University
lAan Yearss (calendar year 1958) Patient Deyss Mcne
Totals 2
Professionals 1
Others 1 -
Project Descriptions
object ivesi To find a new roentgenopaque contrast mediusrs for
aiyelographyo None of the now available contrast media is, in facts
satisfactoryo Heavy metals have a marked x-ray opacity. Several
of those are more radiopaque than barium and iodine, elements
present in the standard roentgenographic contrast media now in
clinical use. Heavy metals exertj boivever., a profound toxicity
in living systems. Chelation is a relatively old method to nsske
Etietal ions cheiitically inactive and therefore possibly non-toxic
for living organisms.. Objective of the present project has been
to find whether or not different metal chelates are suitable as
roentgenopaque contrast media, and more specifically as roentgenopaque
contrast media for myelography.
Me thpd s Employed s The essential properties of a useful
radiopaque rfiedium includes 1. . high radiopacity? 2, iovt systemic
and local tissue toxicityi 3., pharmacodynamic silence? 4. prompt
and complete elimination. In experiments in vitro the radiopacity
of 3 relatively large group of chelated compounds has been tested^
For this purpose a board of non x-ray opaque material with small,
equal Vv'ells to hold the test solutions was used. This board with
the different solutions vjas x-rayed using standard technical
factors. A standard dark room technique was used. The chelated
compounds v.'ere tested against commonly used iodinated contrast
mediao Different concentrations of the various chelated compounds
were tested in order to determine the concentration for the optimal
opacity. Once determined the radiopacity in vitro, the radiopaque
chelates were used in vivo and so administered to dogs and rabbits.
The compounds were injected intravenously, given per mouth, injected
intyacardiacally, and injected in the cisterna magna. Urographies}
phlebographies, gastro-intestinal studies, angiocardiographies, and
myelographies were so obtained. The quantity of the chelated compound
injected was varied mainly according to the concentration of the metsl
in the chelated solution, its x-ray opacity previously proved in vitro,
and the animal's weight. Accurate toxicologic studies have been
carried out to date only for few of the substances under study. No
autopsic study was done in the experimental animals after the x-ray
studies had been performed. Here follows a list of the compounds
tested!
1. Lead EOIA
2. Lead Cyclohexyl EDTA
3. Lead DTPA
4. Cadmium EDTA
5. Cadmium 5^-^^
6. Cadmium Cyclohexyl EDTA
7. Zinc EDTA
8. Cobalt EDTA
9. Cerium EDTA
IC. Copper EDTA
11. Nickel EDTA
12. Barium EDTA
13. Bz'smuth EDTA
Abbreviation's EDTA -= Ethylenediaminetetraacetic acid
Cyclohexyl EDTA - 1, 2 -Diaminocyclohexane (N, hi ~
tetraacetic acid)
DTPA "- Diethylenetriaminepentaacetate
54^ A »« f,!y fjl. » (2 - hydroxycyclohexyl) ethylene"
diaminediacetic acid
Concurrently, a group of experiments concerned with the metebolis
and distribution of injected lead EDTA was carried out by one of the
investigators to study the general problem of the quantitative fate
and tissue distribution of the administered Ketal. Lead EDTA adminis-
tered at a dose level of 10 mg/kg appears in the urine to the extent
of 85 ■= 8^ of the injected dose within two days. The fscal excretion
of the lead EDTA following its intravenous administration is below 2%.
The lead retained in the animal (some 10 - ]5% of the injected dose),
is deposited in its major portion (about 5Q5£) in the liver. Some '
2C^ of the residual lead EDTA was found in the bone marrow. Small
traces of ti^e injected lead EDTA were found in other tissues and
organs* kidneys, hsartj lungs, muscles. This group of experiments
shows that at least for what the lead EDTA is concernedj the retention
of the metal is not to be discounted despite the high urinary excretion.
This is particularly true if ive consider that the quantities of metal
chelates th«t are to be injected for contrast purposes are high.
Ma,1or Findings s As it could have been expected several of
the tested metal chelates have shown good radiopacity, useful for
diagnostic x^ray purposes. The higher the atomic number of the Esetal
present in the chelated solution^ the higher the opacity Vv-as found to
be. The experiments in vivo show that studies of good diagnostic
quality may be obtained, as far as x-ray contrast and detail are
concernedj with radiopaque metal chelates. 10 ccc of lead DTPA 1?^
injected into the cisterna magna of dogs gave excellent myelographic
pictures with outlining of the spinal cord. However, the acute
toxicity of the tested metal chelates in myelography as well as in
most of the other x-ray examinations carried out proved to be high.
Death occurred shortly after the chelated compound had been injected
into the cisterna magna. Vthile therefore the metal chelates seem by
the present study to be interesting as far as their radiopacity is
concerned^ on the other hand this study shows that the systemic and
local tissue toxicity of thes© compounds is too high» We cannot
therefore at present recommend for clinical use the metal chelates
which we have tested.
Signf'.f icance to Neurolop.it^al, Researehs Despite their good
radiopacity and other characteristics which would make the metal
chelates tested good contrast media for myelographyj their general
and local toxicity is too high to suggest them to be used in clinical
myelography.
Part B included? Yes Txl No
Serial No, _JiiiiNDB5-Jl9(c^=
PHS--NIH
Individual Project Report
Calendar Year 1958
Part Bt Honors J Awards and Publications
Publications other than abstracts from this project*
Rubin, Martin and Di ChirOs Giovanni? Metal Chelates as Possible
Contrast Mediao (To be published in the Annals of the New York
Academy of Sciences, Section of Biology with reference to a
presentation at a meeting held by this society on Radiopaque
Contrast Agents j October 24 •= 25, 1958.)
Honors and Awards Relating to this Projects
None.
NaCional Institute o£ Neurological
Diseases and Blindness
Clinical Research
Medical Neurology Branch
Section on Clinical ,^plled Pharmacology
Serial Nuaibers of Projects s
NINDB- 20(c), NINDB-21(c)j KINDB- 22(c), NlNDB-23(c),
and NINDB- 24(c) „
Sstimated Obligations for FY 1959
T®tal! $39,500
Direct; $32,400
Reiiabursaaents $7,100
Serial Mo= i^Km~-20 Cc)_______
1. MediciT'°liurol.ogy
PHS-MIH 2= Clinically .Applied
liadi vidua! Froj©ct Report Fharms-cology
CalQsdar Year ISSS S. Bstfeesda^ lid=
4., Coatilisuad C^SUDB-l Cc) )
Part A.
Project Title: Blood SBd tissue choliaeat@?a@®@ In
neuTomascnlBx blockaddo
Friacipal lavestigator: Bichard L« Irwin
Otfe@r Investigators: Jay Bo Wells and Haary J.
&ith
Cooperatisag Baits: Noaa
Man Years Ccalend&r year 1958): Patient Day® {calendar
Total: o66 j®&t 1958}
Professional : . 33
Other: .33
Project Description:
OBJKiTIVEg; To determine the fimction of specific
and non-specific cholijuesteras© in relation to tfee
blocking of transmission in between nerve and auscleo
MITIOPS EMFLOYEDi The standard Warburg saanosetric
TicSn'iqis®' "ls~5ied to estiiaate Muscle and blood
cholines tera@e@. A cross circulation preparation
i@ used in ^^hich the blood of one animal is m&d®
to perfua® one leg of another animal „ The blood
and ti@isua cholin@@terases of the two anissals are
saanipulated by use of either di~i®opropylfluoro-
phosp&ate (DFP) or isopropyl aethyl f luorophos-
phate (sarin) so tSmt neuromuscular blocking action
of other coiapound® can be observed irith high pla@3Ea
esterase activity and low muscl© esterase activity
or low plassaa and high aiuscle activity^ Activity
of the following three type® of coaspouad® have been
studied la relation to aeuro®usciilar block and in-
hibition of cholinesteraeej Ca) aon-depolarizing,
(b) d®polari2ir&g-not aetaboliaed by cholinesterase ,
Cc) d®polari^iag~ffi©tabolised by choliBe®t©ras©«
MJOE FIWBMQBz The neuroHUSctilar blocking activity
oF°a"Hon^^^pQXariaii^ compound, d-tisbocurariae , i®
reduced or prevented by eelective inhibition of
" 2 ™
s&uscle choli]3@@ter&@@., IshibitioD. of tli@ pl&anB, choliis-
esterases greatly prolongs the netxroBiwsciil&r blocking
oi depoiarisiag compouads w&ica are destroyed hf plassaa
estsraseSo Tli© seleetiY® iisliibitioBi of au©ci© cliolis©®-
t@ra@e prolongs th@ xaeurorauscular block produced by a
d@poIarisiQg compou:sid, d®csus@tho£iiuias which coBtaias
no e@t@r groisp &nd thus is not cap&bl@ of destruction
by the mu@cl@ cholin@stera@eo The selective Inl&ibition
of iBuscle cholines terase also prolongs tli@ block produced
by succinylcholinej, a blocking compound which contains
@®ter groups, and thus could conceivably he metaboliased
by fflu8cl@ cholines terase. Since the prolongation of the
blocking activity of succinylcholine occurred to the same
estent as the prolongation of the block produced by de~
caraethoniism, it appears that succinylcholine is either
i^ot metabolized by muscle cholinest erase or metabolized
at a rate which ^<|oes act influence its blocking actions^
The e^eriments of this project emphasise that
inhibition of plasioa cholinesterase can greatly prolong
the neuromuscular blocking activity of depolarizing coi@-
pounds ^hich are destroyed by plasma esterase. In this
situation, inhibition of muscle cholinesterase plays a
minor role in relation to the total neuromuscular block
produced. These experiments also emphasize that vhen
neuromuscular blocking compounds are not metabolized by
plasma cholinesterase the inhibition of muscle cholines-
terase markedly modifies the blocking activity of all of
the types of blocking compounds investigated. Blocking
activity of non-depolarizing compound is decreased. On
the contrary, th® blocking activity of depolarizing com-
pounds is increased.
SIGHIFSCMCl TO FRCK5BAM OF INSTITUTE: This project will
furnish additional information on tEe metabolism of clin-
ically useful neuromuscular blocking compounds and cholin-
esterase inhibitors. It may, in addition, contribute to
an increased understanding of the physiological processes
related to myasthenia gravis and its treatment by use
of inhibitors of cholinesterase.
PEOPOSEB G0OES1 OF S^OJECT: This research is to b© coa-
t'im£il"«niIT~the objectives stated above have been ful-
filled.
Part B included: Ho
S©;i">
1„
PHS-HIH
2„
Issdividiia.! Project Eeport
Calendar Year 1958
3o
'Sn
Part Ac
Medx GiX"liurology
Cliaically Applied
Bethesda, Mdo
Project Title: Th© actios of saewromusculas' felockiKig
drugs oa directly stimulated ismer-
vated fta<i daaerv&ted muscle.
PriBcipal Investigator: Hichard Lo Ir^in
O-fclier Investigators: Jay Be ITelis aad Hesry J-,
SBith
Cooperating Vnttu: None
Mass Years: (calendar year 1958) Pati©at Bays:
Total: ^^S (caleadar y©ar
Frofessioaal: .33 1S58)
Otfeer: »33
Project Bescriptioa:
OB JECfXlTES'. To study the decrease ia costractil®
respoese of directly stiiaulated skeletal muecl©
wfeicia follow©' the administration of certain quat-
sraary ±osm<.
METEQBB SISPLOUD: Alternate direct aisd indirect
itiSEI&Hoi^oFTfe® gastrocneasius or the anterior
tibial msiscls of tiae rat has bean usedo Tfe© effect
of aerv© stianlation has foeea eii^iaated by adaia-
istratioa of a aeuromuscuiar blockiag cospound
that doe® aot affect the contractility of the
dirisctly stimulated muscle. Contractile responses
t^ere recorded either froat a damped strain gauge
ifhich si&rlsediy restricts external shortening or
froa &n optical system activated by an isotonic
lever, lecor dings were made with a cathode tslj
oscilloscop@o
The optical- isotonic lever isystesB was con-
structed by Jay B, Wells, The depolarizing prop-
ertieig of th@^e compounds have bean investigated
u@i:s£g the isolated frog @artoriua muscle and a
travelling fluid el®ctrod®o
mjOE FIHBIIC'S: Previous findiags were stated ia
iSiii~aii"£mi the 1957 MIWDB Annual l®po:pt C2Cc))o
MoE'e recent ©sperisjeats have shown that the block
"- g "
to direct stimul&tioa wkicii occi^r© with ©iscciaylcholi&e
aad d®eainetlioEiym is not due to merely iBcreasing mvEScl©
cojapIiaae®c Added, coaijli&u'jsce ia B®rie© with the »uscle
doss aot giv© aoatrastil© rssposas© siasiiar to those
obtaiaed wkess block to direct stiaulatioa is issiduced
by @ucclnyIolioIiiie or d®camethoiiim!i«
Wh©Q the block to direct etimulatioa which oc©ur®
i^ith sueci»yIcholiai@ is observed with isotonic recording
of contra-ctioQ, a method vhich reduces to a great ®xt@at
the role of the elastic eleiaeut of the iimscle, less
shortening and a reduced velocity of shortening occurs o
Thi@ &lBO indicates the reduction in contraction is
not increased compli&nceo
A prolonged latency between stimulation and the
start of contraction has also been observed following
the ad&ainistration of depolarising blocking compounds.
Cc^pounds which reduce the contractile response
of directly stimulated muscle also depolarize the muscle
membranes. The depolarization has a spatial distribu-
tion in the frog sartorius muscle which extends through-
out the muscle except for a small area at each end. The
depolarization could occur from widely scattered end
plates throughout the muscle or fr^m temporal spread
of the depolarization from the motor end plate.
SZGHIFICAM^E TO THE mOGRAM OF TEB SHSTITIJTS: The drug©
investigated in tije course oi this project are in com-
mon clinical use. They also serve as valuable research
tools in clarifying physiological processes related
to transmission of impulses between nerve and muscle.
This project may contribute to an increased
understanding of the physiological processes related
to myasthenia gravis.
PROPOSED COUESS OF PROJECT; To continue to investigate
the contractile Feclucing and muscle membrane depolaris-
ing properties of th® neuromuscular blocking compounds.
Part B included: Yes
gerial lo„ lllBB-llCel
PEE-Wm
ladi'/idasl Project Eepc-rt
CaisKdar Tear 1058
Part B; HosaorSj Awards, and Pubiisatioas
Public&tioffi® oth@r thaa abstracts from t&is project:
"TIaa Coatractll© Resposs® of Directly Stimulated Musci®
after Administration of neuromuscular Blockii&g CQmpound^'\
Ace®pteci for publication ia TJag Jouraal of PharBaacology
aad EKperim®Qtal Therapeutics, fef chard "L, irwin Siud^lm Be
Wells, ~~-^:^:^^— ^^-—
loaors s,tjd A'^ards rslatiag to this project: Mon&o
PES-HIH
Individual Project .-..,...-■,
Cai®adar ¥©ar 1958
Serial .Wo.liroB-12Cc>
la Medical Meisrologi-
2, Ciissically Applied
Pharsaacologf
4o lew
Part A«
Project Title: Study of lauscl© clioliE®®t©ras® aad
its iaijibitorsc
Priacipal Imvestigator: Eicha,rd L, Irwla
Other Inveetigators :
J&y B. Wells And Hes&ry Jo
Smith
Cooperating Uaits:
Central Intel ligeQce Ageacy
aad Uo So Army, Office of
Surgeosx Geaer&lc
Man Years (calendar ye&r 1958):
Total: a 66
Professional: .33
Other: -S3
Pstieat Day® (calendar
year
1@5S>:
Project Description:
C^>?SCTI¥ES; The choiiaesterase content of mui&cle is
io^ assd Qot uisifonaly distributed throughout the tis-
su@. Muscle has therefore not been adequately studied
in respect to either th© type of oholiaesteras® it
coista.ias or a® to substrate and inhibitor specificity »
A® August iHssEoa has recently pointed out (Method i.a
BiochesEical Aaalysis, 1S57)5 the results oFtilaiH
wItE^TiSIHitori"*i5il substrates are dependent upon
the anaya® preparation used, both specieis and organ
specificity being of iiaportance* la ^iew of these
coasi deration®, on® of the objectives of this pro-
ject is to adequately characterisse this important
sauscl© ©isssi^-a® as to substrate specificity^ Another
objective is to essasin© the activity of the enzyB©
in the pressac© of th® well-kaowa .inhibitors which
are in wid© use clinically and to correlate this
activity with their usefulness . This '©ould forss
a ba^is for testing newer compounds havi».g a poten-
tial in th@ treatment of myasthenia gravis o A
further objective is to determine th© inhibitory aad
depolariaing activity of compounds either used or
proposed as useful agents in the treatment of myas-
theEsiSo
™ 2 ~
MUTHOQS B1FS.!0¥BB: Th© staadard Warburg laaaoastric t©ch-
arqueT!iil^i3"""^r d®ter?it.^atior^ of muscle claoIia©Et@r&s@
activity » Th© depolarisirag properties of cboliasetei'as®
iffihibitors are detesniBiaad hy as© of tise travelling fluid
©lectrod© sjst©® n&&d by Fatt, Jo of Pl^ysioi, , 111: 408 o
MAJOR Fim)lHGS; The choliaesteras© of muscle homogesmtaa,
S3i"Ii^ muscle previously perfused with ©aliae to r®-
saov® blood t hydrolysis ac®tylcholiii© mor® rapidly t&aa
it hfdrolysss ^Qtfeaclaoliae aiad markedly bios'© rapidly
than it liydroiyaes benzoylcholia® ot fetstrylcholine^
The enapee is laliibited by excess of substrates aad
with acetylcholiK© shows as» optisaum substrate coaces-
tratioa ia th© raag@ of a 5 k 10"^ mol&r coacentratioBo
Optiaiim coBcant rations for substrates other thaa acetyl-
choline are higher. The fiadiage ar© all ia accord
with th© belief that hwscI© cho lines t eras© is chiefly
of the true or specific typ®. Siace bensBoylcholia©
aad butrylcholin© are hydrolysed by muscle hoiaogeaates
at raeasurabl® rates ^ the possibility exists that muscl®
cosataijs© a ©s^ll amount of non-specific type esteraseo
A choiiaesterase preparatioa of high purity iB seeded
for farther iavestigatioa of the substrate specificity
of latseci© choliaester&seo Obtaiaiag a purified c&ollia"
esterase froia ©uscl© preseats a. formidable j although
feasible s, problssj, iimsisiich &b the esterase coateat of
lauscle is low and' the aaouat of other proteins ia th®
muscle is high.
The depolarising properties of th© inhibitors
of lawscl® choliaesterase which are used In th® tr@atia©at
of my^sthesaia hav® beea exaajiaado Heostigsiia© has beesti
observed to depolarise muscle memhrwueB amrkedly at a
ssolar Goaceatratioa of W^. In coatrast to this, pyri-
dostigasia® failed to show depolarisiag activity at thi©
coacentrattoffio This finding is of interest sine® thee©
two compound© are both effective in aaelioratioa of th®
symptoms of myaatheiiiao IdrophoniMm produces only slight
depolarisatioB at a io~^ molar concentration,
Galanth&iaia© J a rec^astly isolated alkaloid ob-
tained from, galaatfaug Woroaor^i C»^Jsaryilidac®a©) ^ has
been introduced into clinical aedieia© for treatment of
myasthenia gravis and other neurological disorders;
Anaotatsii o Kovykh Lekarstv^aiaykh . (Hotes on Mew Medic-
liaFlgeStsJT"^"" B. "^Sedova, Medgis, Moscoxr, 1956 » This
eompoissd is of particuias.-- int©.t*est siace it is smxksdly
diffareat chemically froja otlier coiapouads currently ia
us® i^ the treatiaeBt of ssyastheais gravis o Galaathamia©
i® a phsamithreB© derivative B.ud ia laot a carbaaatc es-
ter» W© feav© ia^estigatea ta© iaJaibitory properties
of tfee compouad and have fouad that the molar conc®n-
tratioa of galaathaaia© which emus@g 50% iclaibition
of muscle cholinesteras© is 6 3£ 10"" ®o T&e vain© for
the iafeibitlon of piassaa- cfeoiiaesteras© is the sam©„
Treaty per csat inhibition occurs at 1 x 10"® molar
aad 85% at 1 3£ lO""^ molar. Similar dstermisatioias have
been m&d@ using physostigmin0 &nd neostigmine . Both of
tk@S6 compoujEicis inhibit muscle cholistesterase &t lower
concent rations than doe@ galas%thssiin@p Th® in vitro
rate of inhibition, is laor© rapid with galant"HiJ®tIii7"
SIGHSFgCAMCE TO TEE PEOGRAM OF THE IHSTITllTg; This pro-
jict relates" "to the devsio'piaent &nd tebtlig of clinically
useful drugs for the treatment of myasthenia gravis o
gEOPOgSB COUBSE OF PBOJECT; Our inability to obtain an
&dequ&ti supply ofgalknthamine has hindered further
study of this dsug. The ©xperiasentatioa ^ill b© con-
tinued «hen a supply is available. Other compoi2nd@
will b© studied.
Part B included: Ho
ms-mu 2. Cliaicaliy Applied
Ixsdividwai ProJ©ct Reports FSmrMacology
Calassidar ¥®a.i- 1958 S, Bettesda, Mdo
4, M®w
Part Ao
Project Titl©: A study of aatiirally occurriag
choline esters 0
Principfai Investigator i Eicliard L. Irwia
Other la v®@ti gators: Jay B, Wells &nd EeuTj J»
^aith
CoopsratiQg Uaits : None
M&n Tears: (ealendar year 1958) Patient Days:
Total: olO Ccalessdas:
Professional: '05 yoar 1958)
Other: »05
Project Descriistios:
€BJEBCTl¥Eg; jS^ isicreasing aaouBt of evidence is
Weumwlatiag iadicating that choline esters other
than acetylcholine occur as oatural const ituents
of biological systeas. The physiological and
pharmacological significance of the choline esters
other than acstylcholine is lairgely uaJmowno Cer-
tain choline esters produce their physiological
and/or pharsmcological action by depolarization
of a^abraaesa The objectives of the present study
are: <a) to d®termine to ^hat ®:gtent the choline
esters which are found in biological system®, and
related coiapound®, depolsria® tissue mesjforanes
Cb) to relate the depolarising properties of the®®
compouncte to their stimulation and/or blocking
activity of synapses, <c) t© study the aietabolis©
of these compounds by tissue en^y^e©.
HETHC^S EIFLOIED: The travelling fluid electrode
T®cEM^m~lWnB®d to measure depoXariaation of
isolated frog sartor iu® aauscl®©, mcroelectrodesi
^iil b® used t© deteraia© resting ©esibrane poten-
tials. Standard saanoisetric technique® ar© used to
study metabolic activity.
„ :g .,„
-=•3
M4JGE S"iroi^g: In feigb coacesatration® (1© molar)
ButyflcEbiiisa, i3©iisoylGholiffl®j isBi«lasBol.®acrylcholi23i@,
aad isaid&soiepropioiaylc'aolisxcs re®®mbl® ac®tyIefeoliffi@
In their depoXariasirag piropeyti©®, MethachoIia©s a
syattt©tic coaapouad, -^laicfe W'-BBBmbl^B ac®tyIciiolia©
la its biological actioa touit diffsr® from it chem-
±c&lly hf b&vlskg a substituted cholisa® aoisty doss
Eot depolai'is® »uscl© ®@Bitoraa®ss at 1©'"'* molar ©oa-
centration. Plassm from jeyasthsmia patients tsa^®
beeici observed to m@taboIi3s@ iMdasioI®acr:$'lcl3Oii0®
at about the asm® rat® a@ plasma fross ffioi3~mya@th@i&ic
humaa l^eiEigBo
SSGHSFICMCE TO THE PROGRAM OF THE I^STITOTE;
Taifoxiiat'iba "coac@iaing""iSen^ological' activity of
substances which occur ±n asiiaal tissue greatly
esihancas our knowleciga of i&orsial and pathological
physiology-,
PROPOSED CQIMSE CXS* PROJECT; The project is ia aa
eSFtf pIsaeiT'^Turthsr s€u3y ^-ill ba nade of the
coiapouiads m@BtioQ®d above. Other choline esters
^ill be obtained and !studied. Emphasis will be
giv@n to those cosapounds which are natural constit-
^}i@Bt3 of ai^imal tissue. The chromatographic tech-
n±q%z®& of Bannister and Whittaker (The Journal of
FhjsioiogFs His 55) w^ll ^® used to identify the
active compounds preaent in tissue. Patient j^ater-
isil will b@ used where applicable.
Part B included: Ho
Serial MOo MSS5BB-24Cc)__
■'? lediciTTISuroTbgy
PfIS".Mi:E Cliri.ic5i.llF Applied
Individual FxojQti :;..:,.,.-..;,. Pb&rm&colos7
Cal®isd^ Yiaar I95S S« Bsthesda^ Md.
4. I^sw
Part Ac
Project Title: A Btudj to d^teraiae tk© ©f facts
of iJepolas-ljBisig ds'ug® oa muecl©
Prifficip&l Is4v@stigat©r2 Riclmrd Lo Irwin
Other Savestigators: Hosae
Cooperatiag Units: Hone
'M&a Years Ccalsadar year 1958): Patiest Days
Total : o 10 Ccal@Ea«iar
Professional: »05 ysar
Other: .05 1958):
Project Descriptioia:
OS<^CTI¥EB; Rec«0t work toy Dr„ Seaaeth Zierler of
"SoBis© lopEijigj L53a.tvar®itF has sho«tt that aldolase
decreases In jawscle® i?itfo iatact as^rabraaes whea
tJ3©y ar® incubated ta f iwid iilgia in potassiuaso
Tfe® coaceatr&tioffi of potassium u®ed are of the
order which depolarises muscle saeabraaes. The ob-
ject i^e of this study i® to detonaia© to what ex-
tent In v±Yo d©pol&risjatioa of muscle sa@mbraae by
drugs af'iect th® ®f flus of aldols^e aad other ea-
z,ym&B from sauscloSo
ffiTHOEiS lMPLj>flI?; Saa^a© coateat of muscles ars
deiemlSiS bf~tE& m&thodB used by Zlerler^ Asa. 4\
Phygiol> i®5Ss 193:534; lowry ®t al» , Jo B„'"CTf54,
ImTWT aad Sibl®sr aad LsSsiiiger., ^^bTUTTSW, 177;
Ib ffea ia vivo ijortioa of- tis® ©2Ep©rim©atB .
pairs of awacies are "us^ la tootls ©acperiaieatal &2sd
control det@2na,ia!i.tioiii,,
Smm.FICM€E '-fO ,3'^._^^MM g TTO IHBTITOTS : The
sf?ISF^F'1iii"fMea"TrlSrSSic1i¥^r@'"To"^^ actioa of
a drug '^ould b® a &@m} ftndlxkg In drug action ■which saaj
aid iB ■■affidersst&ndisg absaoxiaff.l states ia sauscla.
.«.™_J§S.^„^£!rjSS" '^'^ fulfill tfe® objective
ititteHr""""
Pai't B included: Ho
latlosffil lastiSue© ©f SSasrologicsl
Biseases sad Bliadiii^^ss
Cllaieai g@ses£ch
lledlcal Meurolagy Branch
Section OR Glialcai HairochaalsCfy
Serial ms^QTS o£ Projects:
HIHDB-25(c), HIHDB»26<c), Nim}B-27Ce), KZ£iDB-23(c),
HHffl)B»29<c), HIHI®- 30(c), KIHDB«3lCc), HINDB-32(c)^
HIHDB- 33(c), NIHeS-34Cc)» HIKBB-3S(c), HI^B- 36(c) »
Bgfei^ted (fellsatieas fog FY 1959
Totals 1160,500
Directs $84,100
SeimburseiBest; $76,400
Serial Noo_J|®B;^Ca}
lo Medical Neurology
2o Ciinieal Neurochemist:
3o. Bethesdao Mdo
4o Sarae as 57«NINDB«3CC)
PHS « NIH
iHdi^idaal Project Report
Calesidar Year 1958
Project Titles Electrolyte asad Energy Metabolism iss Norraai
aud Epileptogenic Cerebral Cortex ija VltrOp
Priacipal Investigators Dt^ Donald Bo Tower
Other Investigators s Mr, Eo Lo Peters
Cooperating Uaitss Drso So, Oo Brady and Bo ISo AgraHOffg NINDB
Section on Lipid Ghemistryo Dr<, G<, Ashweil„ NIAM) Labo
of Biochemistryo
Maa Years % Patient Days ; 0
Totals loO
Professionals 0o5
Other; Oo5
Project DescriptioQs
Objectives,? To study ij vitro metabolism of electrolytes
Cpotassiunip sodium^ chlorides^ etCo) 'and of energy«prodi5e :
cycles aad components thereof in incubated slices of cereL
cortex from experimental animals and from human patients
operated on for focal epilepsyo
Methods,; See previous reports on this projectp sumsariae-
in publication ^^l (Part Bo)o
Patient Material; Obtained from NINDB patients ad'»itt-3c;.
for other purposeSo
M.ai or: F i adi nas % (1) Studies on the mechanisffij of aotioo aad
the effects on incubated slice raetabolism of 2«deoxygXuc3se
were completeds The following resalts and conelusioijt: ;
forthcoaisg;
Ca) 2«dsoxyglMcbse prevents cellular glucose utilisation
by inhibiting the hexokinase step primarily due to depleti'
of available ATP required for this stepo The possibility
that the 2«deoxygly.cose's6'=»phosphate formedo which cansot h-:
further metabolised,, also blocks the glucose«6«phosphate to
fructose«6=phosphate step could not be ruled onto ^^"^
appears to be less isportant thau the effect on hexokinasSo
Dnder anaerobic conditions it was possible to overcome the
block in glucose ytiiisatioa bj 2*?=deoxygli2eose mth addition
of either ATP or glucose<=^phosphate to the sliceso No effr-^.'
of these additions on. aerobic metabolism was obtained^,
presumably djse to their failure to penetrate slices ?shish
aerobic conditions exiiibit less membrane permeabilityo
i'-osphate dehydrogessase ox
oxidativs sliU-il: patiiuayo IliiB finding was checked by ,^
incubating controj. and inhibited sliees with alucosewl^C*
and glucoss'^^sC ^ detei-miHiag utiSisatioH„ C ©2 siJf^
C «Iactic acid produetiono The ratios of C Og
and C^'^olaotate froa C«6 compared to C«l samples t^ere
loO in both eases, whereas C^/C«l will be less than 1<,Q
if tlje shunt pathway istilises a sigsaifiesat portioa of the
labelled glucose (©ogo ia liver it is Oo3)o This fiKding
is consistent mth reported ioi5 levels of braia TPN^ I'jhiol?.
is the necessary coenzyme for the shunt pathway aad isidieat;.
that the shunt pathvray is relatively unimportant in eerebr?-:;
cortexo
Cc) 2«deoxyglucose inhibition raot only results isi marked
decrease in glycolysis but also in oxidative ssetabolissso
With giucose=0«C^'^ less (^ Oo^ less labelling of the free,
asnino acid pool and less C «laetic acid are obtained,-.
Under aerobic eoMitioas with 10 nM 2«deoxyglucose glacdse
utilization and labelling of these compoQsnts were reduced
to one«third of control sliceso These findings complement
the previously reported[5?'=I^INDB*=^CC)3 depletion of energy^'
rich phosphates^ ATP and creatine phosphate,, in inhibited
sliceSo .
Cd) From the studies with C "labelled glucose the distrib;
ion of glucose utilised by normal slices to various inter^*''
mediates could be estimated i
Glycolysis (lactate) 70%
Amino Acids Cglutamate) 22%
Bespiratory COo 7%
Other intermediates^ lipids and proteins 1%
From the oxygen uptake (85 filo /go /hro ) it is c3
the 30% of the latter accounted for as amino acid and
respiratory CO2 almost exactly balances the oxygea uptake^,
assuming 6 moles of the latter per iEOle of glucose oxidisec
This eslculation is consistent mth recent reports by
Geiger on C ^sgliicose raetabolisna by perfused cat brain zb sj
wherg.2C^ appeared as amino acids and 3(^ appeared as
•C'^^02o It is also consistent with his eonclusioas and
those by this laboratory from studies on 'f^^minobutyrie
aeid [see 58«^IP©B«30Ce) ] that non^glucose substrateSp
such as affiiao acids^ normally support oxidative metaboiisE
by brain and that they are repleted subsequently by part
of the glucose utilisedo Also our findings in this study
agree well with similar previous studies by others using
labelled substrates in ^itrgo
(e) TIius„ it Kiay be concluded with Geiger that brain
possesses a versatile system for support of oxidative
raetaboiissa in trfiieli not only glucose but non<=^giucose
is sfcili costpieteiy ooissisteui sfith iAs ovsraii. sts.-.
tliat gl'iicose serves as the principal or sole sabstifu
csidativ.re jaetabolism of brainy since lack of glucose
makes rspletioa of the son-=glu0!ose iKteraediates iiapossibJ.e
asd esiergy^productioB rapidly failSo The studies with
2«deoxyglacose clearly demoastrste this not only by the
depletion of ATP aad creatine phosphate but also by the
dsietejrious effects on glataaio acid and electrolytes in
such inhibited sliceso Both these systeais depead lapos
energy production froa glucose oxidatio3o When tliat fai5.s
glutasie acid le¥@ls fail profoyjidly to abOMt 30% of Hormalo
The eieetroiyts picture esn be summarised as follo-=sJSS
Potass i'jja Sodiiam Noa°Chlorid@ Spaee
Wc2iaal«I?iitiai 46 100 "" 3?"'
i hTo Iraeubo 92 59 48
2-4X5^1 hSo Imubo 62 84 ST
ThiiBg inhibited slices fail to extrude esssss sodii® and
rscoaeeatrate pofcsssisjua in the normal maaaero It may be
sigriifieant that this type of defect ia electrolyte
setsfeolisia is also eacouRtered in ineubated slices ef
epileptogenic cerebral eortexo,
Ci) Detailed studies of electrolyte jnetabolism of iBcabate:
slices of epileptogeaic cortex frosi a variety of experisneat;
anisal preparationSp The previously deaoGStrated aljaoriaalil
demonstrated for Irman epileptogenic slices [see 57«s>iINDB=--'
8 CO] were also fouad ia cortical slices froa? cats with
seiaures induced hj 3*34Bethyl=3«3tl3ylglutariraide (Megimide)
aad h^ aatliionins ssjlfoxisiseo Little de%'iations from
aorraal u-as observed for saraplas frees cats i^ith thioseasi^
earbaside seisareSo Sicce ail seisjjre preparastioas inclydlr
the latter are associated mth defects in glutamic and
T«as3iH©batjric acid sietabolisao this diserepaacy in the
electrolyte disturbances with thios^aicarbasicle jsay prove
sigrdficsrstc. Oh tha oas haad it may iadicate tlnat
electrolyte disturbaaces sre seeosidary to other more
fimdamesstal distarbances^ perhaps ic energy metabolisSp
sBd oa the other It suggests tnat disturbances of glEtataic
acid sisd T=aniiacbatyric acid metabolisa raay be prisnary
svests ia the seisare process,, perliaps through effec'^" ■'-""
energy isstsbolissso
Ta© ability to deteriaiae chloride levels accurately
OH aiiqaois of all thess samples by tfes aaperosetric
prosedm-e of Got loire el. ,ai,o CN ^.l-L I o ) has added greatly to
the sigaifieaBee ©f ths studies os ©lectrolyteso Th©
Cotlove iastrisaect aafees It feasible to detersias tlse
relatively 1o?j tissue levels acearateiy aad reprodsioibly
on as little as or£g«tenth of tlse -iotal saspieo so that
eaoh sample can foe analysed simultsnecusly for potassiisso
sodlHs and ehlorideo Froai tissue aad sediKBi eosjteats the
ehloiride and Bora«eSiloride tissue spaees ean b® reps'odacibiy
calsulatedo la noriBal slices these spaces regtslasly behave
dwriag iiieubatioa as followsi
Chloride
Solids, NoBfc€hlorid.e S^aee
Initial 16% ^ 35
I hro Incubation 16 48
Th© final space distribution is alsost identical with that
Sound ioT in.Mi^ biopsy gai'aplsso Furthermore th©
changes canaot account for the electrolyte cliaages
(extrusion ©f sodiunso recosicentratioH of potassiiiin) obserFed
for normal slices o thus providing further evidence that
th© latter are mstaboiicaHy^^deperjderito Fiaally tfeese studi.
again coafirts work of others that the swelling (gain of weif;
of jsorraal and epileptogenic slices daring iiacubatioa is coii^
fined to the sMloride spaceo
Calculatioa of the electrolyte eoacQKtratiois per litre
of -aoE<=chioride spacs ffi?ater at the end of slice iKCisbation
can be derived from the above data and may be siammarised
ss follows s
Po|g3§Jjm SodiijE
Korjaal 180 ^
140 80
The significance of the excess sodiwsi coaceatratioa ira
epileptogeaic slices is aot kaowno The effects of Neabutal
aeesthesia in vivo, apoa s?ibs@qiient behavior of electrolytes
in iaci^feated slices of normal cat cortex have bess st?Jidiedo
No differences from t&e tjefeaiifior in iiaaESSthetised slices
have b©eB foasido This is in aarked coatrast to sigaifieasit
effects of anestlaesia oa sabseqissat slice sastabolisis of
bousd acetylcMoiine and of gl?Jt^iic acido Tls© findings for
electrolytes are contrary to what sisight Isave been anticipate
frsffi SEieh tesalts as Bieli as from J^ vivo observatiosaSn
©ogo hj Koodbary,, that certain Mreotics stabilise nearoaal
iserabraaes and affect ioaic fluxes across theeJo Woodbury °s
most striking results oecarrsd vdtii Dilaatiia aad Diasaos
will ell have »ot beea evaiEated hereo
MMMMJ:P3S&® o|., Prgjeg.!, t Energy metabolisni is the basic
factor in. neisroaal fmictioa sad activity^ aad electrolyte
metabolisso which is clearly depesdeat upoa it^ provides
an important bridge beta'eea cellular ehesistry and tlse
faactiossai actiidty of impulse sond^ctioRo TIse uraderstandii"
of tb® factors iavolved is esseatisl both for aorraal nejjsrom
tissise as i?ell as in hyperactivity status like seisiaresc.
Progjl^d , Ccmrs e ; To coEtiirae the abOTre studio
pv>oeedijres developed in the coets® of the 2'^dea:'''yg.;.uC':o:.-^
studies ?jill be applied tG epileptogei;uc saapies to
ia¥6sti.gat© glucose utilisation aad eraergj prodMStiou sna
aaintenasceo TIse electrolyte st?^dies will be aaiplifisd
by studies of effects of hypoxia and of Tasrioiis anti'='
eonvijilsasit agentSo An attaapt to studv ion f,l,^ges between
iscssbstioa aedisim.aQd slices^ using K-'* and Na'*"" and a
T«2"3y speetroBieterp tsill be madeo Ii successful witli
norsal slices, applisatiosss to the defect is epileptoge-aic
slices will follo^o
Part B Included s YeSc
Serial NOo Nli^B^iiSlcj
PI-IS - NIH
Individual Project Report
Calendar Year 1958
Part B:
The Effects of 2«Oe0xy"D<=^liaeose os Metabolisa
of Slices of Cerebral Cortex lumbnteA in jTitgOo
Jo lleMgocIsOTto 3 s 1e press o
2o To^jer, Do B<,
Tbe Evideaee for a Neurocheraical Basis of Seisares-.
ppo 301«348 in Baldwiao Mo |i, a|o CEdSo)
T^BMSl XiSfee SgilgBi£.g Spriagfieldo ThoaaSp '-
So Towsr„ D<> Bo
The Nsarochemistry of Convulsive States „
ini Foleh, Jo (Edo) Chemieal PatholOOT. M
' IM NogJI^ Sygtjgo CSrd lateraatioaal NewoelieaicaJ
SymposiiM) Loadoag Pergamon, in presso
, Do Bo
Glutaraic Acid Metabolism in Matasaliars CeHtral
N©rvoas Svstesio
im Brackeg Fo CEdo ) S^eiMsiuK ^ Bioaheii.lstrx
of the Ceatral Nervous Syst^a CIV lEt®rraational
Biocheisical Congress)^ LossdOQ„ PergsmoHg in press,
HoQors and Awards
Appelated to Editorial Board of Biogheai'sal Pharaa^J^Mxp
Serial No.>_WIMI'B°26vt©}
lo Modical Necrology
2s CliBicai Neusrochetaistry
3« Bsthesda^, Mdo
4o Same as 57 NIWB«16 CO
Isidi¥idual Project Report
Calendar Year 1958 ' _ ^
Project Title? Csaparati^© BlQchmistry ©f Smooth Masele
and Striated MKsele^
Principal Investigators DXo Beat Horvath
Other laves ti gator ss Mr„ Jo B= Proctor
Cooperatisg Units s Kooeo.
Man Years ; Patieat Days s ,f ^2
Totals 0,6 . _ ^'
Professional; 0c3 ' ' -■ ,,
Others Oo3
Project DeseriptioQs
Obi ectives s To characterise tiie aetomyosin of smooth Cuteriue?
aad striated muscle in" physico^^hesaical terms preliminary to
study of actcsayosis synthesis in muscleo
Mttfeods-; See tsudsr Project »58^INDB^35Ce) and 36C®)
In addition to raissele ssaaples from normal aniasls,,, samples
from a strain of tniee in t^ich a disease reseHsbliag msascalar
dystrophy is inherited are being usedo Controls for tJaess
aice are provided by normal littermateso
None<,
Major Findinggg Because ©f the physic»=*©hemieai ©haraetsrist:'
observed for the control aed '"dystrophic" moss© saEples
[5T-?JINDB'=5l5Ce)] it ®as dsesed advisable to carry ©sit sitroger
determinations ©n the variojas fractions ©f eobs® misscle
hoaogenateso as in 58-*iINDB^ 36Ce) before proceeding further-
Changes in the ianscl©s of tlie "dystrophic'* @ie© ar© similar
to bat of lesser magaitade than those observed in biopsy
specimens of human dystrophic raascieo These is &n iracrsai©
of total solids aad aoa-proteis solids„ indacati^© of fat
replaceaiejfit of rausslSo There is also an increase of Na
and decrease of K„ as found in hiaaan dystrophic ejsscI®.
Th@ relative propartions of water-^solaMe proteins „ ayosia
and alkali -soluble proteins do not show remarkable changes «
bat so far only, rauscles in the early stages of degeaeratioa
hafe been stHdiedo
Daring the smmet a brief visit ^as made to the
Eossse Be Jacksea M^aoriai Laboratories at Bar Harbor,
^he'£& this strsia ©f mice ^as dises^sred sad is being
aaistaised asid studied genetisallfc The ©bser^atioss ©f
i Investigators tlis?© relative t© tbe genetic aspects ami t©
successful propagation were gos® iato ia detailo €o@p^^atioE
of the Bar Harbor group was streagthessed by this isterchaage
of iafertaatioSo'
Siaaifioaace ef Prolect; Studies os format ios of actojsyssia
may have an iapostant bearing on the loejis ©f disease ia
muscalar dystrephy aad GtMer myopathies,, Developmeat of
suitable mi cr methods aad aaisaal preparations is a aeeessary
prelude to eKtension of experifaestal animal data to hrms^n
specimens p
Proposed Cotasse; To extend these studies to iaclude mice
in various stages of their disease iE order t© obtaie infor=
matioB OQ the dynasics of suscle degeaeratioBo Such iHf©r«=
tsatioQ will subsequently be applicable to projected stmdies
©f muscle protein formatioa CsyBthesis) ia uterine ai?ssle
jiader hormoaal stimalatioEo
Part B Incltideds Yes,
Serial Ho..,_MjMJJL2M^l
Individual Project Sepsrt
Calender Year 1958
Msisele Py©te-ins is Dystrophy
Eaners, a^AmsM,' ^^m.
Serial MOc^WT?CT:-5|7^ifi.'l ^
1,, Medical Kearology
2o CliElsal Nejirochesiistsi'
4a Same as 57=liI!^DE<=='5«CC}
Individual Project Eeport
Calendar Year 1958
Part A;
Project Title; Aaino Acid Metabolisffl ia Norraal and ^llept@genU
Cerebral Cortex ij yitro,.
Principal lavestigators Dro T}OBald Bo "^^er
Other latest i gator ss Hr= E,, L<, Peters^ Dro Mo Bald^iHf, Dr. Co
Ajomae«MarsaHj Dr, lo Klatzo ;;. ' .:" ", - ':-^ : -._:,.
Cooperatieg Ouitsi Dr„ So 0, Brady„ NIfDB Ssctica 00 |,ipid
Chemistry (Liquid Sciatiilatioa Coaatiag)? hr. So W„
Albersn NI^B Lab„ of Neasroasistoraical SeienSeso
Man Years i PstieQt Days i 0
Total loO
ProfessiOEal 0<,5
Others 0c5
Project DescriptioQ:
,^lectives ; To study the ia vit?tt metabolism of affiiao
acids sad related compouQds in brai& tissue samples txQm
experimental aniisals and froa hmEan patients operated on
for focal epilepsy,,
Methodse See previous reports oa this projects sssasaariaed
in publication »1 (Part BJ,
fatiest Material ; Frois NM2B patients adsaitt®d for other
purposes o
15
series ©f studies ©a slices of cat cerebral e©?|ex
iaeubated Mith L^-glataiaie a©id"0=€^^(, L=glMtEBaiR®=^IM]'^'^„
r==aEiiiobatyric aeid»l-C^^, L-aspar|i© acid«l3=€^^^ DL-
asparagiHe=2«3=C*^o D«glwc©se--'SJ-C^'^9 Sodiaia PyrOT®te?3---C*^,f,
a-pyrrelidinoae-^a-C^-, md a-feetogliatarie aeid«lo2^^'^ h&s
been carried ©Mt,-, Fifjdisgs by previous investigators sssisg
soffls of the ab0¥@ isotopisally^Iabelled cempeiaEds ha^e beea
confirmed asd extesdedo Startiag ^ith either glMeose or
pyrwatSo the order of labelling in aain© acids is first
glsataEsic acidg follo^sed by gl5ita!Biae„ t'=^aiainobi!styric aeid,,
aspartic acid^ Oralaaine and serineo a^ketoglsitarate
equilibrates so rapidly with glsataisie acid tliat its EBet-aboii\
behavior is equivalent to the latter for all. praetisal parpes
GlMtasjie aeid is rapidly Mstsbolised to glMtaraiJiSo r-mim'^
butyric aeidp aspartie aeid and ««alanin© in tfekt mdeto
Glutaaiae is srapidly coBTertsd to glutaaaie aoid and
thea thru the same pattoays„ T-sstisaofouftyarie a©id is
rapidly aetabolised mtk lafesliiEg appeariag in ©spsrtie
acid^ glutaffiie asid and glutamine indieatiag its aeti¥©
entry iato th© Krebs eyele, Pyrrelidiiioae is slewly
hydrelysed t© r^aalEObsitjric acido Similarly aspafaglE®
is slmlj laetabolised to aspsrtic acido Aspartie acid
is rapidly Mtiiiased aitli sctivity appeariag in glsitaiBie
acide laetie acid^ gliataairae aod T='SBainobatyrie aeid ia
that erder<i Metabelism of aspartate to laetie aeid is
compatibl© with th© folloKlEg rs&etion sequences s
Aspafftie Acid -»" Oxalaeeti© A©id
Laetie Aeid ^ Pyruviie Aeid
This pathway is knoivn in bactsria„ plants and eertaia
animal tissues but has not been reported before in braii5c
It has considerable significance since it would tiseafi
that aspartic acid could prime the Krebs cycle by prouiding
both oxalacetate and acetyl-coenzyme A (froiB pyrtsvate) in
the absence of the latt©r from glycolysis. The foregoisag
studies indicate ho^ active the components of the glataraste-
aspartate ataino acid' group ar© ia metabolic particiDatioti
in the Krebs cycle. Concurrent detexsai nation of C O2
liberated during these experiments confirmed this conelesioa..-
C2) Glutamic Acid^ glatamine,, Y^-asinobutyric acid and
free aBEsonia metaljolisa was studied in incubated slices
frtMB various noffi--e0rtical areas of cat Mraisi;! subcortical
white matterj, thalsaias CtotaDp caudate nucleas^ and
cerebellar cortex., Levels and metabolic behavior in ail gray
©r neuronal areas were similar to that previously obsertred
ia cerebral cortex^ but eshite matter exhibited extresselj
lOH i©¥elS;5 aM little change on incubation for glutamie arsd
T-^aminobiatyric acids while whit© matter glKtaaine mas set
greatly different from cerebral cortexo These findings
©osapare fairorably isith levels for iiaany siailar areas in rat
braia deterained ■ by other methods by Waelseho
C3) Levels of glBstamie acid, glutaasiae and Y^assisobatyric
acid in subcortical sshite aatter and in cerebral cortex ©f
the cat brain ^©re detenainedo Ssiisg the calculations of
Elliott and Heller it could be estiisate^ from these levels
that at least 8S per cent of cortical glmamie and Y^^amiiio^
butyric acid content raas associated with |e6jroBs while only
about 50 per cent ©f the giutasEine appearld to be neuronal
^.-^n'locatieno (This finding is consistent lith interpretations
by Waelsch based on in vi^o isot'i>pe studies for a different
"compartmentation" for glatamineo) In addition ©at cerebral
cortex was fractionated by the.Brody and Bain technique and
the cellular loci of these amino acids determined by aaalysis
of the fractions ©btainedo The majority of giutaraic and
T«aminobutyric acids wis associated with fraction R«
; P /
or the mitochondrial frae-tioUp ishereas glutamine was
distributed almost equally between that fraction and the
combined S, 4 Eg fractions erfiieh contain cell debriSj,
axon fregmeBtSo nuclei 5 ©tc^ No ccntest of aaj of the
three amino acids was detectable in B^o the mierosomai
fractioHo The presence of most of the glutamic and
r^aiainobistyric acid of cortex in mitochondria is compatible
with their close assoeiatioa «3ith the Krebs cycle and
oxidative ssetabolisa which are mitoehendrial fiiactioBs
exclusiifelyo Ih® significance ©f considerable glutamine
in a ROE-«aitoehondrialo aon^aicrosomal fraction is not
clear and sill require farther studyo
(4) Exteasiy® studies were carried 03st oa the nature
of the inhibition of glutaains synthesis associated mlh
fsethionine salfoximiae intoxication of catSo The preliminary
findings reported la 1957 were confirmed that slices ©f
cortex from cats with seisiires induced by Esethioniae
sulfoxitaine shorn decreases in levels of glut^nie acid
and T'^^aminobutyric acid during incabatiom both of ^hich
can be corrected to noraal levels by added L-^methionio®
10 ngo Such additions ha^e no effect on inhibited glutamine
synthesis in sach siiceso Addition of samonium chloride
10 rM to normal cat cortex slices during incubation caased
two to three==fold increases in glutamine levels at the
expense of glutamic acid («=an in, vitro effect entirely
comparable to in ji¥0, findings recently reported fro®
Greenstein*® laboratory in NqCoIo) A pressaesably secondary
effect in these incubated slices was lowered le¥els of
T-arainobutyrie aeido HoBever„ addition of NH^Cl t© slices
of cortex from methionine sulfoxiffiine^iatoxieated cats failed
to stimulate any rise in glsataraiae levels,, which remained
sear sero^. But ^hen both L^^ethionine and NHXl were added
to such slices a significant increase in glutaraine levels
of these slices was obtaised» It would appear that the in-
hibition ©f glutamins synthesis by methionine SMlfoxisaiB®
Is prisaarily ass interference isith the aiwonia ©oietyp
possibly by the imine grosp on the toxic compotsndo hwt that
adding only NHXl is ineffective in overcswaing the block
unless adequate lesfels of gltatamic acid ar© a^ilabl© to be
amidated to glutaraia©o These findings do not shed any light
oa the meehaaism producing levered glutamic acid lewis exc<?pt
to indicate that they relate in s&m mf to iahibiti©n of
methionine metabolism by its antim®tgbolit©o
(5) The effects of 3^etliyl«3«ethylgliitariside CMegiraid©)
©n glustaElc acid jsetabolism ©ere extended to imclrad® T=afflis®=
butyric acid-s ? Slices from cats with seiaures indsEced by
Megiraid© shewed levels ©f r^'astiaobntyric acid sigaifieaatly
lower than normai„ Siiailar studies ©n slices from eats with
seiaisres isdaced by thiossmicarbaside Can Isactivator of
pyridoxal phosphate) showed not oaly very low r^aaJiaobBtyrlc
acid levels bst also very low levels of gliatamie acid.
CKillaBi had j-eported the fOEsaer fiBdifflg„ bat Md arat
fQuM the latter to be trssej presmaably beeaes® fee faiJed
to quick-fre^Siis brai^ biopsy samples frosa ssish aaisaiSo)
C6) IncubatioH of aorraal cat cortex sliess with 40 vM
maloaate prodaced soae ^©ry samsisal ilMingSo Maleaate
is kaoHE to Gompstetively iahibit- succiisic clehyds-ogesjase and
the effect of this coracsatration oa oxygea ceastasptioa of
slices ■ 'CSO^gxedsaetiQW) ms reeoafirmed her®. Tb.es@ sa®©
slices exiiibited HOTraal prodactioo of la©ti«j acid so that
no iaterfersnee wth glycolysis or glacos© Mtilisatioa
occurred, la tSie pr©s©Bc© of asaloaate slice levels of both
glutasQlc acid and T'^amiaobnatyric acid r®se t© desble isenssl
valsuesr This observation is iceportaiit fr<Ma tifo staadpeintSo
First Weil'^Salhesrbe had fouad raaloaate caused Efedmtioa ©f
oxygea uptake in both hoffi©genat@s and slices of braio tet
aa aosiuaialatioQ of succinat© only in hosaogenateso At tSie
time it was isot clear why slices did not also show siscciisat©
accismulationo Our data sssggest that in th© whole cell
preparation it is gliitamate arad T^arainobMtyrate ratfeesr thaa
SMCcinate ^ieh accumalate aad require a study of th©
reiatioEShips amoKjg these thre© conponeEsts of Krebs ejel©
Eetabolissr. Secondly sirace studies in another project
fffoffi the ssctioa C58-^INDB=^30Cc]) indicate that y^aaisobatyr:?
acid may be aa important Krebs cycle SMbstr0te„ the effect
of m iahibitor t^dch blocks the next step b©yoiad in produaiag
aa acciaaalatioB of r^aminobutyrate aad its preciarsor glutasaie
acid is most suggestixre eoafirssatsry e^srideHceo
.SigBlf^caBcg of £¥o,^eet; The prosecution ©f this project
is of fuadaiQeQtal isaportaace to an understaQdiog of the
roles of the glistaffiie aeidwaspartic acid group in neiirojiai
metabolism aisd ia the seisure processo
pgQBOsed Ceairseg To coHtiais© th© above stiadieSo Particular
attention will be paid to aspartic acid metabolismo utiiisi:
a aewn unpublished microaaalytical saethod saade a¥ailable tc
us recently by Dro Oo Ho Umsfo ¥ery little is known about
its metabolic role in brainy bEt the prelirainsry indicationc
obtained in this project during the past several years
suggest that it asay be as interesting and important as
glutsasic acid has proved to beo
PartJa^JlglMded; Yeso
S®«fial KOo_S|^i:;2Jiel
PHS 'Mli
Ifidi^idaai Froje<et E@pofft
Cal@r«dar ¥ear'l9S8
fMLR-
lo Towers Do Bo , ,.
Th© Sffests of a-Beojsy-D-Slaeos® on SSetafoolassi
of Si lees ©f Cerebral €osftes Iiisiibated in Iill£»
I. MMSSMSo 3; ia press.
2o To?3er,, Do' Bo
The Svidesjse for a Nearoehemical Basis of
Seisares; . s
pp„ 301 - 348 in Baldwin, &1; et al CEds.)
Tgmijgggl Lp!:^ Epjleps^o Springfield, Thosas,
l958o
3o Towers 0; Bo
Di8@assion [Cliaisal snd Pathological Aspects
of -fosieitF. ^^^^ "Ageniaed" Proteisss aad Metfeioai!!®
SwlfosisJifflelo
pp, 288-295 in Ibid, l958o
4; Tower „ Oo Bo
...Glutaaie Acid Metabolisa in Maianialiara Central
NerTOUs Systeaso
ia BrSekeo F-o CSdo) S^iaposiMm on MoeMgAitry
Bio@lseg.igsl CoMgiM). Londoa, P^rgaMs^ sra P5»@ss.
5. Tower, Do Bo
Th© Nearoeheaistry of CoiiTOlsiw States.
iu Folsh, Jo CEd;) Gh©i|©ai Fatfeoloffl Sl lit
1^1213 Sistgs C3?d late^atioSii M§MIi^
eliesLieaJ Symposiaa). London, Pes-gaiBOH„ isi Pre.-,
6o Tower, Do Bo
Th© MiESiMilstry of ejMiliLt sM MsiiafMt
Ik Bradjf, R. Oo and T©w©r, Do Bo CEdSi)
SiSMsHS MS. msS^SBMrj of m&MSlMSM.^MA
Amiao Asids CAsaeriean A©ad©sy of Nearologj),
nm YotK Wiley» isJ Press;
Appoiated to Edites-ial Board of Buofffleiiial
Phagaa©olow,°
Serial HOo
lo Medical Nearelogj
2o Cliaieal Ksisreeiissaisr&i
So Bethesds,, Md.
4o Sasse as 57 KIlDB'-^CCl
md 5T-^lfIM)B-5CC)n
iQdividual Pr©jeet Msport
Caleadar Yesr 195B
Project Titles, Cliaicai Evaluatioa ©f Various Mim Aeids
aad Selated Gompounds in Control of Seizures iraclssdiag
Studies of their Metabolism in VIto.
PriBcipal Investigator i Dro Donald Bo Towero
Other Investigators 3 ©To Peter Bosjleys Dro Guy McKManp
Drc Bushaell Smithy Dro Co Ajsone«Marsan^ Mr.. Eo U PeterSc
Cooperating Units ; Noaeo
Man Years ; Patient Days s 75
Total? loO
ProfessienalsOoS
Others. 0o5
Project Descriptions
Objectives s To assess the effeetiveaess of -various Bmim
acids and related compounds in the glutamate-aspartate group
for control of epileptic seisares c«aaplemeated ?jith stadies ®e
the in S-JM. aetaboiism of the various compounds in meo
igthodj,; See 5?'=IIIM}B-4 and 5CC)o
Fatieat latgrial; Admissions to NIM)5 wards specifically for
this projects Pli^s patients adsaitted for otheKpi^oseSo
Maler.FJBdlaqs; As indicated in 5T**'3INDB«3CC}(, patients raaaiffiikg
oa L'-asparagiae have bees or are beiag discoatiaued or ahee
appropriate s^tehed to y^^amiaobutyrie acido Data froa the
cooperative stEdy of patients on L-asparagia© ha?e been eolleetp;
frtsa all bat one clinic and are being pr©©essed and tabsslatei- '
The fiisal report on this study ?fiil be drafted early in 1<?59,-.
Pstieats oa r^sminobutyric acid have contlnsted to d© wsilv,
Oas ease became seisare free after three months on the csaspoHsd
compared t© ssaltiple daily seiaisres previoKSly,- On stoppisig
r-aaisobutyric acid her seiajures retiariaed and have again beea
abolished bj re=starting r-arainobsatyric "acid. Several other
patients ©re experiencing raBich better control on T'-affiinebatyFie
aeid than on L«asparagiBe.. One of these^ follosred for feisr year-
on the latter with sustained impr©¥©Eent over his previous level'
of control^ is now airaost free of seistareso One patient has
experiesBCffid little change in ssisare frequency aftsr startlag
©a T'^asaisobutyris aeid^ Ml has bg@H abl© to redsic® ©tfeer
eedicatioQ sigaifleaiatlyo Sefiaite iKpro¥®seHt is cojatr©!
has bisea ©bser^ed iu patieats ?dth petit aal abs@j@® type
seisures aa«3 is these with gsaeralissd eoavaisiess eqiaaily,.
It Is clear that ia this SEall greup of patients T^^asaiaotatyri©
aeid has proved relatively effeetiveo The sigHificaac© of this
ebservatioa is difficult to evaluate antil studies sow ia progr-ess
ia amth&t project C58^E®B=30Ce} )claFify the qsaestioss of
fghethsr &s not systesic Y^amiaobatyric aeid ©rosses the biood°
brain barrier and Mhetker it famctions as an importaat 5?sbstrat@
for cerebral ©xidatiire metabolisHo
Iq oq® case seireral att^apts Tser© made t& administer
Y-'amin0batyric acid istraveBOsssly disriug EES recordiagSc f ?ae
solution had beea aaatoslaved, check for pyrogens and rapidly
iBjected iEtraveasusly iat© uaanesthetissd dogs in a dose of
4 isM/kgc, bedy weight Kith m uatoward effects » Peak blood
ls¥els ia ©ae dog were 10 i4a„/Eilo of serisao Previous stadies
OH affiesthetized dogs 1]S6=NIM)B<=97(C)3 d^oastrated a© sigaifieaat
changes ia BoPo^ respiratioaj, S£G ©r EES follcming SMCh injestioas.
¥et ^m 1/200 of this dose of the sam® solatioa Mas sleisly
iBjseted iratraTCaously iat© the patieat there sas iimediat©
agitatieas, f lEshing„ hyperpsiea^ and drop^ia diastoli© blood
pr©ssar©o. On repeated study,, with iujections ©f saliae
interspersed withosit effects the sara© phenosena e©ald be
repreducedo EecoTCry oeciirred ^thia 5-?10 ®isut©s„ Despite
reports by Elliott that such occurrences can be igsored asd
large dose safely isjectedp it woisld seeta that pssrsait ®f this
type of stMdy is too potentially dasgerous t© b© jusstifiedo
Studies OS a patient with "pyridoxiae dependency'' aad th©
interrelationships of pyridoxin® and r^asainobtatyrie aeid
EsstabolisE and fanetion in the ceatral nen&m systOT are
sufficiently naique that they forta a separate project (see
58=I^IN0B»29Cc) ).
Ostil data on r-^aminobjatyric acid have been ssssre fally
stiJdisd„ trials of 2"pyEi^«iidiaone and P-^alanise h aw beea
deferredo Ho^feFer, stadies ^th 2»pyrr©lidiaoa©=2=€^^ with
slices of cat cortex iscubated ia vitro indicate that
cerebral tissues can ©pen the pyrrolidiaone rlBg to yield
r-aaiB©bat5?ris gsid„ hmt th® rat© is relatively slm ^ about
15 per ©eist of a 4 /iM/g. dose of specific activity 1=2 ^/p^
was hydrslysed daring 1 hoKir'"s inenbation of eortiesl slicesn
This fiadirag cosspled ^ith the pr©baW® effects of .blood-
brain barrier in ^j£ suggests that 2-pyrr©lidin0ne is sanlikely
to b® a practical prscarsor fors ©f T=®«ainobJstyric acid to use
clinically. In ji%TQ studies with p-alania® deiaonstrat© a
definite inhibition of oxygen uptake hj cortical slices incubated
with Ito Albers has suggested that if p^alasin® iji setabolised
hy braiE it srowld yield maiGaic acid ^ich is a kKOwa ialiibit©-f
of SMCcisjic dehydregeuaseo Despit® a fa^orsbl© report bf
MiiiiaiES at Eiery that p^alaaiae ®x!iiMts cliaieal effieaey agaiu,-y.
seisijresj the jj, jitrg data mskes its use qsaestioiiableo
Si qBi.figaBCQ_Ql-g^Lgigct i TSsis px®ject is part ©f a U&q tem
stadj of the biosh^ical basis ©f §©isur®So Cliaieal applieatioas
©f promising leads developed is tls® ©xperimesital pliases ©f t!i©
study ar© important both as peteotial coapliia©Btary clinical
validation of ©jq^erimertal fiadiags and also as potential aew
and Qore rationally based therapieso
Proposed Cgsigs®; To contimi© studies discussed abo^eo
Serial KOo^Jiwa^Mri^^
y
PIS ^ MIM
Caieadar Y@ar 1958
fublieatioMS;
lo TeweSo Do Bo
Pyridsxine aad Cerebral Activity
2o Toiifer„ D„ B,
The Nearoehmistry ©f CootuIsIvs States
ia Folclio Jo CEdo) Cheijejl. Pa.tholoav
Si t|e W^.MMS. Sistm (^ IgterMlioM-i
Neuroohepicil SgrjBogJM) L©sd0ii„ FergsEaeBp
ia press.
3o Towsro Di, Bo
The Ke«roch©siistry ©f Gliatamine aad Asparagira©
is Brady(, Ro Oo asd Towers Do Bo CEdSoJoSroposij
IB Brady(, Ko Oo asd Tower^ Do Bo CEdSo^o^gosjjm
on_Neiargchemi_atry of Rueleoti^es aad..Affi.i.ia^,^iB§..
CMerlcan Academy ©f He^rology) Nss? York, Mi ley,.
in press c
Serial Ko„.Nl^B^29ife
i„ Medical Ms^affO^ffiyv
3o Bethesda„ M
4= New
PHS » NIH
Inditriduai Project Report
Calendar Year 1.953
l»rojec5t Titles The Relation of Pyridoxine C¥itaain B^) to
Certaio Seizure States,
Principal Investigator; Dr„ Guy M„ HcShann,,
Other Invsstigatoxi ; Dr, Do B<, Tower? Dro €■» Ajmone-liarsaiao
Cooperating Daits? Br. Louis Sokoloff^ KM!! Section m Cersbral
Metabolisms Dr., S„ W. Albers« HINDB Lab. of Neure-=
anatomical Scieuces? Dr„ Olaf Mtckelsen,, num lab, sf
Kutritioas Br, D. B. Coursis, St= Joseph "s Hospital,
(Laii©astar„ Pa,,,)
las Years s Patient Days; 20
Totals 0c,4
Profess ioas is 0o4
Other t 0
Projeet Descriptions
Obl^eeti'gesj To eiaberate the role of pyridoxine CVitaffll:
ia' eertaia seisare states hj both ia ^im and ,|j.xli?J,
iairestigatioKSo
6
Methods.; Pj^ridoxine defieiensy eaa be indueed io experimeata
asisals eithsr by dietary meaas or by use of pyridoxine
aatsgonistStt sueh as thiossiaiearbazidec Siase seizures sesul
froa the fsll blossn deficiency states regardless of method
©f inductioa,, thair appearance is taken as the ®n4"p&int tor:
studies o Effe-sts of the defieieaey is et^aiuated by in siMM
determinatieas of cerebral aaaino aeids^ espeeiaily giutamie
aeid aad T^'saiaobsityrie acid^ measuring levels and invest! g
Eetabeiic pathisays of these sotsipounds in normal eoctrols asa
defieieat sampi-as as outlined in 58=WIM»B'=30C©)
She hursaa soaraterpart under study is a ©©nditioQ
origisally described by Huut ^ alo in 1954 as "'pyridoxine
depeadenef D " A haif=d©sea such cases are now reesgniased
in the U^Sc^ representing a condition in which seizures
oee-^r unless large daily doses of pyridoxine are provided.
Such patients are stiidisd by the Krypton method for
measisriag cerebral mstaboiism developed by Sokoloff and by
receut analytical procedures for pyridoxine asid EStabolites
deif eloped by Coar$in„
Patient Mat griaj s ©btaiiaed ttm UimE patients a<fciai'fc?,@d
speeifieslly fer this stsidy aad tJi©s© admitted Ut ©tfeer
parposes^
Ma for Fiadiaoss Use erigifflal eas© of ''pyridoxine depend®rjc.y™
reported in 1954 by Hunt ©Ljic has been restudied= Tlie
patisat nm age 7 years is still dependeatr, ^©galasly
developing seisur@s aithia 72 hours of ©aissioa of 'mt tequlBr
daily d©se ©f 10 Ego of pyriddxiaeo Typieal SEG sbaormaUties
at sisefe tiaes were repeatedly observed aad eoald be abolished
withisa 30^-60 seconds by iatra^eBOus pyridoxiae-SK^I 15 rag.
ConcuTfesst ssibjeeti^e and objective irapro^eaeat ia the
patient "s eossditiea was draaatic and reprodueiblec.
!>i!ring a typieal period of pyridgxioe depletiss esrsb^a'.
setabolisa was measured' by the Krypton teefeniqts-e aad Vm
effect ©f intravenous pyrid®xiae=aCl 15 aoo observed disriBg
the game observation periodn Results are tabulated as follows
Depleted State
CBF Cmi o /lOO g„ /mi n„ ) 63 70
©2 ConstyaptiooC^*) 3o3 4 A
A«V 0„ differesiee C¥ol=%) 5o26
Cerebfal S.Qc ' 0o85
6c. 23
0 %
Sine© this patient is both sentally retarded and swbjeet to seigiss'©s^,
she must be ©<H3pared to analogous groups of ©hildrea ftmimnlf stadi®c?
by Kennedy asd Sekoloffo These groups exhibited halites fei" e@Sie@braS
blood flow and oxygen ©onstjaptioa l^er than those for eOTpa^abl®
norEaai ehildrea,, but in only an ©©easionai instanee was blood flo^ @t
EoQo as low as in this case and ic only one aas 'O^gen eons^mption so
los^o The TOiues obtained after pyridoxin© repletion are siailar to
those reported by Kennedy asid Sskoloff for their groups. On the ©the?:'
hand the decreased oxygen eoasamptioa during the depleted state in this
ease is similar to the situation reported hj Sokoloff for hypoglycfsssitf-
subjects I'iliere an obvioas substrate deficiency exists-., TfeuSj, tfec
interpretation tentatively put upon the data obtained in this ease -
d^rifsg pyricoxias depletion @ deficiency of a sabstrat© for eerebral
©xidatiye setabolisai existed ®hieh isas promptly eorrected by pyridoxi::.
idEiinlstrEtioaa iin©e pyfidoxine defielency appears to affect T=asaiiac;-
butyrie aeid isetabolSss primarily aad giaee that cc^posand appears to bs
a sigaifieast substrate for cerebral oxidative ©etaboiis® [see
58«^JlDB«30Cel Jc this case Eay represent an exaiaple of r=asjinobatyrie
acid deficieaey ?sitM coaseqiient reduetioa ia oxidative tsctabolisea..:.
Ths latter redECtion by about 25% is coapstibl© with ia vitro estiraates
of 15% of total o:^@©a aptaka being due t® Y^aminobstyrie aeid isetabolis;'
if the respective in ?itip, and ^a ^Im levels of saetaboli© aeti^ity
are tak@a lete censidsratfoac
T!i@ mtuTQ ©f tls® defeet in pysidoxia© aatrities ia ©sses of
•^pyridsxine depeadeiie©'" is isaportsHt to aa MMtrstandiag ©f tfee E©de
bj ^hieli pyridoxiae is feaadled in the boiij„ Sasples ©f bloody «^ts©,;,
ete^ fre® this patieat have beea ©btaiaed dsajfiag periods ©f depletiasa
aad repletioH ioT detessaiBatioas of levels ©f pyridoxirae,, pyrid@xal^
pyridoxamia®fl pyridoxsl pSi©sphat® asd 4«pyridoxie aside Th©s® gsiaples
ha¥© beea froseia aad await aoalysis antil the microfijaorirastE'ie pf®-
cedisres developed by CoursiQ have beea ttoroughly pr®tred,
Coraplimeatary studies oa experimental aaimals are uader say-,
Produotion of dietary defioieQcies ia kitteas has been att^ipted
repeatedly by Bt„ Miekelseo djsriag this year, A satisfactory artifieia ^
diet has now beea achieved so tliat suitable aaimals may be available
shortly., Chemically^lnduced deficiencies tssisg thioseraiearba^ide^
have becB produced and preliraiaary studies on cerebral samples^Jj, M-MM
carried ©ut, eompls^enting these in 58=NINDB-30Ce) „ Significantly
low leirels of T=°amiD©btttyric acid and of gltatanic acid have been fo^nd
in such sampleso
Sianifieance of ProlMtj The association of a seisure state with a
specific defect in cerebral oxidative metabolism would provide a long
sought solution to the problem of i»hy no such defect has been
demonstrable in the past and to the possible bases for biochemical
abnorsaalities clearly present in seizsre states Cdistarbances in
electrolyte and amino eicid sietabolisGs) %hieh seemed most readily
explicable in temas of defects in oxidative aetabolisso The inability
t© find such defects in preoesss att^pts raay be dae to CD the
less widespread dysftanction as SoSo in focal seiaares and/or C2) the
mush le^er le^el of oxidative aetsbolisia in, vitro, cosepared to thsit
4n vifo sjsch that a fractional defect coisld be obscMred in the abseaee
®f Ig. .gigg, activity danandSo Thus,, a study of this type eoald provide
a very valuable key to uaderstaading of these problems o
Propos^'d CmTSfjt To eontinne studies outlined above notably CD in
tsrss of pyridoxin© issetaboliss in the bodyj, ((2> ©n experimental
anijaal material jji, vivo and in vitro and C3) if available, with si®ila:c
'"py^idoxine^dependeEt" patientSo
Koo
Ti'Djec i Is: ""i ^e res.-' diet, or -^-^aipj j'lcyi'i's Mid
in V jal " icsje,
C ^ s: 3r. 'j.. l/?5 »e ^dbees, rjn®BL:...
Total: ■ ■0;9
Profess iona'i: " -OiS
Other; - 04
Pi'sjeet Beseriptiofi: .-. ._, -
QMSSMZSS.^. ^o stiidi? the sTietabolsim of Y-asiinobaty^i© aeid
'■■ ia aejiral tissues, to" study fsetors affeatisg sucli astabolisEH
sBd to declase therefrosj tm signif ieonce ©f tfeis eosipoatid
in neurai sstsboliss aad fuactioii.
-Metjiods,: fhe fliioi^isetrie aetsod, for detersjzEatioii ef
Y-asilisObuty^ie aeid is deseribed iis 57-iIKDB^7- CG). Ae':: ■
ensyiiiatia method, osiag ari easj/ae systes im^ EseiJdgj£aas„
is ^el:::- rd"^-^e'% b^pe^ ^n reports by Jakofay, bivl is still
in - : ; /- :.•■:- :;, :'':.'3.;-. ~; ritro. studies ©a Y'^^^-^^^-^^l^F-^-® ' '
ce:,: .;o;::.L :.liG;u 1;^ iiiaiiLoted braia slices and iso-^'-e^''
:::.■:. o:;:.^::ia are carried oLJt by establistied smt: :iee
'■"". Detailed studies of sietaboliv; ;:;;iv.;.„;£ys
-aaiiiofeiityrle acid -<• 1 - C^'^t. _ L-glistasie
aaiu - u ~ \; ■-■ cuiu ff,-l:etogl.a{fc-arie' Esid - 2, S-C~-'ssiiig
sliBss or hosogeaates of btais tissue vdth Ssoiatioii of indi-
vidual .--—•-:---'---,- '— -•-■•• -:eiiaa§e Ei5d GliroMstoQTapliis
teefeai:/ ^^tivity by gas ilQm and liqaid
seiatiiioi,i.;i ojiuiotsro.
iJil,§M,,.,^Ji§£ial* Obtainad from mMM patients adiaitted
Majo?
useQ
•■'-" ■'• - "'\, ■■-; :« aeid as -"■■- ■■-■-■■-:-■'
.''.'. f.ess q£ cc
L.i'.i3 tit-ifa \:;.;itesrt as r"*?" .^ ^w was
ilsrly siien, ester if! c ; pliospS^ate,
Sain techiaiqusc P/0 ratios (moies of phosphate esteg'Sf led
per ntms of ©Kygea eoEsaissd) with Y->a!Bisobi!ty5fie aeid
OT Bm^i'&iQ sesiialdefjyde as sabstrate were idesstical milh
those obtained ^ihea glatasate orpyravat© were. the sabstrsteg»
These f iradiags indieate that Y-aaiaobytysie asid eaa fuKstioa
as a sKbstrate of esrebral OKidatiFS EetEbolissu; Beeaase
of its, positioffl ia a .shant patM^jay s^oeseci the a-ketoglKta^at©
to sueeiaats step of tij© Ktehs ©yele (see diags'ssi), it may
hav0 nn iiaportant role in oxidative jaetabolissi:
Giutaiaie Aeld .4-™.===^ a-Kstoglatarate
- cx)s I* fv 4 - CO3
Suseissyl - Coenzyss A
I
SiscciQic Ssaialdehyds -> Sueeicate
[* - steps ©atalyzed by Vitamin B© as pyridoxal phosphate]
<2) In iscabated sliess of cortesK fron eats sjith seizMS-es
Isidueed by methionine sulfoxisiifcReo SoS-EethylethylglatariisJide
CMegiraide), or thiosessicarbasideo the levels of ysmijaobsityri©
aeld are iavi asid decrease further daring iiieubatioiSo Pr©-
lisiiHary data on hamass epileptogeaie eortea: sliees ar©
similar^ The lei'els of y-aiainobatyris aeld obtained in raoifaal
eoMtrol slices are higJier tisan tMose reported by Roberts
using a ehroaatograpfei© method of estiEatioa. The possible
reasons for the disereparicies are beiag iavestigated using
the speQlfic easyaatie method adapted from Jakoby. One of
the @0K¥elsaats eited above, tliiosemicarbasideg is a Yitaaia
Bg Cpyridosiae) antagonist mM^h has been shown by others
to inae'ti'/sta the ©oeasysE® forai;; pyrido^sal ph6sphst®„- to
prodiiee a cheiaieai defieieney. Pr'elissinajfy e^pferimesjts
indicate ttet in the p^esesce of tfeiosemicsFbasid© the ability
of y-zminob&tyTiQ aeid to support cerebral osidativ© setabolisa
is Inhibitedo
CS) Sinee eontroi ©f seSziires in anisaals due to a vairiety
of eoavalsant agents as well as tlsose in elinieal patients
Mas been obseyved witis oral or pareateral v-assinobatyrie a©id„
the preiriosis elaisis by others that systeaieally-adaiaistered
Y-Esinobutyric ae'id does not penetrate the biood-braia barrier
ai?© being reinvestigated. Blood levels sris® sharply apoa
adsiaistration and fall pvomptlj, indieating rapid distri-
bution and metabolisia eiith little or no spillage into the
urine. In some toaman eases definite rises of cerebrospinal
fluid levels have been observed. Osing C^^ - labelled
Y-sEinobatyrie aeid, preliminary ievidenee for penetration
across tSie blood-brain barrier of siee has been obtaiaedo
asing constant infusions of high doses to eosspensate for the
relatiTOly ssiall eerebral blood flow/'&pati® blood flow ratio
p?eseri'£ in rodeats.
(4) Siajultaneous detersiiaatioas 011 tke_ sauss tissas
sample is ?itgo of the rate of sigtabolisa of ysminobntfTU
acid and ©ssygea aptaks iKdieate that smtsbolisja ofy^asiiao-
batyfie a©ld ean seeeuat for appro^jiisatQly 15' per ©sBt of
the total osjQBn uptake'^ This estiinate ®ospares favoffably
with obssryatio53S repojftsc! in 58 - KIIDB - 29Cp) pm a
pyri«ioxias~d5p@BdSEt patieat where the J^ liio. fs's-stiost of
eereliffal osiygsn eoasumptioR possibly attributable to
etaboliS5i was 25 per eent.
SiqHifi'eariC© of Frojests . .In er®asiQg ©videae© frpa t.Mis
study aa€l r@poi?ts by otises? iEvestigators iadisates that
Y-aiaiKobHtyrle acid Sias an iisportaat r©le ia eerebral seta"
bolisa. The shaat pattesay Cglutaisate - y-aainebatyrate)
sppeag's to be aetive and impQSts&t in eesrebral oxidativ©
metabolism, and is sigeifieastiy iavolved ia certain dysfaaetiojis
of th© braio sissb as selsi!?©s\ The possibility tfeat tills
pathssay for Y~aais3obatyrie aiid rastabolisss, istsielx is tjfiiqa©
t© the braia, aay ejjert a regalatory aetioa oh a ©sfitieal
portios of osidati\''e s^tabolisia and feeae® oe energy prodaetioa
?jarrasts eareful aad detailed iovestigatioa both in tei'ias
of aormal ftsnetioa asd of seissar© states,,
^ugse; To eoatiaae studies owtliised abo^©.
Part B. Ineladed? ¥es
Serial NOo ..B.Lm.mt^OL^— —
[Tidual Projeet Eepost
ZsUndzr Year 1958
Part B„
FabiieatioBgj
MeKhaifiss, G« M. aad Tower„,Bo B.
Gsnjsa-Amiaobatyrie Asid: A Substrate
fo? Oxidative Ifetaboliss of C©r©bral Gorte:!
Asio J» Fhysiolo 1%; in p^esSr;
Honors and At^ardss noa©
Serial No,JSll^Mf|_.,._
2„- CliHieal NeutmhemUrj^
3. Betfeesda, Mdi
4„ Sam© as SV-NIKDB-l? CO
PHS - NIH
Individaal Project Eeport
Caieadap Year 1958
Pgo.1@^t Title; Mieroeteinieal Detersaiiiation of Aiiityl6lioliE2©„
Pffiaeipai lavestigators D?„ Willias Co Curtis.'
Otfesr lB?estigat©rs; Mr„ Lamar Efemnersoni Msf=- J. W<, Phoenix
Cooperstiiig Onitsj Non@.'
Man Years; Patient Dayss 0
Total; 0,2
Ps'ofessioEals 0,1
Other; 0,1
Project Beseription;
Oble®tiv@gs) See 57-NIOTB-17 CC)»
Methods' >
MalQg Fiadincs; All attempts to eliminate gross
iaterferenee eneouEter@d when applying the aietyleSsoline-
boron-flavaaol reaetioa to biological natefials have
proved uQsu@e@s3ful. Siaee sueli iaterf ©rene® resideirs '
this reaction useless for fluorissetrie determissatios?
of aeetyleholine in biologieal saEples this project feas
beea diseoatinued until sueh time as asor® iBformatioR
on the kinetics and sieehaaisfns of the r@a@tioa besoo©
avaiiabl@o
Part B_Iaelad®d° No,
■Serial !Jo. HimB~32^^'^
lo Medical NeMrology
2„ Cliraical Keuroehsaist-ify
So Se£5ies«ia, Md.
4. Sams as 57=^31^'aB«19(C)
PES ^ NIH
iBsdi^idaal f¥ojeet ieport
Caleadasr Year 1958
Part A„
Proj©st_Titles Physico^eheaiical studies @f Hiraaa C©rebi.'9spiaa.\
Fluido
Priacipal lavestigators Dr. lilliasa C„ Curtis
Other Investigators = lr„ L., Keserson^ Mro Jo Wo Bsoeeis
Cooperating llaitss QSo m Ao Zismaa^ Sarface Climistry bM
High E'elymer SeetioHj KBL. ; Dro Joha Ho Seipelo Mto
Alto ¥oAo Hospitals Dro Do Bo CO!Ersia„ Su Joseph's
Hospital CLaacastero Pao)
Mas Years 2 Patient Days i 0
Total; lo5
Prof essioQaJ ; 0,5
Others 10
Project Description;
GMmsMsM-' ^o deteraiioe w2sether specific sabstanees
liberated ©r produced in association with primary or
secondary derayelinating processes in the central aerwias
systffia give rise to altera-iions in cerebrospinal f laid
cuapesition and characteristics which can be demonstrated
by physic0"eh®Eical techsiqaeso
Matheds; Se« under major fiadingSo
P.lj,ieBt Material ; Obtained from NINDB patients and outside
sotarees (see cooperating 'iaits) adsaifted for other pisrposeSo
Pooled sMples coliectsd at randc® used for preliminary sttsdiesc
and specific samples froir. individual patients ?dth verified
aeraro logical disorders us^d for sisbseqiaent worko In additioa
fresh hmsan brain samples obtained at aatopsy^^ as soon as
possible after death, ase<i as sources for various coapoisests
under studyo
Mai or g'indiaosg PrelimLiary investigations ©f saitabl®
p^'oeedures for iseiatioBj analysis and their control have been
necessary o IWo approacfef;s have been adopted? CI) study of
a model proteimn guinea pig serum asparaginases a globisdin
with easily assayable SEir.jEse activity as an indicator cf?
<■ of the effects ©f isolation aad analytical procedures upm
the natural state of the protein; and <2) application of
S!irface<^ha3iical techniqaies for separation and isolation of
micro quantities of lipids and proteins in cerebrospinal fluido
The studies en giiiEsa pig smam ssparagisas® detailed
is previews reports [sse 5T=-^IKDB=i9CC)3 liave besn esispleted
except for a fes additisal coafiraatory studies ©a ©lestro-
phonetic aad alts'aeeiitirifKge data new in progress, TMe
partially purified eBS|igs prepa2'a'&i®a ceataias at least tw©
aasroaoleoalar C?pr0tsis) coBtaaiBaBts ®hicfe liaire defied
attempts at sepa?ati®a by eieetrophoretie ©? ssltracestrif^gal
sseaaso Tks easjiss preteia eaa be qeaatitati^ely adsorbed
OB iaodified oellulsse and ia carbsa di®Kide foaiSo Parifieatl?
by these seaas is esrreatly beiiag atteaptedo
Th@ difficsalties eaeeaatered with this relatively slsple
profoleia iilHStrate the aecessity fer saer® ssBsiti^e aed dis=
criffiisatiag isoiatioB techEiqsaeSo particularly fer eerefer©'^
spinal fluid where proteius aM lipids sre pfeseat is trae©
amoQBts SQd the moBists o£ fluid available a7@ 30 lissitsdo
Hence attentiea hss beea directed toward applieetios &i
surfaee^cheaical tscSaaiqaes to these problessaso
Progress is fractioaatioa of separable siarfsee eeti^®
fluid constituents in uschanged form has bees saade^ tssiMg
selective adsorptioa in foass or at interfaees between
iraaiscible fluids. Appreciable quantities of fljsid protsis
are definitely removed in carbon dioxide foams „ Air f©a®s
proved such less effective. If xaathoproteic aeid deterffiilQat-
ioQS OH trichloracetic acid aad pliosphotangstie a©id filtrates
of the fitfiid provide valid estimates af polypeptide c©!iteat„
tfeea polypeptides are alse r^aoved ia the f 0^0 Sigsif icaat
amounts of lipids EOt bosrnd to protein are reaoved by
adsorption at beas9ii8'---fli5id interfaces sfeile proteiE=b©aHd
lipids are apparently not reso^edo All fluid lipids ar® eo5i=-
centrated at ehleroform^fluid iaterfaceSo
Coasiderable time and effort has beea devoted t© devising
suitable procedures for ideatification asd qiiaEtitatiois ©f
lipid constituents so isoiatedo Qaalitativs ideatificatioas
of total laaboiand lipids^ free aad b©M!sd cfe©l@ster@l„ eerebr©"
sideSo sphiBgosyeiiu aad the various phospholipids have b®©Q
achieved by the chroaatographie methods of Hack asiag
s'jJ'tabl© color reaetiosso Qaaatitativs estissates have beea
possible in soaa© .cases aad are in proesss of developasEt nmi.
SigBlf JeaHcg of groleci-L This prsject represesta a |OBg.t=
range effort to solve the analytical problems presented by
small saiaples available fros patieats i?itfe deayslisatiag
diseaseso Traditional approaches aM setliods failg s® that
the development of n^^ sensitive aad specific procedssres
uould not- ©aly-be-of -grefet-practieal value but ?iouM also
psKsit fresh attacks on promising leads in the pathologieal
chmistry of deaayelinatiag diseases ia sBaQo
Proposed Course; To^^ continue stadies along liE©s ©istliE©d
above.
Part B laclMded; NOo
Serial No„_JIf©i^33ieL____
l"o Rigdi©eJ. Nearology .
2'o Cliaieai NeuroeMS'aistry
4o Ssae as ST-NIM)B=18 CO
, FHS » NIK ■ ,
Indi^idaal Project E@por'c
Caleadar Year 1958
;t Titles Tlje asrfaee-Cheinieal Behavior of OriQe.ia
Relatio.pi to its Surface^Astiv® Ma©romoleealar
ConstitaeatSo
P'riBcipail Investigators Dr; Williaa €» Cartis
Other Investigators; fiSro L; Keisersoni Mifo Jo Wo PhoeaiSo
Cooperating Ofiits; Dr; Wv A'„ ZisffiaJSg Stsrfaee Cfe©aiStEy
asid High PolJEser Sestioa'^ WoBoLo
Man Years s Pati'©rat Dayss 0
Totals loS
Frofessionalj 0;5
Others liO
Pfojest 9®3@riptioa3
Obl^ti^s.; To de^'elop suitable physieo-sheaisai methods for
isolatioBj ideBtifisatioa Sijd ©Iiaraeteriaation and of masso-
EoieeiileSc sueh as polypeptides, pyrogens bM the like, whieM
oeeis? in HFine aad otSjsr biologieal fluids but s?bos© ehgtiBiesl.
isidi^idualityo physiologisal sigaifieasieen ssd iBsshaaisms of
aetio55 resaia obssar©o
Methods 2 S©« prsvioHs i'©poi?t 56-riIl\DB-=95CC)
f-SJ^-tM— iatjgglai.g Obtained fron PSINDB patients adinitted fsr
otlie? purposes o
Mja.lpr FigdiMsg SEsrfa®© tesisioR of aria© in relation to Qtket
parasffiters hss been ■e¥alijat®d<, A stroEg ti®®"dep®sd©B©@ i^as
fottnd with eliaraet©i'isties stsggestiag at least two ph^sisai
protesseSj oae asi initial rapid ©a© and the otkeir a l©Bg©r
and slox-jsr proesss, Fros this observation it is obfioas tliat
in ai'iaa tfeere is no saeh tlJiag as sisrfae© teasiosi gf£ s£ bsat
oisly sarfE'S© teiisioa at a partisalar tise. Effects of tespera-
tare (not signifieasrt)^ of pH Cobs©?^atioa of misiiiaa);, asjd o£
adding ^arioiss pressaa'.ed sarfaee-aeti^e agents smSj as slbiiiasjs
Coo effeet) asd biie salts Cpsfoaoeiased lowering of tesisiosj} wer®
s tad led „ ^ Ife^sTer, it was obvious that sasffae© teasioa seasar©'*
sjents aloa® as"® too gross to repeal .Eor© thas th© otJ^erail
phesioinesjon of adsorption and in addition ar@ diflietdt to
repFodiEee from os© operator to snoth®g b@®aiis® of th© tiis©-
deDead©a@@o
The possibility tliat serfas® teasiora 3®asur®ss®ats ©©
^•©plaeed by optieal asasaresentSo whish iHtrijisieai|y ar® isor®
eiosely related to tfje ultrsstifEctur© of as iist©?fae@e Mss
bsesi eKpioredo PrelisJiKary ©rad© aejjsareasejsts of . this sort
haiir© bees <nads at tSis Watrai Eeseas'eh Laboratories aad imdieats fe'
isethod has iserit„ Expl©ratioia ©f this approseh is
costiiiuiisgo
Meanwhile atteatios has boen turaed to fraeti©raa£i©Q
of the difficultly dialyssble uriaary eeHstitussits,, ^hieh
contribute to sarfaee acti¥© fesfea^iorg by adsorption ®b
solid siarfaces (adsorption column resins aM the iike)^ in
tomis Bud St interfaces I^etweeQ urine aad ^ater^^issisiseibl©
liquids, Coavenietst^ simple analytical procedures ha?@
beea developed to evalisate the effectiveaess of these
surface«chani©al ssethods of fractioaatioDo Such work is
n&m in progress a
Siqnificasice of Proje.ct; Many of the complex polypeptide
lEscromolecules which normally are excreted in the uriee are
cossidered to be by-products of In nm degradation of
proteioso In pathological states^ notably netsrological
and neuromuscular diseases, urinary output of these materials
increases raarkedlyo The increase may b'S due to substances
characteristically associated mth a particialar disease state.
The lack of any successful attempt to characterize these
i §obM®«®®.oSakes this project of importance in this regard..
Proposed Course i To continue investigations along lines
outlined aboveo Progress mast necessarily be expected to
be slow because of the requirements to develop suitable
analytical and control methods^ to explore a large nuiaber
of possible surface^chemical techniquess and to clarify
the theoretical bases for observed data and phenomena,..
•■
Part B lBelud,ed; No„
Serial No. NI»B-34Ce>
1, Medical Neisrology
2^ Gliaicai Nsarochesaistry
3f. Betliesda„ Md.
4^ New Project
PHS - NIH
Iradii^idual Project Report
Calendar Year 1958
PasL-A^.,
Project Title; Cersbral Protein Metabolism and Turnover in
Tisswe Slices iracubated in vitro »
Principal Inirestigator : Drc. JohnL„ Wherrett (Guest Worker)
Other Investigators; Dr„ D^ B„ Tower« Mr„ E. Lo PeterSc,
Cooperating Onitss Dr^ Heinrieh Waelsch, New York State
Psychiatric Institute (Advice and Iso-
topically=-labelled Materials).
An NIH unit for Mass Spectrograph Analysis
(to be arranged later )o
Man Years ; Patient Days % 0
Total; 0,4
Professional; 0^4
Other s 0
Project Description;
Ob[gct_ivgs; To determine whether slices of cerebral tissues
will' incorporate labelled arairjo acids into the protein fractions
of these tissues during incubation in, vitsLO„ and, if so„ to staady
rates of incorporation,, turno¥er and factors affecting them in
samples of normal mammalian cerebral tissues.
Methods.; Standard incubated slice techniques developed in this
laboratory will be used for cat cerebral cortex. Multiple slice
samples of pooled weight lo5-2.0 grams will be incubated ^sith
L-=Gliitamiae'4J-C^'^ and L-=GliiatamiEe'=^mide N^^^ separately and in
combination isith L=Glutamic Acid-0'=C^^ and with L=Aspartic Acid-
V}-<}^^ Slices and incubation media will be separated^ the siiees
homogenised in trichloracetic acid to yield a stspsrnatent fraction
containing the free amino acids and a precipitate or erisde protein
fraction^
The free amino acid fraction Mil be assayed for glistasai©
acid by methods previously applied in this laboratory to obtain
total content of each of these free components, Goisnts of C^^-
activity in each free amino acid pool will be obtained after
chromatographic separation by methods now in wse in the laboratory
to obtain the specific activities in each free amino acid pool^
N^5 specif icVHtivities will be obtained by mass spectrograph
analysis (arrangements to be concltsded later for this phase of
th© study).
Tae CiTude proteia itB.otion t-dll be pm'ified bj tlse saethod
of Sieksvits sad Pottesr as adapted by Wsslsch to resole Bi?-3Ssie
acids and lipidSo The resulting pur® pi'otein f^actiori Kill b@
vneighedg clissolisred i-& tisiogljeollie acid aiid/or.MsOH Cas specified
by Waelscli) and repreeipitated to shesk total C oseti'^fity as
remaining coBStaJst aad heaee truly presejst ia the proteisso Tiie
protein fractioa otII tfees be easyaaticaiiy liydrolysed nsinq a.
preparation from bog paJicress^ as developed by Barry speoifieally
for gip.taEsal;e and aspartate milno acid gjoupso After dialysis
and coBceHtratioH by iyopjiilisatioH^ tiie free asaiso acids derived
frora the proteins will b© separated' chrosaatographiealiy aad
counted as abo^e, ajid aliquots of the free smina hydroiysate
will be assayed f©r glatajsine^ glutamic aeid,, asparagise and
aspartic acid as aboveo: Thus specific activities both in terms
of protein and isi terras of specific amiEo acid eoHtesits of the
proteiH esa be cslcwlated^
If preliminary experiments on slices iraeisbated i<"2 tioars
pro¥S siiccessful^ rates of aaiao acid insorporatiom cam b@
obtained hj using a series of iracubatioa tirassp From these
data plus the activity in the free pools, tursiover rates of
the araiso acids in the proteiss caa be estimatedo'
Mai ox Firadlggs; This project has just begusi,^ since Bta. Wherrett
case to the laboratory ira. Septeaberp, 1958o Basic tecliaiques have
been mastered in reproducible fashiora on slices isitfeOMt added
isotope down to th© purified proteia stag©o It has proved feasiL"
to carry out the assay of free amiHO acids io the slices osi th©
tricSiloracetic acid superaatent after removal of the latter by
ether eiCtractioK so that these data can be obtained ora the same
samples from ishieh th© proteiji fraction is obtainedo. Proteia
yield of 1% of the ^et ?i?eight of the slices ha^© b©@ra obtaised^,
in good agrs^aeat with theoretical yields hj ealcMiatiojSo,
35
Sigajfieaiie© of ^ Proj ect, ; Studies, ^y Eiehter idtli S ^aethioHine^
by\ Waeiseli wit^'^C^'^i^iae and C "eleacisie sad by Sporra aad
Dingiaaa with C "^proline iudicate that i^ ylvo amis© acids are
v©ry rapidly incorporated isto c©r®bral proteias arad that the tssrH"-
over of the latter is coaparable ia rat® to that of liver proteinso
It is practioaliy not feasible to.eosduct such studies in raae djae
to limitations on the level of C ®hich csk be admisiis teredo
ThereforOy a sueeessfHl deaorastratiora of ^ vl/iro, amino scid
iueoi'poration isto csrabral tissue proteirss aud their turnover
ijsisig, asiiaal samples would make it possible to apply this teohaiqae
to'SiraaR-'iJstEJTOsiarigical brsiss sasples aad thus provide soa© data
on mmt both for normal samples asd for those srlth disease proeesses
sjich as seiaureso Siace so data oa the saost prevalent aad
aetabolically active amino acids Cglutasaie aeid^ glataalQ© and
aspartic acid) are available eveira in yis&„ it is felt that the
use of them isjitialiy will carry additional value of itself o.
Proposed Course; To eontiaue the stJsdies as outliraed usader
rfetliods, above o
Part B Ificladgjjs Noo
1« Medisal Neurology, ,
3-„ Bettes<la„ id.
4= Sas© as 57 -nimB-U CO
■.;::/PHs - ^m,,. .
Imdividaal Projeet Report
€al slides Year 1958
Projest Titles Distribution of Aetis. aad Troposyosia Ib
•-. :; Morasl aad Diseased fesele,
Priscipal; Investigators . Bt, Beat Horvatli
Gttses- iBviestigatorss Dk-o Igo? Klatso; Ms-, J. B. Proeto?,
Csoperatiag Units; Dr. Ko Laki,, LPB„ NIAMD.
Maa YeaE-s s Patient Days ; 0
Totals 0,,6
Pi'ofessioBals 0,3
Othett 0.3
Projest Bessriptioas
Obj^tives; To obtain additions! iniotrnQtlosi on tfe® GWleealag'
areliiteetare of lauseie, to stady the distribution of fissetioBaliy
isjportarat proteiss in Kormal aad diseased massleo asid to ■
establish imaiusologieal properties of faastiojially isipoE'taat
masele proteins.
Methods s Aetin is p^epas'ed by tfe® proeedare of Rlos!i3a®i.'ts.
Ti'opemyosiH k is prepared by the snethed of Laki frosa Claa
musele and hj the raethed of KoiniKs from rabbit B^siHc
Trapoayosiis B is prepared by the ssetlsod of Bailey. Myosin Is
prepared by tlie sethod of Saeat - Gyorgyi. Tises® siias©!© pro-
teias sr@ obtaised in a high, state of. .purity. Rabbits ar®
imsiKsised by repeated iajestions of.tSiese proteins with .m4' .&i
skiB tests. to assess tiie respoisse., of .tli® aaissals., ,,S@ra are
.solle^ted frosa immunized aninals aad'sr® y.s©d la pr#©ipiti®
. r@astioi5s. Tfee globijlija frastloa ©f siiefe s@ra is .sonjagatigd
with £lmt-QS®®in, and, the eorajisgated antibodies are,«as@d as
spseifi® staiBs for siasele proteins Caatibody-aatigea eomples)
on seetioas_ prepared -f roE biopsy' s^aterial aed- stadled tisder tM@
fl(2oresse&se siisrossope acsordlEg to tlie teeteiqae ©rigiisaliy
deirised hj Coons.
PatieBt isteria„i s Musele biopsy speeisBeas are obtaiaed froiB
NIfUB patients admitted for other purposes.
Ma.lsr Findings; Sera of rabbits issusEsised agaisst .elaia
fropesyosia A precipitate samiEaliaa Myosira. Asjtisera to liiisissi
and eat %osiE precipitate elas Tropousyosin A, No siicli eross-
reaction »jas fo«Ed betsaeea aatisera to efeiekea Tropomyosis B
OE the ou© liaad ©ad elani Tropomyositi A or ssajaiaaliaa Myosins
.011! tfee otfeer. Astia lasd pre'/ioasly been foisad tob®
isQ-nntigeniQ^.hf Boriratls aod al,so bj Kssztfisk. ^ This
fimfisg Was reeonfii'sed using Aetia iiihieh is 99%,,pt5fe» Im
addition Tsopoajosin A, Trdpomyosia B aad Myosia ^©r® sow
also, foimd to_be iso-aHtigejiie.. . Scjies of the asisisls issffia-
siissd witiS'-mjisele psf&teiss api^eared to isIjow weakness , aad
stiffness of .aaseles asid exhibited sas^l© lesioas 00 siei'o-
seopie ejsasiKatioB.
Using antibodies to lyosia 'eofijugated taitfe^flaoFss^^'iSo
Myosifi in sestieas of siorasl MuniaBi .'ffliissle mns. ele.sglj aad
distiaetiy ■dstaoBstrated -wMer tli© flaor©s©©Hce .si®ros©op©o
PrelifiJlRasy seetioas of ..djs trophic aaseie siBiil^Flj; treated
skQvied layosia isi residaal, islarads of aiasels and-.a ^siiggestioa
'tfeat ia' areas, of a©tive .degeneraties mj?'osis~rea©ti¥e ssaterial
vfas p^esesjt ia aaero phages,
.SlgEif|earige_oiLF.fgJ--t:g^° '^^^ Is'-Huaoiogieal findings are
sonsisteat with pyesesjt concepts of the Myosin sjolesule as
cosisistlKg of subuQits - TropoBJyosia A, Troposayosis B aad
AatiB. Tfe .latter eaa be prepared-. ia a higter state ©f puritj/
than Myosiffl itself 0 so that th©y are sore saitaMa for iKivesti-
gatioaal .purposes. Siaee these proteins ai's is©~aBtigeaic\
issHiunological, respoases of the orgaBis!?! say be. iaportsat in
'so£"3ditioi3s whei'e dest5:uetio3 of laiisele coa'id pes-iBit .these
.proteins ts essspe frosiths issaal ©onfiaes of the aassl© aisd
enter tfee general fo©dy Qircaiatisa.
.,^£S2lM...£2lllliS» T® estesjd aad ©©afizfas t&ess fiedisgs by
qaarstitatiye. iamiHoehesiical s©th©dSo To i aires ti'gate tis© aatsir©
of tl58 EKsele lesioss. observedo And to OFaluate tfee sigai-
fisasise of issiiQe sreaeticns Ih patieats ^^itls nsi^roamsesslsi?
disoasss by skia tests atsd hj qaaetitative presipitia reaetioBs
of their ssra;
Part B I ae laded; Yes'.
Serial No,__Ki^j3»3DCc.)_
PHS ~ MIH
Individtial Pi'ojeet Sepoi?t
Calendar Year 1938
P|sM?,,gatioasi
1. KiatsOs I/j Hoi'vatfe,, B. atid Satnasrt, E. W.
BesjoRstratiott of Erlyosin ia Haaaa Striated [fesel©
by Fiao£^scesit Antibody.
Froc. So So &5£. Bi£l. ,W, 97 J 135, C1958)
2= Lakij Ko^ HorFatho B. and Klats©,, 1=
Or the Helationship betsjeen Myosin asd Tropoayosia A«
Bioehisu Biophys. Agts 28; 656 C1958)
Honors and Awards; None.
Serial m^mmB-^i^}
1. .Msdieel Neurology
2. Cliaical Nsuroeheaistsy
3. BethesdSg Mdo
4o Sams as 57 NiNOB-iSCC)
PHS - KIH
ludividuai ftrojeet S©por£
Caleadar Year 1958
""""^^^oject Titles Alteratioas of Aetcsyosin Tensile Strength nM
Muscle Proteins la Neuromase^iar DiseaseSo
Priacipal lauestigators Dr, Beni Eorvath
Other Investigators 2 Dr. Go Mo Sliy„ 8ir„ J, B„ Proetor
Cooperatisg Usits ; Noae
Han Years % Patient Days ; 0
Total! Oo8
Professioaal i OA
Others Do 4
Project Descriptioas
Ob|eetives; To eosapare aonaal and diseased humaia muscle
biopsy specimeas for astsmyosin content^ tensile strengtis
of actomyosin threads prepared from swch biopsies „ aad the
physico-chemical characteristics of protein therein^
Methoti^s -See 1957 report [Nir®&='15CC)] for msthods previously
developsdo Additional procedures in use are the foliowiago
Muscle samples are hcssogenised in water and aliquots of the
hotnogenate are extracted respectively with CI) trichieraeetie
acid„ (2) water, C3) Edsall's solution C0„6M KCl with
carbonate^'bicarbonat© buffer) ^ sad C4) Co 05 W NaOH^ Samples
of the hoffiogenate diliated with water serve for determination
of total solids (by a microisethod standardization on normal
muscle) and total nitrogeac Nitrogen content of the four
extracts and total nitrogen gre determined by direct
Ness lerisat ion (a speotrephotemetric procedwr© adapted for
this purpose and standardised in this laboratory on normal
muscle)c Extract C2) is also utilised for determination
of K,, Nsa and Clc These procedures permit triplicate analyses
for all sospoaents on 0o3 gram of dystrophic masclec,
Mai or Findings s The determinations detailed under Methods
permit calculation of the folloising compoisents for each
specimens CI) total solids^, C2) total proteing C3) non-
protein solids Cfat)^ C4) aon=^ollageaoas proteias, (5)
©ollagenj C6) Kater^'soluble proteinsg CT) myosin^, (8)
alkali^solubie proteins ^ (9) non»protein nitrogssig CIO)
electrolytes CNa« K^ Cl}„ and (11) tissue ®atero Comparison
of nonaal and dystrophic rauscle analyses reflect the
misting of isaseie and replacement of muscle by connective
tissue and fat.es indicated by increases in CD^ (3)^ C5)
and Na and Ci in dystrophic specimens ahile all other
craoponents are loi?er in dystrophic %pe©imeBs« Values
obtairaad ©n dystropliic specimens for p^oteiB fractions
(6)b C7) aad (8) iadieat© a relative imcrease of myosia
aad decrease ©f alkali«sol!sble proteias in most eas©So
These chaages sem t© be iadepeadent of remaiaing musele
saasSg estiiaated from i4}^ md also of the kind ©f tisss®
replacing muscie^ Water-^solable proteins appear to be
increased relative to other proteins in most dystrophic
samples and as infers© relationship is indicated between
remaining nsMsele mass<j estimated from C4}o on the one hand
and the percentage of water=soliable proteins ia the muscle on
the othero It is concluded that saa^les of dystrophic
muscle not only contain less muscle and more connective tissue
and fat than normal rauscleg but the protein composition of
the remaining muscle is different from normalo
SioBificaace of Proieets; This project is part ef an
integrated effort » involving many disciplines^- directed
toward elucidation of the nature of myopathies such as
muscular dystrophy <> myotonia and myasthenia gravis o One
possibility is some abnormality of muscle proteiOn and this
project is designed to investigate that possibilityo
Proposed Course g To continue these studies on biopsy
specimens obtained from patients with other types of muscle
disease^ and to extend the study of muscular dystrophy
samples by physico-chemical investigations of the protein
fractious here determined by the general approaches previOffiSlj?
adopted [See 57 NINDB^lSCOJo
Part B included; Yes
PHS - nm
Isidividaal Project Eepoyt
Csiesdar Year 1958
Publieatioas?
Horvath, Bo aod Ps-oetorg J. B,
th© CoEpositioa of DystJfopliie '
ProSo Assoe. Ees. Werv.
Studies OH
DiSo in press <I95C
Hoao5?s sad Ai'jardss
none.
MHIM, REPOK Calendar Year 1958
OPHTMlMSLOSf BMNCH
NATICmL INSf rtlJXE OF HEimOLDSICiL
DISEASES MD BLXIIDNSSS
\
The past year broisght several changes in the staff of
investigators in the Ophthalmology Branch with ttee arrival of
nev visiting sisientistgSg, Dfo vsn Alpheng, DTc faisl@y, DCo lale
aaxd DSo fsaakis, and the dspartssre of ©ther sesaarcte worke^t,
AS D7, Dodt md HXn goes-fSSo Or. Dodt^s six mdntfo st^y ended
all to<9 early in W&bvimsy ii»f |thls ^ear sad wss felt ss s serliOHSs
loea,, the severe sfeirsin ®n Ihe ©nly seni^ir f^sllelroe «jpferh?»lRH>log5,9t,
s strain resulting fi?<9m ^he gtrsuth ©f tise ^lisiisal p7«g^^ia!>
wss consideff^bly relieved by the appolatffiait ©f Dsv, vati Alphea
&» Aas&&iMte Ophthsls^l^gist-, His eseperience in G»di@.%l snd
surgieal aphthslmcrls^y sod his interest lis resesrsh pT^^ided
ftfee possibility to assist ym&vig ©pfettelTOilsjgjists in th^, e®K-
pstient eefviee, in th^ ®pe^@t&ng roem ^nd sm the wss-dSs, '^'^
also to disetsss ce?t®ia Isbsisr'stoiry proeedta^es witl^ thetSo Hi.s
&nd ny t^sks in patient e&ze ma ^ell ss the tmk& ®£ the
nursing stsiff, wese gyeatly facilitated by the eoneeattratlon
©f ft^je partsntis @n ©ne fla©?,, ffee advsafeisgea ©f fcfeiK system
will be fuii ly realised when the neeesasty orgsnizatien®!
ehsaoges hsve been eonpleted, the help of Htc O^Rowrke^
Consult seat to the Brsndis in this endesvor^ i@ greatly
appreci^.tedo tka c&asma&^le perforiamige ©f Cliniesl AssocL^teSi,
feheir iij.m;>sual .oadical bsekgsound snd their devotion %o assign""
ments laade it possible to sibsorb s eonsider^ly ina^sssed
pstlent' loadj, to es|>loy net? titse'-xoneuming nsethods of SKSi&im-.
tl@D smd to ineresse slinissl investigative worko timt theee.
men hsve time to esifjry ®!st these tmltiple d«»ties m.d Imh^v&tm.j
•tidies is midR to theiic eseditc
The iisdlvidtssl projeet sepos't® £s^v@ ke©!!% m&x& detsil.ed
thsin ia pirevloas yesrso It is tiot neeessas'y, then^ m eite
specsifie results ®€ the reported iavestig.%ti^is, is^>ortsnt m.a
they say beo b«t convey sn overaLX is;>re$siono l£ is obvi<@^i3
fif@ffl the repots ihae fehe general trend ®f lafeoratery asad
ellniesl inva8tig@ti@a pvsieeeds along $ steeply ^seeading Il«&v
ffeis obsexvjfetloa refes-g b®th to the qaaalifey mad quantity ®f
resQ®reh s&ativltleSo ■=>=' 'fhere e®tt be little doubt in say
sdnd'.thi^t Dr, Fuortes snl the eo'-^o^keirs in his seeti@n &t&
leading the vsy in their eseploring ^ork on pr^blsms of vision^
This seisms ttue in regsi^d to the <p»&llty of the ceatribations
&s well £s their be®rtag ea basis gaaeyal pffoblsFJS of sensosry
physiology 0 It is always amazing to w& that sadi exeellent work
coosld ba aee©Bm'il!.'#i« in so short ^ tisae„ -«■ In a seli&tad field
«2»
Dr^ Dodt^s resefireh in sp@et7«I sensitivity o£ ehe ?ot:i&a
by physiesl methods of esuKoination is outstmdiQg imd tha
offspring of his ehcughts ®7ft reflected in studies on color
vision so ably eonduc&ed by Dr» Copenfeavsr @nd 3ro ©ssdesl -^'^
Os-c Kffitharina T^aBley^, eid^ajug physiologist f^om th& Xnstitaee
of Ophthsltcology in London, is ae w@i?k to ®£ilise ^® .
facilititts fos aaaiainsaion ©* yp^efml 9»n«'i?t's'i»:y «@i fflie^ff
fusion fi:«4|«i«ney «} maiaml species »ith ptsre consi srstinj&e
in order to d®ia»3QstX'st« eleairly ® separstion of pho£opi©
responses froie contsxsinsting seocopic infl«aen£es« —^
The cllnieslly impos'tfint studies on patients with various
types of degeneffstive retlnel disease by electrffiretinogsaphy
is being continued in the Bsm& laboratory snd sdvanead by
te^hnic^I dev@l<^iDen£8 of ^t&^taw&tsy «nd periimtsrie: light
sense studies {Us a G«£nkel)a These physical amd pisyehopl^ysieal
nethods eooibined uith spectral sensitivity studies @lIo» for
a most eocpprehensive vorktsp of diagnostic probleioso In t:he three
other ausjor projects of the Br^ich laboratory investigations on
@ basic level have also been linked to ellnic^l p^'oblesisia
These projects are (l) physiology and pathology of the
intraocular pressure, (2) cataract, smd (3) uveitiSo
<1) Physiology and patholOi|y of the iatgaoc^lar preasMgea
Of great promise, is the elegant technique origiaally devised
by Dra Lele for studies on sensory receptors in the cornea when
applied to investigations of afferent discharges trisvalling
in posterior ciliary nerves in response to small intraocular
pressure changeso Dro Lele and Miss Grimes have already deoon-
strated that such discharges can be obtained from all mlKed nervea
in isolated eye nerve preparations and that positive results ara
n»re frequent than they have been in the in vivo wor k reported
last year. It was also shown in this study that the pressure
induced discharges differ distinctly from those produced by toucbe
The whole theory of nervous regulation of the intraocular
pressure has received significant support from these results <.
— In vie» of this, an anatocoical examination of posterior
ciliary nerves started early this year is of particular
interest and tiaiely» This study vas greatly facilitated
by the nodificetion of Christensen's silver techni<iue by
Miss Grimes for gross demonstration of nerves so that it can
be extended to systematic examination of species differences o
It was pointed out last year that DTo Maeri^s studies on
the elasticity of the eye, the effect of esKtraoeular ttuscle
on the elasticity, and his aqueous htmox ojstflc^ experiments
have been helpful in evaluating the rigidity factor in
tonographic studies » Systematic recordings of the pressures
in venous ch«iaaels near their exit on the surface of
the eye carried out by Dro Kaerl are intriguingo Ihs
&hmb@x itT^h&cwliss ax&&} sad spt&cl&tsl veins mnA she inter »
c®nn«€ti@a8 bet^e@n these v^seislsr b«ds sad @tlker oeuls?
ven@ffls &y9t&m sh&ssiM hmse. is®p&t&usBl&n@ in debs^ee on
aqaeoffis feKior dyasjalcSn Tfcess prslisdr^ry resttlts as veil
£8 D^o MsiSE-i°8 €»bserva£i©ns ®n eh® l@w@S'ing of tfee psreesura
in ehe vosrgex vein ®f eafes fef Disaioss rasy well te-sve elinlcel
io^lieationso ■^ -^ ^^ Cs^ssn's spp:^&ad% £@ syise 3ngiig>gr.%pky
for dcc^nstragion of ixttis^nmlme vessels slso inigkt: ps^ovida
infoffffistian ps?£®.tnii8g to pTobleras ©f iaeys©«SKlsr pyessaife
reg»l£tion. He eammlstes the sm&eriox' miliary veii% mtd i-ajacte
r^io<=apaqtsse maserial in ehie vessels, With Imolnsigrspfeds
fi^echni^es he obtained eles? pi€t«ites @f ^e ixttTsxumLws
venoffits system in the liviisg snimmlo
St is pisstly ®ia the bssis of S'ep©trt8d lsb<§ir%t@xy re»<yilt@
that the eliniesl glaueoEOEg prisblem h&s been es£psszid@do C^^3ss
feztilisstlsn between lsb@¥St®3ry «nd elinieaX stisdies is
esspeeted t® insresse issltien sufficient pli!%fB@@olegi@al dats
sre srrsiisbis .tnd c©nfiTif(ted„ At preearAt tfec ainlqsse app®^anigy
t® admit glau^oms) p^stients £©7 p!?ol«mged periods of titae with
the sv^ilsbility of neeess@sy inetxtsments peimit di^gnostis
steadies in do«sbtfttl esses sad the detes«ain®tion of the relative
vslse &t disignoatie ps-oeedtares (O^o Paton and eo^f^orkers) ^
Cataract o ffce c«t®r««s£ ps'ojeet reeeivad gresE lu^et?;?®
from the eaosellent work of mc Wmko ^d ms9 ^sivin» ti^o
shcxred by eleetron nicroscepie studies the fine snosephology ©f
noretml lena tissue elements snd of changes in sells ssad lihf^tB
In initi.«t stsges of eatssact development. It Is tl^ tltut
tims that the taltrsstwsetsss-e of the lens«-® nsise diffi@5!»lfe
(gissme to hsndle^^ss been investigated with s reliJible
tscfenl^e^ mA it seems that the infosmation ®fet®ined will
ghsnge the eoneept @f ^ytopathology bmied on ligl&t saicrosesc^plc
©bsesvsfcionso Of considerable general tote^est srs ©bsesvstiuus
of & \ti& density element in the epithelial sells and the lens
fibetrs sand the analysis of thes@ fine strar^ttutifsil pss'tieles
by ultracentirif^gatisfn snd (shemi&al cKSthods,:. this v?@rk,
sondmcted by D^-.. Resnik @nd Dr<. W«nko, w^ptmrn^^s %h.& s«:^£3
in ^i(sh moii>hol@gi@@l elements might be chemist ly identified,,
fhe stnsdies in the elestti^i mi^sroseope Isbos'stosy hsve been
extended to otheir areais ©f the eye^ p:atitie«ilsarl^ the elliaiey
pvoieesses^ but this investigation is in ^n initial stage.
F^rthexmore, the ssase investigator's condasted studies on
biopsy si^terial of no3fm@l snd dystrophia 4keletsl Tms&l@8o
Itse shsnges observed in a^tesl^l obtained tv&m p^stients with
myotonie dystrophy have been descffibed ®nd tlmu&. apiaefci'ins ®f.
other nsa-rora^sealsir diseases will be ©labjeeted ft® eiee-tron
miero8@opie esssmin^tiono
Llg&£ ssiearoeci^ie esesBoimttioss hi^ve been smxtsied &it oa
two typ&8 of expesriffleatsl cs£€x<tsts ^hich deososistifsted dir^sa^leslly
the diffetresitse of cstis^se^ fo^mstlon ps^od^seed by dif£e-f@nt
e£££r®s£@sex&ie sgants when sttsdled on s eelMl®? level.
Althougli the v&Lue of sueh obss^vstioas fo? the dif fereatlal
diaignosie of Smts^n cstrnxmst is definite, £s^ vivo «ad histologisal
findings do not give mty cl^es for ssadicsl thesr^y in these
t«ro t^es of Qatm:mst >•> It is possible th&t siaeh «l»es sire
fortheooiing i^en tisstsse and oirgaa culture siathods h^ve reedeired
more reproducible res^slts thsn obtained £t presents The tissue
cttlture tgotk on the lens last ffith diffioslties d%e to the G&s^lesi
nutriti<ssi®l requireosnts of this B»terl£lo b^t efforts «re
being made to proceed in this line (MSo Csrsvsggio) ,
Belated to the e^sts^ast prc»blem sre outstanding studies @a
proteins of the lens hj Dr» Resniko Ultris^entrift^ation,
eleetrophoresis, spsetr«^&©tc5iBtry» equilibriwsa dialysis
and viseosimetry are esiployed to establish the ehM:s&tetiBttc8
of alpha eryst&llino Dr. Resnik^s values of molecular ^aiglis,
diff«S8ion co^efficient snd spparent speclfie vol^Moe %re.
neost sstisfsctorily confimad £b the studies of OrelSiovicho
The future vill sho^ whether the ^hecdciil d^te on slph^
cryst^allin and the inflaaence of environasental faictors on the
proSfiin have s^plicabillties t© stssdies on transpareacy of the
Dr. l^lsissa tfrorked with elaborate laieroeheodeal teehniq^@@
on the enzycsatic systesss of the lens, but the oesin part of this
iGqportant investigation deals vith the eomesl epitheli%imo With
regard ' to the lens problem he eonf inaed thst hexokinase *nd
gltatsthions redKetase was unaffected by the essposure of the eye
to lj,000 S>rsys, despite the presence of s^rphological changes
seen at the tiise intervals esiamined. the t&mlts obtained
on corneal epitheli^isa were particularly noteworthy with
respect to the lactic dehydri^ensse activity in the rabbit i*ieh
exceeded ten tisaas that reported in other tissues or species o
«=>» Xa radioactive tracer axpsrisssnts Bro Ssshlasm demonstrated
the importance of lactate for corneal tsetabolism ssad showed
that the stroma is capable of oxidating laetate better than
glucose. The direct oxidative shunt plays an integral part
in corneal ostabolism as shoem previously by other tachniqiises o
(3) gveitiso Uveitis is one of the Esost frequent ocislar
diseases, of which still very little is fcaowtio the investigstiw
efforts of the Branch in previmss years have been geared t® the
tosoplasasa problem and this situation has not changed essentially
in the last year. This is explained by the apparently high
-5-
ineldence of toxoplasisa infections as a. e«use of w^reitis
and further by opportunity to have the undivided cooperation
from Dro Jaeobs" laboratory, who has eonts'ibuted so greatly
to the knowledge of this disease^ It seeais, then, that
eiinieal and laboratory studies on this subject promise to be
most rewarding. -• Dr^ Kaufman, in cooperation with 'Ow<- 4eco- s"
laboratory, has carried out studies on the virulence of
strains of toxoplasma gondii and shown in a beautiful study
the dependence of invasiveness on virulence and that of
susceptibility of the organism for Daraprim aetion on the
grofwth rate of strains; that is, slow growing organisms
are more resistant to the chemotherapeutic agentrii than rap>.d
growing strains, -- Distribution studies on Dar«^rlm
(Dr,. Kaufman) conducted on humans and laboratory animals
show that the drug does scarcely enter the »queoas humor from
the blood but that it reaches apparently levels :.n the iceeias
cooparable to those in the serumo Dr. Kaufman £.so ha^
demonstrated that an initial high dose of Darj^rim res«alt.<
rapidly in a satisfactory serum level, which- then can be
maintained with the ttsual smaller daily doses. The elln^^al
implications are obvious. The problem of increased toxicity
by a high loading dose requires ft^.rther study^ -• Jit^ O'Rourke
continued his studies on the association of recurrence rate of
«tvei%is and abnormalities in the peripheral «j<tiJ.lzation of
thyroid hormone. It seems, on the basis of obEsrvations oit
about 30 patients, that uveitis eases utilize the cireislatlng
thyroid hormone at a shcwer rate than normals out the eli'lcal
material is not sufficient to draw more defin:Ue eonclusiims
or to proceed to ther^eutie trials » -- Dr. van Alphen's
project on imounologieal relations of ocular cissies, nr/7
conducted with lens capsule and lens protein, might lefi to
isasmologieal studies connected with the uveitis prob^^m, fhe
results obtained on antigen antibody 7esponr>es on an^^cals
innninized with lens capsule and lens protein were negative so
far, Inassuch as cataract could not be produced unc'sr the
conditions of the experiments. It should be strersed that
Dr, van Alphen's experience with immunological work fills a
need of the Branch and has many potential ramifl<;;ations.
A small nutober of investigations cannot oe grouped into
the four main areas of research efforts. (1) Dro Kaufman
collected a nunber of patients with the main ocular sign of
cottony vitreous opacities interfering with /ision but without
signs of hemorrhage or uveitis « Vitreous aspiration provdd
that the opacities were due to amyloid depr/aits. These
observations carry considerable weight as &hey suggest that
the disease, diffio^lt to diagnose when utiaxpected, can be
recognized on the basis of a sickle biomiaroscopic examination
which might point to the diagnosis of other members of the
family afflicted with primary familial aiciyloidosis.
.6»
In another clinieal study DTe'CRouxke deroonstrstcd the
ffelstlve reli<ablli£y ®f eysjiselersl siMSittng ®e selected eseposed
sites of the globe of P^^ emissions for the diagnosis of w&Li^"
tisat walmoismk of £h<£ shox&ldo The %}susl tsxasssmjomsciv^l
@otsnts of the llsabois xmy give negativa resislts,, <°'» To ehis
giro^p of Investigations also belongs Dr, vim Alphan^s study on
interrelations of optls eleiosstts in the h^issn eye ss m basis
for a theory of refraetion Knomalles aod ssa experimental
st«)idy to detemdne the role the tensicm of the choroid tmy
plsy In the growth ©f the globe c Msa«iffleea»ents In the peri-
ehoroidsl spsae indisated that the pressissre there is severial
nillliBftters less th^i %he intr^fiogoslsr pressssrec Su^h d%ta
mi^t slso be of interest fo2° ^siderstandixiig the de^;%lopB)ent
of choroid det^hment.. '-<- Finally, in the tlS8«ie ^mltusre
laboratory $ Dr.. Itolf mid Drt, Aronson eoqpersted in a etiutdy
on staining living pigmetnt isells of the eye with j^rldine
orange o The results are ©f great interests sinee they esra
provide a way to distinguish between living snd dead sells
in tissue eultiarco Other aspects of this ps^bletn are being
diseussed by Dro Wolf in his project report «
The attached list of ptabli(i£a£ions em&ld be supplenffint@d
by seven p^apers whl<sh are e3q;>eeted to be cleared and accepted
by pr<^fes8ioQal Jousnals within this year^ these p^ers de&k
partly with subjects presented at the E/iiStem Section meeting of
the Association for Researish in Ophthalntology November 2l«°229
i938n The list does not include t^ertain clinical studies
which are forthcemlng as for instance " Retinopathy in Hypo»
albuBBinemia • (Br. Aronssn); "Skin and Choroid Melanoa®"
CDTo Paton)^ ^d "Angoid Streak:^ and Sickle Cell Disease''
(Drr, PatonK
Participation of members of the staff in scientific
laeetings was gratifying c In fact contributions by the
Ophthalmology Branch as a single unit rsiked first in the
overall activity of three meetings of the Association for
Research in Ophthalmology this year„ There arc several
reasons for the apparent ^pst^lng in the research activity,.
The most decisive factor seems to me is the continuity of
a program in laboratories headed by eminent section or
laboratory chiefs who have a permanent status^ Seecnd,
the stiraalffltion from vlsitli^ scientists (Drsp Bomschein
and Dodt) who introdaeed areas of research ^ provided fertile
soil for further worko A third eatsse of a favorable score—
which is f^lly reallsed-^ls the capability, diligence and
unselfishness of medical officers as well as the sound
knowledge 9 mi'-'li&'yslty ©f a highly qualified technical staff
=.7 =
@nd lasfEs ast i«s«ts the senSiaiaeaa «^yppkoif<£ in ®ll adminls£rs6iv®
and seere^sri^ laslcs by cesipafeentg efficient sad deva^ed ©ffiee
One deeasrenfe ^9 the efforts of Sha Br®neh rasasarefe ^tt&vts
is well kniKm; thm is ^li^a short ta^n ssslgiUBent of Clin^s^l
AssQcsisteso An@£hev rastrslning tsi&t&t is seen in the
ini8£@:r£t!sne thmt it wsia iii^tiassible. Sow vestBs^s beyond She
Ssii8el£«ii&e''3 m>ntvyils to reeimife s section ehlef i&x the siss^e
eultnare laborsitosyo It Is l^ped th^t Ss^rtlier sttes^ts vill
meat vith si^eeeesc Ast £be @nd of this report I T^^aald like k<&
express my deep grs^itnsde f^ar the oppoirtMni^y £® be S8«i®simted
wi^ 8U€h la selected gse^p @f qu^ified^ d@di@s€ed %nd h@aes$
trorkars ^nd b@ing sdvised smd helped in sa^^tss'S p$ri£ inane to
£he pr«»sp@rlty of th@ Btsskuh imd she welfare &S it» wesbexa
by the Directsrs @f £he las£i€«tte„
Lcsdorig vea Ssllmsmn^ M, Do
Chief, ©|>hth^l!aol©gy B^^eh
Kat.ionsil Instietsfte ©f Nsaiiffologiesl
Diseases and Blindness:
PUBLICATIOTS
(^thaliBolo^ Bratjch
1958
1» AroosoTij Samuel Bo, II, and Shaw, Richards Corneal crystals
in BJultiple iryeloma, AoM«>Ao Archo Ophtho (in press )o
2o Dodtg So 5 Copenhaver, Rol'.os and Gunkel, RoDo» Photopischer
Dominator und Farbkoirponenten iir. Xenschlichen Slektroretino»
erairar., Pflugers Archiv«, 2678^7~507« 1958 =>
3o Dodtg Eoj Copenhaver, Ro^'o» and Gunkel, RoDe: Electroret=
inograpiiic measurements of the specti»al sensitivity in al-
binos, negroes and whites, AoKoAo Archo Ophtho (in pr©s3)o
ko Fuortes, KoGoFo: Electric activity of cells in the eye of
limulus, Amo Jo Ophth.^ 46s2l0-223 (Pto H) 19580
5" Fuortes, KoGoF, : Generations conduction and transmission
of nerve impulses, Archo italo Biolo, 2§*285-'293, 1958=
6, Puortes, !4oGoFo: Generation of nerve impulses in receptor
organso A summary of the anrmal Bi^op Lectureo EEC
Jo\umal (in press) o
7o Fuortes, IT^GoFoS Initiation of impulse in visual cells of
Limulxis, Jo of Hrysiola (in pres8)o
80 Goodman, George, and Gunkel, Ralph Do i Familial and adapt©-
metric electroretinographic studies in retinitis pigimentosao
-Amo Jo Ophtho, ff6:l42-178 (Pto II) I958.
9o Gourasj Peter: Slectric activity of toad retina, Awo Jo
Ophtho, ii^i59-72 (Pto n) 19580
lOo Gouras, Peter* Spreading depression of activity in am^iibian
retina, Awo J^ Physiolo, 195:28°32o 1958,
11a Kaufman, Herbert E,,: Primary familial aByloidosis, AcMoAo
Archo Pp^tho (in press )e
12<> Kaufman, HoSo| Remington, JoSo| and Jacobs, Leon: Toxo-
plasmosis s Ihe nattire of virulence, Am^ Jo C^tho, ji6i
255-261 (Pto II) 19580
13o Kaxifman, H<,Eo; Jtelton, K«Mo5 Remington, JoSoj and Jacobs,
^oi Strain differences of twcoplasma gondii, Jo of Para-*
sitology (in press )o
l4o Remingtons JoS.j Jacobs, Lt.; Meltons ^^I'oS and Kaufman, HoEo*
Research Note: Chronic toxoplasma infection in a human
uteruso J» of Parasitology (in press) o
2 -
15o Herringtons JoSoj Jacobs, Lo? and Kaufrnanj Ho So: Studies
on chronic toxoplasEosis s The relation of Infective dose
to residual infection and to the possibility of congenital
transmission^ Awo J« pphthoj ff6s26l-268 (Pto II) 1958 o
160 Ktihlman, RoEo, and Resnik, RoAet Quantitative histochemical
changes in the developroent of the rat lens and comsao Aaio
Ja Ophth,, ii6s^7-55 (Pt» II) 19580
17c I'-acri, F. Jo? WankOj, Tej and Grimesg PoAoS The elastic
properties of the himan efye, A«K„Ao Arch* pphth (in press )«
18» l^crlf FoJoS Outflow patterns of the cat eyog Aaio Jo Oohtho
(in pres3)e
19o Ilacria PoJej Vfenkoj, To; and Grimes, PoS The effect of
extraocular muscle contraction on the elasticity of the
eyeo AohoAo Archo O^itho (in press).,
20 e Ilacrig FoJoS Some aspects of aqueous dynamics. Glaucoma,
Transo Ihird Confo Jano 8p 99 and 10, Josiah Kacqr Jr*
Foundation,, Sdo Frank Jo Nejfell (in press )«
2I0 Dekaban, Aoj C'Rourke^ Jo 5 and Comman, TaS Abnormalities
in offspring related to maternal rubella during pregnancy.
Neurology 8:387-392, 1958.
22a von SallF^anng Loj Fuortes, KeGsFo} Macrig FoJo; and Grimes,
PoS Study of afferent electric irspulses induced by intra-
ocular pressure changes. Amo J. Ophtho, 4^:211-220 (Pto II)
19580
23o von Salltnann, Lot The role of the central nervous system
in the regulation of "Uie intraocular pressureo Transo
Glaucoma ^ytnpositan, ^Aegs, Belgiian, Septo 3=5» 1958 (tn pres3)8
2^0 von Sallinanns L,,: Studies on morphology, physiology ami
pathology of the lens epitheliurao Trans* XVHT fctemation^^.
Congress of Oph-ttialmolo^s Bnissels, BelgiUBg ^g^embar ,
7-12, 1958 (in press).
25o von Sallmann, Lo : Early lenticular lesions resulting trrm
ionizing radiation. Trans. Ahio Aead„ Ot^th. and Otol. (in press).
26. von Sallnann, L^: Aspects of ^rvous influences on the
intraocular pressure. Glaucoma^ Trans. Third Confo g January
8-10, Josiah Maqr Jr. Foundations Ed. Frank J. Newell (in press )o
27o Wahko, Theodor, and Gavin, KeA.: The fins structure of the
lens eplthelliano An electron uilcroscopic -stu^ye A„KoAo
Archc Opbth.s, 60j868»8799 1958 =
. 1958 Fresansstlons
Syc;>@8itsa &1X Eleeer<9pb.y8i3l@s^ &t the Visual Systmxa^ Ba&fefisds^
Msrylmd, Mtsmty 16^ 17 » 1958 g
©aMrssg Peta?o ElesSriesl AciElvl^y @f fe®d Re£ln®o
Sieedaan, Gs^sg®, Fasilisl Adspt^mee^ls »iui El«e£S!»X'etlzti3g%'^^l@
Sgodles la Reelnitic Pigsiant^eSo
Hedtg Ehethmrd, F^ysissl fmt@v» In the Cds-iralssi^ci o£ EBd:
Sensitivity Curves i^ieb Visual Fi^senC,
Easteim Saetion Meeting, Ass&ei&tt&a S®« Reses?i&h in Opfeel8€alQ»logy,
BethesdSj, m.tyl@skd^ Smm^xy 17 » 18, 1958s
Maeri, Fyank J, ffea Distensibllity @f £he SsBan Eye,
Kuhlmsn^ Boberfi So Qwmeltstive Histo^asdgsl @^0ges in tM
D&vel'spvo&nt ®i £hft Rat Len« i^nd Coshes o
W@sito» TfeeodsiS'o El@@tir®n Mi€7es€®pe S£»d^ <»n tfee Lena Epieheliim.-.
fliird Conferenee ®n (SI^&^iSsbs 9f the Josisls l>!»^y» J7o fsmnd^ttisa^
Prineeteoe New Jersey g JaaKSsy 89 9, 10 2, 1958s
Ms^i^is frsak J. S@me Aspeets ®f Aq^eaxi&s Dj&.smi@8o
von Sallinsan, Lo Dissttasisn @f Afferent Disefe.^?ges in Posteyieif
Clllsisy Kerves in Response to Ey® Pircssure C&angei!
XVIII Sneesuseimial G<9ng¥«ss ^f OphehmlmDlogy;, Bs^sselSs Belgiosat,
Septo 8=12, I958g
van Sailfflsnn, Lo Stasdlas ©a Mosphologyj ^yai&l&gy «nd Pathology
of «he Lens Spithelimo,
Oeodn^n, George, Elestroratlndgrsphy in Slight ^Blinding Dises«S8<,
@lai@3iois SyiBposimaa LiegSg Belgima, Sep£„ 3»59 1958;
v@n Sallnsmis h, tk6 Role &f the. SenSrsl He'Tvt^s Systen in the
Reg®l®£i®n ©f 6he latraoeislar Pu-esswreo
Catsrsee Syoposimats Sts'sabousg, Frsnee, Septo I5^16g 1958s
v@n Ssllnsnn, Lo Infoanasl disetssslon 9n Len» (^emiastry^
^1"
Ws»lsing«®n S®el<sty fox El®eS^®a me'somopyg fourth Meefeing» ISmdti 146
1958s 8 W®sMngfe®ffis Do C:
W<^k®g to The fine Se«s€t!aiife of Laa» Epitfeali®®.
0et®be!? 12I»16, 1958:
v@Q Sallia^m, L^thrigo Eas'ly L&nti&&ls,T Lesions Resulting item
loBlziog Radlattoao
Eas^esia Assselatlon for £lee£r9en@^ksL@gs<spka?Sg, Ste. Adeleg, Canada^
febo 27 " iSsreh I, 1958-
Goay^Ss Peter <, The Slnilaifity Between Loais's Pfeeaoaanon
and Spreading Retinal Dep^^esslonc
24Eh Proeeedings ©f the Gevmm. Physiolegleal S®8letyp Jfenieh, ^enEStiy„
May 27-31, I958g
Dodtj Ebesfessdo Ffeofeopissfeer Dominator wad FaiPbk®Bp©nenfeQn
Asmmal Bishop Leefeuse, Sfec Louis, Washington Wniveieaity^ Ap^il 11 » 1958s
F«®rte8g M, So F» Seneyation of Herve Sn^sslaes in ReeepEoy Osgsass,,
Naei@n®l Meetings Association for Rasesueh in Ophefeffilnelogys San
Franeiseos California^ jane 23»27s, 1958;
FTOrseSj Mo So F. Eleeferigal Aetivit^y ©f tise Cells of the Eye
©f tb,e LiHB<aSc
Maeris Pranlc J. ffec El®8fcie Psopesfiiea ©f £toe JfeiBM Ey«o
Eastern Sesfeion Meetings A880<gia£i©n fos" Reseafftsfe in Oifefchalmologye
New loifks New to^k, HoVc 21 -aa^ 1958s
Kasifc^n, Hr. Bo PhSTOScology of J)mss.ftiaio
Rssnikj, Robert Bo Smsll, Cytopl^mis Elements in Lens Fibers.
Integ?e®ted Bioisheiaiig^l aad Electron Miegeseopi© ObseirvationB o
Resniks Robert Eo Lens Pg>©teins !%„ tfee Effeet of pH on Alpha
Csystsllino
Wank©a ni5e©dorc Elee£z©n Mieraaeepe S£«dy on Htarmsl Lens Fiberso
Coperihaver, Richard Mo Spestral Sensitivity CDiatribwtion) ©f
Color "Defective Individiaals Determined by EleetroyeSinogr«ipfey<
Goodaten^ deorga. Elaetrore£ln@gMpfey in Higfet-BIinding Disorders™
*Only tlfee first anatfeors are Banti@ned„
Clieleal
§«rial ^ua8>«ff® @f Fsej@e«:s;s
HU©B»37Ce), KIHBB-38(c>, Siai®-39<c), SI®B»4«>(c>,
KIMDB"4iCc), HIli!»-«2(e), H1[Sl»-43<c> , NIKS®-44Cc),
BIiaffl=45Cc), ilIlBB-»46(c), SISBB-47(€). BIHI»-48(c)8
8ISfflB-.49Ce), HIKKI^SOCc), HHflHI- 51(c), SIlim=>52(c),
SIHB»-S3<e)9 HIBI®- 54(e). HOT®"55<e), B5IMI®»56<c)«
SiIBBB-57Ce), HIHI®- 58(c). HIHBB-59{e), sad
SI!©»°60Cc) =
SfttiBafced ObllgattoM for FY IfM
Mr«cts $335, :^K)
:s 154^,700
Serial Noo NIHDB^37 (c)
lo Op8ithalm©l©g3r Braneh
2o Cytology atjd Hist©-
path®l®gy Seetiai
3o Ee-ttiesdaj, KarylaM
ko Same as KINrB=.^ (c)
PHS - NIH
IMividual Fi=ojeet Report
Calendar Year 19^
Payt Ao
Pi=®ject Titles Studies ©a Diet and Drag Ihdueed Ejcperimental
Cataract
Principal Ih^estigators Ludwig ¥C65i SalltnanMs Mo Do
Other Im'^mstigatos'®? Pats-ieia Gpimesj, BnA.
Leo CaravaggiOg MoS^
Eleanoff" Ko Collins
Cooperating Units! I^o Kary Elizabeth Reid^ Laboratory ©f
Nutrition and Endeerinolegyg NIAl^D
Kan Years (calendar year 19^ )s Patient DgQrss 6?
Totals olO
Pr^sfessional: 0O5 Outpatient Visits? Hk
Others ,05
Pk'oject Descriptions
Ob.ieetiveais To extend the knowledge of eataractogeHs^ia
first hy studying t>pes of experimental cataract either thaa theses
examined in the paste SeeorMs h^r vtging on the material new
methods of examination as sterecadcrosespy at law powers electren
mier©3e©pyE, mi^r«8he;.±c.al proeedus^s and other techniques o
Specifically to folios? the clinical and cytol®gical changes
produced by a toxic agent (raimosine) and these resulting from an
amino acid deficiency dieto
It is expected that the data collected from these two
forms ©f experimental cataract will be useful in the under=
standing and prognosis of the human complicated and ■vitamin
deficiency cataracts
u= 2 -
Methods Einpleyeds
MimoslKe Cataract i lite experifflentai ireported last year were
supplemented bgrTl) administrati©mi ®f the toxie ©ompsund locally
instead of hy feeding | (2) @t«dl3ring th© effects ©f high p^ic=
doxine and aiacisi levels ©f tlie toxicity of the mimosiae diet|
(3) subjecting the experitneatal lens to eleetroia micr@gcepie
examination CEfe'-. Wawiko)? and {H-} applying Ujmy*3 his tcchemical
technique to small portions @f the lens (Dto Kiihlmr^n}., The yov.rn
rat was the experimental aaimalo
Ta^tophan Dafieiengy Gataraet; &5inea pigs were used wl;:ieb
were fed ®ia a synthetic diet devel@ped by DTo Ko Et, Reidc. Tlie
animals i^ceived either a diet eontainisng as a basal cosjistityteat
Ool/2 tryptophan or diets with additional small' quantities of
ta'yptophan rangiag from Go 2 t® Qol'^n The eyes were stadied
biomicroscopically and used for histological examination for
sixg eight amd fourteen weeks after the five-day=®ld animals
W8E^ put @n the diet.-,
Ma,i©r FindiaiB;s?
Mim@gine Cataracts 1318 unique histologic pict^ire of this cat=
aract with the seleetiip® damage ©f the cells of the germinative
aone in the earlier gtagss and the proliferation of these ©ells
in a circumscribed area have been described last yearo The
ceOTbinatioR ©f lens damage with conjunctival » corneal and sr;,-
teri©r uveal changes suggests the local use of the ccsnpoiaMd
in the form ©f frequent instillation ©f -f-!^ solutiai int® the
conjunctival saco Thig treatment completely fails to produce
any of the aurxace changes or signs ©f lens dainage as they are
seen in mimcsine-fed animals o
The chemical . structure ©f mim@gine shows a certain siro=
iliarity with that @f the vitamin pyridexinej, and to a lesser
extent,; t© that ©f niacino To examine the psssibility that in
this case cataract foMnatien is connected with an antivitamin
effect ©f ti^e toxic eompeund^ high levels of vitamins are ad°
ministered in an effort to protect the experimental animals
against the affect of mimosinea Such treatment d©es in no waj
alter the ociilar and syst-amic mim©sine effects -.
Electron microscopic examinations (DTo Wanko) shos? con=
apieuous development of the r@u^ surfaced endoplasraic reticwlum
in the equatorial cells.; distention ®f ^-sti^^ space tetween the
membranes ®f the reticulum^ dispersien ®f aggregates of rib@=
nuelecprotein grpniales aM accairailation of abinsrffialg fine
granular material in the cell nueleuso The departure fr&m the
normals theas implieatesboth nucleus a«i eytoplasmo
» 3 =
The investigation ©f cei^ain ©n^srtnes (dehydrogenases)
have not led t© esnelusive results and reqtiir© further stiadieso
Trrpt&yhan Defieieney Cataraeti Centrarsr to &th@r studied types
of SKperimental cataract all' tissue components @f the ©3\iat®rial
zone ranained mnaffeetai l^ the eataraetoias p^oeesso The architecture
of this area is well presein?sd even *ere lesis cortex aiod l&nm
nucleus are destr©y©do T© a great esctejit the first changes are
seen in the peri-nuclear gone aronmd the aaterior pole of the
lens aBeleuso Progressive deeoniposition of fibers spreads later
t© the snarfaee ®f the lens al@ag the sutures o Superficial lena
fibers succumb t© the destruction weeks after the changes in the
deep cortical l^ers have appsaredo Whereas the epithelium does
not undergo degeneratiw changes initiallja in advanced stages
it proliferates at circumscribed sites to foKio'-Giultilayered
plaques or knobs o These changes reaembl® thm eaitiielial js»o=
liferation observed in galactose and alloxan cataracts
Further observations were made in a series of guinea
pigs whidi were fed in I>o Reid's lab®rato5?y diets supplemented
only with the D^isomer ®f tsyptophano Anisalg wsra compared
with others 9 fed equivalent levels of L=tryptoiAan and ©f a
D L mistturso Although the sappletEent levels covered a w3.de
rangej, the clinical examination did not reveal elear=@ut dif-
ferences in the utilizatien of the D=is©mer Is^ itself or «hen
fed simLtaneously with ISi® L ferwo The histological exami-
nation carried out on all lenses confirmed th® clinical resultso
Significance to IVogram ®f Institute? Information on
eataraetogenesis based on histological changes aM the sequence
of the development of these changes of varioasly induced leas
lesions is limitedo Many types of escperimeatal cataract (other
than radiation cataract) have not been subjected to a coinpf®^.
hensive st\jdy utilissing net-jer methods of examination^ Cataraet
therapy as a medical aM a surgical problem cannot be contemplated
without the knowledge ©f the cytopathol©^ of cataracts The
two exaraplesg mimosin© and tryptophan deficiency cataract^ show
clearly to -i^ieh extent cataract format? on might vary from one
to anoliier type irxiicating that the ps^^osis (recovery or
progression) depends on the involvement ®f the geirminative
epithelium,, Clinical observations of experimental cataract have
an obvious application for the differential diagnosis of lens
opacities in the humane
Proposed Course of P^^^eets It is planned t® continue
the studies of ultrastructure in incipient stages of experimental
cataract and make concentrated efforts t® investigate pertinent
^ k ^
aspects of the ^rtologie problem fc^ tissue and orgasa eult'ore
teehnS^ueso Ifiaman lens roatezdal will be studied histologically
and egrtologieaLly whea it bseoiaes available in qtaantityc
Part B incited Yes !W No /^
Serial Mo, MmDB^37 M
PHS - NIK
IMividual EVoJect Report
Calendar Tear 19^'
Part Be Honors 9 -%ards aM Publieations
Puiblieatlons other than abstracts fe-ess tbis projects
¥©a Sallraarm^ I^dsrtgs Studies on mor^ologys pbysiol©^ and
path@l®gr of the lens epithelitame Transo XVIII Ih-fc@raato
Congo Ophth»g Bnasselsg Belgiums 19^ (in press)
Honors and ^ards relating to this projeets None
Serial feo^MMB^-^ Co)_
lo Ophthaliffiologr BraKdi
2e Cyc@l©sr and Hist©=
pathology Sectioa
3o Bethesdaj, Kaiyland
^o Ssffi© as NINEe-?i!f (e)
PHS =. KIH
Individual Pmjeet Report
Calendar Year 19^
Eart^o
Projeet Titles Study ®f Suteaieroseepic Struetiures ©f Lens
Tissue Cosiperaents bff Kiase Contrast Mies'oseopy
Principal la'^restigatsrss Lud^g vcii Sallmaan? Mo Do
Le© Caravaggios, Mo So
Ssmuel Armssosag M«De
Other Investigators s . Jlcoe
Coopsrattog IMtss Kene
Man Years (calsadar jear 1958) « Patient D^^ss Mem©
Totals olO
Fi<t>f ©ssional s 0O5
Others .05
3'roject L'tescriptions
^^©et^es?
1) To establish the gjfswth pattesti of rsanal leas epi-
theliiJHi of varimia sp©@ies in tissue and organ eultwe after
a technique has he&n de-^^leped Aieh is bstter adapted t® the
rajuiremaits of this tissa© than that preiriously us©do
2) To study in contin^Kg ex!ltsa2>8s the effect of mediwm
coEipositioHs eataraetogenie substassces astsd ssutri.timal defieieneieso
3) T® iavestigate the nature and pg^e^entiea ©f injwious
effects ®f this tissue »
= 2 »
^) T© expand the investigations t© ®ther tissues @f
the eyeo
Ifethods . Einplqyisd s A Paul cshaater adapted t® phase cGnr-
trast mier©sc©py atxi rat tail eollagens an optieally wqt® homo-
geneous substrate than plasma elot^ are used in sjorphslegiesil
studies on 0utgr<s^ftli fr^m lens epilJielium explants of einbx^onic
ehiek^ newborn ehiek aM rabbit sxid adult rabbit* Ih® major
poartion of this work has been directed tosfards the study of
variation in cell form ©eeu3?ring in vitro <> The medi^M used
has been a combination of balanced salt solution (Hanks 855^) g
horse serum (10^) and ehiek enibiyo ©sctraet (5^)0 A stuc^ of
the effect of various media combinations on the growth of lens
epithelium in vitro has recently been initiatedo It is hoped
that a medium will be formulate! in ^ieh lens qsithelium will
grow in a manner more predictable aM analogous to that in vivoo
Preliminary attempts have been made to cultivate lens epithelium
without the benefit of a collagen substrates, i,©. directly ©n
glass surfaces e The success of this technique limld pejrasit th©
growing of a continuous culture and the use of replicate culture
techniques for quantitative determination of the eff eets of
cataractogens and nutritional deficiencies =.
Ma.1or Findings s Thus farg the cultivation of ratg ehiek
and rabbit lens epithelium in th© Paul perfusion chamber has
proven successful in regard to the initiation and maintenance
of outgrowtho In adult rabbit outgrarth from the ®splant usually
begins in about seven dg^So Cultures in the Paul chamber have
been successfully maintained for periods up to one months, while
cultures on cover-slips in roller tubes have been maintained
for up to two msntts in a chemi©ally defined medium (Eagle's)
containing as little as 3l° horse
]jidivid\ial cells in cultures \ander F*iase contrast micro-
scope ^camination have showi a great amount of morphological
variations ranging from totally round forms to extremely elongate
and multi°pr©cessed cells c Although the study on the growth
effect of various media is in a preliminary stage^, there is seme
evidence to indicate that the elongate forsns are mor© constantly
produced in cultures containing chick einbjr^o e^rbraet,, while
cells grmm without this media component grow in sheets more
analogous to the in vivo eonditiono
Significance to I^ogram of Sistitutes It has been es-
tablistoed that certain types of cataractous lenses are due to
the formation of abnormal lens fibers ty physically damaged ©r
metabolieally altered cells from the genninativ© zone of the
lens epitheliUHio This inf orsmtion has been obtained from -Oie
stu<^ of fixed and stained tissue from in vivo ejsperimentatioRc
. 3 -
Vfiiile this type ©f iramstigatiea has si^ifieaatly inereased
our understanding of the eataraetogenesisg it has ©ertain ln=
heremt liznitatiorsso
The use of jfease snier©se®py aM tissm® eultm=® teehniquas
enable the direct obs@£^ati«i ©f the ef f eets ©f cataraetegesiie
agents &a ih® liiring eellso itcrtifacts ©f the histologieal method
ast@ eliminated and the cell responses that escape histologieal
detection are recoa?ded ^ tim©=lapse einematograpf jgr for further
studtjTo
Pro.ieetg Ctoee a basically sound
gzwrth pattern has bem determinedj, it will be possible to
subjeet cultures to i»pfusion with cataraetogeaie agents (sogo
dinitrophenolp mimosinej, alloasan^ galaetose) and s= irradiation
for st^^ ^ tiai©=lapse cinematographsr and histoehemieal raethodso
Ih a eoeatisming study ©f media ©ff eets ®n growths an
attempt tsill be made to adapt lens epitheli\an to ga'ow in a
chemically defined mediwiio Recess in this technique w>uld
enable nutritional studies t© be^rried out for l®as epithelium
as have been done for -HfeLa and L= strain fibroblasts o Th©
study of tryptophan deficient cat^^act wotjld also fall In this
categoryo
Pasi; B included Yes JZJ ^® ^F
Serial No« MINIB=^39 (©)
1» pphthalnKjlesr Brmoh
2e Phgrsi©l@gy Sectieja
3» Bethe^ag MssylaM
*o Sasaie as UINrS=57 (e)
PHS « NIH
IndividmL PyoJe@t Report
Calmdar Year I958
£sl»A°
Project Title X Studkss m Central Ken^ois System Conts-el of
Ihtrsoeular JVossureo (Anatoaiy of Posterior
Cilisry Nerves)
Pi'ineipal Investigators J lasdvdg von SaLlaianng M,Do
Patricia CMmes., BoA«
Other Investigators s Nefie
Cooperatijag Uhits? Y>me
Man Year's (ealeMar year '-958) s Patient Dayss Nosae
Total* olO
Pr®f essionaO. i « ©5
Others 0O5
Project Description:
Ob.1eetives8 The iss ^vo studies reported last year
dealing with the sfianeh for lOP (intraoeular pressjare) receptors
and for afferent pathways signalling changes in lOP show that
in positive experiments discharges eoald be obtained fvm
eithe.v ©nee a f ewj, or all isolated and tested posterior ciliary
aeries t Althots^ d^anage ©f indi'^idual ner^^s dwing preparation
can esQjlain the irre^-^arily of these resultsj, it is felt that
the employed surgical, procedttr© does not p?©mit differentiatioa
of somatict, parasympathetic or ^rmpathetie nerwes assd that for
interpretation of reaalts@ an anatosssical study is necessaryo
The scope of this w«'k then iss
=. 2 -
1) to define a teehaique whidi allosfs a seleetive aM
stable silver iinpregnatien of the nerves in the orbit for
ya.tisfactor3r disseetion. The extresaiely small dimsasiojs of the
■sn&sUmotis brandies oblate ihsir dsmanstr^illx;^ in uaastaiised
preparatioBSo
2) t® study the esstent of fusion between fifth news
and post«ganglionic thi»d nerve fibers and t© deaionstrat® hista=.
logieallyB the fiber distribution in the s@»called short and
long ciliary nervese
3) to coBipare the anatoaieal and histelegieal char@et=>
eristies of posterior eiliary nerves in various speeieso
Matheds Bapleyeds Ih© orbital contents are removed
completely including the nerve sw^ply as far back as the fifth
nerv® gangliono The eocfcraocular rauscles are carefully dissected
away before fixation in f osanalin avoiding damage to the undert--
lying nenreso For stainingj, the fised material is wa^ed in
water for at least one hours aiKi is then placed in an 0^55^
solution of silver nitrate for two hours » At the end of this
timeg the eye is transferred to 10^ formalin for 10 minutess
andg f inallyg to a 5^ solution of sodium thiosulfate f (mp approx=
imately 30 minutes^ Treatment with silver nitrate and the
subsequent steps are carried out in darknesso In "t^ss resulting
preparation nerves are stained a dark brown i^il® ganglionic
tissue remains idiiteo Other tissues are unstain@d«> Dissection
is accoraplished under water using the Zeiss stereomicroscopeo
and photographs are taken as warranted.
Hh some instances fre^ material is dissectedj, aisd the
relationship of cert^^ Ranches to 13ie ciliasy and posterior
ganglion is deterraineds These branches are then removedj) fixed
in 1^ osmium tets«3d.ds9 ©snbeddedg and eross»sectioned« Ifeita
on fiber»sise da.striJ:^ticn in the variGus nerves thus may be
c^tainedo
Ma.icr Findings t With the original Osristensen technique
yreparations are obtaimed whidi l®s© dif f espentiation after
a sh^i^ period of times) since esposure to lig^t causes all
tis^ies t@ gradually tus% brotme The selective staining of
nerves according to the new technique is stable to light and
the material may be worked with for masy hours before loss of
differentiation beginso Such preparations are well suited for
photographyp as the pictures show clearly the ^ol© course
and ramifications of the ciliaxy nsrvesg and are excellent for
demonstration o
» 3-
His pattersas of nerve distribution differ in eats and
monkegrso F^eparatioas from these two speeies have in ©emmon
the fa@t Ihat the nerves close t© the glob© are generally nixed
nervess that isj, th^ contain fifth and thiM nerve fiberso
In the eats hossevere. fine branches ean be derisnstrated «hieh
travel along a tortucais sourse t® Vae posterior pole of the <^^
as disttoet burkOLes without fosion with asjjjr of the post=>gaBgli«mie
brsindhes of the ciliasy gangliono Sach isolated •*long" ciliary
nerves are not observed in the iiK>nikegr osbito The ciliaxy gang"
lion in -Ujis species receives three er fotir veiy fine br^ches
of the fifth nerveo Vfiiethar they undergo synaptic cosanectie^s®
is not kRosm» Such toanches joining the ganglion are absent
in the eato Pasion of fifth nerve fibers and "short" eiliaz^
nerves ©eetsrs at various sites and at various distances from
the ciLiazy ganglion in both species d Bie nun&er ©f theanast=
omoses also differs greatly from animal to animL and from ^e
to eyeo Kerves arising in the ciliary ganglion t^ch travel
directly to the ^e without joining wi-^ branches ©f the
fifth nerve have not been observed in either species 9 but the
segregation of fibers within mixBd nerves md subsequent branching
might give rise to a few nerves ^ich enter the scleral coat
as purely parasyinpathetic nerveso Aecon5>as:iyiGg the long eiliaiy
arteries in the horigoatal meridian of the globe course nerves
tahieh supposedly contain only fifth nerve a»d :^yi!?>athetic fibers <>
It is shoim that these mixed neirves als© c^sy p®st=ganglionie
parasg^mpathetic fibers^
^aificanee to Pro-am of Ihstitutes The renewed in=.
terest in nervous influences &n. the intraocular pressure srd.
a possible central nervous B»chanism playing a part in the
regulation of this pressure has resulted in studies reported
from this laboratosy in the past years and repeated in lab=
oratories abroado Demonstration of discharges led off from
ciliary nerves in response to small lOP changes brissg int©
focus the importance of identifying the nervous pathways idiich
conduct such signalSo The present stus^ provides iaiformation
on the nature and disteibution of these nerves in tifo speeieso
P^posed Course of R?ojeetg It is planned to t^ctend the
anatemieal and histological studies to human asatop^ materialo
Part B included Tes fW No /T
» i^ «
Serial Ho. NIKDB^^ (e)
PHS -. NIH
nhdividual Project Report
CaleiKiar Year 19^
Part Bo Honorss Awards atid Publications
Pttblications other than abstracts from this projects
vGXi SaLlDiamio lUxdid^s The role of the central nervous sfystem
in the regulation of the intraoeular pressures) Ts»ans. Glaueosa
Symposiiang LiegSg Belgiume Sept« >5j, 1958 (in press)»
Honors and Awards relating to this projects None
Ssi'ial Noo NBIDB=.iKD
lo %hthalm@log3r I
^c Same as KINIB-.J^ {el
PHS - NIH
3ad3.vidual Pr>^Je©t Iteport
Part Ao
Project Titles St»i(^ of Afferent Eleetric j^cpulses Induced tgr
Ihtr^ocular Pressure Qiangeso
Principal Investigators Po Fo Lele^ KoD-,
Other ]^vestigat®ipss Patricia Gri5r.es9 Bo Ac
Cooperating Units. None
Kan Years (calewdar year 19^ )s Patieat
Totals clO
Professional ' o 03
Others 0O5
Project DgseriptioKJ
Objectives s The role of n&r^am mechaatsi-ss in- the
regulation of iRtraoeular pressure isg as je%, not clearly
under stoed. The existence ©f such a loechaiiisa casi h& poatul=:t-ad
©aly if both the efferent aM the afferent pathways eaa be
determinedo Electrical gtiiisalatiow ef certaia rsgioEis ia the
diencephalc«a of the cat has bees reported t® cause ®ceasi®®al
alterations in the intraostilar pressure without c@irieejnitasat
changes in sjrsteisic blood pressure aad pupillasy reactioaaio
On this evidence an efferent pathway ©riginating frmi &r passing
thr©\xgh the dieneephalon has beew suggestedo "The preset'jee ®f
an afferent pathwsQ^ signalling the level ®f » or the changes
ittg the intraocular pressure (lOP) would strengthen the concept
of a nervous snschanism fer the regulat±>n of lOPo von Sallmaai .
et alo obtained some inf ertnative data in their study of af f eresit
impulses induced try lOP changesp but a large prspertion of their
experiments gave negative resultSo
"La this present investigation^ this prdbl&m is being
re-examined using different techniques 9 originally developed
- 2
for a study ©f the sensory receptors in the cemea of the eato
la view of the eomplexity smd the vascdabllity of the anatesnieal
arrangement of the nerves supplying the glob© of the eat and
the necessity of adequate exposure ©f the nerves in an untrau-
matized eosditiong the eyeball . togethrar with the nerves atai bload
vessels is reseeted from the animal and used as an isolated
organ preparationo
Methods Emplcrsreds Ihe animsaj, under light pentc°)artoital
anesthesias, is sacrificed by air-euibelisiBo Craniotouy is per-
fonaed and the roof of the orbit resee^cedo The caateats ©f the
orbit 9 with retrobulbar tissue and the trigeminal ganglions, is
carefully and quickly dissected frem the animalo
The ■ isolated ey© is kept immersed in ?jarmj, ojsygenated
Krebs-Ringer=»Glucsise (KoRoGo ) s®luti@no The extraocular muscles
and connective tissue are eiSDcisedp (srefully avoiding trauma
to any of the nerveso The dissaetion of the cHiasy nerves
from the aptie nerve and connective tissue deaths is facili-
tated ty injection @f a solution ©f %alur©nidase in KoRoGo
solutioQo
Tao 22 SoWoGo 'hypc>dermie needlesj, connected by Short
lengths of saline columns to a pressure transducer and a micro
injection apparatus r&sp&s^t'rcly are inserted at the 3 aad 9
o'clock positions at the lirabus int® the anterior chamber o
Th® eyeball and the needles are securely mounted cm a trans-'
parent plate of acrylic resino
The nervous activity is esaminsd by sucasssively placiBg
each of the dissected nerves onto a pair ©f platinum electrodes
^ich feed through an R«.Co coupled preaaplifier into one channel
of dual-beam eath@de=ray=»©scillograph CC,RoOo)o The pressure
transducers, ccuneeted to the needle in the anterior diambers is
fed through a carrier amplifier to a pen recorder and the ss-^^
cond diannel of the CoRoOc Photographic records ©f the ©scil=
loscope traces are made at different film speedso
Major Findings 8 Branches of the ophthalmic division ©f
the fifth nerve going to the globe (isasg oiliaxy nerves) did
not show say spontaneous electrical activiigr» but in ©very
instances, affer®:it is^julses were evoked yten the cosftiea and/ or
the bulbar conj\mctiva were mechanically stisnalatedo Neither
spontaneous nor evoked activity was recorded from short ciliary
nerve originating from the ciliary ganglion and entering the
globeo Responses obtained from ciliary nerves of mised origin
were essentially similar to these from the long ciliary nerveso
The preparations remain active for at least 12 hours if the
SoRoGo solution is kept ossygenated and maintained at pH 7o^
and betaeen 30® and 3?®Co
All long and mixed eiliary nsrvss tested showed a re-
sponse to increase of lOPo Ih each iastanee the^e is a sha:^
but transient increase in ttie frequent ©f ispulsesg lasting
as long as th© lOP risesj the aaxiuMin frequent being proporticaal
to the rat© and the height ©f the rise ©f lOPo 3a approximately
60^ of the nerves examineds) th© activit^^ was sustained i&ile
the lOP i?as maintained at a steady elevated leirel» The fre-
quency of sxistained activity wasg in general? dependent upon
the inerease in lOPo Every eye tested has sh^jn this type ©f
sustained response in one or more of the nerveso "Riis impulse
discharge is quite separate and distinst from responses t®
acute raeshanical defennatien (teucb) which can b© seen as
momentajy discharge superimposed on the sustained discharge^
Similar daservati^ESs ware made in an ecreball excised from a
n^nksyo
Significance to Program of Institute g "JRie results ©f
the present ^5)erim©nt5 have isportant implications in the
stuc^ of intraeeuLar pressure regulation and may be useful in
Interpreting the mechanism ©f some types of increased lOP in
man Cglauecana)^ The isolated eye js^paration with functionally
viable n9U3:'o=muscular apparatus would be ideally suited for
studies on the ciliary body (effects of drugSs, nerve stimulations
etc) 9 elasticity of the eyeball and evaluation of local anes»
theticso
Proposed Course of Projects A systematic quantitative
investigation of "Uie effect of lOP rises on the afferent nerve
activity ty isolating single active units is l^lr^ laadertakeno
A oeaiparative studty of the effects of raising "t^e lOP In dif-
ferent ways C©ogo external pressure^, injection of fluids into
the ophthalmic artery and injections into the vitreous) on the
afferent iiqsslse disehergess, is contemplatoia Efforts will
be made to localize the site of origin of the sustained afferent
activity <= Attempts will be made to ci^iria the essential
findings in experiments with the eyeball in situo
Part B included? Xes jW^ No rf
PHS ^ NIH
Jhdividxxal J¥oject Report
Calsndas' Year 1958
Part B., Honorsj, Asfsrdss, arad Publieations
Publications other than abstracts from tliis protests
von Sgillinannp Ludwigg Puortesg Michelangelo GcFo| Maeris Frasik
Jo J and Grdmess Patricias Study of affesfent sloctrie iitipulses
induced ty intraocular pressure ehajigeso Asio Jo C^tbo p -k^i
211=220 (Pto n]
Htmors and Awards relating t© this pifojects None
2o Rssxnaacolo^ Ssatioira
^o Sam© as !mnB»60 (c)
PHS - KIH ^
Calendar Y®^ 195S
Part Ac
Ppojeet Title « Effsets of ^?uscle RslsKsnt® <m IOP« and
Exte'aocfular S"^4at© Kusclss ■
Psdneipal 3Jw®stigatos'S SV@nk Jo I'aexle HioDo
0'fe@i= 3is'iy©@tlgst©r8 Patg«loi® Gfliaesg BoAo
Ckioperatii^ tJhitss Non®
Ksn Ifeas's (ealeisdag' J©®? 19:^)s Pati^t D^®s Noa©
T©tal» loO
f*rofe®sl«mals ,,5
i^rsject Itesoripticnas
C!b;js8tlv®s3 To detsCTdn® ^@ meehaaiaa ^ is^ioh d^ain©^
thonl%m and sucoini^ldsolln® aff®@t th@ lOPo
A probablo mechanism tgr vMj^ th® ©lastieiV ©^ th© cat
cgre ^ms ^t«r>®d \^ ®li£i«r decanethonitsn or syj3cis^ldaoliif&@
(as reported Isat year) appeared to b® en® aigdlst©d by th® spsasr,
of esdraocislar sn2sol@@ indu@®d \^ thesa agents^ III© p-^oblssii
vas studied bgr izidueli^ ©^ts'aomlas' mt@cl@ spaasns ^ oth^*
1) 8tiBSiIatic« of th@ third n@s?v© ijatrfficramislly sad
2} rigor ifiortiso
* t&itraoeular
Ihe elasticitiss %r®i?s deteraiinsd imder these scsr*ditic
and coEpared t« those ebtainsci la^ dnig aetioTS-j Th® e^v w,,; ....
then enusleatsd and «ir';41ai:» rcefflg\2j:»@i««its and aoirparisc«@ siadso
Ih oX'vier' to d®t®"mina -^©the^ s diffus® outside prsssm'e eevQ-d
sijfculat© th© ©ff«cts produced 1t^" ec^tffaotlssi of e^rfcrffiocpxlsr
Tiiuselsffi, a rubber chainbeis' was consrferJioted sudh that constant
outside pr©8sar®3 ooijld ba ®sj©srted on the ©mielsated ^©o
3iaaftlcity mtssjguirCTi^nts ^@g»© mad© under thea.® cossditions skI
eoniparsd ?d.th thoss® pra^ousl^r' obt®to.©do
Fethods 5?npl<!!yads Cats m©3th®tis8d with psntobs^bital
IJa tuaop© «sad in this atudyo ^en th® effect of ri^x? sfiortis
on the ©lastieity of th© ^s was studiedg th® aaiiEnjals w»$^
killed V msstis of an overdo®® of Va® anesthetic agento
!Ih9 ©laaticsity of th® ^® was dsteradaed \w naxltipl®
infusions of fixed voluiaes of salin® into th® aaat^fior ebawber
md lay reoopdiag tJje intraocular pressEur® Isc^sls bsfora saad
aftsr ®soh infasioao Ihs d&ta %«ar« th»n plottesd aa volxme
"^ersfus lOPo %©©« elssticsit^ cw*^«a fona^i tlis® b®isls for com^=
parison of th® ®la©ti3ity of th® ^® imdsr th® ifarioas! osspeav
ijaental conditiosiso
Madoy FiitHHn^® i %)ai5m of ths eodraoeular etriat® tmascles
induced bjr -Baix^ n«n?a stSjEUlaticsi or x^or mortis produced
ohsttsges in the ©Isetloity of ths sgr® sisdias' to thosws obtaisjCKi
aftsr tho adjsdnisfcapatioa of decsirAthoiiivan os- s53s«iT^lisholln®o
This ©f f eot eould h& sabolishM \^ r®s®8ti<sa of th© ®st.resK5«la3r
utriate tnosclss or esi'tjelfestioKJo Sise® •&© thy©@ diff«r©at a©tbcci -
of IftduciiTg ijiusel® spaOT produced similar ehar«g®a in th® ®las=
tici^ of th® ^®0 it sppe®r®d »ai3lik®ly that fsctors ©th®f
than the iraisieleis ■would \^ involvsda Th@ Rormal ^lastie ^alu®Q
obtained aftiar s^seetion of th® spsstlc imsclcs cjt sftes" §nu-
clisation mq>p©rt®d this cosaee^to Ihii prijnasy isiflu©n©@ on
tha ejfe b8.">cmght about tgr inuscular eaatraetien '^aig cai@ of a
dif ^M®ly «®«Brt®d ©atosid© p8?©®awr®o Ihis was d«®Tistrat®d
by- es^jsrteeats to istoicfc th® ey® was ^.selossd to ® dsssttbor fill®d
%d.th salinso 1h® prsssur® ©f th® ds®a&« wa® rais^ t«- vsri-ou®
l®wlfs and th© ©lastiel^ detsrsninedo Th® ®ff€(«yts tsn ©lastiei^-
^ li« i|>ie m^®r these «5oaditi«mSi, war® pj-astiosiliy id<mti<sal
MM m^^ mUlmd \w rottsele spsisno
I) To Bti^'im ®a a g«id® for esAtimis ©llvdosl vm® of
th<s3® iraascl© relaxants la iBitrao<jtilfiar wsFgssfs^o
2) T© inersa^ C3^ laio-Kiedg® of the j^ss=Eiss©legy of
these ag^itso
3) T© add to the W5d®s"®tsEdirig of the pi^^siolo^ of
th© ©xtraooulss' stslst® inusclss sn3L thsi? effsets on ih© intra"
ecular fxressui^o
^^8@d Ckaigge ef ^oJ^8 T© dstsrste® .if th®s@
ag®its aff ^t th® irastafl-atur© of the ^® \w th© cjOB^pressxvs
action of the imiieleso
Part B iml«ds8di Yes f^ Ho £^7
pHs - imi
Calendar Year 19^
Part Bo Heaors^ Amspdse and Fublieatieiss
Publications othss;' thsp abstracts f^'^^n this pfojsott
Kacris FoJog Wahlcop To^ and Oissass, PoS Eff@ots of estraoeular
m^cla contractlcsi on hi® ©lastiei^ ef th® ^®o Ael-IeAo P.^n<,
<^htho • in presso
Bmors snd ai^srd® s^latiiEsg to ^hUa js^jssts Kcs^®
!<, Gphthalmology Branch
2„ Pharmacology Section
3. Bethesdaj, Maryland
4. Saaifi ae ];nND3-59 (c)
PHS - NIH
Individual Project: Report
Calendar Year 1953
Part Ao
Project Title: Study on the PharEoacodynaaiics of Various
Agents Affecting Intraocular Pressure
Principal Investigator: Frank J. Macri, Ph„D»
Other Investigator: Ludwig von Sallmannj, MoD.
Cooperating Unita: Kone
Man Years (calendar year 1958): Patient Dsys: 334
Total: loQ -
Professional: o5 Outpatient Visits: 59
Other : o ^
Project Description:
Qblectives; To teat various drugs includiag the new
atraaics as well as older coH^ounds for their ability to in-
fluence intraocular pressure. Thi© approach may contribute
to the aedical treatment of patients with gleiicoiaac The
results of this investigation caay also shed some light on
the mechanisms by which the intraocular pressure is regulated.
Methods Siaployad; Various factors kno??a to be of iia-
portance in the maintenance of lOP are messured and recorded.
These are the "facility of aqueous outflo-u?" (flow espresaed
as cmm/min/aaaolig) , venous pressure in different vsins of the
eye, elasticity, and aqueous inflow. In addition to the
Hseasureassnts of lOP and the local venous pressuraSj the
systemic arterial blood pressures are also recorded to deter-
mine the correlation between these functions. Cats and laonkeys
are used.
- 2
Patient Material; Patieats x^tth ^ide and narrow
angle glaucoma^ parfciculerly borderline cases, are admitted
in greater nusribers thaa in previous years. Laboratory ob-
servations made on aqueous outflow? sechasismSj scleral
rigidity emd the sctioa of locally or systemically intro-
duced drugs are utiliiizad in the clinicsl studieso
Major Findings; It was reported last year that a
method for the deteriaination of aqueous outflow devised
in this laboratory produced results cotapatible mth those
reported in the literature, i,e«, outflotf was proportional
to the lOP. The flow rate in the cat was approaimately
0.30 cmm/iain/imaoHg. Since that txjse, ho?jever» a much larger
series of determinations has been carried out. It now
appears that there is a second, biphasic outflow pattern
which is not proportional to the lOP throughout the pressure
range examined (usually from 20 nan. to 80 ssa, Hg..), The
biphasic curve was characterized by a very fast outflot? et
the lower lOP levels (spprosimateiy 1,5 cisa/min/sEa,Rg) which
then inflected at pressure levels between 35 and 50 isn» Hgo
to a rate of approxiiaately 0.3 cssm/tain/iHa, Hg<, Such an
outflow pattern could ba induced by the parenteral admiais-
tratios of Diaasoso Here the outflow ^as changed from a
t^nophasie to a biphasic one.
It has been reported that the pressure in the veins
to vhich the aqueous huinor floors is essentially constanS
and independent of lOP changes. This hypothesis lias been
accepted for the situation in hujaans mid haa been apptied
by many workers to that in the cat, rabbit and laonkey. The
outflow pressure is the difference of pressure values between
the two ends of the outflow cliannsls, i.e., intraocular
(trabecular) and venous (episcleral). It becanj© quite i2i-»
portant, theraforCs ^° study the venous pressures on the
surface of the eye.
«Hiree veins (anterior ciliaryj losg posteiior ciliary
and vortes) in the cat can be cansulated for pressure deter-
mination. Plastic casts ^^re made to illustrate the complicated
anatomic relationship of the various venous vascular beds. The
cast material was injected into the anterior chsisber under
coatinuQus pressure until many of the eplssleral vessels were
grossly seen .to be filled. The material was allotted to hardenj
then the tissue was digested a^ay. The aqueous outflow channels
were thus d^ioastrated in their course from the trabecular ares
to the Circle of Hovius, Pressure readings in the anterior
ciliary and vortes veins, and those of the intrsocular pressures
appeared almost identical tssdiSS restiag coaditions, Ho^Bjevar,
if the lOP was either raised or lowered, the venous pressure
fello Tlias, it is shown that induced chsages of the lOF
cent alter the venous presstsrc. A suHsaary of ths conditici?.s
affected fay changes in the venous pressure follows:
1) The biphaeic outfloir patterns becasne smsch ii^re
pronounced when outflow pressure was calculated as lOF-
venous pressure.
2) Arterenol, scetylcholina^ histaainSs hesamethonitass
Br, Bsmipsthetic and parasytapathetic nerve stimuiatioa all
produced changes in the venous pressure which paralleled the
changes of the lOP.
3) Trauma of the eye induced identical eleirations of
the lOP and venous pressure.
4) Diamox lowered both the lOF and vesious pressure.
The effect; of Diamox on the vortex venous pressure
was also determined in the siKsnkey. In this species there
arc no anastotnotic connections between the vortex snd epi""
sclerel veins (into which the aqeuous veins feed). The
pressure in the vortest vein ^shen measured under resting
conditions was found to be 10 tc 20 tsm. Hg. higher than
that of the lOP. Disiaox was capable of loweftisg this pres-
sure as veil SiB decreasing the lOP.
Significance to Pro^rom of Institute:
1. (a) Ihe calculations of "facility of outflow" in
man are based on the assumptics that outflow of aqueous is
proportional to the intraocular pressure and also that the
episcleral venous pressure is II .0 naa. Hg, is both noraal
and glaucomatous eyes. The effective outflow pressure then
equals the 10? minus 11.0 v^x, Hg. In the experiments on
ffinesthetised cats neither of these t'so conditions appear
fully met,
(b) GlaucoT&a is thought to be caused in z^st
instances by a diminished outflow and a relati^mly constant
inflow - the net effect being an increase of lOP. In cat
experis^nts the intraocular pressure and venous pressure
were nearly equal to each other, a result which casts soEse
doubt on the correctness of this hypothesis. Hcweverj the
difficulty to interpret the esperinsntal restilts and to
epply thea to the husaaaa eye is fully realised.
i} ^
2, Diaiaos has been Bho??n to lo^er £he 10? in glaucosa
by the reduction of aqueous inflow, Tha£ this drug is also
capable of reducing the veaous pressisrs in the eyes of cats
aad lEonkeys indicates that anotheir lajschaaisia njay play a part
in this Icsweriag of lOPo These observations poiat oace sKjre
to the necessity of studying carefully the role of the
vascular bed ia the maintenance of the lOP.
Proposed Courae of Projeet; It is plasmed to continue
studies on the lOP »i£h other ph&rssacologic agents, A ■miT&
intensive &ttei:B|>£ %7lll be tsade to investigate the znechanism
of Diaiaox action on the eye.
Part B in<sluded Yes i^X^ Ito f^
Serial IJOo NIIJDB~42 (cl
PHS - NIH
Individual Project Report
Calendar Year 1958
Part B^ Honors g Awards and Publications
Publications other than abstracts from this project:
Maori J, Frank J,: Outflow patterns of the cat eye^ Aia= Jo
Ophtho (in press)
I'lacris, Frank J„p jJanko^ Iheodorj and Grimes j, Patricia Ac'.
The elastic properties of the human aje^ A„M„A, Archo Ophth,
(ia press)
Maoris, Frank J,: Some aspects of aqueous dynamics j, Glaucomas
Transo Third Conf.p Jan, S^, 9 and lOg 1958s, Josiah Ilacyp Jro
Foundationo Sdo Frank J„ Newell, (in press)
Honors and awards relating to this project: None
Serial Hc« MBHB^^-^ fe)
lo Ophthaliiislogy Branch"
2o
% Betfcssdap Marsrland
4o New Plmjeet
PHS =. NIH
Ibdividual Project Heport
CalsBdar Tear 19^
Part A,
Project Titles Clinical GlausoTsa Study
Prdneipal Issvestigaterss
David Patens Mo Do atsd
lAMwig iTiSK Sallinatms McD<,
Other l&ivestigat©rss
Herbert Ksuftenj, MoD,,
Richard Cepenhaverg M,Do and
Bruce Cohans Mo Do
Man Years C calendar year 1958 )s
Totals ,10
Professional % „ 05
Ofeers 0O5 y
Patient Dsyss 33^
Outpatient Visits? ^
PS?oject Descriptions
Ob.lectivest
1) !Si© chief objective ©f this stiady is t© ©'^aluate
suspected cases of early glaueoaia to detersaine ths^oogh Ktaltiple
clinical obseirvaticsis aM testa those ^ieh are most helpful
in the diagnosis » prognosis » aiid adequacy ©f glaucoaia therapy o
Becaiise of th© intraocular teEssissB visiual field lisnits and
indiiddual response t© test situati^nsj) a diagnosis' of glaucoaia
is often withheld or qtJite uncertain with present day techniques 0
Although one single test wHl ujjdoubtedly not be found ^^jon
which a diagnosis can be based© it is the tsbjeet of the present
stud^ to determine ■^lish jroeedores are Eost ^lsef^il9 what is
■their reproducibHityg, and t^at retspospeetiv© inferaiatio^i can
b® gained in regard t® prognosis \^ carefully f ©ll<K?ing these
patients o In particulars the study is coace^ried with the in-
fonnatien gained fr-om tonogra|fey in borderline cases of glaueomao
2} The Btxsdy is als© eonseraed with pharsiolegicsally
distinct forms of glauessna su©h as "lew tesasiea"© hypersecr©»
tionp pigmsntaiy nasirow aejgle^ @r infLasanatery forms @f glau-
e@inae Ihese cases are studied as e^teissi'^ely aisd in the same
manner as the bosrderline eases ifiiieh most often eensist ©f chroni©
simple glaucomao Here again 9 the stisdy is primarily coa®emed
with the diagnosis of the disease aad the means of different=
iating &ae f&sm from the ethero
3) Beeaias© of its e&a.eesn with tiie value of tonography y
the protest also ineludes the perfomnance of ^atever tme&saasy
surgical procedures are indicated to reduce uncontrolled intra=
ocular pressoreo The adeq\xa<^ of surgical Iherapor as well as
medical means t© reduce the pressure is jiadged in regard to
day curve t®riop»aphy snd prevention of fteth^ visual field
losso ,
Methods ^Etepleyeds Each patient aeseptabl© f®f' the stuc^
is admitted t@ the hospital for a minimum of several days in
order that an extensive glaucoma worktqp can be performed under
rather constant ew^lronmental conditions and at all times of
the d^ or nl^to In addition to history and general physical
escaminationg specific tests incl\ade tonoinetiy wi'& day curve
determinations 9 applanation tes^ossetiyg measurement ©f d^th of
anterior chambers) bioBieroseqfgr and gonioseopys tenograj^j,
at frequent intervals ?> and prevocativ© tests also incorporating
tenographyo in addition^ visual field studies are carefully
perf ermedB usit^ the Goldmarsn {g"o jeetion perimeter and i^ie
.Gunkel tangent sereeoo Wherever possible g j^otograj^ of the
optic discs are obtained^ 'Sfeen the patient's isntreated intra=
oetilar pressure has been observed over a time int^val suffi»
cient to obtain either a positive diagnosis ^f a sizable amount
of datat) the response to glai^oma thars^:^ is subsequeEitly eval«
uated and the patients are not discharged until the intraocular
tension is satisfactorily centirolled 00 a twenty-four hour basis..
Subsequent t@ discharge^ attesspts are made to follow each patient
at four to six month interwa^.s? at ^licih time tsaomstryj, ton©=
graphys and visual fields are Ji^aiia tecludedo
IJa^jcy Finding;sg At lai© present time^ the number ©f
patients ia this study is t©© small^ and their period ©f follow
too short t® drasr matsy c«melusi<^as fswa. this long-tes^ projecto
On an individual basis g. w© feel that we have encountered some
©jctTCTiely provocative cases illxistrativ© of a broad spsotrum
ef glaueaaa formso 3a oae os' tw® eases of see^idary glauoema
■^e diagnosis was suspected on the basis ©f t6Kogra^3y--si@'BS
and aubsequently validated when episodes of increased tension
oceurredo la other cases repeated provocative tests and ton©=
graplgr have failed to incriminate glaucoma duriag an initial
= 3
admission arai the diagnesis made ia subsequent inoathso At the
present tdrnes the cases ©f borderlia© glauosma ha¥<s not beea
followed for a sufficient tiraa t© judge the value of the data
obtainedo
I¥ep@g@d Csfurae of PSi'o.lects At the ps»©s©at tini©^ efforts
are being made to instruct a teehxsician in the esqpsrt use of
the electronic tonoaietere If this dsjectiv© is realiaedg it
is hoped that the project size can be inereasedj) that more
egtbensive tonography can be undertaken and that the increasing
size of the follcfwup patient population can be adequatel^f
handled b^ outpatient 'visitso Furthers^rea the need for identical
€fosenrations on normal control subjects of the glaucona age
group is apparento
Significance to Proayam of Institute s The described
project is of timely inportance to the Public Health Sw^ce
due to the growing awareness of doctors and laymen alike qw&'
ceming the significance of early glaucoma diagnosis » Through
clinical e^erinents with glaueesiaj laboratosy iasvestigations
are further inspiredo It is esiphasized that this Hiistitute
offers a unique opportunity for such detailed ^Aser^ations ®f
glaueoBia patients » for routine laboratoa^ studies can be repeats
edly performed under a consid^'ably longer period of hospital^
isation than is a'^ailable to private institutionso Through
the quality of prolosged study ra^^» than throu^ the quantity
of patients esgamir^da this project should offer information
iihich cannot be readily gained elseidiereo
Part B included Yes £1 No /^
Serial Ko„ JffiflS=MXsL„
lo Ophthaliaology Branch
2„ Microbiolo^ Section
3 a Bethesdas Iferyland
4-0 New Project
PES - NIH
Individual Project Report
Calendar Year 1958
Part Ac
Project Titles
The OciLLar Diagnosis and Treatment of Ocular
Cor5)lications of Primary Familial AnQrloidoslSo
Principal Investigators:
Herbert E„ Kaufmans, MoDc
Gerard van Alphsnj, MoDo
Other Investigators
None
Cooperating Units s
None
Ifen Years (calendar year 1958)
Total: olO
Professionals 0O5
Other; 0O5
Patient Days: 2^
Outpatient Visits: ^
Project Descriptions
Ob,iective3: (l) To detaimine whether vitreous opacities
reported in occasional cases of primary familial aa^loidosis are;,
in factj diagnostic of the di^easeo (2) To elucidate their histo-
chemical characteristics, (3) To evaluate the possibility of
vitreous replacement in the therapy of blindness caused by these
opacities ,
Methods and Materials; Ihe appearance of the vitreous
opacities is carefully studied by biomicroscopyj and the presence
or absence of systemic deposits of aD^rloid is determined by skinj,
muscle and gingival biopsies. The histochemical appearance of
anqrloid is determined by various histochemical stains o The
electron-micro scppic appearance has been studied by Dr, Theodor
WankOo
An instrument has been developed which will remove
diseased vitreous and replace itj, after lavage^, by fresh donor
vitreous without appreciably changing the pressure within the
eye. Vitreous replacement and lavage has been attempted on one
patient with this instrument »
Ma„ior Findings; (l) In addition to one patient with
primaiy familial an^rloidosiSs, who was admitted for study of
loss of vision and was subsequently diagnosed whec systemic
symptoms appeared^, two other patients with primary familial
amyloidosis without systemic complaints have been diagnosed
because of the characteristic appearance of their vitreous
opacities „ In one of these patientsj, autopsy specimens from a
brother on whom the postmortem diagnosis was diffuse arterio-
sclerosis revealed thatj, in fact^, the cause of the brother's
neuropathy J, gastro-intestinal distio-bance and heart failure was
due to amyloidosis „ The other patient j, who was confirmed as
having unsuspected amyloidosiSj, is from a family in which one
brother has neuropathy with muscle wasting^ and a third brother
has bouts of cardiac syncope <> A single skin biopsy of the
brother with cardiac syncope was negative,
(2) Vitreous aspiration was carried out on six eyes of
four patients with the disease. In four <i)f these^ opacities
were obtained at the time of aspiration. These opacities were
found to have the staining properties of amyloid^, whereas control
vitreous and opacities in bank vitreous were not found to have
these properties,
(3) On one patient, vitreous lavage was attempted with
the newly developed instrument. In this patient there was a
transient improvement of vision^ but this was neither dreinatic
nor sustained^ and was accompanied by transient inflammation.
Patient Material; Patients with vitreems opacities are
obtained by physician refeiral, (Usually these patients have
been diagnosed as having uveitis,)
Significance to Program of Institutes Primary familial
anyloidosis is often readily diagnosed once it is suspected. The
demonstration that the vitreous opacities present in some of
these patients are actually comprised of amyloid points up the
diagnostic importance of their characteristic appearance. The
demonstration of angrloidosis in one familyj where it had neither
been diagnosed clinically nor on postmortem examinationg suggests
that the disease may be more widespread than was previously thought o
The discovery of two patients with unsuspected amyloidosis confirms
that the ophthalmol(^st is in a position to suggest the correct
diagnosis in a syndrome otherwise difficult to identify and mayj
thereby, permit the correct diagnosis not only in the patient
under observation but in other members of the family who may be
- 3
ill J but may aot have the vitreous opacities o The •usefulness
of the instrusaeiit for vitreous lavage in cases with blindiiig
vitreous opacities must still be evaluated. Certainly it is
potentially useful in conditions other than amyloidosis o
Part B included Yes
Serial No, _Ji]ra^-44(c)
PHS " NIH
Indi'^id'aal Project Report
Calendar Year 1958
Part, B ; Honors j, Awards and Publications
Publications other than abstracts from this projects
Katifinanj HoE,; Primary Familial Amyloidosis^, A.M^Ao Arch,
Ophtho (in press)
Honors and Awards relating to this projects None
Serial Ho„ HINIB-4.5(c)
1„ Ophthalmologj Branch
2o Microbiology Section
3^ Bethesdaj, Maryland
U^ Same as NINDB-68 (c)
PHS -^ NIH
Individual Project Reports
Calendar Year 1958
Part A.
Project Titles Study of Toxoplasmosis aad Its Therapy
Principal Investigators Herbert E„ Kaufmanj, MoDo
Other Investigator: Lee A, Caldwell^ BoSo (Investigator
for Part III)
Cooperating Units; Studies described in Part I acd II were
done in conjunction vdth Br^ Leon Jacobs'
project on Studies on Toxoplasmosis in the
National Institute of Allergy and Infec-
tious Diseases,
an Years (calendar year 1958) j Patient Days: 2„201
Total: olO
Professional? o©5
Others o05 Outpatieat visits: 202
Project Description;
lo LABORATORY STUDIES
Objectives: (1) To investigate differences between
virulent and avirulent strains of toxoplasma gondii and the
effect of some parameters of virulence on therapy„ (2) To
study the efficacy of present thers-py with Daraprim and sulfa
in vivo and in vitro o (3) To explore the metabolism of
Daraprim in man and the details of its toxicityo
Methods Employed: Various strains of toxoplasma gondii
were maintained in chick embryos and haarvested from ground chick
embryo chorioallantoic mea^raneo The organisms were counted in
a Keubauer-Levy Counting Chamber and inoculated into standing
roller tube tissue cultiires of monkey kidney epitheliim and
human amnion cell monolayers, man^tained in appz'opriate nediao
Ttw-3 tiMe of aivth.ahiii&nij of. the. oi'gs.nlfsn)-': cowJ-d 'a co0.OrG J J.ed oj
¥».f)h:?rift' 07.it the inoculum at desired intervals o The organisais
were permitted to multiply^ but cultures could be fixed and
stained before cell lysis had occurred. In stained preparations
microscopic examination enabled the nuniber of cells invaded and
the number of organisms per parasitized cell to be counted „
Other cultures were permitted to go on to lysis and the time
required for lysis could be correlated Tjith dnvivo virulence o
The effect of Daraprim on toxoplasmas in vitro was investigated
by exposing infected cultures to known concentrations of drug,
and then grinding up culttires and inoculating them into mice to
determine whether viable organisms could be recovered.
Major Findings; Strains of toxoplasmas that were most
virulent for animals also destroyed tissue cultures most rapidly.
In the strains studied the virulent Ri strain invaded 4--5 times
as many cells and multiplied 2-3 times as fast as the less
virulent 113 CE strain » The S-5 straia of intermediate virulence
was also intermediate in invasiveness and rate of multiplication..
It was expected that Daraprim^, which is a metabolic antagonist^
would be considerably more effective against more rapidly grow-
ing organisms, iJhen cultiares were exposed to varying concentrations
of Daraprim for five d'aySj the slow-growing organisms were shown
to be much more resistant to the Daraprim than the rapidly growing
organisms. Further studies of a similar nature reveal that appre-
ciable time is required for the organism to toe in contact with
the cell before invasion takes place, and that chronic infection
of tissue cultures can be produced with slow-growing strains. In
this chronic state j the organism and the culture seem to be in
balances, and damage to the culture is not apparent, ./hen organisms
are incubated with serum containing a high titer of dye
test antibodies J, these organisms axe killed by the serum^ suggest-
ing that the dye test antibody may^ in fact, be toxoplasmocidalo
Patient ^!aterial: Cases of granulomatous uveitis are
obtained by physician referral. Active cases that suggest a
possible toxoplasma etiology by hlstorys morphologyg akin test or
dye test can be selected.
Significance to Program of Institute; In the past^ seme
cases of ocular toxoplasmosis have been reported that do not
appear to respond to therapy with Daraprim and sulfa. The
differences between strains elucidated can explain possible
differences in the clinical course of toxoplasmosis 5, as well as
the apparent resistance of some infections to therapy,
IIo EFFICACY OF THERAPY
Ob.1ective3s To determine whether Daraprim j, as used
clinically J, can penetrate into the eye 5 and., in addition, to
~ 3
determine •whether it is effective against the proliferating
form of toxoplasma in the eja.
Methods Employed; To determine whether Daraprim pene-
trates into the retinaj, monkeys were given Daraprim.5 and after a
suitable period of time the serum and the retina was analyzed
for Darapriffio In additions, guinea pigs were inoculated with
proliferating toxoplasmas into the vitreous and were then fed
with Daraprim and sulfas Controls that were inocolated with
the organismj, but not given drugs ^ vere also maintained. After
appropriate periods of time^ the eyes of all animals were
examined and the ferains and inoculated eyes were ground up.
The suspension was inoculated intraperitoneally into mice to
determine whether li¥e organisms persisted. Dye tests were
done on control and treated guinea pigSo
Major Findings; The concentration of Daraprim in the
retina of monkeys vras comparable to that found in the sorumo In
guinea pigs treated with Daraprim and mlf&p in almost all cases
there were no organisnfe recovered from the brain and the dye
tests were negative <. In untreated guinea pigSj, organisms were
recovered from all brains and all eyest, Chorioretinitis was
seen to develop .in controls oixLjg and dye tests were positive „
Si^fiificaace to Prpgram of Institute; Despite the fact
that Daraprim has been, used in the treatment of ocular toxoplas-
mosis for many years^ many dispute its efficacy. It is surpris-
ing that no studies have yet been done to determine ifhether
Daraprim actually penetrates the retinaj, and no other studies
have been done confirming its efficacy on intraocular infection ,
IIIo PHARMACOLOGICAL PROPERTIES AND TOXICITY OF DARAPRIM
Objectives; (l) To investigate the pharmacology of
Daraprim^ ^^"^ to study the cause of Daraprim toxicity 5 its mani-
festations and the best method to safeguard patients from it.
Methods and, Patient Material; Serial serum Daraprim
determinations were done on patients being treated with Daraprim
and s\0.fa on various regimens. Serial hemotology observations
were obtained and were correlated with earlier heiaotological
studies in the records of patients previously seen in the
Institute wiser© th© studies %'otq doae,
Ma.1or Findings; Patients had been treated with 25 mg,
of Daraprim orally per day^ but on this regimen it was found
that when the Daraprim was stopped the compound could be
detected in the blood for up to two weeks. This slow decay
- u
and slow rise in drug levels suggested that a loadtcg dose
would be desirable followed hj a maiBtenaace dose„ A regimen
was therefore developed which gave rapid stable levels approxi-
mately two weeks earlier than the previously used regimen o Wide
differences were found in the final concentration of Daraprim
from patient to patients When the drug waa stopped,, the fall in
drug concentration from a given concentration in all patients
was similar o therefore g since elimination was the same ^ the
difference in drug levels must be due to differing absorption
of the drugo In those patients who developed hemotologic
toxicity^ th© Daraprim level was significantly higher than in
the non- toxic patient o
Sipnificance to Program of Institute; Patients have
been treated for many years with Daraprim and siilfa drugs and
yet the phaiToacology of Daraprim in humans maintained on the
drug has never before been studied o It is obvious from these
studies that therapeutic regimens different from those previously
used might be desirable p and ftothermore it appears that the
differences in drug levels obtained are due to differences in
absorption of the drug from the gastro-intestinal tract. Since
toxicity appears to be a function of the absorption of the drugj,
it may be possible to obtain stable high uniform blood levels
with minimal toxicity and minimal variation in absorption by
using different salts of Daraprimj, such as succinate or glutamatec
Early studies by Dr„ Leo Gaudette in the National Institute of
Allergy and Infectious Diseases suggest that this is 5 in factj
true, la addition 5 because of the toxicity of Daraprim it is
important to determine whether one parameter of hemotology can
be followed or whether it is essential to do complete hemoto-
logical worloip in all cases of patients on Darapriffio There is
no data on the precise nature of the hemotologieal toxicity in
man 5 and there is no indication in the literature how long the
toxicity can remain and how serious it may become.,
Prpijqsec^ .Coiyse. of Pro,1e<?ti Continued studies on the
hemotologic toxicity of Daraprim are in progress. In addition^
it is hoped that an evaluation of the efficacy of Daraprim in
patients treated for chronic ocular toxoplasmosis will be
possible.
Part B included Yes ^ No £J
Serial Koo NIim=A5(el
PHS - NIH
Individual Project Report
Calendar Year 1958
Part 3o Honors., Awards and Publications
Publications other than abstracts from this project:
Kaufman;, HoE^^ Remingt'iap JoS,;, and JacobSs, L=: Toxoplasmosis;
Ihe Nature of Virulence,, Am„ Jo Ophtho^ i:6i255-'26l (Pt, II) ^ 1958,
Kauftaaaj, HoSoj, Melton^ lUlUs Remington^ JoS,^ and Jacobs^ Lot
Strain Differences of Toxoplasma Gondii^ Jo Parasitolo (in
press)
Remington^ J0S05 JacobSg L05 Melton^ MoM„j, and Kauftaan;, H^E,?
Studies on Clironic Toxoplasmosis s The Relation of Infective Dose
to Residual Infection and to the Possibility of Congenital
Transmission^ Ma Jo Ophthoj, 46??6l-26C (Pto II) j, 1958,
Remingtons JoS„g Jacobs^ Loj Melton^ MoM,, and Kaufman^, HoSoS
Research Note; Chronic Toxoplasma Infection in a Human Uterus j,
Jo Parasitolo (in press)
Honors and Awards relating to this project: None
Serial Moe HIMra-^,lj6^ (c)
lo OphthalRioloQT Branch
2c.
3e Bsthesdas -''asylard
ho Same as RIT'1DB"66 (c)
PHS » HIH
3hdi%'idiial Probst "Report
Calendar Yess* 195^
Fart Aa
Project Titles Thyroid H©g^;oR© Tm«sio-t?sr in IVsitis
PHjieipal Iswestigators Jarnes Fe O'Roug^css V^Do
Otfeer Im?@stigat®ra8 Richard Copstiha^ers I'oDe
Herbert KasafmatSs I'oDa
Dsifid Patcasr !-'oDe
Coop©patiKg IMtss MvlQB &rd. assistence given 'by Dto Eo
Rail of t&© Cliiffiiesl Eadoorfnolog^
Vm Years (calmdar y-©®r 1958 )s Patient D^ss 2^31
Totals 1,0
Professional § „ 5
Other? o5 C?itp®ti@nt Visits i 206
Project Dsserlptlont
Interiir: IMgs -As iodicated la tfe® 3?©p©rt for 195?e th©
ercphasis in this projsct has been plss^ m two pax'ssr.etersa
Fisratj, the utilisation or tm^no'^er ©f r^^-*- tsgrrcKisaSs adinimi©-
tered ©xogenoijslys in patients ^.th ehroniss relapsing uveitiso
Seeesadp dets^miastlon ©f similai' factors in nosnnal son-
trol patients for cotnparisoa wi-fe p^li^ed iralu@Sa
Qb.jectiveas To d®t@?mia© ^©ths? ther® is an association
between ebroKieity sskI rat© of s-eetara^no® in uv-eitis with ab-
normalities ia ps2»iph®ral utilisation of tigroid hormen®©
?'®thods Einplqy^@dg Patistits aeespt^i for tfe® atuc^ ha^s
been scrssnsd to dstemiiRs ^e statas^f ihjr-^M t^motlon (basal
metabolic rata^ mnm cholesterol » I^'-^'^ tsptak© stsd ssruss protete-
bou^ iodin@)«.
«. 2 "
from th® rate of disappsarane© of I -^^ hotv.on® fr<m th®- blood;,
fbllosdijg tetpa^enous tojsotiono lis® radioastlrlt^ injected
is 50 iricr©s«rt0S,, Bleod ssmplQs are ds^swR dsily f©? 1'^' day®
and pp©pas*ed a© 2o0 tnl© comitiBg sssjpleso Hadioaetivlty is
as@^^' ixi a wsll-scixttillati^ ©oiiater and eoasparsd lAih known
standasrds of tb® iMj®et®d dos@o Half tim® values @2«© caleti-
latod Srcvci th© eountii^ f©s^lts @jid coBpsi'ed for sccurse^ with
valuess d®ri-v®i linearly from th© slop© of s s©rai«log plot of
th® datao All plott^ results ^® fitted l^ th® ssisthod of
least ^|uas'©So
By eoitparisoB of eoanting ?@sults with ©srum l®v®l@
of prot©i»=-boiffld i«Kiin© th® following ^alvissa are dsrivgd for
Gdch osti@i^t s
Xo ^iljT i^Fc@nt of thjr^o%in@ ix)oX dsgic^Gdo
2o Eixtai>atliyroldaL iodine poolo
3» l-:ic2^grfflms of iodin® dsgrad®d daily.,
is-o Correction of all data for sg&g ®&Xv weight snd
surfsc© sr©ao
?S^^|^iJfet^^;.,^.,^5^.^S^Ml« Kadiothjrostee
• stiidiss have be®n coEpl®t©d ia 30 uveitis patientis
wd 5 wtsemal controlss
Dsta available indicate that in ths laMa t^® wsitis
psti«nt8 #io(w 3?©ta?^d®d K-at© of utilisation of oirealatiBg tfe^«^
roid honuon^.9« owp@s?®d Mth noznaal c^ats^lso Cos^pmi^ims
based on agse ssk^ bo(|f frsi^t @nd surf sc@ @r@@ do not sff @et
this r@gjat signifieaatlyo %« sajor dif fee^sc© lies in th©
dsily s°3t@ of dsgrsdstisss of 1. -^ til^£%siS.si@ @sid of ooursdgp
in tiis%@"«r@r hsslf tiin©«
Trestwemit «iUJ 12^g©id hofmm© hasp iJs s f®? patisatsg
resultai ia eosev^xt&jm of th@s® gSx^os^msI s°@stalt@o
Stfsdi®3 ca3id)i6t®d esi 3 siss^stsl o^its;^! pstlsists bsic^®
giifCTi 3?®salts that sgi?©® ©l©g©l^ id.'fe th©s© of se^eafsl oihm>
issv^stigstaiPSo
SSfflaifieaaee ^_to^ F^ts^oB?^. of ^ ^Ifastitut® 8 Psti^its with
r&s&T&nt uveitis eosistitut® s sisjor eliM-Qsl piraibl<^ xn th@
C^thalBsolo^ Bfe-aneh ss«g?8m« IMs p2>o^©st is d©@i@s®d t©
60BpX®i!@nt oth^r stiKii@s thst as^ don® in th® routiu© wos^c^
tqp ©f Bi^^tis pati^tso . It is felt llsat hesiissmaL i^alane®
Bi^ b© an iB?H»^snt eo^fgstor relstsd ©itiifsr t@ sasc^tibility
of patisats t© th® disgeas© or pessSbly te their pattern© of
th@rsp@iEtie
Hsisr© is seed f os= fte'ther
\jSsSo
this it t-dll ba of int®2»©8t to dst^isSs© ^lat effects tg-sat^
Beat «ri^ tigroid hoi%»m@ has oesi
lo Tusraovsp yeaalts
2o Cli?il«sal f©gt^sr@s cjf uveitis in patl^its pr^^l&Qsly
Serial N®o KimB°»7 (e)
lo O^thaltnology Bpaneh
2o F}Qrsi©l@Qr Seotleo
3^ Bethesdap Masylasd
^o Same as KINIB=65 (e)
PHS = NIH
3bdividual Project Report
Calendar Year 19^
Part Ao
ftr>©jeet Titles Detection ©f QsuLar Tuai@r by Isotope
Tracer Methods
JViRcipal 3hvestigat©f J James Fo O'Reurkeg Mo Do
Other IswestigatorsJ David Patesss M<,Do
BsvLQB Eo Cohans W-Do
Richard Mo CopenhaTer^ MoD,,
Herbert Eo Kaxifmans Mo Do
Csoperating Units s Noie
Man Tears (caleisdar year 19^)? Patient Dayes 313
Totals 1.0
I^ofessionals ,5 CHatpatimt Visits? 98
Others o5
Projeet Dessriptions
Interim N@teg Am iadieated in the report for 1957b the
covirse proposed for this project was to contiraa© the study ©f
tumor detections) mainly at the clinical levels
Previously^ basic sfe^di^s indicatei that anlaaal tujasrs
may take \jcp relatively more Zn^ ©r I -* than P-^^o These reisults
are not at once appiieabl© t© the clinical problem of tumor
diagnosis for the reason that a reliable method for localising
foci of gamma ener^s within the eysg has net been foundo The
result is that we continue t® depend on tracing the beta emissioos
of radiophosphoruso
= 2 -
tlves s T© assess the accuracy ©f tracer metheds
used for diagnosis of ocular tumorg, eoupariBg trans^scleral
and trans=conjuncti'^al methods @f eeuatiago
Methods Employed g RadiophosjSiorus uptake studies continue
to be done on patients lowing evidence of intraocular tmrnvo
The technique presently used is t@ inject >3C ricrscuries (adult
dose) of sodium radiophosphatej, intravenouslyp and t® measure
the relative radioactivity ®f each eye after tw©nty=>f©ur hours»
using an ead"=window Geiger tubSo
The initial counting is done trans=>CQnjunctivallyo Fol-
lowing this9 the appropfiate portion ©f the surface ©f the
globe is ea^josedjj surgicallys and tran®»>scleral counting is
dene over the area of the fundus lesieno
Major Findingas Th.e advantages ©f trans=seleral countings
done as a surgical procedures, are indicated by results obtained
in sevefral recent attempts o Biere is evidence thatg in the four
patients most recently studiedj, the trans-scleral eowiting re=
suits were correctly positive? although results ©f th© traa8=
eenjunetival method were negative or ©quivoealo Th9 forsier
method seems t© be the present one of dioiceg alth@u^ patients
available are to© feu t@ support this as a definite conelusii^o
Sifinifieancfa to Pgof^
CJjphthalmelogy Branch a
am of Institutes Patients referred
to the CJjphthalmelogy Branch as turner suspects often represent
doubtful or unusual eases s the need f©r continued atteitpts to
Imprewe P=.32 counting methods is indicated by several false
negati^"® results obtained in this group of patientso
Proposed Ctourse of Projects
1) T© continue P^ study on patients available with
suspected ocular maligpaneyj
2) to attempt correlation between the locus ©f highest
radioactivity axyS. the position of intraocular malignancyg on
histopathologic sections o
Part B included Yes /"T No /^
Sarial Noo MIMDB;=>48 (e)
lo OphthaLinol©^ Branch
2o
3. Bethesda^ Maxylatsd
^o Net? Project
FEK - KIH
Individual Project Report
Calendar Year 19^
Part Ao
Project Titles Ikssiunological Relations in Ocular Tissues
Principal Investigators Gerard Wo van Alpheng Mo Do
Other investigators Sarah Rebinettss, Bo 3=
Cooperating Units s None
Kan Years (calendar yesc 1958)? Patient Da^ss None
Totals oio '
Professionals 0O5
Others 0O5
Project Inscriptions
Objectives?
1) To determine the possible antigenicity ©f the leas
capsule I
2) to produce cataracts iramunologically by iBiiraaniaati^s
with lens capsule and l&ia proteins?
3) to determine labether various tissues of the eye are
immunologically related!
k) to find imaiunological clues f©r ocular involveiaent
in patients with various allergic dermatoses c
Methods Bnplojyeds Lack of sufficient animal space
necessitates the us© of guinea pigs throu^out the esperiisentso
Guinea pigs are i-nsmmiaed with guinea pig lens capsules aM
^ 2 -■
guinea pig lens proteins hoitiogenissd with i^etad^s adjuvant
to enhance antibody produetioiSo IhjeGtions are repeated everj
two weeks for several iMnths and the eyes of the iimnuniged
sasimals are observed on the slit laspo Blood asai aqueous
humor sre tested for antibodies by the Ouehterlosagr Biethodo
Other guinea pigs are immunized with calf corneal epithelium^
calf vitreouss, calf lens capsulej, and calf lens protein^ respec°
tiveljo Hie sera are tested for antibodies xd-th partietxLar
regard for reactions of identity or non-identity with the var=.
ioxis antigens mentionsdo
Ka.ior Findings g Ihe sera of guinea pigs imnamized with
guinea pigs* lens capsule atsd guinea pigs* lens proteins or
with calf lens capsules and calf lens proteins show cross»
reactions with corneal epithelium aM vitreous and do not react
with donor bloods «.irisj) -retina,) or »■ squeeuso teti<=calf
vitreous sera show strong cross-reaetlons with calf blood and
none with the other c&Lf antigens o ^ti«ealf corneal sera
react with corneal epithelium only and not with calf blocd ®r
ocular calf antigens o 3h none of the animals immunized with
lens capsules and lens proteins did cataracts appe^g although
repeated paracenteses were carried out and some ier^es were
traumatisedo
Significance to ftfogram of nastitutes Hie conplete
knowledge of related antigens in eye tissues is theoretically
intriguing arai necessary for a better understa^Kiing ©f the
response of the ^e to diseaseo
Proposed Course of Pr©.1eots The nature of the antigen^
antibsK^ response between various eye tissues has to be further
investigatedg particularly ^e-Uier one deals here wi"Ui true
cr©s3»reactions or reactions between identical or related pro-
tein jnoleculeso The lens being the priaie essirsple of organ spec^
ifieity suggests testing of oilier ©eular tissues en organ and
species specifi^gTo
It is Imowa that subcutaneous inplantation of donor
skin in recipient animals will lead to opacificati«i of a
corneal grafto It would sseirip therafores that both skin and
cornea have related or identical protein rooleeulesp but in the
above-mentioned easperiiaents anti-cos^eal sera did not react
with skin= Neither did anti^lens sera react with skino The
obscure isamanological relation between skin and ecmea and
skin aisi lens seems well worth investigatingo If the results
of the animal experiments so justify^ it is hoped to study
patients with certain dermatoses showing ©eular manifestations
of cornea and lenso
Part B included Yes /"T No fWf
Serial Noo MIKrB^^9 Ce,)^__
lo <^hthalHiol®gy BraHsh
3o Bethesdas Mafylaad
k. N^y Pretest
PHS =o N2H
liidivldiaal Project Report
Calerai.aff' Yesr 1958
Prejeet Title? Basic Factors in Hefraetion A^oinsaies
Principal Itwestigaters Gerard van Alifiiens M^Do
Other 3hvestigat@r3S None
Cooperating Ifeitss None
Man Years Csaleadar year 19^)? Patient Dagrgs None
Totals olO
Pt-ofessionals 0O5
Ot^ieys 0O5
I¥©jeet Dessriptions
Ob.ieetiyes?
1) To eoHrplete a statistical asaalysis about the intert
relations of the five optical elements ia the h^aaan e^ as a
basis far a theosy on refraction anotaalieso
2) To test that part of the theoiy ^idhassmies that
the tension in the ehoroidg bgr reducing ihe pressure ©n the
sclera 9 is a factor in detes=miniag the sise of the globe o It
is clinically known that ^«herev^» the diespoid is abseatp the
sclera becomes ectatico Quantitative ecnfinnatioa may b© ©b=>
tained by measuring the pressure in the supraehoroidal space
and coE5>aring it t© the intraocular pressureo
fethods molm®ds
1) Statistical analysis c According t© preliminasy
estiisatess three factors will esEplain the correlations between
=> 2 -
the five optical elements = It seens desirable to dheck these
estimates on a computer as so©n as a f aetor analysis program
for an IBM 650 l^eomes avaHableo
2) Ihe s^toseleral pressisre (SSP) is measured tgr i»=
sertiag a 27 gawge needle in th© sR^^'ashoroidal epaeeo Th.&
needle is connested to a transdiaeer assd a pressure head allows
the intrediictioi^ of a djroplet of fl.uid at the tip of the needleo
As soon as equilifariuin is reached between the SSP and the pr@s»
sure in l^e needle^ the outflow stopssi and pressure changes
xaay be recorded fr@m pg^ssur© changes in the droplet. The SSP
is compared with the lOP measured tgr a needle in the anterior
chambers
Ma.1or Hndingss
1) As the lens power in Stenstrom's data was calculated
from the four other optical elements in the human &^s pseudo»
correlations v.s^ M'^e been intrediaced in those correlations
in idiich the lens j^swer is contained as one ©f the variableso
Since ihe amount of pssudocorrelation cannot be directly esti^
mat^ig c^relation calculations were repeated in Sor^drf*s
material in %*iich the lens power was measure^ and the ^dal
length eoenputedo As correlations in Ste»stre«i*s data chedc
well with "Uiose of Sorsbys the amount ®f pseudoeoss^latiois so
introduced must be negligibleo
2) Th@ SSP measured to 12 eyes appears to be lower than
th© lOPo The difference aatounts to between 2 and 6 mmo %»
Para^rmpathetie stimulation (ciliary ganglion) leads to de=
creased pressure and ^nrapathetic stisnulatlon (cervical syn^jathetie)
leads fre^uentlya but not alwaysj, t© an increase in pressuroo
Several investigators consider tJie choroid as too fc*agile
to be able to stand pull aM pressure o However 9 by trephining
scleral windor^s in the posterior pole 9 the ehes^id bulges out
and is considffiE°ably retracted on para^rmpathetic stimulationo
A large scleral window causes a large herniation of the choroid
but even when overstretcheds the choroid is able t@ stand 90 i™°
of lOP before rupturingo
Significance to F^o^aa @f Ihstituteg The reduction
of all correlations between the optical elesaents of the human
eye to three basic factors demonstrates a link between corneal
power arai asdal lengthy asKi at the same times) between lens
p©w^?9 depth of the anterior chamber and axial lengtho An
adequate ©asplanatisa of these factors will mean a step forward
towards a tfceosy on refraction anomalieso
» 3 -
Propesed Cmrae of Pro.ieets
1) To eomplete statistieal ealeulations 1^ a factor
analysis ©f SteastsSm's datai te deteiroine the manbes' assd lead
of the factors tjMerlying the optieal elements c
2) T© eonfiwm ppeliminary pressure measureiaeiats in the
suprachor^idal spaee by an extx^mslgr fine solid type ©f ps^essur©
gauge o
3) T© caifty out a ntHiibei? of tests on v^opiQ and hype3^=
Bietiropie patients to obtain infennatien <m ^sapatheti® aM
para^rmpatheti© acti^i-^ in s^fi'aetion ansmalieso
Part B inelMed Yes /77 ^® IW
PHS ■» KIH
Calendar Yesg^ 19^
ftpinsipal Investigator' $ Bsnas© Eo Cdhmg I'oT)..
CJoopspatiRg tfeitsi Koa®
Km Years (catsndar year 19^ )s Pati«t I^ss Moa®
Totals olO
PS^ofssalsssals =,05
Pro^eet Deseriptiojii
Cbj[^tig§ss R©s©nt stisdl©® p^ml&aslj s?^©rt@d hav©
d»on8t»'at©d th@ ijrsittal patlsissg's of a^^aas htsssosf oatflow ia
the rabMt l?^ raessis of ?©diogs?'Eifcs ef ^® ^® t®k©a aft®? 5!?©=
plaesrossit ®f equ®oaa husBJor Is^ tiatesp seltisOL© Midi0psq[tt© soluticmso
It i» thwj^t that !^ nodifSrisig these t^lmiqties aasd d0^isirjg
nm m®89 father Infomatioa lasy fc© «^tg3sr@a ©mesrairsg ®«
queous outflow eb^mels &M iatf aoedlsr vessels Ib sq sssqss^
tially intset e^o
K^ods S^ly^8
1) %© atnJ@o!2s hvsmr of th® rabblfe w& is s^lased
^:^ th© 8mm ^mbm® of ©oditis diatadsoat® C%psqi«®) md pilet
®ti«ai©s with eon^g^Rticmal laisinagpsf^ls siM ©attaet lasal«!!®«.
gpajtoio t©8hM.«§a©® as^ cearted «Kat u®is?g a Oo3 e»o fosal Ep©t
tub«o
«. 2 -
2) Aftesf esseision of lidsg orbital sfteg prijcarily nas=»
alVs, and tfe@ oassl ox^ital v&Lls fe® anterior ©lliasy vela ®f
th© eat*g ^® is emaulatedo Radiogr^s ag^ tsskesa disriKg
hsod in5@otimi of sodiun di&tf°i@oat@ a»d els® duri^ infualcsi
st vsisyleg speeds with a sepsaK-^p© ©l^tg'ifial inf^ision puspo
3) Ba© sann© tedsasiqii© ®® lis 2 is tised osi ©ats*^^s
wi'&out r®5ioval ©f lid® and b«a©o Cllnieal axsd histologic
studies of th® ^©s ai?© raad® at ^as^iiig intgrval® eftes? th®
xp'Tss^ sjp© al3tslE2@do A nuHts^E* of s%diogj?i^io ,t®chni.<|ii©® as^
©Bpl(^ed to ebtsia th® latyaoealar ^^joas patt^Ttss in tra?ious
pj^Jsstionso
k) Th® @s!R@ pr^aration @s in 3 is us@d in pilot @tadi@3
of laaduagr^^ of th® oat ejjr® dsuriag th© s^tpo^ed© psrftoion
of th® aiitfsd.os' @ili@^ v@isio
LtJ^
Findig^@8 Th® t@s!m34u©® »!©iitioa@d sbo^e bar®
resulted in th@ sucesssfal derEonstrntim of th@ ro©ntg«n-anat©w^
of th® intfsooular v@nous ^stons in th® @8se8%tially intact
oat ^®o The ©rsti?@ eiliaxy v@nous s^st®m ineltsding the iriag
oiliaay body^ ehoroidal and rslated intrascleral eomponents
ar@ ol©srly visible «m the radiegraphso 3hfu®i«m at vss^v^
rates has s;'.sd© possibl® fractionat© intraeeulsr venogrsmso
A nuaSsss? of asseeiat®! int®r®8ting f indijsgs hsw® h^si ©btainedo
^^iB^^JsJSS^WM,^^^l It is h^ that
th© atudi@3 Bisntion^ ribcwe m& lead to better uudfiSK-etasiding
ofs
1) %® assatoagr of th© iBtspasclss"®! v&na^iB please ^ich
is intiB8at®ly related to th® sagehssissi ©f @^®o«s mitfl<^ andg
th®r@fo(r©9 glsaQSBfio
2) Th® anatoaie pattwR md int©j^^leti®sship8 of the
intraselsral pl&sas ^th th© v®s8®ls ©f th© e^Hspy bo^ md
choroid %Moh or© ±ssp9^mt in th© st^ai^ of mew diseases of
the €!^o
3) The d^Qanics of th© intraoeuLssr -rasstslati^reo
k) Th© posslfoilitieffi f og' f^tep© possible iswestigation
of the intraocular v®5ism in m@n %^ mgi.o^^%e Bi®thodSo
th® st^sdies oesoribea in Ketnea j ana %nen t.o ©^seasMs© wixn
« 3
the lisrdnagraphie ss^loratioa of th® latf^soamls? ^©asgrsphic
patternc Aa attescpt vill b© ®sd© t© tstllls® this t©ebsikiu©
to stac^ th© ©ffset of pfesrmacologleal sg©ats ®ad surgisal
l?Et®?iriBisfci©iis on th© lis.ts'aoeulss' vassulatssfgo
Pai?t B inel^eds Yes O" ^'«^ ^7
Serial No,
1» Ophthalmol©^
2o ecology and Hist®=>
pathology Seeti^i
3o Bethesdag Maryland
H-o Same as NINEB 70 assd ?!•
PHS =. NIH
Individual Project Repoi^t
Calendar Year 19^
Part Ap
Project Title 8 Eleetroa Mieroseope Studies on Epiiiieliuaia Capsule
and the Fibers of the Lens and on the Epithelium
of the Ciliary Bo^ and the Optic Merve
Principal Ijwestigators Theodor Wankoj, MoDo
Oth©r 3westigators8 Maiy Ann Gavin^ MoS, aRd
Ludsfig ■van Sallmannp MoD.
Cooperating Units? None
Man Years CealeMar jear 1958)8 Patient Days? Nme
Totals 1
Professionals o5
Others o5
Project Descriptions
Objectives s l) To investigate the normal characteristics
of lens tissue elensnts as seen in the electron mieroscopegE) to
investigate the ixltrastructure of the lens epitheliusi and the
lens fibers after cataraetogenie agents had been administeredg
3) to study the morpholo^ of the ciliary epithelium -sriLth the
electron microscopeo Information on the ultrastraeture of the
epithelium may lead to an understanding of its ftraetions and
partieularlys) ^"^s role in the fos'siation of aqueous hiamoro
Methods Brnplgyeds An RCA electron microscopes Model EMlt
3C9 is used in these studies o Control and comparative exami-
nations are carried out with "Sie aid of i^ase contrast and light
microscopy a
« 2
"Bo® lens of the anesthetized animal ^as removed f^m
the 6!7e in situ a»d imnediatel^r isoaersed in the fixation nediuino
It was then dehgrdrated in a graded series of alcohols 9 d3.s=.
sectedj) and embedded in methaezylateo Mequate seetiens were
cut on a Servall ultramierotoma. JM2d transferred on collodion
filmed grids covered with a fine carbon layero
Ma.lor Findings 8
lo Normal Lens
Subsequent to the study of lens epithelixnag as reported
last years the cortical la^rs of lens fibers were investigated
on rat, rabbity monkegr and calf lenseso The fibers represent
elongated^ prismatically shaped cellss, outlined bgr dense menSsrgnes
and separated from each othrar> bgr a small intercellular spaceo
From the transitional zone of the lens epithelium^) •vbldx is the
equatorial region^ to the fibers and hence to deeper layers there
is variation in amount and configuration of intracellular elements o
A description of these cytoplasmic stmetures follows o
Nuclei 8 In the lens epithelium^ the nuclei often show marked
indentations and the presence of one nucleolus which can be
recognized as an irregular aggregate of dense granuleso In
the bow region of "Uie lens fibersg nuclei are somewhat larger?)
show fewer indentations 9 and commonly possess two nucleoli,,
Her© the nucleolar granules are disposed in thick coilsj, embedded
in a lifter matrixo In more central partsg structures of a
size between 1 and 2 ax are found in the cytoplasmo They are
composed of opaque granular material massed into a boc^3r ii^iich
is outlined by a dense bordero The occasional presence of a
double membrane boundary and association with elements of the
Golgi CoBples suggest liiat these bodies might represent remi'.
nants or fragments of nuclei in a state ©f regressive meta=>
morphosis in deeper lens regionso
Mitoehagidrias Mitochondria in the lens epithelium gradually
increase in number from the center to ihe equator and range
in size from Od to Oo? lie 3h the youngest lens fibersg they
attain a length around 1 f arad are fr^uently seeno In deeper
lying fibers and in the sSdal regions of the lens cortexg they
are sparse and occur in the form of large long structures
measuring up to 3 P in lengt^o
Endoplasmic retigolrmg Endoplasmic reticulum of the rou^,
surfaced type gradually decreases from the central zone of the
lens epithelium, towards the periphesyo In the youngest lens
fibersp it is found in the form of dense profiles with r©la-=
tively large spaces between its meisforaiaous pa&'tso In sectiews
of deeper lying ssellSs it appears smaller and more vasiculars
^ ■}' _,
assd is present neither to the deej^st imvestigated layer® n@r
in th ? p©ri;2«lal i^glmm of the Imw ©ort&xo There is a gradual
increase c-f small elttstg-'s @f s^ibaKMeleaproteiKi granial®® frora
the jTvjmngegt to the elder lesis fiberso Tb@ eliasters are dif-
fuselj dispersed throiaghaiat tte i^^plasmo
N® changes in the appearane© ®f the smooth gitarfased
"typ© @f ©tri@pla,imie retiewltm are R©ted in this i«westigati@R
@f differesat areas @f the lenso
Gelgi Complex? The G®lgi eesnplex is similar in the varimas
regiC'BS ©f the lens inwsstigatedo However? in the fibers©
it appears mere segregated from the auelear baiEsdair^'g and in
deeper eells its indi'^dual e@tnp®nentg are seme^at dig@@eiated
from one another^
Lm? density elements g In the letis epithelimn as well ao to a
hi^er degree ±n the leRS flberse qyteplasmie elements @f
spherical and filameiratoias ghape Kith diameters rasjging trmn
100 to 120 % are obsew'edy. They are eharacteriaed bgr la??
density to electrons and eossatitnate the preponderant eyts^lasmic
stmct\ir© in the investigated, lens fibers o
Ih a joint in=?festigati®iPi witii T)to Ro A, Resraikg leases
were fractionated l^ t£Ltraee!Btrif'62gati@n in order t® find a
residue which contains tJiese low de^isity el©-msnts eorclusi^elyo
F©r this porpose sisc different fractions were obtained^ aM
after f iscatiena entoedding and segtionisngj th^ were studied witii
the electron microscope o The supernatant of a fraction ®b=
tained after centrifugation for l6 hours at l©5j,000 x ge ©oKf-
tained these low density elements exclusively. Ihie fraction
was ftarther subjected to morjcfeological studies and to chemical
analysis o
2.-. Exj^riraental cataract
Studies on the effect of catsff-aetogenic agents on the
fine structure of the lens are being carried aatc At present
the f®ll@M.ng preliminary results are availableo
X^Rgjy Cataracts s Rabbit lenses were observed after e^os^jre
of the <&^B t@ 1300 rad »=rayso " Four d^i after treatment, a
few cells Jji the peripheral aone of the epithelium contained
rather l©ng (2 ^i) latranellated str-' atures which after & 1@ d^
interval seemed t© be transformed int© mltochondriao Another
finding 10 d^s after ii'radiation ceiisists ©f structural changes
in mitochondrial ttiey appear as ballooned?, club- shaped « and
el®ngatedo In one cell a dense congloauerate of granular and
membranous configurations around a lighter homogenous core
^ fy ^
was seen opposite the aueleuso In se'^eral ©ellss Buslear inat=
erial is located ©utsid© the boundary ©f th© ©ell nueletago
F&arteen da;^s after irradiatien seyere changes ean ®ci«jr in
both auclei aM qyt@plasino Kuele@plasin appears agglomerated
into dense masses in a lighter mateixo Ihe nuclear tnembrane
appears as a wide bansdj, without dojtole membraHsd e®at®m's arid
with its eontiauity interrupted bjy as'^eral largSg circular
epeningso Ih the eytcplasnip profiles ©f the eBdeplasmic reti=
Guhm appear larger thaai normal aad a eensiderable inerease
in ribonucleoprotein granules ©ceurs siHiuLtanesusly with aa
augmentation ©f lew density filai^ntso Other «grt@plasinie €°«©;o
ments appear reduced in numbero Generally^ it may be noted
that the transformations in the lens epithelium after irrad=
iations, ©csur in some areas alternating with apparently normal
Eyleran cataraets C2-^^6^ weeks)? Rat lenses were prepared
from an; mals fed from a diet eoataining Bcrleran for 2g ^9 and
6 weeks o Ihus far 9 observations have been on lenses ®f animals
that w^:»e sacrificed after tw® weeks on the dieto Ihsy reveal
a great quantity of low density filaments in the s^oplasm ®f
the lens epitheliumo la areas beneath the epitheli\im se'^ere
damage can be observed in the f ®rm ®f conplete loss ®f lens
fiber eytearehiteetus?© and the deposition of a dense am®rph®us
substaReeo Oeeasionallyg posterior parts of lens epithelial
cells are included in these focio
ttoosine eataractas Rat lenses of animals subjected to a diet
Gontalning mimosine for % ?» and 14 days ha^e been preparedo
Observations on lenses of the first grm^ ^sw that the rough
surfaced endoplasmic reticulum ig transfosmied to large eystemes
which present a vesy se^irplicated three dimenaional conf igurationo
At the same time mitochondria appear swollen with a displace-
ment ®f their internal crista© t© th© p©rii*ieryo In the nueleig
dense;, irregulars deposits can be t&m& at th© site of the
nusleolio Ihe regiosnss observed in the seeosad and third stages
of this es^erimente in the lens epitheliucig do not display
©easiderable ehangeso Extensive changesg howe-f^erg are seen in
the region ©f the lens imv in the form ©f Intracellular vac^
u©lizati<Ki5 destruction ®f several lens fibers with irregular
deposit© of a dens© mat^'ialo
Maiy Ann Gavin in cooperation with Bo Jo Hoyda Jro (NCI)
developed a method to iuqjrove seetiessing of tissue for electron
microscopy investigationo In this procedure a knife broken
from l^or brand plate glaas is used instead of the conventional
kinds ©f plate glasso ^sreor rssewMes fused quartz in its phys-
ical and mechanical properties and has the advant> %e of cutting
satisfactory sections of brittle apeeimens and generally main-
tains a usable cutting edge for longer periodso
° 5°
Sigaifl©anee t@ I^ggaai ®f Institute? Tne completed
investigations ®n th© e®rtieal lenm fibers together with the
pre^eus study on 1©ks epitheliuin provide itJiformatioia on the
general fine arshitectuff^ and eellular ©rganiaatiem @f the lens
@f the normals, mature animalo .This series as a basis fer G&m-
paris®n in stxidies ©n e3g>erini©ratal ©ataraets whidi h.&w& been
initiatedo
Th© jeint investigation with ]>o Reanik led to th©
ehemieal definition ©f qytoplasmie eonirtituents that are tnier@=.
scopically visible and represent the soluble lews proteins o
Exposed Ceurge of Pro,1eet8 Observations on experiments
ally^indueed eataracts will be continuedo The joint investi-
gation with DTo Resnik will be extended t© determine Aether
solt&le lens proteins san be identified as individual m©rpho=
logical entities o Stu<ty on developing lens and tissue culture
material is eontemplat^»
3o Ciliary Body
Kethods Bnployedi An RCA electron mies^iscopej Model
EMlI=3Cs) is used in these studieso Control and comparative ex3m=
inations are carried out with the aid of phase contrast and light
microseopyo
An extensive study on fixation methods has been init=
iated in order to establish the most suitable means of preserving
this structure in vivo and in situ during functional changes
induced by sympathomimetic aM para^Tnpa*^.©mimeti© drugs s easi»=
anhydrase inhibitors and the enzyme chymotrypsine, For this
puarposeg the eyes of anesthetiaed albino rabbits have been
infused throu^ the eannulated anterior chamber with buffered
osmiiim tetroxide of vasyis^ concentrations f@r 30 "to 60 minutego
]h another series the same infusions will be repeated on ^es
^ere a eoloboma has been surgically introduced to secure pen-
etration of the fixative into th© posterior ehantoero A parallel
study of histological dianges is planaedo Ihe speciirssns thus
treated will first be ©xamined under the light mieroscopSo
Ma.1or Findings? None
<=. 6
^^£J:g,Q»g® t@ Progiiram of Jaatitutes Developmsat ©f
an adequate preeediire tar such a st'ucfy tdll als® allow a sim^
ultaneous investigation of other regieas in th© ^e bordering
the anterior and postafior chamberso
Proposed Course of P^:1sets Stuc^ of l^e cilia^ epithelium
under the aspects indicated in the meth^So
IIIo Optic Nervec
This project has not been continued in the last yearo
It will be resumed as eeaipletien of other projects pax^siito
Part B ineltided Yes i^' No £^
Serial N©o N^.: —
PHs ^-> wm
Individual Project Report
Calendar Year 19^
Part Bo Hcsiorss Asrerdsg a'od Fublieations
Publications other than abstraets from this projeeti
To atsd Gavinj, MoAoS Ihe fine structuy© of the lena
epitheliuHio iki eleetrea sdcffoseopie studyo A„MoA« Archo
Honors arKi ^ards relating t© this project J
Serial Ho. NINIB=.S2 Ce)
1-. Ophthsltn@l@g5r BrsRch
2o (^ii©l(&gf and Histo-
path®l@g5r Saetion
3o Bethssdag Masylarai
ho Same as KINIB-.72 Ce)
PHS » N3H
Individual Project Report
Cal©adar Year 1958
Part A.
Pr©;3ect Titles Electron Kicroseop© Studies on Bicpsies ®f
Humaa J^cle lUseases
Principal investigators s Th©odor ¥ank@j, KoD^
Go Milton Sliyg, MoDc
Other !&w©stigat®rs8 Masy Ann Gfa-via^ Ma So
Cooperating Units § ' J^Ione
Man Ysars (calendar year 19^ )s Patient Dajss
Totals olO
Pi'ofessiDnals 0Q5 "^^^
Others ^0$
Project Dsscriptions
Objeoti^esg T© deteritsine \^ means ©f the electron
microscope detectable changes in ^arionxs human c^strephi©
Biuscle diseases as cess^ared with normal tissuso
Methods Eroplqyeds Ifeiterial obtained from traisele biopsies
on htiman sabjects wjth muscle dystrophies ®f ■various kinds are
iinmersed imsjediately after esccisiess in V^ soditam tetread.de for
fixationo iSiis is relieved by ddwdration sxyis in s®me eases 9
by additional impregnation with 1/6 ph©^h©t%mgstie acid;, iffi»
bedding in methacrylate and sectioning on a Servall \jltrarniere=
tomeo
Aa RCA eleetrtai sd.eroscopsg Model EMU-3C9 is ns&d in
these s'tudieso Control and cosjiparative ^caminatimss are earried
out with the aid of ji^as® contrast and light aiieroseopyo
■=" 2
Ma.1©g Findtogss
1) NoOTial muscle •= Spseim^as ©f feur noOToa), hmiats sub-
jests have been investigatedo Ihe results generally are com=-
parable t@ those obtained from ether ■vertebrates wiUi a ©lug--
gesti«m of the presence ®f tw® types ©f ngr® filaments (primary
and seeesKiaey)o Special atteratien has been gi'^s^en to the ©rgaxv-
iaation of the ir^yofibril at the Z baiwl levelo It is felt that
a double nsmbraned structure with a tortuous configuration
separates the individual aareomereso It seems t@ eoiAead h^©&5.
the limit of the nyofibs'il towards the sareolemma andj, on toe
other sides, t© adjacent EgrofibrilSo Korphologicallys it is
barely distinguiahable from the sarcoplasmic reticulums at the
present time it cannot be decided xiheth^r these two elements
are parts of one systemo 3h mm respect the human samples
appear different from animal tissueo All observed normal ss^
pathological biop;^ speeteeas of adult pea°s©ns craatain a nsmber
of cytoplasmic inclusions in mwscle eellsg capillary endothelial
cells and to a lesser degrees in p^Acyteso These ^jiclusions
vary in size between 1 and Zjx and consist of an agglomerate
of vacuoles of various densities and opaque granuleso
2) %©tonic cystropl:^ ==> Four cases ®f uyotonic dystrophy
trare studied and the f olloadxig characteristics ebservedo In
small muscle fibers cjiains of central nuslei are seene Th^
are frequently surrouaded by f ilameatoxis structures arranged
in bundles which sometimes display cross bandings that are
similar to normal striated muscleo Sonetimes they appear in
directions perpsndieui^^ ^^ °^® snotherg suggestive ©f ring
f iberso The small muscle fibers also coatain numerous mitochon-
drias, sareeplasmic reticulums ribonusleeprotein granules andj,
in a relatively hi^ suiriberg d^ise agglomerates described be-
fore in s^rmal muscle c> ^ other muscle fibers of apparently
normal siae the follosflng structural traasforroations are ©b=>
served! Increase of s?ib©nueleopr®tein gramiles. sometimes con-
centrated under the ssircolemraa eomparabl© t© the sarcoplasmie
pads knoan fTom li^t microse^se findings o Rarefication of
mitochondria assd changes in their internal «»'ganiaationo A
pattern of cross banding at the Z disc v^ich differs from the
normal and Qicea<9i®nally assumes the appearance of a double Z
disc with a lifter space between two dense contourso Ifyofil-
aments can be more widely spaced than normal which might rep-»
resent a reduction in isumber^ In e^-d ease ring fibers were
dbservedo
3) Werdnig^Hoffinan's disease - Preliminasy observations
on a cMld shoa? elub».^:iaped mitoehendria of a large siaeo S©m©
present an aoparently normal configuration while the bouadariss
of others seem ' disX'iended awi s!H=r©u'?Ki areas #ii©h lack organic
aational c'etailso
» 3 »
Slgnificaace to Trop^axa of Institutes It is p©s§ible
that sigaificaat mospholegical differences between nonnal assd
dystrojiiic rmscles rtsight yield s€M3ie insight in the qytopatholcQT
involved in these diseases o
I^pesed Course of Projects Studies on Rormal musele
will be continued with special attention beii^g givsK t© struc-
tural details at the Z disc levels presence of tws inorph®l@g-
ically distinct t^pes ®f nyofilaments and identification of the
dense s, heterogeneous agglomerate in inusele fibers and blood
vessel walls o
Observations on inyotonic dystroj^iy will be centismed
Further studies will be carried out on samples of Werdnig-
Hoffmann's disease arad subsequently on other spaeini^ns c©l«=
lecteds of familial periodic jsaralysisj, progressive muscular
dystrophy and other neuro-muscular diseaseso
Part B included? Tes FJf No fW
Serial M@o NIMB^g^ {s^
lo (^tha3.TB@logy Branch
2o ecology aM Histo-
pathology S@etioa
3o Bethesda« Ma^^laM
ho Same as UINEB-?^ (©)
PHS => NIFI
Iisdiiridual Fi^jeet RepoH;
Calendap Year 1958
p^u^
Project Titles Stut^ of Submicroseepie Struetijires of Qcular
Pigmetjt Cells. C Staging of the Livittg Tissue
Cultur© Pigment Cell hf Acridin© Change )=:
Principal Investigators s Ko Ko W®lfs> HoD, and
SsBRiel Bo As^OBSOHs) KoDc
Other Investigators 8 Le© Cayavaggie^ KoSo
Cooperating Ifeitss Dr. Ko K, Wolf^ Lab®rat«fy ®f Ke^sr©-
anatOTical Sciences^ NINES^ "The Sig-
sdJficasise ©f the Aeridiae Orange
Staining of Neus^Bnes 3h Vitf=® and In
Kan Years (calendar year 1958 )s Patient Dayss None
Totals clO
Professionals 0O5
Others <.05
Project Deseription $
Objectives s
1) t© establish the fluorescent iisage ^ich th© living
tissue ctjltm°© cell will display when stained with AO andp
2) to define changes in the image produced fcgr injus?yo
Kethods Emplc(yeds Tissue cultures of chick heart fibr©-
blastsg choroid pigment cells and eHiary body pigment epithelial
cells were raised In the Paul CJhairiber in the preserasse ®f staining
coneentratiens of AO arid the fluos=eseent image ^served at
regular intervals thrms^ the lifetime @f the isuLtureso The
granulo^kinesis of living pigment eells pro'ijlded an is^wrtant
eriterion for distinguishing betweess liirisig astid dead eells in
these experiments o
Ma.lQr Findings? AO is toixie in tissue euLtur© at the
concentration ©f Is 100^000 3 but pennits continued growth and
good cellular health of eultiares in a concentration of 1?
IgOOOgOOOo The AO has a photodynafiiie effect in stained ©ul=
tures rendering them more susceptible to light in^iay t^ian
unatained controls o Healthy AO stained cells ^en first il-
luminated show green (orthschromatic) fluorescence predominantly
of the nucleus and nucleolus o With continuing illwnination
the cells become brighter 9 and red (metachromatic) granules
appear in the cytoplasm^ At this stage light injury is still
reversible o If illumination is continued the nucleolus and the
entire qsrtoplasm acquire metachromatic fluorescence© and at this
stage Sie cell is irreversibly injuredo ^Oie metaehresnacy of
irreversibly injured cells is like that stained in fixed AO
stained preparations at controlled pH and probably is due t®
RNAo Ihe metachromatic granules in reversibly injured cells
probably are not RNAg because the cells ebsers?ed do net e©n=
tain granular aggregations ®f RNA large eneugh t© preduee the
image observedo
Significance tc PSrogram ef Ihstituteg The ©rthoehromati©
and metachromatic fluorescence of AO are knmm t© cesrespond t®
monomoleeular and associated states of the dye respecti^elyo
Study of the staining ©f living eells may provide important
information about the chemical state of the components ©f living
eellso The use of the Paul CSiamber in these e^q^eriments per»
mitted more rigorous definition ©f criteria for cellular health
than has been possible in previous eatperiments with this c^eo
Proposed Course ©f Projects The eatperiments are @©ii?>lete
and are beir^ prepared f®r publisationo
Part B included Yes Fl' No /W
Serial m. ^IMB^^ (&}
lo Ophthalussiogy Branch
2o Cheaaistry Section
3, Be£besda, Maryland
4o Same as KIHDB~75 (c)
PHS - NIH
Individual Project Report
Calend&r Year 1958
Part Ae
Project Title: A Study of the Proteins of the Lens
Principal Izrvestigator: Robert A. Resnikj Ph^D,
Other Investigators: Theodor Wanko, M.Do and Edith Kentoas Bo So
Cooperating Units: Hone
Man Years (calendar year 1958): Patient Days: Hon©
Total: 1.0
Professional: o$
Other: o5
Project Description:
Ob.lectix'^es; To study the chemistry of the lens proteins,
the structure of alpha crystal 1 in, its interactions with other
molecules, and its properties in solution.
Methods Employed; The techniques of ultracenferifugation,
Tiselius electrophoresiSj, and spectrophotopstry are used to
characterise these proteins « Other techniquesj, such as equil"
ibrlum dialysis and viscosimetry also axe employed to obtain
information concerning the properties of these icolecules.
The sise axid shape of the lisjlecule calculated from the
diffusion data indicate that it is an asymsstric molecule vxth
ascial ratios of 1:15 or 1:20 depending upon whether it is repre'°-
sented either as a prolite or an an oblite ellipoid of revolution^
The viscosity coefficient of 12.1 is in agreeii^nt Mth these
general results, indicating that it is & thlnj, elongated sctoleculec
2 »
The effects of pH, ionic sSreagth and solvent have
been studied aad sisggesfc that alpha. erysSallin isjsdergoes
trsnsfoztaatioa ia solution as £h©Ee factors sre varied »
For eKsn^lSj, two Gonip©nen£s are present in the sedisEsntafeioa
pattern at pH 3ol in 0„1 M glycine btiff®r„ At this same pH
in 0,1 M sodium chloride only one eos^onent is se&tXo In
additions between 4.5 and 7.8 the ©ediasntation is asyimaetrlc,
while ebovg pH 8 it is syasB&trical. These data are inter-
preted as indicating the presence of subunits which under
certain conditions exist in equilibrimio
Ma;ior Findings; Additional studies indicate that tha
value for the sedimentation coefficient of alpha crystal 1 in
is 17 oO s 10"i3, ^ig value is slightly lower than that
reported previously but is bas®d on additional, data. The
HBolecislar weight of alpha crystallin is 900,000 to SSOsOOOo
These values and parameters^ previously reported @s the
diffusion eoeffieiest, and the apparent ^ecific volumes are
similar to those reported by Orektevich ^ al«
la collaboration ^ith Dr. T« Wanko electron micro-
scopic observation of isolated preparations of this protein
and the lo^ density eleiaents seen in sectioned lens fibers
has also been carried out. At the present tims it is not
possible to state whether the elongated structures in prep-
arations of theb^ density elemssnts are alpha crystalling
one of the other soluble lens proteinss or the results of
interactions between these tsolsculesa In any events the
chemical aaalyass of these low density structures indicate
that they are proteins aad that the soluble lens proteins
alpha, beta, and gaaaaa crystallin are present.
Sigaific^aee to Program of Institute; It is ikjw
possible to assign a definite siz@ and shap® to alpha
crystallin. Additional data for beta and gansia crystallin
are sot yet available due to ths difficulty of separatism
various coa^onemts of thss® two groups of protein,
Ifee infonnation now avaiiSible for alpha crystallin
may provide some basis to correlate chemical changes in th@
lens to its transparency.
The low density elemeatSj visualized by the electron
microscope^^ill provide important information about the sise
and shape of alpha, beta, and gass&a crystallin. This may
penmit an evaluation of the soluble lens ptoteins, as related
to the transparency of this tissue.
Proposed CourBe of Project; The effects of pH^ ionic
strengthj and solvent will be ixrvsstigated in greater detail
in order to bbtain more infonastion about the effects of
these variables or the sise and shape of alpha cryst&llin.
Electron microscopic studies, in collaboration with
Dro Wanko, on electrophoretically isolated lens protein
as well as the low density system will be continued.
Part B included Yes CU *> .M7
Serial Woo„, MINIB^^g ,Ce) ,,.
lo Odithsiiriologjr Bram:^
2o Ch©ffllsribsy Action
^o Saw© as NMI®«76 C«)
VES « KIH
Individual Pro jeet Heport
Calaidsr I©&2? 1953
Part.A,
Project Titl©« An Investigation of th« En^matic patens F^@®nt
in the Lmsjj Cojraea and Aqueous KuiBor and their
Relation to to vivo Tlssu® Ketaboliswo
Principal Isvestigators Robert Eo S^laans MgDo
Otfeor 2m®stigators3 Robert Ao R^snikp Bio Do
Cooperating Chits* Nonso
Kan Tears (oaleadar y©ar 1958 )« Patient D^@3 None
Total* loO
Pjpofesaicmls 03
Otfeeg'J o5
Project Desoription*
^TO^gsJ Itei® pr©5s;«it ha® b©®a dlvidsd tot© few
partgJ
1) M investigation of tia© variatim of laotie d©hy=.
drogenase In the corneal ©pitheillusa of diffes-snt ^ecieso
SMymos closely related to lacti® d^drogmes® in asstabolie
pathway v&^9 also @tadl@do
2) Radioactive trsoear i^p@rlm«tg ^'bs^ xmdfsrtakm to
gain an xmderatanding of the ®ns^B@ conte§:at iis th@ eor-n@a in
relation to it® acstual glucos© eatabolissq
3) The ©ff®ct of tnod«s^t® domBs 1000 r-. of s-ragrs
D^pon sp®oifie ©nsyaass in ^® l©ns aM eor?j®a was flrthes* iii5»
v«stigat^o
- 2 -
k) I'leroehemical pswcedures f«p th© assajr of ©is^roe®
and metsbelites in miero samples ©f squeoiss htKusr snd t®sa?
fltJld as?® te«l83g de^elepisdo
Methods Enplcsy^g. 3« psBfts 1 and 3 of this project
th© activity of vario«s sn^mes wasi det«r!B5jiied in diss®8t®d
areas of fS?os®n dsded sections of leas and cornea W8l«g
Bicrotfiiainioal tedaniqueao
Part 1 • Lsctlcg aaliCg glixso9^6-ph<Jsphat®e md lso»
citric d^^rogmm®, h®2okiaas©» aldolass iSifid glutathica©
r®do«t^'9 w®r© issfs'estSgat^a in eomsal ©pitfeeliam S^em ratp
cat and rabbito
Part 2 - ^© o^dstiv© metabolic pathwsgrs of tfa® #iol©
and d®«epith©liali8@d eemea war© ©tudied W ^s «©© of^^
glueps® l«^sr-gl«oo®© 2"^^ 9 sad glucos© 6^0'-'^ ss^ 1-C^ o
2«a'-\ and 3«cr^ labellsd lactatso l^osetto^s laodified fsota
those of Bl©«Ka and St©ttm w«r® t^ed, and the isotopes tser®
eo«at«d with th® liquid seintallation co^mt^?.,
F&et 3 «> The lesis and eomaea of anijnale ©^essd to
1000 V. of x«rssr wesp© assayed for their ccsjteiat of gltstathion©
reductases heisbkinases, and Iswcitrio d^isrdffog®sa8»o Whole
lens hoffiogenateso aa^l dissected areas of frozen ck^issci etiiVit^^l
epith©llum %!SE« usedo
Psrt h » Fluorcmstrlc ass^ irethods for lactlCs malic
and gluco3©=6-ph0Sphat© dehydrogenase and lactic acid are
being adapted to stiidy biopsy ^ecimensp aqueous hwior and
tear fluido ^bdifled Levl-Lang constriction pipisttea and
special tnicro=»glassware are being utilissdo
Ka,ior Findings 8
Part 1 =- Of th© three species iavsstigatedg ratj, rabbitj
and cats the rabbit corneal epithelium had th© highest general
levaO. of emymatie activityo Two enzymes of th© citric acid
cycles, malic and isocitric dehgrdrogenas© ^-ere fcsund in all
three groups to be present at levels equivalent to those
present in th© cellular aspeas of the brain and retinao wall©
all species also contained glucos©-6=phosphat9 d^^fdregmase
at a level equivalent to brain and retina^ sadolas© and hesoo
kinase were looero The rabbit corneal ©pitheliuBi was ©oat
unusual tgr virtue of its vei^ M^ lactic dehydrogenase act°
ivity which was 20 times greater than that in the ©at or
rati th© values were 10 tiims greater -^an those reported in
other locations or in other specieso InTOstigatimss of the
"ichaelis constant and pH optiroum for lactic d(^:^clrog©ns,s©
from corneas of th® three species^ irviicat© that th©r© is no
slgnlficsmt spseies vssdation in the csharaeteristics of this
Part 2 = The i^ol© cornea 03d.dis@s glucos® at & rat©
approodmately one°half that of the rat li's'sr or diaphrsgsno
1h© prominence of the direct ojcidativ© ^skimt in corneal
metabolism is conf irmedo In additiost^ it has be@i% d@monatrat@d
that the cornea is able to oseidise .liictat®9 and m^ do so ®v@n
in th© presence of glucosso Retfiotring the epithelium reduces
the ability of the ccmea to oxidia® glucose in ranges from
88 to 97^8, depending en tW© pc^sition of th© C^*** lab®lp ^ereas
lactate-^S^C oxidation ls\tr&d^c<&d only 275^9 indicating aomsal
stroma has tnor® capacity w^' the osddation of lactat® than
for glucosso
Part 3 =■ IhotJg^'' jnarked inwrphological changes have been
not®d to appear in -the l©is and corneal epithelitsan following
irradiation^ there was ho alteration in the ^ol© lens content
of hexokinase or glutathione reductes@o "in addition^ ptxte
cellular areas frorc irradiated corneal epithelium ehot^ no
alte(rati«:i in their ccjntent of hexokinasep glutathimi© re=
ductus or isocitric del^drcgsnaseo
Part H' - F!r©li)inlBary iiavestigstions of lactic and
malic ddigrdrogenase activi^ in th© asiueous huiaor froir a small
gr«sp of rabbits Indicate much indi-^doal "\rariati0n9 but
iralic debydrt^enase is the more active ensyaie (average of
23o9 rxH'/lE) and lactic d^bydrogenas® to be less active {lo96
mf/lH), Volumes of 20 _^1 of sample were used for atu«^ and
this can be reduced te^foldo ftfocedures for glucose=>6=»
phosphate dehydrogenase and lactic acid asa^ are being de-
velcpedo
Si^nifieanee to f^ograro.of Institutes thrcugh an
understanding of the biochemistxy of aqueous huxrsor and ooxneal
tnetabolismg information pertaining to the xnschanism of corneal
hydration may be obtainedo Sadh information wight b© valuable
in th© understanding of some rare forms of corneal dystrophyo
3h part 2 of this work the importanc® of lactate to corneal
metabolism is desBonstratedg as is th© relative d3.stribution
of its oscidatien betwarai ©pithelium and stromso
PurtSierj, the failure to find significant enayme damag®
to result from therapeutic levels of X'-rays is iuportant in
the consideration of th© mechanism of radiation cataract f©3Hn=
ationo
» it
Proposed .Cotars® of .Py^jogts Both tbe study of SEiaywe damsge
resultixjg from 3C"irradiatton to ths lima and csomsa asnd th« in«
vestigation of tJi© speci®s variation of laetio dsbginap^enas©
and oth®r ©nsyniss pp®s@nt in th© eoraeal ^itheliam laav®' fe««^
coneludsdo F^s'th®®* in^estlgaticai using radioaetiv© l®b®ll^
glutamats and nc amino butyric asid will bs trndessftskaai to easplor©
th© poseibllilgr that these sabateat®s also m^ b® isaportmit
to corneal metafcolifflEo
An effort is also being snad© to cosrslat© the cajfboiqr*
drat« Ketabolisin of corneal stroma vi^ that of other tissuss
ridi in eollag^io
Niero methods for ensiytne assssys end th@ dsterminaticn
of intemffidlatss of cascfeotoiyxirat© inetaboliian in biopj^ spsoim^ss
of eom®a snd ssinplsa of tsar flrsid sisd aqiieoaa) b^m»r of 2 to
10 vl volvsm are being d»v«lop©do Thes® earn then b© applied
to available patient materials and th® altered rostabolism of
pathological states roay b® iJSV©stlgat@do
Part B inolud®d8 Yes /§7 ^ fT7
- 5-
PHS - NIH
. Calendar Xesr 1959
Part B 8 Honorso A^aifds and Pwbllcations
Publications cliier than abartpscta tram thia pnj;}©ct8
Kahltiusn9 HoEo mi Resnikf, ReAoS Qosntitativs bistoebeoieal
chmgss in th® dsvelGpnsnt of Um rat l@ns erA eom®&o Anio
Jo c^thog M8^7«555 19580
Honors and awsrda relatiag to this project? Notie
J^rial :io„ NIWDB-56(c)
lo Ophthaloology Bz-anch
2o Ib.ysiolog7 sSection
3.1 Bethesdag liai-jland
4o ijame as IIZ4IB-II (c)
Indivichaal i-'roject Reports
Calendai' lear l'/5fc
i-Toject Title? wiectrophysiology of the %■©
Friacipal Investigators I!oGoFe PuorteSj, lUD,
ether Invest igatorss Peter GouraSp IWDos and
.vyoji Tasaki, II<,Do
yoopei-atixjg Units: "Ion©
Ixin "fears (calendar yoar IV 5C) i'atient Dayss Hone
Votals 3
Pixjf CiDsional s 2
ethers 1
Project Desci'lptions
0^j,gcy^2^t The scope of the present investigation has
been expanded from a study of the f eattu'es of the activity of
visual aerv© cells in the eye to the Bore gensral probiaci of the
transducer action of sense organs o There is reason to believe
that the processes leading to generation of impulses in receptor
organs aay bo appropriately subdivided in two groups s specific
transducer effects <. whereby external energy is traaefonaed into
a change capable of stixa^ating nerve cells j and n,eig'aJ|. processes r
whereby nerve cells discharge iiapulaes ±1 a manner related to type
and intensity of the etiimsluso In some cases (for instance in
free nerve tanainals) the transducer action is accoiaplished hj
the nerve cell itself j in other cases (for instance in the eye)
it requires activity of specialised receptor eleffisats e^ermtl to
the neurones producing impulses o
l^^^JS-^M'S^^S^M' .-.spsr-liaents are xjox-fox'^isd by Dr„ J'larotos
on invertebrate . eyes (LiimJAis) and by Drso Goui^as eaad I'aaaki on
the vea-tebrat© retina (toad and f iah) o In addition to tfce con-
ventional tecimiqixes for intracellular i-Qoox^ingp a nothoa peradt--^
ting intr-acGliulai-- application of electric currents is routiriely
employed. It ia therefore jx>soibl0 1) to teat the effects of
light? 2) to stisdy th© stisaalatiag effects of ol®etrio eurrocts?
3) to analyse tm interactions bstweoa Ilgbts smd eurTsatso la
this way direct infonmtion can be obtained on i) passive proper^
ties of visual norve cellsj 2) physical correlates of the proeessse
leading to their activation « In additions, cojj>j;)arlfJon of the effects
of light (wliieh stisialate ners/e cells thrcugli a pi^tochojiical
action) and those of cujfi-ents (which stii2sxlate newc colls directly"
provides indirect infonnatioa on eom© properties of th® transducer
effect of visual receptor organs o
f^gj^^^j^^lgggs Dro Pt. Gom^s has brotight to a sonelusion
the work initiated laat year oo the rslations bstwean sloy el®C"
trical waves and iw^ulso activity produced fey lUiunination in
amphibian retina „ Potentiala wore recorded by laeans of rs©tal or
glass EJiGS'oelsctrodee either from th© surface or froa desp layers
Of the retinso The slow electrical activity recorded fj'oa the
surface is sinilar to_ the eloctroretiriogramg presenting a sharp
surface negative wavo'j (siiiiilai' to the A^i^rave) followed \rj a
slower surface poaltive wave (siQi3jar to th© 3-uav8)o Depth
recording shoi^s that both negative and posdtive %ra.vos have
vaaxiEJal uEiplitiKie in the ganglion cell layer and that the negative
wave is closely related to ii^olso discha:i:-ge froB tiaess qq11s<,
,ihen the ©lectrode is noar the receptor layer only a slow positive
vffivo ia recorded o These results jmggast that both ths ganglion
coil3 and recoptor cells produce electrical potentials dui-ing
illujaination and laay both contsdbute to th© electroretiaograme
i^ilo pei^foraing this researt^hp hVo Gouras ofeseryed tiiat a
phenoaeaon coiaparabXa to Leao^s "cortical spi^eading dspresfiioa"
ocCTJTB in tlie esxsiasd amphibian rstinao A color change occurs
in SOBS part of th© retina and spreads slowly to gi^oater and
gi'oater areas o Ganglion cella at the front of this uavo present
intense impulse firing yhish cfeanges into profousid dopreesion as
the wave pi^ogroasea upon themo This process is spontaneously
revere iMsp recoveiy occui-ring in ^-l? minuteao ISTo Gouras
points out thEt there are aoss© Jjateresting aiaiilarities fe^tueen
this phonojasnon and Jacksoniaa epilepsyo
In the ©ye of Liiaulus it i& found that f roqueney of
4Epj3„3es discharged in response to nattiral stiJmLlation (light)
ia an approxlEate linear function ©f the logarithm of light
inteneityo The frequency of diocharge of tte Bass© cells in
response to depolarising electric current iSg however^ a eiapXe
!,«4 '?'.S !..■•?«•>>;?;;! f-^,Kr< r^T . '^\'rp^.P'^ffr«»>i- p. f • '>S3':^.i".fii'^s-.+<r'r
3 -
linear function of cm-rent intensity. It uppeca-s therefore
that the logarithriic tronsforaation which ia tyi^ical in
perception (Fechnor-.iebor lay) is oscei'tcd in this case b/
thfj photochenical proccaseSo I'ollowiiig otiriulation with lights
of supraliulnal intonsity^ one records fs-oni vieuol cells a
sustained depolei-isation with suporiiaposed iaptJlaeSo ^Jith
subliuinal intensity of the nattiral stimulus only a sustainod
dcpolai::ation is recorded « It can bo thoiight therefore that
light evoiiQS firing of nerve cells by depolarising their
cieiabrGiieo It becomes then important to find out how the photo=-
chexuical pz-oceases initiated by light in the photoreceptoi"
stixictures of the eye evoke depolarisation of nerve cells o
x'inalyais of interaction between light and electi-ic currents
giv^s indiatjct Imt convincing evidence that the depolaz'isation
evoked h^j light is the consequence of a change of conductance
of the nerve cell's juerobi-aaes and isc^edance ia&aaurea©nts per«»
forraed >?ith the bridge-balance method^ using slow frequency
alternating current Ss> show directly that a change of meffibraae
conductance occurs during illuniinationo By contraatp no
conductance change can be measured when electric currents of
either- direction are passed through the meitibraneo For these
x'easons it is thought 'that the change of conductance occurring
in nerve cellajy following li^t stiiaulatlon^ is induced by a
chemical agent and it is suggested that this chemical agent is
libei'ated by the photoreceptor structures under th® action of
light o
It has been fotuad (Svaetichin) that certain structm-es
in th© ©ye of fish respond with a depolarising potential change
to lights of certain wavelengths and with a hyperpolariaing
potential change to other wavelengths c This icroi-^iiation has
groat inportance for understanding the functional organisation
of the retina and th© processes underlying color vislono Un~
fortunatelyg vosy little work has been done on this problem ao
far, and natui*e of the Qtructures generating these color responses
ave still uncertain o DTo Tasaki plans now to perfoiai experiments
aiiaed at identifying loocfcion and nature of the sti-uctures respond-
ing to light with hyperpolariaing potentials „ Dro Tasaki Joined
the \mit only last ^pteiuber and has obtained no results so faro
ri^(B4f4ffi^?^ t9„.Pyosyaij^,.9.g.;i^s.tiai;ts: The worU described
Eiay further our understanding of the following processes! 1) I-cchan"
ieni by which rhythmical trains of impulses are generated by nei've
cells following sustained depolarization of their moiibraneo It
appeal's likely at the present time that different parts of the
sasi© cell may be specialized for production of either graded and
sustained responses or all-orniono iEpjlses; 2) I'echanisas by
which photoreceptors evoke nerve cell depolarisation,? 3) Ftmctioa
of the retinal eleuents interposed between receptors and ganglion
cell layers f
4-
£'xppo^<$dOem3e_()t Prciject i In the -biork on Liimlus an
attaupt will be iiade to identify tho proportioa of ecssntric
cells (iri-th a largo axon) arid retin\jla colls (ijitix a sjaall
axon) 0 Kiubliainai rusponsoa to light will also bo aacdyaed
with the pui'pose to widorstand sosao foatiu-es of the activity
of pbotoreoeptorso rhe work on fioli will continuo as plooaed
as long us Dr« Tasold. j'eaaina with the section.
Fart B included Yes ^ IJo £7
PHS - NIH
IMi^dual Prejeet Report
Calendar Year 19^
Pjart Bo Honorsc -Awards aM Publicatioias
Publications other than abstracts from this ps^jeets
Fuortesa l^GoFoS Eleetrophysiologsr ©f ths visual ^stem? a
syraposium C Editor Is Amo Jo (^hthc, h6zla.l82 (Pt, H)? 1958o
Pu©rt©So KoGoFo s Generation^ conduction and transmission of
nerve iiapaLses^ Archo italo Biol.^ 24*285=293s 1958o
Fuortess MoGoFc s Th© Annual George Ho Bi^op I«etur©9 Generatioa
©f nerve impulses in receptor organs^, EEG Clin^ Neurophysiolo ©
±in-73, Suppl. 10, 19580
FuorteSg KoGnFo? Electric activity ©f cells in the ^re of
li.ffial.us9 Amo Jo Ophthag 46s2l0=223 (Pt. IDs 1958o
ruos-teso KoGoF. s Sjitiation of impulses in visual cells of
limuluss, Jo ?^^sl©lc.v " (in press)*
Gourasp PoS Spreading depression of activity in anjfcibian
retinas Abo Jo Physi®l=,e 121*28=319 I9580
Gourase Po s Electspic activity ©f toad retinas M^ Jo Ophthot,
f}6:l»182 (Pto 11)9 19580
H®nors and Awards relating to this projects
Puortess MoGoFc. Deliveay of the Annual Bidiop Lecture at the
Washington University Medical Center? Sto Louis & Kissourig
April lly 19^9 entitled "Generaticm ®f Ner^e Saipulses in
Receptor Qrgans"o,
Ssriai Roc.^^KTJ^LiEi^37 Cs)
i, OphtlialnKslogy Branch
2. Physiology Section
3. Bethesda, Maryland
4. New Project
PHS - NIH
Individual Project Report
Calendar Year 1958
Part A«
Project Title: ERG Reactions of Pure-Cone Mananalian Retinae
Principal Investigators: Katharine Tansley„ D,Sc„ sm(
Richard M» Copenhaverj M.Do
Other Investigator: Ralph D. Gtmkeij O.D.
Cooperating Units: None
Man Years (calendar year 1958): Patient Days:
Total : ,10
Professional : 0O5
Other: 0O5
Project Description:
Objectives; To study the spectral sensitivity i, dark
adaptation^, and flicker fusion fsrequencisB of various jsetDbers
of the squirrel family, these having the only pure-cone retinae
known amongst msmcnalsc
Methods Employed: The apparatus » consisting of a Xenon
high pressure laoip and a set of double interference filters to
produce nearly moaochromstic light stiisulis already in use here,
will be employed to saeasure the spectral sensitivity curves of
various squirrel species <. Both flickering and single flash
stimuli ??ill be used to study flicker responses an ark adap-
tation curve s»
Major Findings; i^one to date»
Sigaificance to Profjgaai of Instifcate; (Me of £he
great difficultiess encountered in studying the human. ERG
i3 the separation of the photopic (coae) respoaeQ from the
scofcopic (rod) response. IVaat members of the sqaiErel family
possess pure-cone retinae so that a good knowledge of the
reactions which can be obtained from such anitisals should be
i&ost helpful ia providing iaformatlon oa the reapcases of s.
purely photopic mechanism uncontsminated by those of the
scotopic mecfeatjism,. Investigations already carried out in
Europe have shown that the reactions of the retinae of two
species^-'-a tree squirrel and a ground squirrel"-are in some
vays quite unlike those of the more usual taiited rod and con®
retinae of which the huiaan is an esss^le. Tiie tree squirrel
epectral sensitivity curs'e ia naich aarrois^er aad appears to
iPeflect the activity of one only of the three postulated
mechanisms for colour vision-^the "green" mechsnism. The
ground squirrel apparently has two^ the "blue" and the "gsresn".
It is hopedj therefore, that a more complete study on several
species of both trgs and ground squirrels %7ill provide infor-
mation about the fxindamaiatal raechaRisias for colour vision.
Dark adaptation has^ so far» only been studied on a tree
squirrel which gives a curve similar to that found by psycho-
physical methods for the huaoan fovea.
It is believed that the photopic and scotopic responses
in man can be separated by Bseans of their reactions to flick'*
ering stiniulio She squirrel responses to flicker have not yet
been systeajatically studied. We intend to repair this oiaission
aiid hope to discover whether in this respect also the reactioms
of the squirrel retina reserable those of the huajsn photopic
mechanlBm.
Proposed Course of Project; See Objectives,
Part B included: Yes / /
io Ophtlmlioology" Branch
2o Physioiogsr 3ectioa
3<. Betkesdap /•iai'ylaad
4o ckijoe as IiiIIir5-»63 (c)
vm « nin
ladividual Project Iisports
Calendar Year 1,$B
Vsa-t /v.
i'rojeot Title: Punctional iJtudies in retinal .Vnoaalies and
biseasea (Jlectrorotinograpliy, ViCiantometr/y
and Pex'iuetric Light iieaae studies} «
Pj-incipai Investigators; :.ichard lU Copenhaverj MoD.j, and
ivoipfa Do Gunkelp C„Do
Other lavestigator-ss Hone
Cooperating Units?
llan Years (calendar year lv56)8 Patient Deysx 280
Totals oio
J-rof^saionals ^05 Outpatient Visits? >6
Others 0O5
Project Bescidptioas
8 This stii«^ ^jMoh ia a continuation of a
previous project is coaeerned with th© inv©stigatloa of visual
function in patients with retinal a^no5Balitj.®s utilising xnm
adaptoi2>et3:'ic9 perimetric and ©lecti^rstinographio tests^ in
addition to clinical Gssminationo i»p9oial ®nphaei@ is placed
upon thos® conditions idisro there is a S9X®eti¥© affection of
th© scotopic or photopic retinal proceesase iviaan possibles
other affect«d assbars of th® family ar® stisdied.,
Th» objectives of this study ar® as follovst
1) To aid ia the differential dia^osiSs prognosis and
genetic counselling of patients td.th s^tinal abno3rmliti©So
2) To in^iaetigato th© clinical us9fula©ss of adaptosastsyj
p®riii!i®tric li^t sens© studies and ®l@ctror@tiaograp^«
.^GtU^ 30X0
3) To study the physiology of rod aiid cone vision in
these patients by means of perimetric and eJ-ectroretinographic
tech'^iques 'whteh permit a separate evaluation of the photopic
and SCO topic response »
Methods Employed; After clinical ophthalE3ol<;^e
esamination the following special tests are performed;
1) Adaptometry s The course of dark adaptation is
determined for a paraj2s,cular retinal area on the Goldmann
2) Perimetric light sense testings The absolute light
threshold is determined for red and blue stinroli from 0 to 40
degrees over one or more meridians in tho visual field. The
thresholds for blue light afford a "rod profile" of the retina^,
and the thresholds for red light represent the modified "cona
profile" of the retina., Special attaehrisats have been added to
the Goldmann adaptometer to make these ritudies possible,
3) Slgctroretinography s The SHG's are obtained by means
of contact lens electrodes and recorded on an SSG machiaeo The
use of an intense light source supplied by a xenon lamp in
conjunction with double interference and neutral densi&y filters
makes possible not only the separation of scotopic and photopic
function but allows the study of variouf« photopic mechanisms
associated with color vision ,
Major Findings; These studies have in the past yielded
significant information of value in the diagnosis of such diseases
as retinitis pigmentosa^ cerebro-mactilar degeneraoionsg congenital
night blindness J, total color blindness and other.'io In addition j,
these studies contributed basic inforuiation about the differentia-
tion of scotopic and photopic function which is necessary in the
study of retinal physiolo©r„
Recently these methods have been applied to oolor-defe-^tive
subjects who comprise 8}o of the male populationo It has beca
possible to ealoulste spectral sensitivity curves froin the electro-
retinograpliic data friileh ix;>.v© given some insight iriLo the photopic
mechaxiis3is which are related to color vision o The VJsf ects
responsible for the typical color anomalies are de constrated to
be retinal in location rather"* than in the optic pa'hways or the
cerebral cortex. The spectral sensitivity cuarves jf several
types of color defectives were found to be sigaifZ-cantly different
from the normal which supplies a method of distinguishing between
normal and defective color vision which is purely pt^sical and
does not require any subjective response on the part of the
patient o The electroretinogr-aphic method allows a determination
of the type of defect and to some extent the degree of deficiency „
By f indiiig the difference between the electroretinal spectral
sensitivity cui^es between norsstals and color defectives it is
possible to obtain inforaiatioa about the color mechanism losto
The difference masimum in the protanops agrees well with the
peak absorption of the red-sensitive pigment erythrolabe which
Rushton . (at Cambridge^ England) has found to be absent in the
retina of this type of color defective „ The sensitivity loss
in deuterenopes agrees well with the gre^ia-sensitivQ pigment
present in noanaals and also in dsuter&nopss and hence suggests
an interruption of the electrical impulses from the green-
senaitive cones at a retinal level rather than a loss of
pigment. As previously stated this represents a new and
practical method of investigation of a large group of patients
serlotxsly handicapped in a n-umber of occupations whsre color
vision is a prerequisite o
SignificaacQ to Program of Institute; The f^mctional
study of retinal diseases may lead to a farther clarification
of basic problems in retinal physiology which will result in a
better understanding of numberous clinical diagnostic problems ,
Proposed Course of Project; To extend the method of
spectral electroretinography and other tools for functional
investigation to a study of acqtiired color defects<, various
congenital anomalies, glaucoma and optic neuritis,,
Part B Included les W Wo [J
B± Jl 8evlcto9'its& •soloo bos slassnca jcasw^ad eevnro Y-^lvii-isxiea
- 4. -
Serial Ho, NTMBB=.'S!^C«'5
PHS " imi
IndiTidiml Project Report
Calendar Year 1958
Part Bo Honors o Avjards and Publications
Publications other than abstracts from this projects
Dodto Eoj, Copeiihaverj RoM,g and Gunkel^ RaDo! Photopischer
Do2iinator tmd Farfekomponeaten in. measshliehen elektroretino-
graiaBip PfMgsrs Archo^ 267; 4.97-507. 1958.
Goodiaans Go and Q-unkel^ RoD<,s Familial Slsctrcretinographic
and AdaptomeMc Studies in Retinitis Pigmentosa j, Aaio Jo Ophth„^
i^sl42-17S (Septo Pto II) » 1958o
Honors and Awards relating to this projects None
serial Ho. MINm.59. (o)
1. OphtMIt!S}logy Braxich
2. Physiology Sectioa
3. Bethesda» Maryland
4o Sss^ £8 N]!M)B-62 (c)
PHS - NIH
Individual Project Report
Calendar Year 19 5S
Parfc A.
Project Title: ERG Spectrsl Sensitivity Curves on Caucasians j
Negroes g and Albinos
Principal Investigator: Sberhard Dodt, M.D.
Other Investigators: Ralph D. Gunkal, O.Do and
Sishard M, Copenhaver, M.D.
Cooperating IJnlts: !Sone
Iten Years (calendar year 1958): Patient Days:
Total: aO
Professional : e05
Other: 0O5
Project Description:
Ob^lectlves; To study the relative spectral sensitivity
curve In deeply pigmented and alblnotlc hucoan eyes by means
of electroretioography.
Methods Enaployed? Colored light stisasli are produced
by uaing double interference filters in conjunction with a
Xeiffin high pressure laiapo To measure the photopic seaaiti^ity
curve of albinos and negroes flickering light flsshe© at &
rate of 32 per second are delivered by a rotating disc.
Patient Material; Kegro patients exhibiting a low level
of baseline activity and albinotic humsn eyes will be selected.
2 -
Ma.lor FladxTga; The relative syr.ctT&l sensitivifciea
for wavelengths lonj^er than 583 were 'iigb ia albinos and low
in negroesij while d&t\ Caucasians aai subjects with "blond"
fundi showed intena@diivt.is 8ensitiv.^<:ies<> The maxitoum sensi-^
tivity ia albinos occurred at 610 toji as ccB5>ared with a peak
sensitivity at 533 siji for .'.eucr.aians end negroes.. The dif-
ference in spectral sensitivity in albinoo and negroes was
due to the reflection of ligits; ly blood in thcs forsner.
By trans- scleral illumiaati->a it vfa0 found that the
only selective absorption of light in th,- tissue coats of
the eye is due to bloodo 7;t was also detsnained that the
blood volume in the sclera and cho7.-oid cinnot be ascertained
with this method. This work demonstratei the itoportant
effect ^skiich the density of. the pigEsenfe vpithelium has on
the electro-retinal spectral sensitivity, With ssnall area,
intense light stitaulation In individuals ^ith a thin pigment
epithelial layer, of the retina and red saisitivity is markedly
increased due to blood reflection.
Significance to Prcr^ram of lastitL'te; The d&fea ob-
tained in this project coE;«:ribute materiilly to the present
knowledge regarding the fcutors affecting the spectral sensi-
tivity curve of the human aye as iseaaured by electroretinogrsphy,,
Recent experin^nts in anicmls by Bodt havj shown that the
spectral r^^efiectivity of the blood may ceise distortion of
the ESG spectral sensitivity curves, By the conqt^ison of
psychophysical data and E'lG spectral sensitivity curves in
patients j, it is possible it:o evaluate the :t!^ortance of th@
spectral reflectivity of blood in humans o
Proposed Course of Froject; Since £he investigation
is cosaplefeed the project is now terminated o
Part B included? Yes f?K/ Ho £_/
- 3 -
Serial Eo. NINDEU^Ce)
PHS = NIH
Individual Project Report
Calendar Year 1953
Part Bo Honors, Avards aad Publicatioas
Publications other than abstracts from this project:
Dodt, Eo| Ccpeaha.v®rj R, M„ ; aad Gimkel, Ro D^: Sieetro-?
retinographic maasurements of the spectral sensitivity in
albinos^ Negroes end whiteSj AoM,Ao Arch, Ophth.^ in press»
Honors and a^simrds relating to this project: Hhns
Serial Noo HIffl)B^60 (e)
1-, Ophehaissjlogy Braach
2. Physiology Section
3. Bsthesda, Maryland
4. Sasae as NINDB-64 (c)
PHS ~ MIH
ladi-yidual Pro j set Report
Calaadar Year 1958
Fart A,
Project Title: Design and Construction of Ophthaliaic Instruments
Principal Investigator: Ralph D. Ounkel, 0,Do
Other lave St igs tor: Hone
Cooperating Units;
Man Years (csiendax year 1958): Patient Days: ilona
Total: olO
Frofessioasl : 0O5
Other: .05
Project Bescription:
Obiectix^as; To tnake ii^roveiaeata of instruEsents used
±n ciinicsl snd laboratory ophthslE^logic research .
Specifically: To further taodify the GoisfcisEn-Weekers
Adaptoai©ter to facilitate psriaiatric jEeasuramsEts of light
sense thresholdSj and in particular to laeasure pure cone
function bs separated fr«Mn rod function in the retina.
To design sad construct & ccKabiaed head-holder and
electrode-holder for use in electroretiaography.
__ To design sad/ or constnsct siaeh other devises as are
required or suggested hy current projects.
Methods Sgs>loyed: Esteasive testing with the double
interference filters, elaborated on hy Br, Dodt for another
project, has shown the need for using red light of a longer
wavelength than that formerly provided :ln fcoth the fixation
- 2 -
light asd the test spot of the adaptometer, Apparestly^
evea at threshold levels any 'fe-aveleagth shorter than sboijt
672 s^ is per<$eived by tha rods as i?ell as by fchs coasss
although in varying propsrtioas by each. Hsncss the para-
iBSCular r@tisie» containing rods, sees th© ixspincs red light
batter than does the sod-free maculaj which would obviously
discoursgs good ceatrsl fisatioa. TItis situation has been
remedied and the test spots have been iti^sroved with Wrattem
filters, which appear to be adaqasta.
The size of tha test spot ^as fouad to ha^e uaespectcd
is^orteacs ia that a beam subtending about 10 miautes of arc
gave ^hat appears to be s characteristic cone curve, ^hile
the con3K3aly-used one degree spot did not.
SEsall electric oastors have susw hsietk moantsd so as to
BKJve the fisatlon light sa angular distance of 45 ia either
the vertical or the horisoatal lasridian ia 15 minutes of tisus.
This persnits an essentially continaous profile instsad of the
plotting of five or six points as was fornsrly done.
Sines it ^ffis found advantageous la spectral electro=
retinography that the subject be seated in a chair instead
of lying, on his back, it ^ss necessary to devise soiae mgans
of prevent isg mpveiasst of the heads ^?hioh would introduce
various artifacts '.in the record i- .This wmb achieved mth a
double yoke arrangeiaent of plastis tasterisl attached to a
stsndsrd chin-rest o Adjtssteble pads restrict Eovement ade-
quately, and bt3ilt-is alectrodes touch the forehaad and
bridge of the tubs&s, thereby simplifying the procedureo
Patient Material; Sxamiastioas of dark adaptation
and retinal profiles are done rontisaely on subjects referred
here for electroretinographic differentiation of retinitis
pigmentosa, night blindness, defeciE:ive color vision and
certain degenerative retirial conditions.
Major Fiadinga; Borsal values srs being sstablished
for retiaal profiles using the sew color filters aad smaller
test spots. Certain losses of sensitivity at and around the
iBsculs of color blind persons have been ebserx^ed. These and
tentative data from patients with other eye diseases indicate
the potential usefulness of the tastijag procedure for a
variety of diseases entities, Barli adaptation curves obtained
with the Hsodified instruiaent have been found quite satisfec£ory„
•^ 3
Sipalficance to Proggaia of Inetifcate: It is believed
that tha modifications of the GoIdisaBB'-I^eeksrs adaptomster
and the device of iamjobilising the patiaat's head contribute
either to the infortaafcida based on praviousiy nsed tschniqoes
or to the accuracy of the testing procedars.
The study of retinal psofiles has already been ahovm
to be helpful in evaluating and corroborating photopic mid
gcotopic electroretiBographic data. It i@ possible that
it tEaay prove to be a sensitive method for detsctiag early
retisal and optic nerve damage.
Proposed Course of Project; It seems to be of benefit
that the present freedom and flexibility of the instrument
project be maintained. This would serve the development of
promising ideas.
Further studies should be laade of perimetric light
sense thresholds in dlseassss other then those studied. By
this technique information aight be gained supplementary to
that obtained by the conventional visual field measurements »
Part B included Yes /"~7 No„ i
Swrgieal Keuarology Bramch
Diseases and Biinds^ess
During ela® pesfieui of t&is rep©sfe, 211 pa£i«mfe® fesw fessea sKudled
&& £he wards, whils In thm Qatpati&nt Separ€a»m£, 214 pagiaats were
^xamified in a total of 324 sefe®d»l®d visits. Use aeurosurgi^sl op@ra&ing
room saw fcfee ©etaplctioffi ©f 106 pr©cedur«s Of Khesc, 35 w@ra s©a«s®fftt(ftd
wieh ehe surgical £r«alsesKt and iave»&iga$ioa o£ epileptic s^ehanisitas,
wMle 32 wer® psrforased for space-oeeupyiag l®®i©«s of £he aervous
syseeia. H&e patients who eane ^o ward aad eliraie tnvmstigatitm, m
w®ll a© felieise who eatss eo ©perafeivs ereataaene, for® 4he basis for
reporeg aad ralftvant lab®rae©ry work, fhefe wer« 21 reports prepared
for pufelieattioa during fehis year, /
As is the pass, £he ssajoriSy of ehese pa£i«se@ were epilepsies,
aad @£ ghe epileptic population, tls@ greater proportion owed their
epilepsy to disease of th@ tesuporal lobe. Utter® were also &as®s of
seffitrefficephalic @si^ur@® sM probleias involving diffuse or ieuieipl<s
Issioffis.
FroR reseat studies of teaaporal lobe epilepsy, it is mow a.p^aT<mt
that OQe of tbe signifiean&t eauses is vascular saalforraatiozn. Such uj&l-
formations eannot be diagnosed by ordinary tests, using contrast as^dia,
nor is there any partieular elinieel sign ©f tbieir e^istenee. 'lW@iv®
of these lesions have been disclosed at operation. In @a©k ease, th@re
was some alteration of the surface vessel pattern snd sosts change in
the sise, €olor, and sonsisteney of the surface gyrio these alterations
were not remarkable and in secae eases appeared only as subtle change@°<=
changes so subtle that their signifioanee was missed in the initial
eicposure. the surface ,eh£Siges are related to a lesion in depths f&is
is a small vascular tumor eharaeterissd by racemose vigssels, and a fins
network of these vessels is surroui&dsd by gliotic tissue whieh tsa
usually peri°in@isural in locatiim. fhe study of patients afflicted
with such lesions has provided further information concerning <gpiieptl€
meeha^sms of temporal lobe seizures, these are unilateral l^^tsmm,
yet their expression is often bilateral, fhis bilatsrality disapp«a«'s»
a£t@r excision of the lesion a&A thus im relevant to a ptimatty epii(«*pti:i<g
fosus, which in turn has activated distant structures.
fhe ward study of patients was concerned with epileptic s%ehai%im«^,
pain problems, space°occupying lesions, transmission of iaf action in
the ward environsasnt, observation of abnormal individual srA goclal
behavior patterns, the effects of urea on is^tracraniel pressur®, th^
effects of hormones on seisure processes, the K'slationsMp of horss^nal
activity to spoataneous s@i£ur@@, the autonomic concomitantsi of t@^s;rfil
lobe epilepsy, the clinical characteristics of aut^isatisms, language
studies, as well as the effacts ®f hypopfeysectosasr aad depth stimilafitcs
responses, fhe observatioit of seizures has provided n@^ infosaatioa
concerning the iK^tor phenomena of automatic states aad other se^usls
of tea^oral lobe discharge. Miss Tinai® has successfully photograpk^d
1721 mimuwe slgis®. Hies® p&ofeogrsphs Issv® h&eu corr®lae®d so &e m
provide a pane^assic @uirv@y of ^e vaotoT md wit&mi^e ph@noafsna of
ttsapovel lob« epilepsy. In addition, the voealisaeioa paeeesms acd
various verbal esspreseiono which aecsa^my ssisisires have been s-eeerded,
Tba iaeerie^al behavior of paelem^s feaa occupied a ec^siderafele eime
in ehe ward eeudiea. "Oie language charae^eris^ics, the eharaceeris&lcs
of hierarchy asid social ineeraceion, as sail as the capabilities for
perceptual fu&etioa have been studied. There have been certain teeh°
Eieal developasenta on the ward, as well. Hiss Pimi® has worked out
a prototype for a nes' neurosurgical bed, and Miss Piraie and Ur. Edgar
are studying the characteristics of air>=b®m infection in the ^srd
have be«B certain developjaents in the aeurssurgieal
operating room as well. Br. Fritehard has developed a new dural
scissors and reports on the history of this forra of cutting instru°>
!^nt in intraersniai and other neurological surgery. £n conjunction
with Mr. liggle. Miss Lm±@ has developed a new eosamsnieating system
for the operating rooa. Also, Miss Lewis has designed a new workrooas
plan which will be incorporated in the n&f surgical suits. This pro"
vides for greater aseptic precautions and increases the work capabilities
of the nursing staff. In addition. Miss Lewis has beg-^n the first
phase of our training course in neurosurgical technique. 'Ihis course
is designed to provide the house officer in neurosurgery with a baek-^*
ground in basic and specialised surgical principles. She is training
Doctors BucknsBi and Lewis in the nursing techniques of neurosurgery.
In i^leisentation of this training, ©r. Laskowgki is teaching tissue
handling technique and- feei^statie methods in the prissate surgery. In
the future, all house officers in neurosurgery will be trained as neuron-
surgical nurses and will receive techniee^ training in the operating
WQom of frimate Meurology. ISiss Lewis continues her work in neurosurgi-^
eal .technique and is gradually accuBEuIating an esstensive photographic
record of the techniques used here, sany of which she has developed
herself. 2n the past yess she has spent iQug hours working on the
desist probleas for the new surgical suite.
"She laboratory studies continue. In Meurostsrgieal fhysielogy.
Dr. Li has continued the application of isieroeiectrode techniques in
tissue culture, and has studied neurological, as well as tausele elee^ats.
It is hoped that this cosi&ination of culture and electronic techniques
say be transplanted to the new operating facility so that tissue
freshly excised in the operating xocm can be cultured and studied in
whole or part in a readily accessible laboratory, ©r. Li's studies
during the past year include studies of cortical neurones, and in
combination with M. Ortise, study of neurological transssission charac°
teristies and neurological regeneration. @r. Ortis has joined the
Branch as a Visiting Scientist on loan from the faculty of the
University of Mexico where he is Professor of Neurosurgery sod Meur&<°
pathology.
Xn neurosurgical Matcssy, Dr. 7sn luren has reported on the
anatoiaieal effects ®i tea^oral lobeetoasy in the huisan. Ihis wosk,
carried out in conjunction with Br. fakovlev, est^lishes the first
struct'aral studies of the effects of tes^oral lobectoa^ in the
Ee has eoati^ed M® studies of the visual patte^ays in various
3.
hi@&®i©gi@ai gs@h{££fu@So Ais©, hm £@ soagiffiuirag his 3t\id±®a ®n thm
a&AteM.®al @££@@&s @f @pa€<B°©sisupyitsg Itssistss a^d is gradually aceuatu°
laeieg isl£i!i£€al Bua£@rial £®v tMs pus|i>«)S®o le hss bacsB abl@ t@ do
sesee H@?k ia fehe pestiasrfegm room with feis tmt 8€@r@®eKslc davie® which
&s, as yee, ustri«d £b th® ©linieal operaigiag s-@em. In addlg£®n, hs.
has e@Bplefe(sd s®ss ®f efee eell eeantg ®f p£tuit&asy sps^inaiKs derived
at hyp®plsiysee«®Bif aed eonrelated ehis materiel with eliffii@al and bie^
chesieai finditags £& the pa^is&ISs @ubj@@g@d g@ this pr®e@dur®<,
Is S®v@l@pa^ffitsl Mat@sQr, DTo Stskabaa eeatinuas his studies @£
hrain Ifssiems tn €as@© e£ SG°eaIl«d esrebral palsy. Sn eollabGraSleia
with various ssetrcaBiral invest igaterg, he is gradually a©€usaiating
a eoasidtsrabl® stor@ <»£ paehelegi^al material, the study @£ whieh oray
light @B various and eos^lieated lesloas whieh ar@ new
by th® lab<sl, ©erebrjil palssy, 1@ has bsguQ a study of
th® @s3bry©l®sy of th® huraaa tesperal i©bs m&d this study, in turn, has
a natural outgrowth @£ soae ei^ry^legiisal investigatiens ®f the
brain. %@@e first investigations eeneem remnstruetioa of
@arly soadte d®v@i®pmeat in cerenal seetionso "Ske temporal lobe studies
will b® undsrtaksn in sagittal seetiens.
In the Lidtoratery of Heuropathology, Sr, Klatee has pursued a
variety ®f interests. With th® use @£ tissue €ultur® teehni<|ues, he
has stud£(sd intraeellular ©haraeteristics ©f astreeytes, and with th@
use e£ pethelegisal spesiie^ns referred t© him, has ©utlined the
eharaeteristies of luru disease. In this laboratory. Or. Miguel has
developed a new method for the (fuantitat&ve study of presipitin reac-
tions, and Or. Laekowski studied the relationship of csreE^ral edema
to e^iirimental brain injury. He has also studied the effects of
hypothsnaia on injured and normal brain tissue.
the Laboratory on Pain and Hauro^anesthesiology lost its @hie£
this year through the resignation of Or. Kenneth Mall. Or. Hall left
the Iraneh to a@eept appoints^nt as Associate Professor of Mesthesiology
in Chars@ of gessargh in Ouk@ University fil@di@al S@h©ol. Aft@r
Or. Mali's dsparturs. Or. Fritehsrd has sontinued his pr@j@€t on flue°-
thantg, and is eos^ajelpg tMr, new anesthetie agent to the various
a®€@pt«d inhalation agents sueh as €hl©r®f©rm, sthsr, and syelopropane.
In addition, I^. Pritehi^ is extending the h#art°lung putsp teehnique
so as to provide an ea|>erii^ntal design for th@ study of anesthesia
effects @n the eerebral eireulation. In addition, h@ is saaking observa^
tions on patients und@r gensral anesthesia and hypothermia, as well as
on ehinqpansees in various anesthetic states. Ihsse observations as9
made with polygraphic t@ehni(|u@s.
fhe Laboratory of Clinical Psychology saw the arrivsl of a new
€hief , Or. ^rbsrt Lmisdell. Under his direction, this group has
reviewed the previous wosk of the laboratory and has begun an energei:ie
eao^aign for the development of. successful testing of tesqporal lobs
pati^sts. In addition, they have organized & projeet for the further
study of the effects of ablation and/®r stiiaulatioa ®f Hesehl's eosvalu-
tion. Or. Lsnsdell himself is developing a prototype for the testiag
of chis^ansees «lio@s teoporal lobes have b@en ablated.
fh® La83i©raie@Ey ©f Firimafe® Meur©Il@sy (genetotiss! ifes a6udy ®ro
feallueimiaggnis drug®, eh® ©ffegigg @f igerapcffaX aod frcsagal ablael®©
in She ©hio^ange®, and aer© iregeisEly, f£ls® effegfeg ©f specific ©©rfiieal
©xsisicsK ©o ®©B8Huniea£i©n sapafeiiis&es ®£ Shes© animals. Dir, Ner^i®
is ^isgiiQi&ing the seudy of depeig elaetsrode effects in ths mesial
gerapegal w®s,imi, asrad Mi®s Lewis, Dr. Norris and Dr. Baldwira hav©
txsguE Sh® M©=a®@ay @f ga£«(sh®l ismi&s preduei^ion affegr £sjnp©ffal l©Ibis
sSlraula^ien. Has egiglgabl® ©©E'lEesK ef (th© cMnspang®© has beats iav®®-
Siga^sd ared the effeefes of pffeeeiseral essisiea era s(elaj«la£i®n paesemfs
have been ©luserved. P@nieill£ni seiguse pafee®ims hav® feeeas sfeudisd.
fhfGugh a »@w £e€Mique, £h®s@ studies hav@ b@ee snad® ©n b®Kb hemis"
phefffis sinsulfeaaeoussly. I£ £® £8®w possibl® e© st^udy afe suE'gleal. @p®ra=
£1016 eh© enfeiE"® eoavescity aad polar surfaee® ®f b®(£h feemisphes'es.
Wish this esefetsifus, ehe effecfe ®f passive EsovemenS on efee piropsrgies
®f (6h® ©seifeabl® <g@rfees has b@ea s!6udied, as &av® th& cfeaffas£erf@ti<ss
®f spread @f een^esral i®be seizures, fhta s@gial hies-airshy and ^©s^ii^aQi^^
eaeicta s^udiss in the €hiispai%s®e e®l©ny are being €®a£iQued by
Miss Lewis.
Mr, Meillsff has developed a phofeegs-aphie €©ebni(|»3e whieh will
peoaie (g®l©r aisalysis ©f the surfa^a vascular psS5:ems ®f (the bfaits®
®f £h®@e aniiEais as espesed at ©peg"a£i®n, Sush a ®esfeni(|u@ eoay penaie
fuff£her study ®f vaseular (S®neoiniSane® ©f eertieal saisuffes. Dsr. OsrSis
teas joined the Laboffae®gy in the study ®f th© ©f feet® ©f th® Mssiisaro,
sssishr®©ia ®n the behavler @f the shis^ara^ge, as well as its effeets ©a
the @th©rwise n®sjaal eloct^agsam, bl®®d pr@s®ur©, ESS, and ®tlh@y vital
sigES in this asaimaio th® first phase ®f the lyssergis aeid study v&s
@®mpligt@d during this y@a^ asud the Mexiesn laushr®®^ p7®j@@t loarks ttit^
feeginffiiog ®f a s@€©nd phase whieh will itie®rp®rate ©the? hallusin^gsni'.'::
substances, fhe apparent siosilarity betwaen the €linl@alL effects ©f
thsse ehemisals and the @lini@al @Kpres3i©ra ®f tao^eral l®be epilepsy
semain® an intriguing and stimulating shallenge t® further inve@tigat£gs!)<
Diseases mud Zlin^snQse
CliQic^l Essearch
HBIDB-6l<c), HIHSB-62<c)» MI@DB-&3(c), HI!SD3-64Cc),
NimsS-65(e), S3IMDS-66(c), H£MDB"67(c), IIIHDB-6S<c)»
KI!a)B~69Cc), HHSB®- 70(c), NHSDB- 71 <c> , HI!i»B-72Cc),
•73Cc),
Estii^ted miizatUm for FY 1959
Toeals $773,000
M?aceg $271,000
:§ $507,000
Serial No. KBg£@-61<e)
Ic Surgical Neurology Branch
EE6 Branch
2. Neuropatfeo logical Section
Primate ileurology Section
4. NIND(B°>25(c>
?HS-KIH
Individual Project: Report
Calendar Year 1958
Pro jl eel: Titles Epileptogenic Mecfesnissns in the Brain of Man,
Frincipal Investigator; Maitland Baldwin. M. D,
Other Investigators; C„ Ajsmae Marsan, MoD,, Bo B, Towers M„Do
J, M. Van Buren, M,D., I. Klatisoj MoDo^
E, Edgar, M,D., So A. Lewis, R.S^o and
S. A. Bach, M,S,
Kan Years (Calendar Year .19581° Patient Bays,
Total; • .25 (Calendar Year 1958),
Professional; .25 3749
Other; .25
?ge^eet BescripfcJon;
Objeetives;
a. To stud^ causal meshanisms of epileptic seizures
in man.
bo To stu«^ the electrogr^hic characteristics of
epileptogenic activity in the brain of laan.
c. To 9tu^ the approved methods of surgical thers^y
for these lesions and develop near ther^eutic methods.
d. To study brain function as it is e:qpo8ed in the
extravagant e^eriasents devised by these lesions.
Methods Employed;
a. Clinical neurological ez^aination.
b. Special radiogri^hic examination.
c. Slectrogr^hic examination.
d. Eleetrocortigraphic examination.
e. Electrical stimulation of the lesion ei^sed at
operation.
f . Selective isolation of the lesion at operation.
go Fhotogr^hic and sound recording.
h. Histological and chemical ezmBination.
2,
MajQg Fiadings; This is a sepoift of observseioia on 119
patieafes. By selection, th.@ majority ©f these pstieots are afflicted
with t€B3p0r«l. lobe seistsres. Ft<m. the most recent study of these
cases of tess^ral lobe epilep^, it is sppasent that ezyptic angioma
is a signific<ant cause of this fo£m of seisure. This deeply lying vas-
cular lesion ii^hich seaas to involve mesial tea^ral st^cuctores close
to the junction beti^een the circulations provided by middle cerebral
and anterior choroidal branches no^ appears as an active agent in the
f>roduction of epileptic activity. I'erhaps it so alters the s^tabolic
requiresssnts of peri-> insular tissue in which it lies as to produce an
epileptogenic lesion. From a study of this year's cases and a reviei?
of previous material, it is evident that this raseosose vascular lesion
is a significant cause in the development of epileptogenic activity.
It casmot be outlined by ordinary contrast studies, and as yet there
is no obvious clinical correlation, ^parently, the lesion can be
related to unilateral or bilateral electrogr^hlc abnonaality, and
it may or tziay not be related to ictal signs ^hich have lateralising
significance. However, in the cases which have been studied recently «
unilateral excision of this lesion produces a good result insofar as
seizure frequency is conceamed. In one such case, the lesion could
not be totally eradicated because it involved the neighboring brain
st«D. After operation, this patient did not have any further clinical
seizures, but lie csaoe to cois^lain of a persisting and annoying per-
ceptual aberration. The objects and people around him seeiaed oiore
distinct, better lighted* and aare vivid. Their vivid and distinct
appearance seemed harsh to him, and from time to tiiae he felt as if
the objects in front of hia were too brightly lighted. His post-
operative el@ctrogr£^hic studies showed residual epileptiform
activity over the said-tea^oral leads. Presumably this was relevant
to the peri*incisural tissue left at operation. It ie^ be that this
remaining tissue has, through its epileptogenic characteristics,
activated perceptual processes in the opposite intact teo^oral lobe
and this activation is responsible for the continuing perceptual
aberration .
Buring the past year, a group of patients with seisures
were admitted for study under the primary diagnosis of t&s^oxak lob©
epilepsy. After further study, it was ^parent that these patients
did not owe their seisures to m. epileptiform process in either or
both tea^oral lobes. However, their clinical seisure pattern was
characterised by epigastric aurae, altered affect, altered awareness,
posturing and adversive nsjvesseats, as well as autonomic changes.
These characteristics are usually related to teo^oral lobe epilepsy.
Yet the si^ificant lesions in these patients were frontal and close
to the anterior cingulate gyrus. These cases are being analyzed in
conjunction with a similar series under the supervision of Dr. David
Daly at the May© Clinic. It is hoped that this analysis will serve
to illustrate the characteristics of cingulate seizures and provide
3.
Major Fiffidiags Cc©nt'd>; .
a meaas iox dlf fsreatiating these seizuires fre^ those of t^^oral
lobe epilepsy. Obviously, this dif£is?e&{:iation has not been clear
in the paet and tha laek of clasity has provided confusion snd un-
necessary errors in diagnosis.
The orator phenomena of tes^oral lobe epilepsy havs been
studied in some detail. It now seoas that there are certain hand
end other upper extremity postures which are characteristic of
epileptic activity in one or both teii^ral lobes. Usually these
hand and upper e:gtr@aiity si^veasents have a lateralislng significance.
They occur on the side opposite to the imast active tea^oral lobe.
The tmvess&nts of head snd neck in a teo^sral lobe seizure are
usually such that there is turning to one or the other side. The
chin points 6mm and the soovezsent is ssiooth and relatively slow.
It is thus different frosa the adversive asovestent which is pathog-
nosK>nic of suppletaeatary s»tor or other posterior frontal foci. In
these szovsments the chin points up and the s^vessent is jerky and
rapid. In ictal sutosoatism, the patient usually looks down and, as
is well known, frequently searches his person or issoediate surround<°
ings. These searching moves^nts can be interrupted by placing an
object in the person's hand, by voice or other sound, and by pain.
They can be influenced in character by the t3rpe of the object placed
in the hand, but neither this interruption nor any of the others
serve to sever the chain of Bsovenients. In autoiiiatism, the patient
uses his hmid as if it were a flipper. He seeaas to "finger" an
object, but he does not do so with precise isovements. At least
in the esses under observation, the hsnd is used with the fingers
en bloc, as if the four fingers opposed the thusssb as a unit.
During the past year. Miss Pirnie and her observation t®sm
have photographed 1721 such phemmoena. These ictal tsovaaents and
their auton^ic conccKsiitants are being successfully illustrated
through the use of a trained t&sm of nurse observers. This is' the
first tJMe photogre^hy has recorded spontaaeous seizure patterns in
the natural sequence of tei^orfil lobe epilepsy.
The autonomic concomitants of these teBsporsl lobe seizures
are being investigated by Dr. Van Buren. During this year, he has
studied Metrazol seizures in seventeen patients. These activated
responses were similar to those which he observed previously in spon-
taneous tes^oral lobe attacks. He noted a hypertension, tachycardia,,
respiratory apnea, fall in skin resistance and skin tsiqperature, as
well as swallowing laov^ents and inhibition of gastric lootility.
Apparently the electrograpbic tracing is indirectly relevant to the
autonosnic or clinical features of these activated seisures. Thus,
autonomic changes may appear coincident with bursts on the BSG or
may appear independent of these electrographic abnormalities. He
eo^haeises the striking independence of electrographic, autonomic.
4.
Major Findlnes (cont'd);
and clinical findings in time. He interprets this independence as
slgnificent of the fact that areas around the third ventricle may
carry on their activity without influencing the electrographic mani-
festations of cortical activity.
Interasittent and paro:^8inal perceptual aberration is a
clinical characteristic of temporal lobe epilepsy. Such aberration
does not always coincide with a clinically recognizable seizure
pattern. It ©ay occur without the other stig&ia of tooporai lobe
ictus. It appears that perceptual disorders of space and color per-
ception are n»st frequent. Such patients do not assess the spatial
characteristics of their surroundings with particular accuracy. More-
over, their surroundings often seem lighter, 2K)re distinct, a»re
brightly colored than usual. The patients are s^are of the unreality
of these spaces and yet paradoxically describe them as being "more
real than real". In tanporai lobe epilepsy, perceptual aberration
is never separate from disturbance of affect. In fact, in disturbance
of the teisporal lobe by epileptic process, the Bsost frequent coasbina-
tion of signs are those of fear and perceptual aberration. It now
appears that the physical basis of fear may be one of the aast sig-
nificant sources of clinical characteristics in tes:q>oral lobe epilepsy.
This prompts the speculation that catechol amine or other adrenaline-
like substances may be increased in amount as a result of mesial
temporal discharge. Recently, Hoff has reported such an increase fol*^
lowing stlsmslation of the temporal lobe in cats. Certainly the dila-
tation of the pupil, pallor of the skin, change in the vital signs,
and other autonomic concomitants observed by Dr. Van Buren are all
characteristic of adrenaline- like responses in the human. !$uring
this year, we have begun the study of effects of catechol amine on
patients with tes^oral lobe seizures, and conversely, a search for
the presence of unusual amounts of these chanical mediators in these
patients o
Perceptual aberration and hallucisatione may be induced in
these patients with temporal lobe seizures by suggestion. Investiga-
tion of these intangibles was begun in the year previous and has been
continued during the period of this report. Mot all patients are
affected by suggestion, but ^^proximately fifty per cent respond to
it. When the patients complain of a psychic aura or perceptual aber-
ration, this is accoiiq>anied by a feeling of fear or nervousness. The
i sling of fear or nervousness can be promoted and seems to serve as
a wedge for suggestion on the development of the perceptual aberration.
If the patient is asked if he is nervous, he may become nervous and
may in turn develop his hallucination or Illusion. Faradoxically,
these patients do not respond well to hypnosis, which has been tried.
In two cases, patients have complained of continuing illusions or hsl-
lucinations without other evidence of ictal process. These illusions
or hallucinations had been previously noted as an aura or beginning of
the habitual pattern. In these cases it was io^ssible to arrest the
perceptual aberration by suggestion, but suggestion seemed to enhance
the aberration.
5.
Major Findings (cont'd);
tiiporaL lot
In teaiiporal lobe spllepsy, there is an ill-defined dis-
turbance of body image 0 The patient does not hsve a usual eppracisi-
tion of Ms anm fona. In the ictus, he often exsmines it or its
^pendageso In £m atte^opt to define some of the chfiracteriatics of
this aspect of perceptual aberration^ patients have been asked to
sketch or outline their owa forois and faces > It appears that there
is an unusual distortion of body image, if such a sao^le test is in
any vsy reliable.
Mast patients with teniporal lobe epilepsy are said to have
"sseiBory difficulty". In the patients here, some tiose has been spent
in further attea^ts to elucidate this difficulty. At present, it
does not appear as a difficulty of mesiory. These patients have dif-
ficulty in relating in space ^ad in time. This difficulty of rela-
tionship is evident in their language which does not use a normal
quantity of substantives, and at its vrorst is a disconnected series
of illogical relationships. One of these patients finds it extreaaely
difficult to describe the fozm of an object. If he can achieve this
description, it is often very difficult for him to relate the form
of the object to its surroundings. Since these patients cannot
achieve spatial and time relationships, they find it extrsnely diffi-
cult to record experience in coherent sequences readily available for
recollection. Conversely, if they are provided with a clearly rele-
vant sequence of events and objects and the relationship between the
events and the objects is made quite clear to them, they can "meniorize"
the series with accuracy and often with excellence. Later they will
recollect the sequence o However, if the relationship is not made
clear to them, they cannot "remember" the sequence and they will
complain of a "memory" difficulty.
Temporal lobe seizures often seem to occur in cycles. Thus
a patient may be seizure free for thirty days and then suffer four or
five seizures in a day. This cyclic characteristic may be related to
some endocrine change. During this year we have begun to test the
effects of estrogens and testosterone on seizure frequency. One patient
who was receiving testosterone went into status during a^inistratlon
of the hormonal preparation. This effect may or may not be related to
the adasinlstration. It is too early for any conclusions, but it seems
valuable to pursue this aspect of the general^ investigation.
Fatients underling spontaneous attacks of centrencephalic
epilepsy are studied by simultaneous six-channel EEG recording and
recording ®f blood pressure, skin teeperature, heart rate, skin re-
sistance, plethysasograa, esophogeal and gastric motility and respira-
tion. They are subjected t© continuous response testing which is also
simultaneously recorded in order to define as closely m possible the
periods of loss of consciousness. (This study is carried out with
Dr. Allan Mirsky, NIMH.)
6o
Major Fiadlnga (cont'd);
To date oaly five patients have been subject to these
studies. The finding found typical of the petit mal "absence"
consists of e3£piratory apnea which may or may not be associated
with tachycardia and fall of skin resistancCc In general, the
findings were gtuch less striking than those attending spontaneous
automatisiBS of tesiporal lobe origin which were studied in the previous
year (see Calendar Year 1957 "Epileptogenic Mechanisms in the Brain
of Han"). Bursts of 3/ sec. spike and wave activity or irregular
polyspike and wave activity bilaterally synchronous and symmetrical
in the frontal regions may appear without alteration in the patient's
motor response to visual stimuli or autonomic change. Changes in
both the latter features tend to appear with longer epileptic bursts,
but still the degree of interference cannot be predicted from the
appearance of the electrogr^hic trace alone. For exai^le, a 3/sec„
spike and wave burst of coaq>arable lengfgh and voltage tssay produce
prolonged expiratory apnea and later produce very little change in
the respiratory rhythm. Itotor responses have been observed during
spike and wave discharges though they are usually absent when the
patient is apneic. In one instance when the patient firmly claimed
to have recall during his 7.^>ike and wave seizures, although he
"could not move", he was never able to recall letters or colors
shown to him. During the latter part of the spike and wave episcde^
however^ without obvious electrogr^hic change, he on occasion was
able to respond correctly to visual stimuli.
Results to date indicate that this is a coscqplex problem
for correlation and will require considerable clinical material.
In the laboratory, the spread of penicillin-induced
seizures has been observed as it occurred over the surface of both
hemispheres. Thus, after siimiltaneous e^^sure of both cerebral
hemispheres in the chis^anzee, a penicillin lesion was created in
one or the other temporal cortex. Electrocorticographic recording
over the surface of the cortes of both hemispheres seemed to indicate
that once the seizure process spread outside the temporal lobe, it
was first evident in the parasagittal region on the side opposite to
the involved teaporal lobe. It is probable that this spread is
through subcortical structtires and is mediated by transcortical con-
nections. As the seizure discharge spreads across the cortex, it is
preceded by discernible vascular change. The cortex darkens, the
veins bec^oie injected, and the arteries constrict. This observation
raises the old question of the significance of vascular change in
the development and spread of an epileptic seizure. If such a vas-
cular change occurs in both hisoispheres as it does following wide-
spread epileptiform activity, both hemispheres are subject to severe,
and occasionally critical, edema. In one instance, this edema could
not be relieved by injection of hypertonic solutions and postural
7.
Mai or Findiags (coat'd);
drsinage. l!he dura could not be closed unCil one frontal lobe was
aq>utatedo This provides sows, daaonstration of the potential
severity of the postictal pheiaoaieQa in the production of brain
daaiage ^snd subsequent neurological sequels,
Sipjalficance to Neurological Research; These observa-
tions may contribute towards further understmiding of epileptic
laechanisms as they occur in tasporal lobes of higher primates <>
Proposed Course of the Projects The various clinical
and experii&ental studies will be continued. Both the clinical
and esperifflental investigations will be strengthened by studies
of catechol amines.
Part B included; Yes /X/ Ko /_/
Serial HOo H£M9B-6lCc)
FHS-KIH
Individual Project Report
Cslandas Ysas 1958
Part Bs Boaorsj A!i?ards, and Fublicatixtas
Publications other thsn abstracts from this project:
Balds^aj M,s Bailey, P.:
Sprlagfield, ChaSo C.
^SLI SS.^^!^5s1.
Balden, M.: Notes on the history of Merican
oiilitary neurosurgery. In Meiroweky, A. M.
(ed.): Trauma of Central and Peripheral
Hervous Syateia. (in preas)
Hall, K.» Baldwin, M., Norris, F,; Succinyl-
ctoline in a^ake ctaaxi&t&ssy . ^esthesiology
(in press)
Serial K©« NB?BB'>62Cc)
le Surgical Neurology
Branch
2c
3. BeChesda, Msrylaad
4. NIN]IB-26(e)
PHS-NIH
I&dividual Project Raport
Calendar Year 1953
IMLA»
Project Titles F«actioaal Representation in the Temporal Lob©
of Man and Higher friraates,
Frinciisal Investigator; Maitlsad Baldwin^ Mo Bo
Other laveatigatQgss John Van Bwrenj Mc D,, Shirley Lewis, Ro Noj,
and Syen Ao Bach, M. Do
Gooperatiag Units; HonSo
Man Years (Calendar ¥e.a£^1958)s Patient Days
- ^ Total; .25 (Calendar Ye^ 1958) •
Fro'fessional; .25 645
Other; .25
Project Description;
Obleetives; To further understanding of ftsnctional represea-
tations within the tea^oral lobe of man and higher primates o
lo Slectrical stlnailation and recording of the human,
chin^an£:e@ and monkey t^nporal lobe@; (a) directly, after opersti^e
e^osure; <b) indirectly by depth electrodes and scalp recordtogSc
2. Ablation of all or parts of the temporal lobeSo
3o Anatomical studies of whole brain after tai^ral
lobft excisions o
Major gjndln^s; During the past year the electrical stiimsl©"
tion of husEsn and other higher primate tes^oral lobes has continisedo
In the results obtained from the hussan operating room, interest haa
been focused on the so»called psychical responseso The majority of
these responses in recent cases have come Irora depth stiEmlation^ yet
previously the saajority seemed to come from stimulation @f the cortical
mantle. In the l^ntreal series, almost all the psychical responses ar©
2.
Major Findings (cont'd);
derived from cortical stimulation, yet in Falconer's series all these
responses come from stimulation o£ the depth. The esperience here
seems to indicate that the response may cotse from either the surface
or the depth. Obviously these are cos^liceted reactions. The patient
is telling us of a cosiplex perceptual aberration or in some of the
aaore "fashionable" responses, the patient tells of a menKsry or dresa.
It is doubtful if such a cojqslicated response should be ^itirely
relevant to either the cortex or the deep structures. It is EKsre
likely that the fimctional relationships include cortex and subcortical
structures of both sides in & functioning unit.
^proziieately 200 positive responses have been obtained by
stimulation of the chin^anzee cortex. The loajority of these are
relevant to what Sherrington called 'excitable cortex' . Thus they are
moveo^nts. imong these responses are many which were obtained through
similar stimulation of the cortex on both sides. When the cortex is
stimulated simultaneously so that right and left motor areaa are acti-
vated in concert, either right or left extreaaity, face, or other
peripheral part m^ move. The brain seens to select the precedence
of activity which follows bilateral sissultaaeous stimulation. Passive
movement of an extrranity influences the effect of this type of stiim-
lation. When the hand is clenched, stisiulatioQ of representation for
fingers usually results in extension, whereas when the fingers are
extended, stimulation of cortical representation is usually followed
by flexion. Perhaps passive movement of the extr^oities on the one
side influences response of extremities on the other. There may be a
contralateral inhibitory process. Such passive movement does not seCTJ
to alter the spread of epileptiform discharge from the t@nporal cortex
even though this spread is not coincident with ictal movements. Elec°
tricel stimulation of the post-central cortex evokes movements. These
movements are not abolished by excision of the ipsilateral motor cortex.
Ferhaps electrical stimulation of the mesial temporal s£ruc°
tures is follcswed by increased secretion of catechol amines. This
seems a reasonable supposition when one reviews movies of a chimpanzee
taken when he is undergoing such stimulation. The autonomic phenomena
which are concomitant with such stitrnjlation are strikingly similar t©
those which follow an injection of adrenalin. In order to test this
supposition, various experimental designs are being constructed. The
first of these followed the model of Cannon for bio-assay of "excitable
blood". More recently, lactic acid has been used as an indicator of
increased catechol ^iines. Dr. Eiorris is composing an elaborate ex-
perimental design which includes capability for mesial temporal stimu-
lation, extradural recording, systematic polygraphic recording, and
biochemical assay. This should provide some further information on the
relationship between epileptiform discharge in mesial taisporal struc-
tures and secretion of catechol amines.
3.
Major Findings (cont'd);
The study of abla&lon preparations continues. This year
soarkg the fourth in ^hich observation of the effects of bilateral
temporal ablation has been possible. Four years after bilateral
taioporal lobectoa^, the chitspanzee is readjusting socially and does
not show any of the acute or issnediate stigisa which were reported
previously. The animal remains more placid than his conten^oraries,
but this placidity is slight and it is is^ossible for an untutored
observer to differentiate between the operated and unoperated animals.,
Such an snimal after bilatersil temporal lobectomy regains his place
in the social hierarchy and seems to continue a normal sexual, play,
feeding and learning development. On the other hand, four years
after bilateral frontal lobectoo^r, a similar animal does not regain
his place in the hierarchy and his individual and social habit patterns
remain remarkably abnormal. There is a rapid recovery from destruc-
tion of mesial ten^oral structures. This takes approximately tf?o
months. As a corollary to such preparations, a 'Weber' syndrome was
created in one animal who was ready for sacrifice. This sjmdrome was
a reasonable portrayal of the syndrome as described in human patients ^
It demonstrated one of the (surgical) risks of tea^oral lobectoaiy.
Mesial t^sq»oral lesions affect cossmmication in the chim-
pansee for approximately four weeks after their creation. Recently,,
lateral temporal and parasylvian lesions have been created in an
effort t© determine their relationship to consuunication patterns. At
present two such animals are ^^vailable for stuc^. In these, excisions
were made in the area comparable to Broca in man. One is on the left
side; the other on the right side. The ^limals are ambidexterous.
Such lesions do not interrupt or obviously change vocalization, hsnd„
upper extremity and face comsamication patterns.
The hallucinogenic substances which are contained in the
Mexican mushroom do not affect the chia^anzees whose tasporal lobes
have been removed, yet these substances affect the normal chimpanzee
so as to make him tsme, relatively unaware of his surroundings, and
somewhat ataxic. As is the case following lysergic acid administra- •
tion, the temporal-lobectomised chii^anzee falls to respond to the
psylicibin cc^ounds of the Mexican mushroom.
I>uring this year it has been established that the ten^oral
lobe of the chimpanzee is relevant to the syndrome reaction which occurs
when he receives lysergic acid. Moreover, it is ^parent that the
lateral tea^oral CQrtex is the significant element in the neurological
chain which forms the background of this reaction. For the chimpanzee
whose mesial teiqsoral structures are Tseaoved reacts to the drwg. However,
his conten^orary whose lateral teo^oral cortices have been removed
fails to respond to the lysergic acid. As has been previously stated^
removal of the frontal cortex does not affect this reaction.
4o
m^ox Windings (cont'd);
Three bs'sias of chiisspaaseea who hswe undergoae t^iporal
iobectoodes are beiixg studied ±0. aeurosurgicsl enatosay by
Sr. ¥an Buren. In addition, he reports on the reconstruction of
two huajjsG fcea^rsl lobe dsfectss carried out at tse Harvard M&°
tcmicsl miBews with Dr. Paul I„ Yakovlev. 0ns c&s® consisted in
anterior tea^oral lobectesjjy for epilepsy. The contrasting ease
consisted of an infarction ^hich involved the most posterior
extremity of the sylvian fissure centering upon the tea^oro-
parieto-occipital junction. With the anterior t^&poral lesion,
nuclear degeneration appeared in the inferior and lateral portion
of the pulvinar» the posterior portion of the medial geniculate
bo^ and the lateral portion of the lateral geniculate body. With
the posterior tes^oral lesion the degeneration appeared in the naiddle
and posterior portions of the pulvinar, the saaterior portion of the
i&edial geniculate body and the medial portion of the lateral genicu'>
late body. There was also thinning out of cells of the posterior
portion ©f the nucleus aiedialis dorsalis. Thus, the antero-posterior
rev^-ersal of the auditory representation froia auditory cortex to medial
geniculate body has been noted and there is suggestion of dorsal
ventral orientation of the projection of the pulvinar to the tec^oral
cortex. The pars or^^lis of the pulvinar showed no degeneration in
eithar case. Tract degeneration studies showed several points of
interest. The stria terminaiis in man appears to arise from the
cortical and ssedial accessory basal nuclei of the asiygdala since it
remained intact when the lateral portions of the mygdala were
destroyed by surgery. The anterior cosmissurs was nearly entirely
degenerated suggesting that the retained zsedial portions of the
aa^gdala and region of the \mcus received vezy little projection from
the anterior coBssissure. A pathway between the saygdala seid the
brainstem which has received very little notice was found which
passed o^sially below the caudate nucleus through the substantia
inooinata, over the optic tract end downward in the lateral-iaost
one-fifth of the cerebral peduncle. It could be followed as low as
the upper pontine region. This degeneration could be followed easily
on the Hisgl sections by gliosis but only with difficulty on the
myelin preparation. This apparently indicates an intermingling of
nosisal fibers although there sea&a no doubt that the anterior tem-
poral region provides fibers to Turck's bundle, i^elin degeneration
in l^rclc's bundle of the posterior tensor al region was sharp ^d
well defined. In the anterior teEsq>oral region, a well-defined
gliosis could be followed into the brachiism of the inferior colliculus but
neither of Che brachia of the colliculi appeared degenerated with
the posterior teasporal lesion.
Significance to Meurologjcal Research; These observations
contribute to the further understanding of functional representation
of the priioate taaporal lobe. Such understanding is in its way a
contribution to knowledge of the structural basis of such abstrset
functions as perception, ssentory, hallucination, as well ss the jeore
discernible functions of the autonomic system which find correlates
in the tsasporal lobe.
5,
gpoposed Ciourse of she Project; This project is develop-
ing because of infosmaCion deri^^ed from electrical, surgic&l, anatom-
ical, and biocb@oical studies. The relationship of the mesial
t^i^ral structures to biocbeadstry of catechol aaines nsust be
clarified and further steps in the study of hallucinogenic substances
may provide smne clue as to this intricate chemical end physiological
relationship, since the majority of these chemicals are related to
the adrenalin coai^ounds.
Part B included: Yes /X/ K© £7
serial Ko, _Hffii&:62M.
PHS-NIH
Individual Project: Eepor£
Caleadar Year 1958
Part B: Honors, ^ards, aad Publications
Publications other than abstracts £ro!B this project:
Frost, L. L., Baldwin, M. , and Wood, C. D.: Investigation of
the priiaate aarjrgdala: Mov<sasnts of the face and ja»s.
Neurology, 8, No, 7; 543-546, 1958,
Serial No, NINDB 63 (c)
lo SurgicaI~TJeuroKgy Brand
2c
3o Bethesda, Maryland
4. Same as NINDB 195? 26(c)
PHS-NIH
Individual Project Report
Calendar Year 1958
Part Ao
Project title; Effect of Tumors upon the Central Nervous
System Function and Structure
Principal Investigator; J. M. Van Buren, M. Do
Other Investigatorat Maitland i^aldwin, M. D.
Cooperating Units:
Man Years (Calendar Year 1958); Patient Days (Calendar Tear
" — — 1958 ) I
Totals o25 603
Professionals o25
Others o25
Project Descriptions
Objective;^ This study has a dual aim; (1) to carry out
physiological-anatomical correlations in man, (2) to evaluate
the effects of newer me"Uiods of treatment for tumors of the
central nervous systems In the present program attempt will
be made to utilize the intrusion of disease upon the central
nervous system of man as an "experimental" lesiono
Method Ep^£J-oyed^
Specific ally J the material is used in three major wayss
(1) physiological observations can be made during surgery s (2)
the effects of sujgery itself can be evaluated, (3) post mortem
material may in certain cases , prove valuable for anatomical
studies o
Major Findings?
In conjunction with the Branch of Endocrinology, NCI under
the direction of Dro Delbert Bergenstal, a study has been made
of the quantitative anatomical and endocrinological evaluation
of graded hypophysectomy in mano Thirteen cases formed the basis
of this studyc. From the serial sections of the sella volumetric
estimation vjas raade of the size of the retained pituitary
fragment and differential cell counts were made in this
fragment post mortera» This finding was correlated in each
case with the patient's clinical course and the response of
the thyroid and adrenal function, the level of gonadatrophins
and the presence or absence of diabetes insipiduso
Between 160 mm. and 0,3 mm.^ of pituitary tissue were
left in the sella in the 13 cases. Initially after surgery
and for a period extending up to 3 months, there was profound
depression of thyroid and adrenal activity, and the gonada=
trophin levels fell to negligible figures » It was dviring this
time that tumor remission might occur and tiiis was seen in
about $0% of the cases. The most striking feature was that the
presence of tumor remission and evidence of severe hypopituitarism
was present in all cases without regard to the amount of pituitary
tissue remaining in the sella. Thereafter, in the case retaining
160 mm.^ of pituitary (which incidentally had a veiy definite
objective tumor remission) thyroid and adrenal fimction returned
to normal, alttiough throughout the patient's 16 month post-
operative course the gonadatrophin levels renained near the
vanishing point. Thus, there is strong suggestion that de-
pression of the Individual trophic pituitary hormones is not
the same for all the trophic hormones. The one feature common
to all cases was- surgical section to the pitviitary stalk and
this may indeed be the essential feature.
The need for posterior pituitary extract to control
diabetes insipidus might or might not be present but this could
not be correlated in any way with the amount of pituitary tissue
remaining in the sella.
In 5 cases where the cholesterol values were followed in
the post-hypophysectomy period the maximum rise of the cholesterol
values (presvmiably an index of decreased thyroid function) were
seen between one and three months following surgery, then all
the values began to fall toward normal limits. Curiously enough,
this initial rise did not seem related to the volume of pituitary
remaining in the sella nor did the eventual fall appear to be so
related since quantities of pittiitary under 3 cu. mm. might be
associated with such a fall. Interpretation of this finding is
somewhat difficult since in liver disease from which most of these
patients suffered, a spontaneous fall in cholesterol may appear.
Histological features of interest showed a uniformly slow
rate of chromophile cells both of the alpha and beta types (10,000
cells counted per case) . This finding was interpreted as de-
granulation of the chromophile cells to increased demand for
pituitary hormones. The pharyngeal pituitary gland was examined
in 6 cases of the 13<. The raeasiirements of the pharyngeal
pituitary all lay within the lower limits of normal and not
the slightest evidence of secondary hypertropl^ was seen
despite the claims of some E\iropean investigators o
This study is now in manuscript form and will be submitted
to the Journal of Neurosurgeryo
During the present calendar year tumor cases have pro^
vided the post mortem material for studies of the visual systemo
Significance to Neurological Research;
The present stxidy on hypophysectomized patients has provided
some basic knowledge regarding the reason for effectiveness of
hypophysectomy in the treatment of metastatic tumors of the
breasto The curious finding that the clinical results of in-
complete hypophysectomy were apparently as good as those of
complete hypophysectomy seems explained by the profound depression
of pituitary honnonal output which seems nearly independent
of the amount of pituitary tissue removedo The differential
response of the various trophic hormones to pituitary injury
had not been previously conjfinned in man with anatomical
control o
The importance of this project in providing valuable
anatomical material for further stucty of the human visTial
system should be eraphasizedo
Proposed coTjrse of project:
Using the lead provided by the above study, the operation
has now been changed to a simple section of the pituitary stallc
without removing ary pituitary tissue <> The effect upon the
individual's endocrine status and tvunor will be evsiuated in
another dozen cases « At present h such cases have been carried
outo It is possible that later in the course of these patients
the area will be re-exposed and the pituitary removedo This
will provide an additional facet for investigationo
The hypothalami in 7 of the hypophysectomized patients
are being prepared in serial section for study of cell structure
and neurosecretiono Whether this study will prove practical
and fruitful is yet to be deteiroinedo
Part B included les /~7 No ^TJ
Serial Noc HIMI)B_6U (c)
1, Snrgic al" Neurology Branch
2o
3o Bethesda.. Maryland
]+o Same as NINDB 195? 38(c)
PHS"«NIH
Individual Project P.eport
Calendar Year 1958
Part A.
Project title; A Study of the Functional Anatomy and Pathology
of the Human Visual System
Principal Investigators Jo K. Van Buren, M, D»
Other Investigators; None
Cooperating Units;
Man Years (Calendar Year 1958): Patient Days (Calendar Tear
'°°'~™~~~~™~'"~~~™~~ 1958);
Total; o25
Professional; <.25
Others o25
Objective ; This study is directed toward a better under-
standing of the correlation of form and function ^ri-thin the htunan
visual systerno It is intended to carry out these studies correla^
ting clinical physiology with . anatoi^y using the best quantitatlTs
methods available o At the present time the study has been
divided into three parts;
(A) Retina
(1) Stucfy of the normal human and primate retina
using the Golgi and chroiaatic methods <.
(2) Study of the effects of the lesions of the optic
pathways upon the retina,
(B) Study of the effects of lesions of the optic pathways
upon the lateral geniculate bodyo
(C) Study of the visual, field defects following temporal
lobectomy „
Method Employed;
The methods employed have been given in detail in report
for Calendar Year 1957c
Major Findings s
The preliminary findings have been given in the report
for Calendar Year 1957 and will not be repeatedo Since it
has appeared desirable to confirm these findings in a greater
quantity of material before publication^ our efforts in the
present year have been concerned primarily with the collection
of more materialo Specifically, material collected has been
as follows:
(A) Retinas 8 casesj (2 chiasmal lesions, 3 papilledema,
2 normal, 1 amblyopia due to life long strabismus) o
(B) Lateral geniculate bodys k cases, (1 temporo=
parietal infarct, 2 chiasmal lesions^ and 1 enucleation of
long duration).
(C) Study of the visual fields following temporal lobe
defects 0 This study is now terminated.. See reference given
belowo
Significance to Neurological Research;
The general aims have been previously given in the 19$7
Calendar reporto In brief, it has been considered desirable
to re-examine the visual system in manly anatomical reconstruct
tion studies which may be correlated with the clinical examina°
tions of the field of visiono
Proposed course of projects
It is planned to use a projector (which has been under
construction for the past fourteen months) for two dimensional
reconstruction of the ganglion cell pattern in the retinae These
reconstructions will be in terms of ganglion cell thickness and
will be plotted with retinal distances equated to degrees of
visual arco In this way they will be readily comparable with
the patient's visual fields, which in most cases, were obtained
prior to deatho The hypophysectomy material has provided a
goodly quantity of normal material which is important for estab-
lishing a baseline o
The lateral geniculate stu^y still suffers from in?;ufficent
cases so that collection will be continuedo
Part B included ^T7 ^^s /""T No
3c
Serial No. /)////li B- & ^cj
PHS-NIH
Individual Project Report
Calendar Year 1958
Part B. Honors, Awards and Publications
Publications other than abstracts from this project:
J. M. Van Bxjren, M. D. and M. Baldwin, M. D. The Architecture of
the Optic Radiation in the Temporal Lobe of Man. Brain, 81: 15-
UO, 1958.
Honors and Awards relating to this pro^ct: None
Serial. No. mig)3,61_(c)
1, SiirgicaTTeurolo^f B;.-;
2.
3. Beihesdaj Maryland
Uc Same as KINDB 1957 27'
PIK-NIH
Individual Project Report
Calendar Year 1958
Part A,
Project title; Studies of Involuntary Movements
Principal Investigators J. M. Van Buren, M. D.
Other Investigators; Maitland Baldwin j M. D.
Cooperating Units?
Man Years (Calendar Year 1958);
Total; .25
Professional; =25
Other: o25
Patient Days (Calendar Year
1958);
Project Description;
Objective; At the present time there is no adeqiaate
explanation of the cause or Tnechanism of production of involuntary
inovementso It is hoped by careful correlation of clini.cal
findings, recording of the electrical activity of deeper structures
at operation and study of the anatomical material may provide new
information on this subjject.
In order that recordings can be made from the basal ganglia
and coagulation carried out here if indicated, a stereotaxic instru-
ment is needed to guide the electrodso Since the available designs
seemed inadequate in some respects, development of a new instrument
has been nndertakeno
Method Employed:
I. Clinical observationo
II. Photographic techniques o
III. Analysis by illustration of movement phases.
Ihe possible use of accelerimeters for the graphic demons tratioi-)
of the directional phases of involuntary movement is being investiga-
ted o vjhether this will prove to be a practical recording technique
remains to be determined.
The initial considerations were given in the calendar
year repor\ of 19576
The acVial testing of the stereotaxic instrument on
cadaver matevial has been severely handicapped by the great
difficulty in obtaining cadaver material at the NIH, After
much negotiation we were peiroitted to carry out our first
stereotaxic plicement on a cadaver in May 1958 and since
this time have been able to carry out our studies with only
5 cadavers o On the whole, the results have been encouraging,
in that they ha") 5 shown that the principle of the arcuate
electrode carrier (please see explanation in previous annual
report) is a sourd one under practical operating circ\3Kist3nces
and that the appaiatus is mechanically accurate o The problem
of obtaining good .ineumography in the cadaver was eventua3J.y
solved simultaneous ventricular and cisternal punctures then
clearing the fluid .Yom the ventricle by introducing air in the
cisterna magna* Thi foramen of Monro has proved to be a
useful sero point foi' the stereotaxic apparatus and initial
localization errors lave been corrected in later stereotaxic
placements c. Due to difficulties in aligning the present base-
line of the ster-eotaxic instrument with the horizontal plane
defined by the anterior and posterior commissure an increase
in the antero-posteriov tilting mechanism tfill have to be
made by a small meehaniial change,, This failure of adequate
tilting caused a number of the posterior lesions to be
erroneously higho
Bie method described for preparation of oiir own brain
atlas has proved economicvl in time and effort and appears
more accurate than the use of paraffin embedding and myelin
sections c Its limitation .lies in the failure of finer details ^
partiCTjlarly in the thalamus to be as evident. This, however,
is not considered of ma,1or importance since gross estimation
of the position of the lesion in the thalamus is easily achievedo
Tlie use of various fixatives has been investigated. It
was initially thought important to provide a fixative which
would support the brain (in order that string suspension would
not distort it) and wliich wouild not cause changes in weight or
size of the brain,, Consequently, all brains were measured both
for weight and displacement at the time they were removed from
the skull and at one and two week intervals thereafter o A
solution made up of glycerin and formalin and water provided
adequate support of the brain but caused excessive shrinking
(over 3^). Thereupon mixtures of formalin, water and mercuric
chloride were used -vjhich again supported the brain in an
adequate fashion but it was found that the mercuric chloride
bleached the gray-white differentiation in such a degree
that anatomical structures became difficult to distinguish
on the photograph o We have finally returned to fixation of
the brain in foarmalin by suspension from the vertebral artery
and find that the degree of shrinkage reaches negligible figures
in two weeks' time (1=2^) which is about the limit of accuracy
of our method of estimation).
The effect of carotid profusion was investigated since
it was thought that injection of formalin into the carotid
system might produce swelling of the basal ganglia and thalamus
since a complete wash through could not be achieved in the
cadaver materials and therefore introduce error. In order to
evaluate this, ventriculograms were carried out^ the 100 cc, of
formalin in^ cted in one internal carotid artery, then ventric°
ulography repeated. Small but definite distortions of the side
injected appeared from this study so that we have abandoned the
practice of carotid injectiono
Significance to Neurological Research;
The present work has served simply to acquaint the
principal investigator vdth the mechardcal proficiency of
his stereotaxic instrument and thus has dealt simply with
technical detailso The, ultimate cotirse of the project is
to study those diseases in which stereotaxic intervention is
indicated on therapeutic grounds o This material would fall
largely in the group of involuntary movements o
Proposed course of project;
Photographic records both by moving pictvire and by multiple
flash stroboscopic photographsj althotigh providing a record the
patient's movements are bulky to handle and are difficult to
analyze. It is hoped that with the use of accelerimeters a
means may be found for simple graphic recording of the movement
which could be correlated on the same time base as other features
(IEj, autonomic, motor^ etc).
Continued use of the stereotaxic instrument on cadaver
material is planned and it is hoped that the quantity of material
may increase in the future » When the investigator is satisfied
with his proficiency in the use of this instrument, patients
suffering from basal ganglia disease will be admitted for treat-
ment by destniction of various portions of the pallidum or
thalamus o During the course of this therapy, studies will be made
of the areas to be destroyed by depth electrode recording and
stimulation^
Part B included Yes f^ No ftj
Ssslai No, WimB 66 ic)
lo SurgicaTTIeOTologr Branch
2o
3'. Bethesda, Maryland
ho Saroe as NINDB 1957 39(c}
PHS-NIH
Individual FroJBCt Report
Calendar Year 1958
Part A.
Project title? Pain Mechanisms
Principal Investigators J. M, Van Buren, M. D,
Other Investigatorsj^ Mildred Blevins
Cooperating Units;
Man Years (Calendar Year 1958); Patient Days (Calendar Year
1958):
Totals olO 211
Professional; ,05
Others „05
Objectives The essential interest of this stu(^ is centered
ijpon methods for evaliiation of pain id.th its ultimate goal being a
quantitative eval'uation of the pain from which, the individual is
suffering o Depending upon the degree to which the primary aim of
the study is achieved the following studies can be undertaken:
(1) evaluation of the standard surgical procedux'es for relief of
pain, (2) evalxiation of standard medical procedures for relief of
pain, (3) the surgical apprcarh to pain pathways may be expected to
provide an opportunity for study. Surgical lesions of the central
nervous system -will be exploited as far as possible as the oppor"
tunity arises o
(A) Psychometric Methods: The patient is subjected to
Rohrsach test and the Minneapolis Multiphasic Personality Invsntoiyo
In addition, he is evaluated during a fojTnal psychiatric interview,
(B) Autonomic functions Sim\iltaneous records of blood pressui^aj
skin temperature, electrocardiogram, skin resistance, respiratory rati
pattern, finger plethysmogram and esophogsal and gastric pressures
are madco
In report for the previous calendar year of 1957 the findings
have suggested that those patients complaining of pain which appeared
more functional than organic in origin had unusually had unstable
autonomic responses.
In the present year examinations have been continued in an
attempt to correlate the degree of autononiic responsiveness with
other features of the patient's clinical picture and the picture
defined by psychometric testing. In sum, the results have been of
practically no valueo Autonomic responses to apparently the same
pain stimulus varied from examination to examination on the whole
tending to decrease as the patient becomes more used to the examiner
and the testing situation. In sum, our failure in achieving ar^
sound information on this study lies in our failure to achieve a
stable response baseline which can be satisfactorily compared with
a postoperative baseline (thus eliminating the factors of adaption),
and of even more importance the failure to establish a baseline
idiich may be compared from patient to patient in a groupo
Significance to Netirological Research;
Any method which will quantitate a patient's pain in an ob»
jective fashion is obviously of the greatest importance in many
spheres of research. Our use of autonomic recording seemed a
possible lead but it has not proved fruitful.
Proposed coiirse of project;
The formal study of this subject has been terminated.
Part B included Tes /77 No fTJ
PHS-NIH
Individual Project Report
Calendar Year 1958
Serial No. NINDB - 67
1. Surgical Neurology
Branch
2. Primate Neurology
3. Bethesda 14; ^^^r-.i^
4. NINDB~43 (c
Part A.
Pro.iect Titles Study of Cortical Intracellular Potentials-
Principal Investigator? Choh-luh Li, M. D.
Other Investigators None
Cooperating Unit t No ne
Mar, Years (calendar year 1958). Patient Days (calendar y ^-
Total: "25
Professionals o25 ^'^'"^
Others <.25
Proj ec,t_Descript i_on ;
Obiective. Recently the dendrites were believed to generate electrical
I^f^ aroely responsible for the potentials recorded from the surre..
Tf S cortex^ 'There wasalso evidence -^f- ^-^^^f ^li^^^^'^tf ^
+ ic:.u6 cultures that the glia element and the nerve cell should be
con idered as a functional unit. Thus the understanding of the acxxv. y
oHhe different components or elements in ^^e cerebral cortex ba.eon
physiological studies may throw son-,e light on the function ox the coi.- ..
Method Emplovedi Cats under light anesthesia were used« The ij^'^^^';^';^^
fgSiifr^^^i recorded with glass rnicropipette electrodes. The respon.
of potential were tested by local application of strychnxneo
Maior Findinosj The intracellular potentials recorded from the co:.::
^g^f5u;;d-i^-'be five in types, il) Steady potentials of "62.9 m^
unresDonsWe to afferent stimulation and local application of st.y^nn.u..
preSbiroriginating from glia elements ^^'^^^]^^l,.
oresu-nably originating from dendrites, (3) Small potentials P^-^^:-^t:
r^Snaptic Regions! (4) Brief spikes with an inHe-- - ^^--
phase presumably recorded from cell bodies, v5) Simple oxiet spiKe.
Serial No. NBiDB - 67 i-z
from astonso These potentials differed not only in their size and time
course, but also in their responses to strychnine. Strychnine showed
no effect on the glia cells and axons but either depolarized or hyper-
polarized the membrane of the cell body. It also appeared to enchance
the activity of the small potential and suppressed the large slow
potential. The results of this study also suggest that the mechanisra
of synaptic transmission in the central nervous system may be siffiiisr
to that across the neuromuscular junctions. Further they also suggest
that the importance of dendrites in the production of electricsl
activity of the cerebral cortess may be over publicized,.
Significance to Neurological Research; This study identified different
forms of intracellular potentials ascribed to different elements in the
cortex and suggested that the spontaneous behavior and responses to
stisnulatiors are different from these different elements. It ivas also
in these studies that depolarization and hyperpolarization of the ceil
roembrane of the cortical neurones by strychnine were first reported.
This observation suggests that there »ay be difference in metabolism
of different nerve ceil^ in the cerebral cortejc. It was also in this
study that sroall potentials similar to miniature end plate potentials
were described indicating that the mechanism of synaptic transmission
in the central nervous system and in the neuromuscular junction may well
by the same.
Proposed Course of Projects The small potential*; recorded intracelluia
from the cortesc will be further investigated. This may yield to some
understanding of the action of the anesthetic, agents which are knovri to
block either monosynaptic or polysynaptic transmissions. The study of
the large slow potentials presumably recorded from the dendrites and the
spontaneous oscillations of potentials recorded from cell membranes vAli
be continued. Finally the action of conv^jtlsive drug^ and acticonvulsive
drugs will be tested with the simple method described above.
Part B included Yes ^ No /~7
Serial No„ NINDD
PHS-NIH
Individual Project Report
Calendar Year 1958
Part Bs Honors J Awards? and Publications
Publications other than abstracts from this projects
Lis Choh-luhs Cortical lntraceil?jiar Potentials
and their Responses to Strychnine» J. Neurophysioio
(in press) « 1958
Honors and Awards relating to this projects
2.
Prinu
3.
Bail-;
4.
PKS-NIH
Individual Project Report
Calendar Year 19b3
Part At
Project Title t Factors Determining the Discharge of a Motor Neuron
in Cerebral Cortex,
Principal InvestiQatori Choh-luh Li, M. D.
Other Investiqator! None
Patient Days (calendar ye
None
Cooperating Unit; None
Man Years (calendar year 1958)
Totals .25
Professionals o25
Other? ,25
Project Description s i:
0.bj.ggtiveg To study the activity of nerve cells in cerebral corte
to changei of the external and internal environments,
Methods Employed; Cats either !.fnder light anesthesia or d-tubocui.
used. The activity l>f nerve ceils in motor cortex was recorded with asicro-
pipette electrodes while electrical stimulation was applied to various sub-
cortical structures and peripheral sensory nerves. The cells which are
intimately related to motor function have descending axons to the medullar
pyraaid and were identified by their responses to antidromic stimulati
and those in the motor cortex which do not have descending axons were
identified- as internunciai cells.
,MlJo^ f^iO^^I^^ As previously reported stimulation of the nucleus ventral-
lateralis of the thalamus activates the cells with descending axons and s\^i
presses the activity of the internunciai cells? suggesting that this thalai:
nucleus may have soEie control over the motor activity of the eKperimsntal
aniroalo It was also found that the internunciai cells in the motor corts:x
could be influsr.ced by the senro.ry volley set up at the peripher/c The
sensory volley, at times was also capable of exciting a motor neurone in
the cerebrum o Furthermore ? it was not infrequent to observe that the
sensory volley roay inhibit or facilitate the discharge of a cortical
ffiotor cello This study also dessonstrated that the refractory periods of
the pyrafBidal fibers varied froK 1,5 to 2,5 milliseconds and conduction
velocity from 8 meters to 95 meters per second „
Serial No. f^r:
S i qoif icance__t_Q._NeJiJXglo.alg-ai..-B6s.aar-Cjb s The above observations further
emphasized the role of subcortical structures and external stimuli in
the function of motor activity. It may be said that while the motor
cortex is immediately concerned in the initiation of movesRenti' infiut
from internal or external sources on the activity of motor ceils c^r
be overlooked. This study provides direct evidence that these
influences indeyd exist.
Proposed Course of Project 8 Further studies of the relationships b&i :
the activity of other subcortical structures such as the corpus striatum
the red nucleus, the vestibular? reticular and subthalamic nuclei as
well as the cerebellum and of the motor neurones in the cerebral cortex
and in the spinal cord may ylL^ld valuable information about the mechanism
of motor function.
Part B included Yes ^ No /^
Serial No» M
PHS-NIH
Individual Project Report
Calendar Year 1958
Part Bi Honors, Awards and Publications
Publications other than abstracts from this projects
Activity of Interneurons in the F.flotor Cortex
International Symposium,, Recticuiar Formation
Henry Ford Hospital, Dstriot. Littles Brown
and Coc, 459-272, 1958.
Li, Choh-luh. Some Properties of Pyramidal Neurones
in the Motor Cortex with Particular Reference to Sensory
Stimulation. J. Neurophysiolo " (in press) « 1958
Honors and Awards relating to this projects
SeiiaJ. No. wxivUp^
lo Surgicai Neurology
Branch
2. Primate Neurologv
3» Bethesda 14, Mar •
4.
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A^
Prelect Titles The Problem of Synchronous Activity of Nerve Cells in
Cerebral Cortex
Principle Investigators Choh-luh Lis M« D,
Other Investigators; None
Cooperatino Units Ncne
Patient Days Icaieiidsr year 1958,;
NONE
Man Years tcalendar year 195Sj:
Totals o25
Professionals =25
Others ,25
Project DescrijDtiqn;
Obj[ectives Since the statement made by Adrian in 1935 it has been ger,3r
accepted that the activity of nerve cells in the cerebral cortex i«;ere
synchronous when the subject Was at rest or when the cortex was synchroiiou:
activated. And if the subject was alert the discharge of cortical cells
was said to bs "dys-synchronized"^ There has been, however, no direct
evidence in support » The present study is attempt to test this hypothesis.
Wethpd,, Employed % The activity of a cortical cell was recorded with a micr:
pipette electrode while that of the others was studied with another micro-
electrode.
Major Findings; In general the notion proposed by Adrian was given support
by direct evidence with the following reservations! (1) Only very £e-«¥ ner-
cells in a sphere of I nun in the cerebral corte^i would discharge precisely
at the same instant, (2) A synchronous volley evoked discharges of nerve
cells with a temporal descrepancies varying from 2 billiseconds to 20 nulij
seconds, (3) Application of strychnine activate about 85% but not all of
the nerve cells. (4) A temporal relationship between neuronal activity
still exist in "aroused" cortex..
^Significance ? It has been said that the neurons in en epileptogenic
cortex tend to fire in unison and neurons in normal cortex of an alei-i
subject randoissly discharge. The present study demonstrated that this
is a generalisation with certain degree of truth, based on logical
thinking but not on facts. The fnethods of simultaneous investigation
of the activity of different nerve ceils may provide additional in-
formation about the integration function of the central nervous systa,
Proposed Course of Study; Multiple recording with microelectrodes frc
single nerve cells will be used in the study of epileptic activity of
the cerebral cortex as well as factors determining the discharge of
motor ceils in cortex.
Part B included Yes f^ No [J
Serial Mo. NINDc
PHS-NIH
Individual Project Report
Calendar Year 1958
Part Bg Honors, Awards and Publications
Publications other than abstracts from this projects
Li, Choh-luh Synchronization of Neuronal Activity in
Cerebral Cortex. SciencSo (in press].
Honors and Awards relating to this project:
PHS-NIH
Individual Project Report
Calendar Year 1958
Serial No, NINDB__-.70
lo Surgical Neurology
Branch
2. Primate Nsuroloi.-f/
3. Bethesda 14, " .
4. NINDB-41 (c)
Part Ac
Project Title? Neuromuscular Transmission in Hypothermia
Principal Investigator? Choh-luh Li, M. D„
Other Investigators s None
None
Man Years (calendar year 1958]
Total ! ,25
Professionals „25
Other* „25
Project Descriptiong ■
Patient Days (calendar year 1958)
None
[i To study the performance of neuromuscular junction in raanaiisis &■'
low temperatures as compared to that in the amphibia,, The latter was repor
last year with the collaboration of Dr, Peter Gouraso
Method^ ^ Employ ed $ The anterior gracilis muscle and the obturator nerve of ti
rat were exposed and the miniature endplate potentials, endplate potentials
action potentials and resting potentials were recorded vsfhile the animal .was
subjected to various temperatures between body temperature and "A'^Co At
these temperatures electrocardiograms of the animal ware also taken.
Major Findings s As in the frog there were also a critical body temperature
below which action potentials of the muscle in response to obturator nerve
stimulation became less frequent. This was i5°C, At about 4-5°C action
potential failed and there were only endplate potentials elicitable by
nerve stimulation. The miniature endplate potentials could be recorded
by body temperature as low as 4 C but not below. The resting membrane
potentials showed no significant change at temperatures between body
temperature and 10°C, below which they began to fall, and at body temperatu;
of O^C no resting potential was recorded. Furthermore, during the process
of cooling some muscle fibers becaiise spontaneously active with discharges
of fibrillation potentials.
SiM!}M^,3S£^^LMMISlSS3:£Sl.,3§3SS:FSh^ This ^udy indicates that th
£ critical body temperature in maHsnals belo\H which the transmission
isiiipulses across the neuromuscular junction become impeded and if thi
tejiisperature is further lowered to 4°C transmission is blocked. This
observation may be of some use in processes involving hypothermia whiui-
are to be carried out in the laboratory or in the operating rooin for
human patients.
Proposed, course of Prpjects In the future similar experiments will t3
conducted with inquiries into the action of some neuromuscular drugs.
The preparation described above is found to be most suitable for this
type of investigation with intracellular microelectrodesi since at iov;
temperatures twitch movement of the muscle was reduced and anesthesia
was not required, yet miniature endplate potentials, action potentials
and resting potentials could be readily recorded.
Part B included Yes ^ No ^
PHS-NIH
individual Project Report
Calendar Year 1958
Part_Bg Honors } Awards » and Publications
Publications other than abstracts from this projects
Li, Choh-luhs Effect of Coolifig on Heurorauscular Transmission
in the Rat. Asaer. J, Physiol„ 194s 200-206, 1958,
Honors and Awards relating to this projects
Serial No. NIIiDBj-_71 (c<
1. Surgical Neurology
Branch
2. Primate Neurology
3. Bethesda 14s Maryland
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Project Title; Effect of Cooling on Conduction of Impulses in Cranial
and Peripheral Nerves »
Principal, Investigator ? Dr, A. Ortiz, M<, D,
Other Investigator i Dr» Choh-luh Li, M«, D.
Cooperating Units; None -
Patient Days (calendar year 1958} g
None
Man Years (calendar year
Totals ao
Professional » oOS
Others .05
Project Descripmons
Objective; Experiments and surgical procedures designed to abolish functions
of the nervous tissue have been priraariiy performed either by ablation or
electrolysis. It is thought that extreme low temperature locally applied
to the tissue may have similar results without other undesirable complicationo
Method Employed I A small segment of the optic nerve and the sciatic nerve
were subjected to -150 C for 30 seconds. The aniraals were than kept for
1 day - 4 months and the impulse conduction was tested at various intervals.
Major Findings i This set of 7 experiments was initiated only 3 v.feeks ween
this report was submitted. Results obtained should be considered inconclusive
and will be reported at a later date*
Significance to Neurological Research s Wiicn all the dates are assembled in-
formation about functional interruption and functional recovery subsequent
to cold may be of some significance in further improvement of the operative
techniques presently employed. '
Proposed Course of Projects This study will be continued and may be extended
froiD nerve fibers to nerve substance »
n
No k1
PHS-NIH
Individual Project Report
Calendar Year 1958
Serial No, NINDB - 72 (c j
1„ Surgical Neurology
Branch
2. Primate Neurology
3. Bethesda 14s Maryland
4.
Part A.
Study of Pharmaceutic Agents Acting on Various Cortical
and Subcortical Structures of the Brain.
Principle Investiqatort A-tOrtis.» ^ia B..
nthPT Tnvestiaators 8 M. Baldwin, M. D. and Choh-luh Li, M. D,
Cooperating Units None
Man Years ( calendar year 1958)8
Total! olO
Professionals o05
Others <.05
Patient Days (calendar year 1958'
None
Objectives Since the introduction of 5-Ht, LSD-25, LSD interest in research
^fl^iP^tll disorders has been greatly enchanced, yet little has been known
about the underlying neurological mechanisms of their effects. Recently a
certain specis of mushrooms found in Mexico was found also to cause similar
results. This study is designed to investigate which cerebral structure is
most effected and how these agents would alter the electrical activi^y and
responses of the nervous tissue.
Method Employed: Cats and monkeys were usedo Multiple electrodes, which
were also capable of injecting minute quantities of the testing chemicle
agents, are inserted into the various <M&p str^ctwres ©f the brain. Recording
of electrical activity and responses to stimulation fx&R these structures
and from cortical surface were made.
MsiS£._5iQSL^SS^ ° '''he results, though interesting, were still inadequgti
for a conclusivs statement to be made.
Significance ti> Neoroloqical Research; to be seen
Proposed Course of Projects This study will be continued.
Part B included Yes [J No ^
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Projact Titles Properties of C
Principal Invest icjators; Choh-
Other Investigators 3 King Engel
Cooperating Unit? None
Man Years (calendar year 1958) t Patient Days: (cale
Total; .10
Professional; .05' None
Others .05
Project Descriptions
Objectives Spontaneous electrical p,/„i-.L>o _.-.-. s c,nd potentialG :....
to electrical stimulation recorded from single elements in the v
nervous system has been carried out in the past without the advc.;... ...
of direct vision. The previous experiments also were subjected to
various uncontrollable factors, e,g.y presynaptic random bonabardments
and anesthesia. With the establishment of a tissue culture laborator,
it seemed to offer an opportunity to study the unit property of the
nerve elements in isolated form under controlled external enviroivmsrit.
It was also desirable to study various epileptic and anti-epiieptir.
agents acting on the nerve cell merabrane. Furthermore agents presu^as
effecting the neuromiiscular junctions may be tested on culture suscle
ceils without endplate organs. Finally the presence of an electrical;
excitable membrane of brain tumor cells may be disclosed. It v;- -
planned that with a similar method study the epileptogenic tissu
removed from patients could be investigated. This project is t!
3 long term proposition and the results will have to depend upc;.
techniauss to be developed.
NINDB -
Methods Employed i In order to test the methods intracellular recording
from heart muscles « skeletal muscles snd spinal ganglia of 2-3 week old
chick embryo were carried out. The results were coiuparable to those obtained
from adult ratsc With this assurance experiments were performed in
skeletal muscle cells and spinal ganglion cells after growing 7-21 days
in tissue culture. The culture technique wss essentially the same as
that described by Murray » Bornstein and P inerat and the recording
stimulating methods were similar to those used by Li and Mcllwaino
Major Results? The observations obtained from the spinal ganglion cells
resembled those frcm the nerve cells of the cerebral cortex and spinal
cord. The results of cultured chick muscles could be summarised as
follows: (1) Cells with slow responses might remain inactive after
excitation for as long as 4,2 seconds and take no part in the initiation
of spontaneous rhyti-unic spike discharges. (2) Cells with twitch responses
had a refractory period ranging from 25 to 35 milliseconds and were responsible
for the spontaneous rhythmic spike discharges. The spikes generated from
these cells might be as large as 100 raV and 2.0 msec. (3) Resting membrane
potentials (66+5 mV) showed no significant difference in cells with twitch
and slow responses j nor was there any change with age of the cells from
7 to 21 dayso (4) Spontaneous rhythmic oscillation of potential could occur i.
the absence of spike discharges; but having attained a critical level of
depolarization J they initiated spike discharges. The spike discharges did
not interfere with the rhythm of the oscillating potentials.
Significance to Neurological Research; In the experiments with spinal ganglio
cells it appears that the results may also be applicable to nerve cells of
maiTBnals. The results from chick cultured muscles are similar to those found
in denervated mammalian skeletal muscles and suggest that the mechanisms of
the fibrillation potentials in both cases are similar. Furthermore 9 the two
types of responses suggested a differentiation of function being present
in embryonic muscle cells.
Proposed Course of Projects A co-relation of the deveiofsrient change and
psychological function of the muscle will be studied. Investigations on
the action of acetylcholine, ClOj curarine, etc. 9 on cultured mammalian
muscles will be carried out. Study of the spinal ganglion cells and
other nerve cells, tumor cells and epileptogenic cells are planned.
Part B included Yes ^ No £J
PHS-MIH
Individual Project Report
Calendar Year 1958
Part B; Honors; A'^aidsj and Publications
Publications ot^ 5r than abstracts from this projects
Lij Choh-luiris Klatzo, I., Baldwin, M.j and Engels K.
Properties of Cultured Nerve and Muscle Cells.
J, Compo Neurol, (in press).
Honors and Awards relating to this project;
National lastitute of Neurological
Diseases and Blindness
Clinical Research
Surgical Neurology Braoch
Section on Clinical Neuropathology
Serial Nudbere of Projects:
KIKDB- 74(c) » NIHBB-75(c), NIHBB-76Cc), HINDB=77Cc},
HIHDB-78Cc), KIKBB-79Cc), NIKDB-80(c), aad
NINDB-SlCcK
Ee£iiaat@d Obligations for FY 1959
Total: $119,500
Directs $55,200
Rsimburssment : $64, 300
Serial No.J!f!!^LlZiiiL„
lo Surgical Neurology
Branch
2. Clinical Neuropathology
Section
3. Bethesda^ Maryland
4. New
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A,
Project Title; Pinocvtosis of Labelled Proteins in
Tissue Culture.
Principal Investiqatort Igor Klatzo, M. D.
Other Investigators s W. K. Engel, M. D. and Jo Miquei,
Ph. D.
Man Years (calendar year 1958); Patient Oavs (calendar
Total; .25 years 1958); 0
Professionals .25
Other; .25
Pro.iect Description;
Objectives s Pinocytosis or "drinking by the cells" is a
phenomenon which has been reported by a number of workers in
tissue culture. Its intrinsic role in cell roetabolism has
been suspected'^ however, these assumptions are based only on
phase-contrast observations of intracytoplasmic vacuole for-
mation. By labelling the proteins with fluorescent component
and feeding cultures with these labelled proteins it should
be possible to demonstrate the uptake of various proteins by
living cells and follow their metabolic fate. The differences
between individual cell types could be demonstrated in this
respect. By changing environment of the cultures influence
of various factors (pH, temperature, chemical substances, etc.^
on the cellular protein metabolism could be studied.
Methods Employed s New-born kitten and rat cerebellum
was grown in vitro. Cat serum albujnin and rabbit serum
globulin were labelled with fluorescein isothiocyanate.
Cultures were "starved" for three hours receiving only bal-
anced salt solution and consequently fed with labelled
proteins in concentrations corresponding to their usual
content in the medium. After washing for different periods
of time? in balanced salt solution the cultures were ob-
served under the fluorescence microscope*
Major, Findings; Our preliminary findings indicate that
it is possible to demonstrate protein uptake by living cells
grown in vitro. A significant difference in metabolism of
proteins by various cellular elements has been observed»
Cultures washed for a brief period of time after feeding
showed abundant labelled proteins in the macrophages and
only few fluorescent droplets in the glial elements^ Cul-
tures washed for several hours in balanced salt revealed
abundant green fluorescent droplets in glial cells, whereas,
the macrophages showed mostly autofluorescence of various
lipid substances. Also, some differences between behaviour
of albumins and globulins have been noted.
Proposed course of the pro.iects It is proposed to
continue this investigation in order to accumulate more
information along the lines mentioned in the statement
about the objective of this project.
Part B included! Yes /~7 No JyTl
Serial Ho. NINDB - 75 (C)
1. Surgical Neurology
Ei-anch
2» Clinical Neuropathology
Section
3, Bethesda, Maryland
4. KINDB 35 (C)
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Pro.iect Title; The Localization of Myosin in Human
Striated Muscle by Fluorescent_.Antx~
body.
Principal Investigator; Igor Klatzo, Mo D.
Other Investigators s B. Hoxvath, Mo Do and E. W<, Emniartj
M, D.
Cooperating Units; NIAMD-E. W, Enraart, M. Oo Project NOo
NIAMD (62303)31.
Man Years (calendar year 1958); Patient Days (calendar
Total? .25 year 1958); ^62
Professional; ,25
Others .25
Project Description;
Objectives,; The inorphQlogical localization of myosin in
striated rriuscie was studied using fluorescent antibody, tech-
nique. Information derived from the study of the normal
muscle was used as a base-line for the observations on the
behaviour of myosin in muscle affected by various neuro-
muscular disorders. Supplementary information was derived
from study of the experimental muscle lesions in the rabbit
and of the erabryonic chick muscle grown in tissue culture.
?/.ethods Employsdj^ Coons' fluorescent antibody technique
was applied for this study. The rabbits were insnunized with
human myosin and the obtained globulin fraction of antisera
was labelled with fluorescein isothiocyanate. Muscle biopsies
Methods Employed (continued) s
from the patients and experimental animals were stained with
fluorescent antibody and examined in the fluorescence micro-
scope. The embryonic chick muscle was grown in the Maxiraow
slides and studied on consecutive days in the phase-contrast
polarized light and by staining with fluorescent antibody,
Ma,1or Findings; In the normal muscle the specific
staining for myosin was observed in A band, I and M bands
appearing unstained and Z band showing occasionally non-
specific autofluorescencec Study of various pathological
processes in human muscle revealed a striking persistence
of antigenic reactivity of myosin in the fibers with far
advanced degeneration. Regenerating fibers observed in
cases of polymyositis and experimental muscle injury showed
similar features to those muscle fibers grown from the chick
embryo. In acute muscle injury and in a few cases of poly-
myositis occasionally few macrophages contained green-
fluorescent inclusions in their cytoplasm. This observation
may be of importance for the interpretation of the possible
mechanism of hypersensitivity due to release of muscle
proteins.
Proposed course of the project; This project is
completed.
Part B included; Yes ^ No [^
Serial No, ^I^™ 75
PHS-NIH
Individual Project Report
Calendar Year 1958
Part Bs Honors, Awards and Publication
Publication other than abstracts from this projects
"Demonstration of Myosin in Human Striated Muscle
by Fluorescent Antibody".
Igor Klatzo, M. D», Beni Horvath, Mo Do and
Eo W, EuimartiiM. D.
Published in the Proceedings of the Society for
Experimental Biology and Medicine, 1958, Vol. 97,
135-140.
Honors and Awards relating to this projects
Serial mo„ WINDS - 76 (C)
1. Surgical Neurology
Branch
2. Clinical Neuropathology
Section
3. Bethesdaj Maryland
4. NINDB 37 (C)
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Pro.iect Title; Study of Pathology of Kuru Disease.
Principal Investigator » Igor Klatzo, M. D.
Other Investigators 3 D. C. Gajdusek, Mo D= and
V. Zigas, M. D.
Man Years (calendar year 1958); Patient Days (calendar
Total; .25 year 1958); 0
Professional; .25
Other; .25
Pro.iect Description;
Objectives; Investigation of pathological changes in Kuru
Disease affecting the Fore people of New Guinea was undertaken
in 14 cases in which brains and other tissues were available
for study.
Methods Employed; Established histological and histo-
chemical techniques were employed for this study.
?4a.lQr Findings? The main pathological findings in Kuru
were confined to the central nervous systeai and they consisted
of: (l) Widespread neuronal degeneration. (2) Myelin
degeneration affecting predominantly cortico-spinal and spino-
cerebellar tracts. (3) Intense and widespread astroglial and
microglial proliferation. (4) Perivascular cuffings with
mononuclear elements. (5) Presence of peculiar plaque-like
bodies in half of the cases studied.
Proposed course of the projects This project is completed.
Part B included; Yes i^J No FH
serial No, "I^^L:Jii£L
PHS-NIH
Individual Project Report
Calendar Year 1958
Part B; Honors, Awards and Publication
Publication other than abstracts from this projects
"Pathology of Kuru", Igor Klatzo, M« D., Do C. Gajdusekj
M. Do and V. Zigas, M. D.
Accepted for Publication in "Laboratory Investigation".
Honors and Awards relating to this projects
Serial N0cJlEELlili2l_
1. Surgical Neurologv
Branch
2, Clinical Neuropathology
Section
3o Bethesdaj Maryland
4, New
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Project Title; Study of Regeneration in the Central
Nervous System.
Principal Investigators Armando Ortiz-Galvan, M.D.
Other Investigators t Edward J. Laskowski, M.D. and
Igor Klatzo, M.D.
Man Years (calendar year 1958)5 Patient Days (calendar
Total:. .25 year 1958); 0
Professional! '25
Others "25
Pro.iect Descriptions
Objectives 3 The problem of regeneration of nervous elements
vdthin the central nervous system is of an obvious importance.
The numerous investigations in this field indicate that the main
obstacle for successful regeneration of the nervous fibers is
encountered in the reaction of the connective tissue which
blocks the pathways. By application of the metal plate at a
low temperature to the optic nerve it is hoped that the connec-
tive tissue reaction will be reduced to a minimum. This
assumption is based on the study of cold lesion produced in the
cortex of the cat. In addition, the intra-cysternal injection
of the prednisolone compound, which is one of the most powerful
adreno-cortical steroids, may further reduce mesodermal reaction
and thus provide conditions for effective regeneration of the
optic nerve fibers.
Methods Employed s A series of cats are being operated and
experimental lesions are produced in the optic nerve by appli-
cation of a metal plate at low temperature. In addition, one
Method £giploved{continued)i
group of animals is being injected intra-cysternally with
prednisolone. The animals will be sacrificed at various
time intervals ranging from one week up to four months »
The progress of regeneration would also be followed
electrophysiologically by photic stimulation and recordings
from various parts of the central nervous system.
Proposed course of the project { It is proposed to
continue this project to obtain complete data based on
histologicalj electrophysiological observations from the
groups of studied animals.
Part B included! Yes /ZJ No fxj
Serial No .J™?JLZii£l
1. Surgical Neurology
Branch
2, Clinical Neuropathology
Section
3, Bethesdaj Maryland
4. Formerly NINDB 33 (C)
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Project Title; Histochemical and Electrophysiological
Observations on the Muscle Fibers Grown
in Vitro.
Principal Investigator; W. K. Engel, M. D.
Other Investigators; Choh Lu Li, MaD. and Igor Klatzo,
M. D.
Man Years (calendar year 1958) ; Patient Days (calendar
Total; .10 year 1958); 0
Professionals .05
Other; .05
Pro.iect Description;
Objectives; Muscle tissue of chick embryo or new-born
rat grown in tissue culture presents an exceptionally suit-
able object for correlations between the appearance of various
chemical substances, demonstrated by histochemical methods,
and electrical activity of the muscle fibers. Specifically,
it is planned to correlate the observations on the nucleic
acids, polysaccharides and contractile muscle proteins with
the electrical activity of the corresponding living cells by
intracellular microelectrode technic.
Methods Employed; Muscle tissue obtained from 14 day old
chick embryos or new-born rat is grown in vitro. The cultures
are studied on consecutive days with the following methods;
(l) Nucleic acids, with methyl green-pyronin, gallocyanin
with controls by digestion with ribonuclease. (2) Poly-
saccharides, with PAS,Toluidine blue, etc. (3) Contractile
Methods Employed (continued) ;
muscle proteins, with specific fluorescent antibodies « Before
undergoing the histochemical procedures the muscle fibers are
observed and photographed in phase contrast and in polarized
light. For correlation, the corresponding cultures are sub-
jected to study of electrical activity with intracellular
microelectrodes .
Major Findings 8 The dynamic changes in RNA content has
been demonstrated with gallocyanin and Toluidine blue methods.
The first appearance and localization of myosin in myofibrills
has been followed with specific fluorescent antibody. Data
on the electrical activity have been obtained from the cultures
several weeks old.
Proposed course of the pro.iect; It is proposed to continue
this investigation to complete the lacking observations for
correlative interpretation of the findings.
Part B included: Yes fl No fTl
Serial No. "XNDB - 79 (C)
1. Surgical Neurology
Branch
2o Clinical Neuropathology
Section
3o Bethesda, Maryland
4. New
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Prciect Title? A New Method for Quantitative Study of
Precipitin Reaction.
Principal Investigators Jaime Miquel, Ph.D.
Other Investigators? B. Horvath, M. D. and Igor Klatzo,
M. D.
Man Years (-calendar year 1958); Patient Days (calendar
Totals ,25 year 1958)8 0
Professionals ,25
Others .25
Project Descriptions
Objectives 8 A simple and quantitative method for esti-
mation of precipitin reaction is of obvious value. By appli-
cation of ant igen-anti body mixtures to the chromatographic
paper with consecutive separation of soluble proteins in the
paper the insoluble antigen-antibody precipitate can be quan-
titatively evaluated by simple calorimetric methods. By
using fluorescent antibody instead of serum in the test, the
ratio between the amount of antibody to antigen in the pre-
cipitate can be quantitatively estimated.
Methods Employed 8 Serial dilutions of the mixture of
antigen-antibody are applied to the chromatographic paper
and run with buffer. The insoluble antigen-antibody pre-
cipitate remains at the starting line, whereas, the soluble
proteins move away through the paper. The paper strips are
stained for proteins with bromphenol blue. The dye bound
to the precipitate is eluted and estimated quantitatively
in the calorimeter. Similarly the fluorescein isothiocyan-
ate bound to the antibody in the precipitate is eluted and
quantitatively analysed in ultra-violet spectrophotometer.
Major Findings; This method has been applied to the
precipitin reaction between antigens of contractile muscle
proteins and their respective antibodies. The quantitative
data obtained with this method were in agreement with much
more complicated and cumbersome Kjehldal nitrogen determin-
ations. The sensitivity of the method was estimated to be
as low as 1 gamma of nitrogen.
Proposed course of , the project; It is proposed to
evaluate further this method in application to various
immuno-chemical systems.
Part B included: Yes /~7 No fxl
„ . _ . . NINDB - 80 (C)
Serial NOo ..
lo Surgical Neurology
Branch
2. Clinical Neuropathology
Section
3. Bethesda, Maryland
4. NINDB 29 (C) & 30 (C)
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Project Title; The Relationship between Edema, Blood-
Brain-Barrier and Tissue Elements in
Experimental Brain Injury.
Principal Investigatori Igor Klatzo, M. D.
Other Investigators; Ac Piraux, M. D. and Edward J.
Laskowski, M. D.
Man Years (calendar year 1958);. Patient Days (calendar
Total: .25 ysar 1958); 0
Professional; .25
Others .25
Project Descriptions
Objectives: The objective is to study the interrelation-
ship between edema, blood-brain-barrier and behaviour of brain
tissue elements.
Methods Employed; In order to allow conclusions about the
chronological sequences of the changes observed} the brain lesion
associated with edema should be reproducible with great uniformity.
This requirement was satisfied by the application of low temper-
ature to the exposed cortex under constant conditions of time and
temperature. Groups of cats were sacrificed following this pro-
cedure at various time intervals.
Sodium fluorescein was used for study of blood-brain-barrier.
Following fluorescence photography the brain tissue was subjected
to a variety of histological and histochemical procedures. A
group of animals was sacrificed for electrophoretic study of
protein patterns in the edematous and normal white matter.
Major Findings; The development of edema was observed
within 6 hours in the white matter underlying the site of
cold application. The area of edesna exhibited strong PAS-
positive staining of astrocytes and less intense PAS staining
of interstitial spaces. Histochemical analysis of PAS-
positive staining In the edematous white matter suggested
glycoprotein nature of the substances involved. The break-
down of blood-brain-barrier in the edematous white matter
as tested with sodium fluorescein followed after approxi-
mately 18 hours. Electrophoretic studies performed at the
time of maximal intensity of edema and break-down of blood-
brain-barrier indicated an appreciable increase of total
proteins with striking elevation of albumins in the area
of edema.
Fluorescence in the superficial layers of the cortex
persisted one month after injury and was associated with
the presence of small astrocytes j lacking well-formed
vascular foot-plates.
Proposed course of the Project; This project is
completed o
Part B included! Yes ^J No [Z]
Serial No. ^^^^ ^
PHS~NIH
Individual Project Report
Calertdar Year 1958
Part Bs Honors, Awards and Publications
Publication other than abstracts from this projects
"The Relationship between Edema, Blood-Brain-Barrier
and Tissue Elements in a Local Brain Injury".
Igor Klatzo, M. D,, Andre Piraux? Mo D. and Edward J.
Laskowski, M. 0.
Publication: Journal of Neuropathology and Experimental
Neurology Vol. XVII ^ No, 4, October, 1958.
Honors and Awards relating to this projects
Serial No, ^INDB 81 (C)
1. Surgical Neurology
Branch
2« Clinical Neuropathology
Section
3. Bethesda, Maryland
4, NINDB 31 (C)
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A. I
Prelect Title; Study of the Effects of Hypothermia on
Injured and Normal Brain Tissue.
Principal Investigator; Edward J. Laskowski^ M. D.
Other Investigators; Igor Klatzo, Mo D.
Man Years (calendar year 1958); Patient Days (calendar
Total! .25 year 1958); 0
Professional; .25
Other: .25
Pro.iect Description:
Objectives; In view of the recent interest in the use of
hypothermia in neurosurgical procedures $ the objective of this
investigation is to assess the effects of the lowered body tem-
perature on the various aspects of brain injury such as edema,
pesraeability of blood-brain-barrier, etc. Also, an elucidation
of tolerance of normal brain tissue to various degrees of hypo-
thermia is imperative.
Methods Employed; The assessment of the effects of hypo-
thermia is based on a comparative study of our standard cold
lesion, as described in Project NINDB 29 (C), in normothermic
and hypothermic animals. Groups of cats were submitted to
lowered body temperature and were operated on in a similar
manner when the rectal temperature reached 26° C. The animals
were maintained at a rectal temperature of 24-28° C for periods
of 4-6 hours after application of the cold plate. The cats
were sacrificed at various time intervals and the brain tissue
was submitted to procedures, similar to those used in normo-
thermic animals.
Major Findings i The most striking difference between
hypothermic and normothermic animsls were revealed in the
behaviour of the blood-brain-barrier. Twenty-four hours
after cold application in normothermic animals there was
intense fluorescence of the white matter extending into
the adjacent gyri. In contrast, all hypothermic animals
sacrificed after 24 hours showed the fluorescence limited
to a peripheral margin surrounding the non-fluorescent
superficial necrotic lesion. At 48 hours there was, how-
ever, an increase in the area of fluorescence in these
hypothermic animals but this was still less than seen at
maximal edema at 24 hours in the normothennic group.
Histological preparations reveal a lesser astroglial
reaction in hypothermic preparations at comparable jseriods
of sacrifice. The PAS-positive staining of the astrocytes
and the interstitial substance is similarly diminished in
the area of edema in the hypothermic animals
Proposed course of the projects These observations are
now based on sufficient numbers of animals to be conclusive
and this phase of the project is complete. It is planned,
however, to continue the study of this lesion followed by
the induction of hypothermia in an effort to evaluate the
use of lov/ered body temperatures in the treatment of brain
trauma.
Part B included! Yes /^ No /™7
„ . , ., NINOB 81 (C)
Serial No,
PHS-NIH
Individual Project Report
Calendar Year 1958
Part_B£ Honors, Awards and Publication
Publication other than abstracts from this projects
"Observations on the Effects of Hypothemia on
Experimental Brain Lesions", Edward Jc Laskowski, M, Dc
Accepted for Publication in The American College of
Surgeons Surgical Forum, Volume IX.
Honors and Awards relating to this projects
Kstioaal lasfcitufie ©f Keurolpgicai
Bleeaees &nd Blindness
Clinic al Research
Surgicel Neurology Braach
Seceioa on Developmental Neurology
Serial Kusabers of Projects:
HIM)B-82<c), NIHDB-83(c), KIK&B-84<c) , inmB
HIHSB-86(c), and NIilDB-87(c),
Essia&ted Obligations for FY 1959
Totals $128,000
'Dlract: $36,800
ReimbuiTiSsmsiit s $91 ^ 200
serial No. NINDB-82 (g)
1. Surgical Neurologj^-
2. Devslopmentsi Neurology
3. Bethesds, Masryland
4. Same as NIKDB-45 (c)
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Project Title; The Investigation of the Site, Type and
Extent of Lesions Involving the CNS in
Cerebral Palsy and Allied Conditions.
Principal Investigator; Anatole S. Dekaban, M,D,
Other Investigators; None
Cooperating Units; None
Man Years (calendar year 1958); Patient Days (calendar year 1953)
Total: .35 1,215
Professional : . 35
Other: .35
Project Description;
Objectives;. Comprehensive clinical, laboratory and genetical
investigation of a selected group of children suffering
from organic brain syndromes and epilepsy and correla-
tion of thus obtained data with the findings in pneumo-
encephalogram. The main objectives are: 1 - Correlation
of the clinical features of cerebral dysfunction with
the site^, size and character of the cerebral lesion «
2 • Classification of a larger group of children suffer-
ing from organic brain condition according to the
etiological factors whenever these were established.
3 - Preparation of publications based on smaller groups
of patients presenting particularly important aspects
in relation to pathology, pathogenesis or response to &
new type of treatment.
Methods ^ployed;
1. Genetic investigation.
2. Detailed neurological esaminatioHs including develops
mental testing and electroencephalogr«BB.
3. PneucEoencephalogram.
4. Other special tests as indicated.
Patient Material ;
NOj_ Aver. Stay in
Admissions: Children Male 28 " 22o5
Children Female 28 20.5
Clinical Project
Major Findings;
During 195S & total of 56 patients were studied in great
detail as in-patients and 23 on the out-patient basis «
Analysis of results of clinical investigations revealed
that in 62 percent of cases the site of lesion was detereain"
ed, in 27 percent the abnorraality was of diffuse character
and in 11 percent the localisation was not possible. In
29 percent of cases the pathological lesion was compatible
with destructive process , in 12 percent with congenital
malformation or hydrocephalus, in 21 percent it v&s of
diffuse character and in the remainder of 38 it could
not be estitaated with confidence.
Special tests as complement fixing antibodies , estimation
of lipid contents in the cerebrospinal fluid, special
retinal studies, estimation of amino acids in urine^
phenylalanine in blood, genetic assey and a very detail-
ed neurological assessment including interpretation of
pneussaoencephalograra allowed us to make etiological
diagnosis in 43 percent of all patients. In 34 percent
of the cases the etiological diagnosis was presumptive
and in the remaining 23 percent only symptomatic diagnosis
could be made.
Final analysis of the material has to await accumulation
of more patients. Results of studies of prenatal factors
iii the etiology, pathology and clinical manifestations
are reported in 5 publications during the year of 1958
and two niore are in press.
Si^iflcance to Neurologieal Research; In a majority of
cases the etiology of cerebral palsy and allied conditioas
is poorly understood. Better knowledge of hereditary
factors, clinical manifestations ^ as well as the loca°
tion and extent of the lesion taay further our insight
into the diverse etiology of these conditions. Full
understanding of pathology and etiology in larger groups
of children with brain damage will suggest eventually
better directed preventive and therapeutic measures.
Proposed Course of the Project; At the present time we have
detailed data on the total of 141 patients but further
accumulation of the material is needed before global
analysis can be attempted. Nevertheless various
important aspects arising from this study have been
already evaluated and this resulted in 7 publica-
tions .
Part B included
Sariai No. HIMBB-82 (c)
FHS-NIH
Individual Project Report
Calendar Year 1958
Part Bs Honors, Awards and Publications
Publications other than abstracts from this project:
Dekaban, AnatolSj O'Rourke, Jaaes and Comraaiij Slllye:
Abnormalities in offspring related to maternal
rubella during pregnancy. Neurology 8^ 387 -"39 2,
1958,
Dekabanj Anatole and Magee, Kenneth: Occurrence of
neurological abnorcnalities in infants born to
diabetic mothers. Neurology Bt^ I93-2OO5 1958.
Balden, Haitland and Dekaban, Anatole: The surgical
separation of Siamese twi.ns conjoined by the
heads (cephalopagus frontalis) followed by normal
develojanent . J. Neurol « Neurosurg. Psychiat, 21;
195«202, 1958.
Dekaban, Anatcle; Mental deficiency: recessive transmission
to all children by parents similarly affected. Arch,
Keurol. & Psychiat. 79i 123-»131, 1958.
Dek£bar2, Anatole and Drager, Glenn: Metastases of the
retinoblastosaa to the central nervous system.
Advisability of a combined intraorbital and intra-
cranial removal of the affected optic nerve, A.M. A,
Archives of Ophthalmology. In press.
Daksban, Anatole: Arhinencephaly, Amer. J. Mental Defie.
In press.
Honors and Awards Relating to this Projects
1, Assistant Professor of Neurology at George Washington
University Medical School.
2. Consultant District of Columbia Children's Hospital,
Surgical Neurology
Davelopmental Neurologj
Bethesda Maryland
Same as KINSB-48 (c)
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Pro.ject Title; Maternal Condition During Pregnancy and the
Course of Birth in Relation to Neurological
Abnormalities in the Infants and Pathologic
Lesions in Products of Abortion o
Principal Investigator; Anatole S. Dekaban, M,D.
Other Investigators; Br= T.E, Cons and Dr, H.H. Hillj
National Naval Medicel Center; Dr.
L.J„ Geppert and Dr. KoL. Siva,
Walter Reed Army Hospital; Carolyn
May Smith, RoN,,
GooperatinR Units ;^ National Naval Medical Center and Waiter
Reed Aroiy Hospital.
Man Years (calendar year 1958);
Total; .25
Professional: .25
Other : , 25
Objectives; Analysis of various abnormal factors oceurrinf
in pregnant mothers or complications of birth which
may cause or contribute to neurological abnorasslities
in infants .
Methods Employed;
1. prenatal care of mothers under research and their
individual final assessment.
2. Recording and evaluating of the course of birth
and pertinent abnormalities.
3. Examination of the nevjborn infants during initial
hospital stay.
4. Follow-up examination of infants.
5. Gross and microscopic essamingtion of the products
of abortion.
Pafclenfcs Material;
lo All pregnant women who were receiving prenatal care
at the National Naval Medical Center and Walter
Reed Army Hospital and subsequently were delivered
in these hospitals between March 1, 1958 and March
1, 1957.
Clinical Project
Major Findings; There are 4,480 products of pregnancy under
study. Up to date we have completed follow-up examinS"
tions on 68 percent of the infants. Great efforts are
being raade to increase follow-up studies to pass the
mark of 80 percent. When this is accomplished analysis
and evaluation of the entire material will be begun.
This will be a task consuming much of our time during
the 1959 year, however j it is anticipated that a number
of important observations vjill be obtained.
Significance to Neurological Research; It is postulated
that various environmental factors acting during pre-
natal, intranatal and early postnatal life may be
responsible for brain damage and the associated
clinical sequelae in infants. This study may be able
to reveal the relative importance of various factors
and also their incidence. Since careful and uniform
examinations are being conducted during all stages
of prenatal end postnatal life, final analysis of
the findings should be significant.
Proposed Course of the Project; Completion of this project
and preparation of the material for publications is
likely to take tv?o more years of work.
Part B included Yes /
1, Surgical Neurology
2. Developjnental Keuroicgy
3. Bethesdas Maryland
4, Same as NINDB-46 (c)
PHS-NIH
Individual Project Report
Calendar Year 1958
Pert Ao
Project Title: Pathological Lesions in the Central Nervous
System Occurring During Prenatal, Intra"
natal and Early Postnatal Life,.
Principal Investigator; Anatole S. Dekaban, M.D.
Other Investigators; Martha Rorlng
Cooperating Units; None
Man Years (calendar year 1958)j_
Totals .20
professional; ,20
Other: .20
Oblectives; The causation and pathology of the majority
of mental defects and cerebral palsy patients are
largely unkno^ra. Detailed examination of the brains
of children who suffered from such disorders and the
correlation of these findings with the clinical data
is expected to provide valuable infonnation for
elucidation of etiology of these conditions and so
to suggest possible preventive measures.
Meth.odg_lro|>lo3rgd;. Detailed examination of brains and
spinal cords from patients Who suffered from cerebral
palsy or allied conditions by means of:
1. Gross examination and dissections.
2. Microscopical study of sections which were treat-
ed with chromatic silverj sayelin and fat stains
as well as by various histochemical procedures.
Material ; Twenty brains from children with organic brain
lesions were processed and studied.
Neuropathological project
Major Findingsj The analysis of petkologlcai findings re-
vealed that in nine children the abnorraality iss the
central nervous system was of prenatal origiUj in five
it wss compatible with birth injuryj in four it was a.
result of intracranial infectiorij in one cerebral neo"
plasm was present and in one no significant CNS abnorraa.il-
ty was detected.
Significance to Neurologigal Research; Such studies are of
great importance ss the number of brains examined in de-
tail in the instances of cerebral palsy and allied condi^
tions is rather small. Studies of these specimen up to
date resulted in 3 publications »
Proposed Course of the Frojeets Further acctimulation of data
is needed before final evaluation will be attempted.
Part B included Yes
Serial No. aiKDB-'84 (c)
PHS-NIH
Individual Project Report
Calendar Year 1958
Fart B; Honors j A^^ards, and Publications
Dekaban, Anstole and Norman, Ronalds Hemiplegia in <2arly
life associated with throsjjbosis of the sagittal
sinus and its tributary veins in one hemi&pher&.
J. of Neuropath, and Exper, Neurology 17; 461"4.'0j
1958.
Dekaban^ Anatole: Is needle puncture of the brain enti.-a-
ly harmless. Neurology 8j_ 556-557 j 1958.
Dekaban, Anatole: Arhinencephaly in an infant bom to a
diabetic mother, J, Neuropath, sad Exper. Neurol.
In press.
Honors and Awards Relating to this Project; None
PHS-KIH
Individual Project Report
Calendar Year 1958
::.i;is5.i Ho.__ Kifl-^;_;o.. ;^ c _;
1. Surgical Heurology
2, Developmental Keux'ology
3o Bethesda^ Maryland
4. Same as NINBB-49 (c)
Part A.
Project Title; The Incidence and the Type of the Central
Nervous System Abnormalities Encountered
in Offspring Bom to Diabetic Mothers »
Principal Investigator; Anatole S, Dekaban, M,D,
Robert L, Balrd, M.D.
Other Investigators; None
Cooperating Units; None
Man Years (calendar year 195S);
Total: .10
Professional: .10
Other; .10
Project Description;
Objectives; Our clinical and neuropathological studies in-
dicated that severe abnormalities must occur not in-
frequently in infants bom to diabetic aothers.
(Dekaban^ A. and Megee, K.: Occurrence of neurological
abnormalities in infants born from diabetic oothers.
Neurology 8: 193-200, 1958). It became important to
evaluate statistically the incidence of these abnormal i<
ties in lai'ger series of offspring bom to diabetic
mothers and to analyse the findings in light of find-
ings in series of normal controls.
Methods;^ 1„ Critical assessiaent of maternal diabetes and
her total pregnancies.
2. Exaralnatlon of all her offspring.
Material: The outcome of 234 pregnancies in 48 diabetic
women and in 249 pregnancies in 48 norinal controls
were analysed. The mothers were personally interview-
ed and the offspring examined.
Clinical Project
Il§l££ ^'-iS^MLi. '^^■** overall total wascege
in'the diabetic mothers was 43,4 percent && coi^pat.id
to 17-6 percent in the normal control. Of the surviv-
ing offspring born to the diabetic mothsrs 6«7 perceBt
showed congenital dalforeations or various neuroIog3.cal
abnormalities J this compares with only 0.48 percent of
the abnormal children in the non-diabetic control group.
>^
G
<^
o
•H
-H
-3
w
O
M!
B
O
•H
ej
i-^
a
^
o
s
1-j
a
So
«l
M
>
O
M
o
w
"O
OJ
•o
Ik
u
«fl
tfl
a.
M
w
CD
M
U
o.
c
00
•ri
c
X!
a)
0)
^
N
>,
«
r-I
1^
«
s
id
<s
m
S_ignificarxe to MaurologicaX Researchj^ To further our
knowledge of the cause of cerebral pslsy and aillsc
conditions „
Proposed Course of the Project; Final evaluation snd
description of the data will be completed within
ten months.
Fart 3 included Yes
Serial No. NIMDB-86 (c)
1 . Surgical Neurology
2. Developmental Neurology
3. Bethesda, Maryland
4,, Same as NINDB-50 (c)
PHS-Nia
Individual Project Report
Calendar Year 1958
Part A.
Fro.lect Title; Measuresaents of External and Internal Orbital
Distance in Males and Fenmle:8 from Birth to
Adulthood „
Principal Investigator; Anatoie S, Dekaban, M.D.
Other Investigators; None
Cooperating Units; None
Kan Years (calendar year 1958);
Totals ,05
Professional; ,05
Other: ,05
Project Description;
Objectives; It has been found that the measurement of the
interpupillary distance in huajans for the purpose of
estimation of hypertelorism and abnormality of the
sphenoid bone is unsatisfactory. It is thought that
either estemsl or internal orbital distance or inde^
thereof should take place of the measurements of the
interpupillary distance.
Methods Employed;
1, Measurements of the above-named distances in human
males and f&nales at progressive ©ges beginning
from zero to 20 years of age.
2, Correlation of physical measurements of a small
group of children with measurements made on
cephalometric x-rays .
3, Statistical analysis in various age horisons.
Material; Measurements of all horizons have been taken „
This amounts to the total of 600 head measurements.
Clinical Project
Major Findings; Tiiis material is being currently validated
and subsequently it will be subjected to the statistical
analysis .
Sisnlfieance to Reurological Research;, To make the estima-
tion of conditions such as hypertelorism more scientific;
the measurements of stable bony structures rather than
movable organ as eyeball, should be performed. As an
example a concotmnitant divergent strabismus can be given;
in this instance measurement of the interpupillary
distance for the estimation of the abnormality of the
sphenoid bone would obviously give false results..
Proposed Course of the Project; The data obtained from the
measurements are currently analyzed. Subsequently the
material will be prepared for pubileetion.
Part B included Yes
PHS-HIH
Individual Project Report
Calendar Year 1958
Serial Mo> riMBB-S'/ (c)
1, Surgical Neurology
2, Deveiopmenesi Neurology
3» Bethesda, Maryland
4, Same ss KINDB-47 (c)
Pare A.
Protect Title; Preparation of the Horizons of the Nom>al
De^eiopmeret of the CKS in Mice and Experi-
memtal Production of Congenital Malforata^
ticns of the CES„
Priacipal Inyestig£.torj_ Anatoie S„ Bekaban^ M„D.
Other Investigators ; Marie J, Kendall j B„A.
Cpo|;egatii;g Units; None
Man Years (calendar year 1958);
Total; '.05
Professional: .01?
Ofcher; .05
Project Description ,^
Objectives: The purpose of this project is the production
and ;analysis of congenital abnormalities of the CNS and
the provision of norms for the development of the CNS
in alee.
Methods ISmployed;
1« Dissection of the CHS of raice of 16 progressive
developmental stages .
2. rreparation and staining of serial sections,
3. Identification and outlining of ssain structures
on the low power microphotographs.
4. X-ray radi^atlon of pregnant mice from a strain
which does not show any significant incidence of
spontaneously occurring abnormality of the CNS„
5. X^ray radiation of pregnant Black C 57 mice in
saveral stages of pregnancy, with similar para-
meters to these for mice in Swiss Albino: the
strain used here show an abnormally high incidence
of spontaneous malformations of the CNS,
60 Gross ^ skeletal, and microscopic ejcamination of
the obtained specimen.
Materials Mice strains NIH stock "general purpose
Swiss Albino" and Black G 57.
Major Findings; An atlas of norraal mouse brain has been
prepared and bound „ It is in current use in our
laboratory.
Careful dissection of brain and brainstem of fetus and
young mice in eleven age horizons were performed . The
specimens are sectioned serially and stained. It needs
to be stressed that to obtain one perfect set of serial
sections for one horizon it is usually necessary to
process and section six to twelve brains. Only those
sets which are in ideal conditions can be utilised for
description,
Production of malformations by means of s-radiatlon.
We are considerably limited in space for maintenance
of tciee. Since only certain age mice can be used^ we
have to harbour them until they attain it. Then- only
about 20 percent of those kept become pregnant as a re^
suit of restricted duration of mating time. In strain
"general purpose Swiss Albino" 98 litters were obtain-
ed from irradiated mothers. Approximately 10 percent
of these had major abnormalities ^ about 25 percent
ffi5.nor abnormalities and the remaining are free of
detectable pathology. Similar parameters of irradia"-
tion and technique were applied to strain Black C 57
and so far 45 litters were obtained.
Significance to Weurological Research; An experimental
approach to congenital malfonoations of the CNS is
necessary to help us understand certain obscure mal°
formations occurring in humans. The provision of
norms of the central nervous systea has to precede
the experimental production of congenital malformations s
as there does not exist any proper guide in the form of
an atlas or of a satisfactory reference during consecu^
tive stages of the development of the mouse. Majority
of the stages In this strain are not yet completed and
final analysis of data and comparison with the findings
learned from irradiation will be done during the coming
Proposed Course of the Project: For technical reasons we
had to stop further irradiation of mice during the past
3 months. Beginning in Jsnuary this experimental pro-
ject will be s-esutned.
Part B included Yes
Matloaal Xastitut© of Neurological
Disssses aod Blindness
Clinical Research
Surgical Neurology Branch
Section on Clinical Peychology
Serial Nussbers of Projects s
NIM)B-88<c), NIHDB-89(cK NINDB-90Cc),
Batiaated Obligatloas for FY 1959
Total; $24,500
Directs |24,000
Eeistbursf^ent: $500
2<, S©cti©E oa Cliaieal
Psychology
4o
Ii%di¥idis%l Project Eepo?t
Calendar Year 1958
Fart Ao
Fg0j©ct Title; Effect of "f®ai'=ps'©¥okiffig" stiawali oa
¥i®?Aal discrimiaatiOB im ps'iiaateSo
ggjacipal l£av©@tigatog; Ho Laasdell
©tla©g la'^estigators; Hone
Goopegatiag Uaits; Mom©
Man ¥@as>s Patiemt Days
Cc&l®adar yeag' 1058) ; Cea^l®adag y©ag 1958);
Total; o5 ' N®a®
Fs-of ©ss ioaal ; o 5
©tk©s>; o5
Project D®s©g'iptioa;
ObjQctiv©; To investigat® the disspuptive effects of
cartaia stimuli oa p@s*formaac© ia a -^isiaal discf iaaiaatioa
task la aa attempt to quantify aad systeaatis© th® EEates^®
of siach "f@a£>-p£>ovo&iag" stimuli „ aad to ias@ such data
to ©valiaat© ehaagss that say h® specific to te^pofal lofo©
^©moval o
iQtlaods Em&l©f@di A Wiscoasia Q@ffl©^sl Test Appa^atw®
tor primates will fe© msedj with the discrisaiaatioa ts-sy
modified so that th@r© is a plastic hoK {Soi^ th@ cSisraptiag
objects) feetw©®ffi th@ discrimiaiatioa cssps.
Major Fiadiags; Koa®, Th® apparatus has bsea bisilti
laboratos-y spac® i® beiagarraagsdi s^^a® preliaiBss-y
traiaiag of moa^@y@ h^ k@gaao
Sigaif icaae® of th© prograia to th© lastittate; The
method, slioiild yiein a. means of mos® preciUXy deseE-ibiag
th@ ffi&tiire of teiBpoical lobe ttmetion ia @raotioi&; tlie
'^el&liorating'* function is relation to perception in
priiaates could be clarified o
Proposed cours® of the projects
esiabiisiB ike uiiiity of this mel
The first efforts to
ithod will use monkeys
rather than the more expensive chimpanzees o
Fart B included Yes A°°°7 Ho /^^
Serial 10. MM)B°8@ Cc)
Branch
2o Section on Cllnlcml
Psychology
S, B®tfe®sda(, Marylaad
4o Sam® as lIMDB-55 (c)
I®cii¥idiaal Project E©p©rt
Calendar Year liS@
Part A,,
Project Titles Psyelaologisal l^alna-tioa of Taiaporal
Lob® Dis@a@@
Friiaeipal liases tigat org Ho Laasdell
Other ZuiTQStigators; 1„ laldwiEj Mo Ble^-ias!,
Jo Weissbaeh mnd Ac Mirsky
Cooperating Halts; MIIH SeetiOB oa toissal Beha^^'ior
lass Y®ars Fatieat Dajs
CealQffidar year 1958); Ccalemdar year l@5@)g
Total; o5 loae
Prof©©® iosal ; o 5
Other; <,5 Cabowt 40 surgery eases j
mora than double for uon^
surgery ea^eS"-- p©r ^®ar}
^oJ@@t Beaeriptioa;
ObJ®eti¥©; To study patieats with teaaporal loto® disorders
with ®mph&BiB is th® ar©a® of iatellectiml ability j, ^isiiml
and auditory perception, linguistic fusictions asd oth©r
more gea©ral "p®rsomalit^" features o
Methods ®spi©y@d; lateiligesi©© aiad personality t®mt@i
aphas ia g a.iidibm@tr ic ^ aa«i oth®r specialised verbal te^tsi
t@st@ of visual p@rc@ptioiSo Tachistoscopic recognitioQo
Contiauous Performanc® TsBto Auditory testiiag durii^g
neurosurgery on conscious patients o
Major Fiiadings; Irso Jc Weissbach itormmrlj ©Ihoeft)
and Miss Mo El©vins i±n cooperation with Sro L« Frost
and iTo So Savard) hs¥@ r@p@r t®d a tendency for patients
^ith left t@iapors6l lobe remoiral® to be "Poor eosnauaicator®/'
the®© patients d® not appear t© differ in other stMsdard
r@©p©cts from other patients «
No ©tSier sigaificaat n©^ findings toav© b©©a ©sta^lisfe
ia th@s@ aareaso A ¥sufiety of new t©sts have r®c®ntly h®@&
added to the standard batt^r^r given to the patients «
Signif ieaisee of the pgogram to the Institute ; Future
analysis of~ToXloW'»up data and diagnostic eff ioiency of
the t®@t@ may help in th® efficacy of temporal lobe
surgery o A careful survey of the test data on previous
and contemporary cases should enable more objective
description of the natiare of dysfunctions with temporal
lobe disorders 0 Such material will elucidate and help
distinguish between contemporary cosiceptions of temporal
lobe function.
Proposed course of the projects Publication of th© study
on the '^'poor CGSomunicators o " Observations ar® yet to be
obtained on auditory function during neurosurgery.
Sufficient objective observations are yet to be obtained
on cases with most clearly-established loci of epileptic
disorders 0 Adequate control data are to be obtained o
l,mt B iE©ltsd®d Jm iJI m i%l
So S©etioa oa Cliaieal
3. lethesdaj Maryland
4. Mew
Indi'S'idiisl Project Report
Calemdar Year 1958
Part A.
Frojeet Title; Body Tefflpeyatis?© im Chimpaazees with
bilateral Temporal Lols@ Daisaage.
Fgiacipal lE'^estigatog; lildrecS Lo El©¥ins
©tlaer la^estigatogs; A. Eow©j, So Lewis » N, Mills
'"'^"^'^'^^^^^^^^^^^^'^^'^"^^'^^^^^^^"'^ and Fo Smith
Cooperating teits; Moae
Man Years Pat lea t Dajs
Ccaleaelag' year 1958); Ccaleadar year 1@5@);
Total: .5 Eosi®
P7@f@s@ioaals Ȥ
Other; .5
Project Desgriptioa;
Objectiv©; To record th© rectal temperatur® of aoriBal
cIiii&paaK@@s aa«S cliispansees witfe daasag© t© tooth temporal
lobes.
Methods Employed; lectal temperature© irere ©fotaiaed twon
Ghimpaiasees, both ia the runs and ia iadi^'idual cages j,
over at least/20«day period o
Major FJBdiags; Th© meaa temperstisres of three aorgial
chiiBpang;©@s were fouad to b® 37„4j 37o8j, and 3?o8®C C®a©
male aad two females respectively) o The mean temperatures
of the temporal lobe damaged animals at least four months
postoperative were 3So5, 36 o 7 and 37.1^0 (male and two
females). The preoperative values on the first two
animals ^@re 37 » 9 aad 37o7®Go A survey of the literstwr®
indicates there are no established normative values »
SignificaBe® @f th© proggam to tiiie Zastitate; loraati'e'e
1;@iisperat^?@ "tallies £ia?® h®®m 6&SS^®^ tor tliis @oiossy„
The valia@s will pro^@ useful in @M@cMsg th@ @oi3£>s@ of
ill«-h@altl£s or the effects of bs'aira operatiosis, in
chiMpa£iis@e@ adapted to tli® proQ@d&r®o
Proposed eoprs® @f the project? Hoa© further j, eseeapt
possible publicat'ibffi of the established drop in
temperature with brain operations of this t:^pe.
Fart B iaelnded Y©©
Mimm-mc}, 13Blll»f2Ce), mffiB'-faCe), «i
SniDB-S4(c)o
Totals $26,©®©
Dlrecas $25,50®
Serial MOo HIMDE-9I (c)
2o Seeti©sa on Pais
4,
FHS»M1M
Iffidi-^idaal project E©p©rt
Gal©mdar ' Year 1958
Part Ao
Pr@J®ot Titl®; Fl«@thais© Studies „
Priacipal Iza^estigatog; Kenaetli Do Hallj !„ Do
Qtfe©g laves tigatofs; Philip feisl©?. Mo Do
Forbes Ho Morris » JToo Mo Do
William Le© Pritehard, Mc Do
Cooperating Uaits; Hosae
Man Years (calendar year 1S58);
Others oiJ^ "^ =—===».
Profess ional : . 33
Totals o3S -
Project Descriptioa;
Objectives; To continue clinical and experimental
oHervaHSJas on the properties of volatile anaestlaetics
^hich promise to be particularly adaptable to tis© ia
neurosurgery. Properties to be determined incl?fld©
potency s rate of recovery ^ physiological @ide>=-©ff©ctS(,
combustibilityp and latent toxicityo
Mettaods Employed; Fluotlsan® was submitted to extensive
experimental and clinical study „ Acute experiments
using subject dogs were performed to determine
physiological effects of fluothane, administered in
concentrations equivalent to 1 - clinical usage and 2 =■
lethal doses o Pathological studies were performed o».
these dogs to correlate morphological effects with th@
physiological observations o
Clinico-^pathological observations were also performed
in the primate laboratory where fluo thane is used as
the primary anaesthetic agent in neurosurgical pro-
cedures on SMtojeet chimpanseeso
0EP-2 CA)
Cliffiieal studies w®r® performed ia the opes^atisg rooa
whes'e fl^sotiaaa© was adiaisiist®s»©d to patieats Msades'goiag
surgsry for a variety of ueuffologieal disordes'So
Clinical ©bservatioas aad electronic polygraphic r©cor«l->
iag of signifieamt physiological parameters were carried
outo
"Fl?s-©th©r" g as asotropic raisttire of fluotha&e was
employed xm ©ssperimeotal studies similar to those
described above o The physico-chemical properties and
physiological effects ±n acute dog experiments were
determined e
Major Fiadingss Fliiothame has proves to b® a poteatj
i&on<°cora&;is tible $ noQ-toxic anaesthetic agent from which
the patient recovers rapidly following withdrawal o It
is a safe agent if administered with respect for its
potency o This is most satisfactory don® by us© of the
"Fluothaa®" ^apor'4s©ro
"Flu-ether" was found to h@ non-combustible and a
relatiirely stable agent by various cheiaico-physical
criteria o Physiological effects in acute dog
©speriments generally paralleled those of fluothanep
the latter teeing- the more potent of the twoo
Significance to neurological research; fluothane
is particularly adaptable to the problem of anaesthesiology
as applied to neurosurgery o
Proposed Oours© g Clinical us® of this agent will fo©
cont in?a® d^a~n© human operating room,
Clinico-patfeological studies will be continued in the
laboratory g ©n subject chimpansees and dogs^
Fart B included Yes
S©r-iai lOo
PHS-MIH
Individual Project Eepo^t
Caleaday Y@ax 1958
Part B; EonorBp Awards and Piafelicatioaso
lo Balls l@na©tfe Do and HorriSp Forbes Ho» Jro
Respiratory and cardiovascular effects
ot fiuothane in dogSo Anesthesiology ,,
¥olo 19 3 No, 3, M&j-Jmi®, 1958 o pp 339-352.
2o Hall^lenneth D. and No^ris^ Forbes H.g Jr.
Fluotl&ane sens itissat ion of dog heart to
action of epinethrine. Anesthesiology ^
Vol. 108 Sept-Oct.s 1958. pp 631-=641o
serial EOo mmu-92 (e)
1 o Surgical' Neiarology
Branch
2o Section on Pain
3o Bethesda^ Maryland
4o New
PHS-NIH
Individual Project Report
Calendar Year 1958
Part Ao
Project Title; Hypothermia in Neuroanesthesiology.
Principal Investigator; Kenneth Do Hallj, Mo Do
Other Investigators; Edward Laskowskig Mc D«
Forbes H. NorriSj, M<, D.
Williara Lee prltchard, M<, D.
Cooperating Units; None
Man Years (calendar year 1958);
I'Gtal; .as
Professional; c.33
Other; .33
Project Description;
Objectives; The current widespread interest in the use
of hypothermia during intracranial surgery and in certain
acute situations encountered in the treatment of neuro-
logical disorders has prompted clinical and experimental
studies of patients to determine and evaluate the rol@
of hypothermia o
?thods Employed; Clinical studies involve patients
»d to tota
submitted to total body immersion to achieve a hypothermic
state o Physiological parameters are recorded; observa<»
tions are made regarding the effects of cooling on the
brain and their relation to technical aspects of the
surgical procedure o Fourteen patients underwent this
type of procedure during the past year^.
Laboratory investigations are underway to achieve
differential cooling of the brain while maintaining
normal temperature in the rest of the body„ The means
employed involves the extracorporeal shunting of the
0^-2 CA)
cerebral felood supply ttooagla a cooling ©tember^ after
which th© blood is returaed to its normal cereteral path-
way by measss of a modified pump or "Hieciiaaieal heart „"
Numerous tectoical and physiologieal ps-oblems pr®s@3it
tlieEis©lv©s for solution „
Major Fiadiags i Cliaical ®Kp®T±®nc® with hypothermia is
as yet to© I'Sited ia nwasber of patieats^ too brief im
loag-term follow-wp, and too poorly doeismeated by
pathological studies to reach fiscal eiraiimtioisso Pre-
liminary observatioas ar© based on objective findings
during surgical procedures j suck as the consistency of
brain t issue «, degree of swelling or bleeding j, technical
ease of manipulating structures o Physiological studies
have outlined the tolerances involved in subjecting a
patient to body coolings The considerable increase in
total anaesthesia time to which the patient is submitted
probably does not sufficiently increase the anesthesia
risk to deny th® patient and surgeon whatever tecfeaical
advantages may accrue thereby o
The @x:perimental work is performed o£i subject dogSj, and
the development of this project is insufficient to
present results.
Significance to neurological research^ The role of
hypothermia in neuroane@lE@slorogy"w7ll be deteriaiaed
by such studies as outlined here in o
The experiment al procedure described above represents
th® development of a laboratory preparation whereby
innumerable aspects of brain physiology^ chemistry^,
metabolism and path® logy may be studied in addition
to our immediate interests th© effects of hypothermia o
Proposed Course of the Project; It is intended to
continue tk® course outlined above to evaluate results »
and thereby determine th© role of hypothermia in clinical
application o
Part B included Yes /^"T Mo /"FT
serial NOo MMDB-93 Cc)
2o S®etiois oa Pais
4„ Maw
PMS-NIH
Individual Project Report
CaleBdar ■ Year 1©58
Part Ao
Project Title; Succinyl Cboline in Awake Craniotomy o
Principal Investigator; Kemffisth Do Hails Mo Do
Other Investigators; Maitland BaldwiiSs Mo Do
' Forbes Ho Norris, Jro^ Mo Do
Wiliiam Lee Pritchards Mo Do
Cooperatiag Oaitsg Hoae
Mam Years Cealeadar f@ar 1958);
Profess ioaal % <> 3S
Other; o33
Project Description;
Objectives; In the surgery ©f epilepsy on humans and
in eiectroencephalographic and cortical stimulation
studies performed on chimpanssees » it is desirable to
have the subject awake » cooperative and able to tolerate
the procedure 0 The development of such a technique as
controlled paralysis by the administration of succinyl
choline has been a sEajor interest of this section o
Methods Employed; Succinyl choline is administered to
the subject hy intravenous drip therapy j, the subject
having previously been induced and intubated by con-"
ventioaal anaesthetic technique o The subject is allowed
to awake from his anaesthetic » Hyperventilation is per»
formed by the anaesthetist „ H© is paralysed » but
comfortable 9 as he is being mechanically respirated
and local anaesthesia is used during the surgical pro-
cedure o The absence of spontaneous musel© activity and
artefacts due to anaesthetic agents „ renders more
successful th© electroeortical studies as performed in
human patients o By sign languages, the human patient
can communicate his reactions to cortical stimulation o
osp-a iA)
I§F titration of tte le^wml of paralysis tii© esperiiaeatal
affiimal dog h® allowed s^flieieut m^ssealar activity to
detect feody movem®nt la rospoase to cojftical stimulatioiao
Major Fiffidiags; Tfeis tecfeaiqii© Sias hm@n developed to
tlje~^©gr®® 'of"pr©f icieacy tliat it r©pr©s©ats as importaat
acSJiiact to th© surgery of ©pilepsjo
Proposed Coar©® of the Project; Tiiis tectoiqia© ??ill b®
us©a^'sTH3Ka£id cSiariag ©poratioas oa hurnass patiemtSc
Further ©xperi©nc@ will lb© derived f.r©m its use ia tis©
priisat© laboratory in physiological and pfearjaacological
@3ep@rim@rat@ la progress o
Paa-t B iaeluded ¥©s /^7 M© /'^^'^
serial lo,
PHS~MIH .
SudiFidual Project Eeport
CalesicSar Year 195S
P^tjo Honors 8 Awards and Publicatioas o
Hall 8 Kennetls D„, laldwiaj, Maitland, and
NorriSg Forbes Ho » Jr„: Succinyl eholime
in awake craniotomy o Anesthesiology , In
Press o
3„ Sectioa on Faia
4o New
FHS-HSH
Sadividaai Project Report
Cal@ndiw Year 1958
Fart Ac
Project Title; Tlie Effect of E^ertonlc UTea Soiutiois
on latracranial Pressor© o
Princip&l Savestigator; William Lee pritcliards Me Do
Otfegr lEvestigators; Eobert Edgars Mo Dc
Cooperatiag Uaits; Uoae
Maa Years CeaI©Etdar year 1©58);
Profess ioisal ; ^ 5
Total? C.5
Project Deecriptioag
Objectives; There is curreat widespread interest ia the
administration of iffitraveaous hypertoaic urea solution
for the redtictioa of intracraaial pressure and cerelbral
edema o This technique proasises to be of eonsiderafele
value as aa adjunct to routine iatracrasjisl surgery ^ as
well as in acute situations often encountered in
neurostsrger^ o Possible side effects of this regimen
await ©valuation o
Methods Employed; Patients undergoing surgery for
suspected' brain tu^or and ward patients with cliaical
evidence of increased intracranial pressure receive
intravenous urea solution o Observations on gross
appearance of the brain at time of surgery j and on
vital signs and neurological states of bed patients
are madeo Blood chemistry and urinalysis studies axe
also performedo Oaly a limited number of patients'
have been subjected to this regimen o
OSF-g CA)
Major Fiadiagsg TIsat feraia volume aad iatrscraniai
pressia^ may ©© r@^uc@6 is apparent in this studjc Tkmm
faff a no seyious sid©-eff®cts lia-^© be©a aoteds altfeoisgli
tfeey May Ij© expected iia c®rt&±n situations o
Sigaifieaace to Netagological Eesearcli; If this tactoiqtse
sfeoiaid fulfill its prosais®, it will prov® an invaitiatela
&djimet to the Management of neisrosm-gical patients «
pgopos®d QovirB® of tfa® Projects Clinical Tifial ot tsr©a
will be continuedo in addiflons as-raagements ar© foeiag
made t© perform tonometsT on patients recei'ying we®&g
to d@t©rjain® t!i© effect on iatraocialar pressure p and the
possifol© relstiossship of the latter to iatraeranial
pressur© o
Part H inelttded Yes f
Mnn^y 1, to December 31, 1958
lOTlOSUCflOM
Th© import aat rol© of tfe® nervous system ia th®
pursuit and achi^T^ssiaBt of aayt!%isig i^ ecianc© t®»ds too
o£t@a to be a®gl©ct@d, or e^ea deisiad. W« live la a
Logical Positivist Society which overlooks the psycho-*
logical processes of scieace in much th® same way that a
Puritanical Society fails to adnit to public accou&tiag
certain fundameBtal biological processes^
Obstacles :£si our path as experimental scientists
are by ao ateasas exclusively technical difficulties: in
lajTge measure p rathe? $ they ax-® cos%c@ptual obstacles
imposed according to the historical moment and philosophical
coffitext of our work. Although ■»© tead to deprecate considsra-
tioa of history and philosophy as aot directly coatributiag
to the task at ha^d, we caxmot escape in our actions as
experimentaliats from certain initial assumptions which m&j
be crucial and yet which we usually have accepted without
careful ©samiaation.
It is say purpose J first, to illustrate how depeadiBiat
all branches of science are upon an insight into the opera-
tions of th© nervous jsysteia. The ultiasat© grasp of any
branch of knowledge depends upon conceptual achievements in
th® mental and neurological fields. Already there is evidence-
that ignorance ia these fields is holding back progress ±n
some others. Second, £ wish to invite reflection as to the
validity of certain pervading assuiaptions which affect r^sea.rd
teaching and practice is th© mental asd neurological fields .
What 1 mean will. I hop©> become cl©a.r tu the course of coti-
sideratioa of thr®® questions: What lis our conception of
reality? How do we consider mind and brain? low do aaswers
to these qu®stio3?s affect undertakings of physicians aad
experimentalists in the biomedical sciences?
^ As in the previous Annual Eeport^ the Laboratory Chiefs ha-v-
provided comprehensive statements of research progress thTOi:ip'-
out the year. In th© following paragraphs I have attesspte .
continue, as last year, the exploration of more general q;,'-
These tend to be overlooked in the iBiaediacy and seemijsg ■.:■.•
of our dally undertakings. Yet I believe they are truly pc ;
tinent to our ultiiaate best achievement.
WHAT 13 OOB COit:^;''riOM OF REALITY?
Manf different coracepta of reality h&v® app«ajr©d
thro'ugSi tls© long spaa of history. Sir@js with our pr®s<asit
degree of technical sophisticatioB tli©r© is ao cossseasiss
as to what coBstitut©s th©'*'r©al" worlds Cont®mporsLxj
theories of reality caa hm distinguished as teelorsgiag to
a rather sixt©iad©d spectriEia which is subJeetivistiCs aiaoist
solipsistic at on& ©rsde aad Materialistic detersaiais® at
the other. Th© experiraestalist asks diff®reut questioas
of nature and treats the materials of his research^ in-
eluding his inBtnaaeatationg quit© differently depeadisag
upon the point of departure he assumes o
Science is first of all a collective^ social ac-
tivity o Scientists operate not only with instrusaents and
language of cossimon understanding but also with certain
abstractions such as charge ^ fields electron^ atoms
moiecule'j, and so on-- -things which do not belong to th©
everyday world but are derived indirectly out of scientific
experiments c. In experiments «,. the scientist must deal not
only with things which can be soiled by the finger and
thumb;, so to speaker but also with things which can be ap-
preciated only by means of elaborate extensions of the
sense organs. We have to get accustoieed to extrapolations
into diKensions where our sense orgai^s fail us^, and where
soaie of the space-time and causality relatione of everyday
experience seeai no longer to hold true, ISoreoverj, and
especially in this extended rang©j, we are already faedliair
with the necessity to resaain tentative about our con-
ceptions of "reality."
Ther® seems to b© no defiBitiv® boundary separatiEg
the finger and thuab world frcaa the sicroworld of electron-
microscope and X-ray diffraction. Yet th® relation between
what is revealed by the experimentalist's instruments and
what is implied conceptually to him through these reve-
lations is by no means obvious. When w© extend ourselves
still further to deal with atomic eystesssp, we encounter
the indeterminisffl of Heisenberg, We become dependent upoiK
probability even with supposedly objectively occurring
phenomena. For exasplej- if we wish to determine the lo-
cation of an electron in orbit around an atom by boMbardiiig
it with a small particle whereby a photon will b© givejs off
which we can then observe » th© finite birth time of th®
photon tales so long in relation to the ©peed of th®
electron tfeat th© electron will hav® made a million pass®®
~ 3 -
arouBid its orbit dariog that Mrth tisa®? ao.d ao m&onmt
of obstetrical scnstiay irili r@dtj.e© asiMgisity as to
wh®r© the electroB was when it was str-ack fey tfe© bom-
toardixag particl®, ThuB the concept of absolute preeisio®
ia physical m®a4B,ur@mmmta at this seal® teeaomes absiitrd,
Itetermiffiisj® is oMt of the qissstion ia the origiaal sessse
of th© wordo
It is accessary iu any tr«satsi@Bt concerning concmptm
of reality to coffi®id@r ±m some further detail tfe® mms.tm hf
which such concepts ar@ achieved. This is especially
important because ordinary contemplatioa fails .us wfee© we
try to explain "reality" entirely in accordance with o«r
©xp®ri@ne© aad fajsiliarity with everyday seissory p©r~
c®ptiojas» As M&s. Bora has said reeeratlyg '"Matter as gl¥©ffl
by owr senses appears as a secondary pfeeaoaesoiSj, created
by th© iateractioB, of out seas© orgsniB with proc@s6®s whose
Esatwr© casi b® discovered OKly iadirectly through theoretical
isaterpretatiOBS of experiase ©tally observed relationships^ ia
other words s throuigh a laeatai effort o To desigaat© th«
result of this op@ratioa by the old word "matter" memmm to
®e wroiago"
Th® whol® realffi that seieac© reveals fii&ds its ratiois-
al order aad meaaiug through meatal activity » It iJieeds to b©
ussderliiaed that mesatal processes ester into the defiaitiosE of
reality at several stages:
First 5 with respect to th® sensory perceptioss
laecessary to obeervatioE.
SecoBdj, ia the meatal processes esseatial to
the establishjaeat of a theoretical aotiouo Her®
it »ust b© observed that 9 whereas a theory caa
be tested hf ©xperieaceg there is ao logical way
to proceed froia esperienc® to the setting up of
a theory o This must be a me3:^tal leap.
Thirdj, since science is a collective social
ei^terprise^ mental processes invade the acts of
coBtmuiiication through symbols and language to
other competest Individuals aadj thereafter^ al-
though suffering considerable reductioa,,; to the
culture of a given era. The culture iis turuj as
a further asd frequently overriding influence j
has already put its own cental strictures on any
conception of reality by virtue of the philo-
sophical liMtations it ha® imposed upon the
scientist prior to the beginning of his experi-
sieatatioao
_ 4 -
It is rare for t.fee propoiseist of &.tm^ coacept ia
any toraach of 8ci@iace to attempt to cosiaiujsicate all of
tfee steps which led him to tise final thBoxj. It is
iiKpoesitole for him to aaalya® all of tlj© assjmptioffis
wMcli weat iato its @Btniili.Bhm.^nt . Tli® asswaptious
which h© accepts on least r©f lectiou. ar® tSios© cosffioa
to his intellectual coiamiiiiity; Moreov©rj, they may aot
even be r^cogixsiased as aseimptions. On top of this ax@
the cosiMOs&plac© limitatioEm i!apo@@d by lack of tiaie;^
iosufficient iatellectual rigor^ ov©r~optimis!sia or by
too gr«at a pride or self-JustificatiOBo
W© concliad@ that isi order to wnderstaiid the
fflatwr® of the ualvers©,. In order to ij&pTOve our coa-
ceptioffi of reality^ certaia defiaite obstacles in our
path r@lat@ to th@ incoaipleteiiess of oiir tazsd^rstaisdizig
of the nervoiis systesa. This in itself coBstitates a
worthwhil® or ev©ss a compelliag reasoa,; to add to all
the nore ob^iotas ojiessj, why it i® importai^t that we
iQcre&se oisr efforts i.u t!i@ pursuit of research basic
to neiarology aad psychiatry.
W© !^ed to learu bow signals ©ater the ner'^oua
systSMs how they ar© distorted fey ojagoisg activity in
the braiffip and how this i© tura is affected by previous
experi@isc®| how iBCoaiiBgj, ^ ceistral aad owtgolag sigaals
are related to mechanisiie of reward a&d puEiishjaeat amd
essotioisal @%p@riei£ce &ud expression; how learali^g
oeeuriii what ar@ the laiisble processes of creativity?
and what are the limit ations of our amei&OffliCs conceptual
aud liffigtiistic response systeiss. These nasolved probleMs
caimot h® settled accordiag to soaie simple recipe o . Ther®
may be shortcuts ^ but few are revealed o These problems
call ismtead for the aiost imagis^atively resourceful eacploi-
tatiora of the anatcmicalg, physiological , chei&ical^
psychological aad sociological chasjjsels of scleucea
Thusj it is evident that the solutlO!:i of profoieass
isi all branches of 8Cieac@j, lEOt Oisly i^ the biomedical
fields J, depends Kipon the furtherance ai^d the success of
basic research ±n the meatal and neurological disciplines ,
« 5 -
HOW DO WE COMSIDSS MIMD MD BHAIM?
Ma!2klD.d has ask®d questions about toiffiseif siace
tfe@ feegimsiisg of reflective tMnkingc Stat it ^eaaiaed
for the Greeks to bring up the idea of '"mind'' for the
first tia®. It. was aroiai&d 400 BoC= that this coseept
first cam© into ejEisteac®, Ho suggest ioa of what w@
coiic@i¥@ to b@ jsiad aad M©sital,j nor of the id@a of tha
separat©ii@ss of miad froai jeatterj, is fo^ad is earlier
writiags throisghout Sgypt^ Babylonia and Greece „ la
fact this cofficeptioffi is siagwlarly unique to Westarsa
civillaatiofflo Although Plato dida't invssmt the idea of
Kii&ds, it is largely from his writiags aad iaflueace that
w© have inherited the fuudajaeiatal assuiaptioia that the
world possesses two iaevitably separate aad iiacoraiKejasis-
rabl® aspects; relating on the one hajad to mind^ s,iad on
the other hand to a&attero
Throughout the writings of both Plato aad Aristotl®,
the peculiar attributes of liviisg things are accouated for'
by a vmry broad coisceptioa of universal paeuiEa or world-
soul a ThiSg In its loftiest forai^ accounts for what w@
identify by th© tmxm '%@!ffital life,'* Cruder aspects of th®
soul J, respoiieibi© for vegetative life^ growth aisd repro-
duction,; is^abit ail living creatures including plaistSc
AaiBsala are endowed with an additionalj^ qualitatively
higher-order aspect of soul which "animates" them^ i. ■.#.,>
which gives then a capacity for movmmmuto Man aloa® pos-
sesses the thirdg liighest foras of soul^ th© rational sowl,
which concerns aaeatal processes = - - -
Plato aad iiristotle ltw@d im ir^m certain oth^swlm®
foreboding aietaphysical considerations that attach to the
material forms of lif©o With respect to dissecting the
6m&dg, they relieved us by authoritatively r#®nforci©g the
conception that on death "the soul flies away with Joy"
leaving the carcass a "disjected me^vmm" no longer
pertaining to lif®^ no longer containing a soulo
The particular flavor of Western civilization which
was contribut^sd by Greek culture played a cossspicuous role
in the developiaent of scientific inquiry through its impact
on the first huiasmists; the scientists and natural philo8»=
ophers of the Eisnaissane© „ miring a later ueriod of
especially rapid development of mGlmnQ®^ during the first
half of the 17th Century j, Descartes went a further step
b®yond Plato ajxd Iristotle. 1@ took the rational soul
.,. ® .„
wteieh liad h&mn proJ®et©d t&rouiglaoiit tli® ¥©atrieal.ar
SfBt^u of tli© toraiiSj, aad d®poait©d it lis tte© piiaeal
glands Descajctes revealed tls® remaiader of the a©r¥Ous
s^Bteas and bodj o£ maa as feeing an i,&&trvm®nta.lltj „
capaibl© of acqtt&irixsg suitable relutlouu witis tise ratioaal
B&ul in tfe,@ pia®aly but otlierwis© beisag ©f as aiitomatic
sort— "lite© tli® feralBS and feodie® of anliials whicli
D^Bcart©© still coasidored to lack th© ratioaai napects
of sowlc This eiaabliffig gestwr© did iu physiology, wliat
Flat© &ad Aristotle had doia® ©arlier In amatoKiji it left
th@ teaiffi (©sscept for th© pineal gland) availafel© for
seiesBtifie iiRvestigatiois sb a "aiiBchaaismo'" Fyom thlm
moment OBward^ sci^sitists eould 1m conc^rnmd with wlsat
aervotas isoclsailsiMS ar© tais'ol^ed ia cosaseiowsuess^, per-
emption^ a.pp®tit©fl reward aad piJssiielMieat j, aad freedom of
Willi, questioais that liad he®n raised ©.arlier hf tli© Qr©@ks,
Dwriag thit is©sct 20© years <, gr®at advaaee® w@r© made
In kffiOwl©dg® of th© gross ajsatomj of tJs® a®rvo«iB system.
Its mecliaiiisi^ w©r® @wce©®siv@ly i?iterpr©t@dp bj aaalog:3rj,
as if tfe© brain aad tfe® a©ry©s w©r® operated by a aecliaalc&i
Isifdrawlic syst©Kij, sisisfe s^ th® eostplsx water fo«ataisi laa-
clsiis@8 ifflT@Bt@d at that tiai®. Later^ they w©r© tfeoisg&t of
sui aceou23it©d for by tto® Bsiv®ri3al a©tfe©r of Sir Isaac Mewto©..;.
tlirougli whicSi gravity was posited to a.ct iUiStaaita®®o«sly at
a distaac® upozs t&e placets o Still latar^ tli© braia was
interpreted on tb© basis of tli® astioas of aijimal ®l@ctricity.
Ivsry age s©#ffi© to mnkm us® ot t&@ most camplmx aaalogy com-
c®iirable in tfeat ©ra. Our ag© aaalogia@s tli@ brain to mi
valsetroyEic coispistero
Se^esty-five years agOp o^r iiit©ll®©tiial gr®at-
graffidfatli@r® supposed tiaat ill© »i®claaais5!i® of thm hrulB. would
becoas© clear a® sooe as th® aieroscopic detail of braias
aaatosiy could be wcr&ed atEt^ . msiiag th® ia©wly acq^ilrfsd t®cfe-
iBi^t6®s for sasUfeifig tfei® slices amd for staining n@Tvoum tlBmum-.
Thim optisaisM was s«cc®i!d®d fey a psssisais® characteristic of
til® sttcceedisag geaerationj; tfeat axaatomy aios© woMld act
SMtfie®: on© xmedBd to wEderstaad tfa@ liviEg brain aad it
was presupposed tfaat th@r@ si©v@r coiald BiHiifisf aetory
t@©hffii<jtt@® for this purposso
Within th© last tw@.aty to tfeirty years j; however »
aore ha® feeea accossplishsd toward a©hie¥i!ffig an usderstaffldisag
of th® great questioa origiaally raised fey tia® Greafea thaa
filsrifflg th© pr®irioia® two ffiilleimiao We shoialdj, th©r#f©r©,; fe®
morm optinistic than oMr iEt®ll®ct-aal grandfathers altho?sgfe
perhaps l^BB optiffli^tic thaa owr int^Ilect-aal great-
graadfathsrso To OTjr enoxmouB adij-aistag© mowy a clisster of
- 7 -
t@clxaical achi©^eM@ssts hsm m&dm possible th@ &xsm±ns.tlQx
of the li¥ii%gf, wakiiiig^ bahaving or^ajSLlsai^ in t@j^@ of
lataroal braii^ sneobaMems „ Thar® is p'resentlj flowering
a treaiendoiss reuaissaisce of all of tlie ®ci©ac®e r®.latiag
to th@ asrvous system. All tfeat is being discoirer«?d In
©©uroaaatomy^ aeurocfeeisistryp meurophaxmacologj, ssg-uro-
physiology J, psyciaology and sociology h&B s^cid®isly foecosi©
far mor@ mesjodngtul ' to ©aoli of tfee otJi©r coiapl®ai@iitary
disciplijiQs.. This rapid ©spasasios SMd increase of coa-
G@ptual penetration into tls®s® important fields has
occurred at a tiai© wfe®a tli© world— aore thaa ®v©r before—
is©®ds desperately to uadsrstaed and to b® afel© to deal
with basic humaja capaMliti@@ &nd limitations.
I© the last five to tea years ^, seisatists l^ave
l@ara@d aaaay crucial features coaceraisig the basis of
coiaseiowsResSj the Mechaaisias of appetit^^ the way experi-
©use forsES asd distorts perceptionj, the limitations of our
a&echaniaa&s for memory^ effiotion^ camnunicatioiij, the ideratl--
ficatioB of ii3tteri3,al reward ai£d pmiishiseBt mymtm»Ss and
have com.® a great way toward understsuadiBg the uaity of
Mi ad ajad teaiRo
For ©xasaple,; it is sow dassoastrated ©sperim@iitaily
that the cortex is not the first step isa seiaeatioE nor do#8
it appear to be the last step either , Iisstead^ sc«sietteiag
fairly coKtiaiaous aad dynamic taJK@® place all along the
ascendijag ("sensory") pathways during wakefulness., There
is aa erosion of impuslses that originally started into the
nervow® system and a® iptrission of additional impulses
into th@ patt#rffl& tfe#s@ alterations feeing condltioEiad by
the pr@¥ioi8s experience of the indi-rid^ial , This im ©sactly
what one might introspeetively suppose woiiid ts^M place
within th® brain: a reduction and distortion froK th©
actual aati;sr@ of the stisaislating worlds of information
relating to perception. Mot only does the ferainstem reticu-
lar formation affect consciousness ^ as Magoun pointed out
soae ten yea^s agOg bMt in this way also it alters th©
coatsnt of conscioiasnesso This effect proceeds froas seajs®
receptors to what@¥©r end point yow wish to choose- It
continues right np to the final motoneuron discharge^ if
yois willo These aaechanisMSp which shap® perception and
pmrtorm&nQ9 are built into th© ¥alv« systems of th® organism,,
into the central reward and pusdshaient mechanisms j. and are
inextrical^ly bo^nd up with the circuits essential to
emotional eaKperience and expression o The brain thuis opmr^t-&u
teleologicaliy. that i@j, it is affected hf internal w&lim
systems which in turn are based upon previous e^olutiaaajry
and individ^aal ©xp®ri®nc@o
.. fe ™
Sosi® of the great questions raised hf tha Greeks
hav@ now feeea provesi approachable by -®xp@riiiffi@ffitatioKo
B^ raeans of r@soxirc@£ul tli.@or@tlcal and @sp@riiasental
approacis whlGh implicates ail of tfe® eoffipl@j»®istary disci-
pliis®s of biophysics p asstatomyj phi^siologyj cbaisistry.)
psychology 8 g®H©tics wnd soeiologys ®©i@Bti®ts ar@
begimiiiig ts> derive principles that will defiase the ra^@s
of ix&variaoc© aiad of ±n6ei@xm%n&cy is th© operation of the
From this bri©f descriptioffi^, it iSj, I hope^
©vidsQt that much is teieg learned through ba^ic r©-
jB@ar€h on the brain and behaiPior^ and that th@ findiBg®
f ro^ &u€h . research ar@ important In relatiosi to our
b@tt@r tmderstai^dii^g the ^@ry basis and linitatioas o£
human ^ffiowl@dg@o Mea^BUirabl® outsids @xp®rie2&ce is not
th@ whol® of ©s£p@ri©iic® o Becaws® of iadsterBiaacyj,
fflechaffiism is aot th® whol© ©splasatioja of a gives reality.
&®BMQU aad £@@liBg ar@ Bot at war froaa their Matures s
th@y are £iis@d ml®m.@nt& which w@ s@parat® in ot«r Muds
only by reflection o Do®® maa hav® fr@®d<MB to asaraipsalate
th® chaisjaels b©twe®ffi his feeling asad Ms id®a®? Can h®
choose his purposes? I sm inclined to beli@v® that
m©iatal and 2i@urological research aow supports th® surest
hopes ±n this r®gard that hav® ®v®r b@e!& put before
ffiankidd.
HOW DO AMSWBSS TO TSESE, QOSSTIOMS AflECT OlDKR-
TAKINGS OF HHYSICIAMS AHD BSPSRIMEKTALISTS IM
T3& BIOMEDICAL SCISMQiS?
W® have loog b©®E accustO!si#d Cintelleetually) to
doubt th® capaeiti©® of our coansora senses ia th© appreci-
ation of stature. He have b@@B obliged to do this wherever
there has lm®n sufficie&t scientific consensus ' ia favor of
scaae other than a comieon sense viewo Yet we seldois reflect
on the lack of foundation for most of our other prior
assumptions and how much they may affect our quest into tM@
unknown. As regards th© scientific frontier under investi-
gatlonj, w@ still tend to cling to a caamon sense view and
to the prevailing assumptions of our environment.
Traditionally J, as biomedical seientistSj, w© thiak
w@ know soM© thing about mindp and something about matters
and that these are iae^c^gimensurately different froM each other
- g -
First of a.llg it should h® recogsis^d tliat this whole
@ci%<^ma is l»a@®d upon aa assumption w@ iKi^©rit@<i £xcm
Plato o Second.^, the old defiQitioia of »atter i@ no^^
knows to he operationally incc«plet®s it fails to in-
clud® th@ sieiital processes inevitably invoIv@d in asiy
conception of asattero St follows^ thevBtOT®o that isstil
the basis and lisaitations o£ tkes® 3@@ntal processes are
ooBpIet@ly understood; any definition of matter aust be
accordingly limited and tentative » Altogether this aiay
seeas hard to accept. Yet the alternatives » over which
so ffiany philosophers ha-F© stnaggiedj, are not particularly
allurisigo Let ts® consider theffi feriellyj Saaie believe
that thoughts^) ideas^ wishes and other i^ental phenoaiena
are essentially ©pipheno:@enap that they grow out otg or
run parallel to^ certain particular material events^
others consider' that such aental events are essentially
incorporeal p peAaps occupying only "virtual spacer"
still others consider that aind is a property of matter »
that there may he a certain atsount of "Mind2i@ss"' extended
in s®all degree throughout all forms of aatt®r„ You may
have your choice o In each case^ it Is still the old word
"aatter" that ia_ teeing considered o Bertrand Eussell on
the other laandTibTds "that whatever we know without
inference Is aentalj and that the"^hysicir"world is only
known as regards certain abstract features of its space-
tigte structure"— -features whichg because of^eir
abstractnessp do not suffice to show whetSir the^physical
world is or is not different In intriasic character from
the world of Mind," This diiMtlsses the prior assumption ~
and gives encouragement to further scientific study of the
whole issue o.
I believe that the assumption of FlatOj, even though
it has a great hold on o^r imaginations » is unnecessary,;
and is unnecessarily distracting in relation to our quest
for further knowledge about the nervous system o Its
admission as an original premise seems to me to be unhelpful
to our everyday perforsiance in the hospital (, classroom and
research laboratory <> I feel this is true mainly because it
fosters professional J, intellectual and conceptual isolation
among scientists who are trying to uncterstand the whole masjo
When people sayg in common parlance ^ that they can
know or recognise matter independently of mindj, they are
really confessing a further assumption that is unnecessary
and may indeed be entirely wrongs What they really mean is
that they consider a percept to li® in some other category j,
presumably not so unreliably mental v, as compared with a
~ 1© ~
thoughts mmiorj or wisfeo The iatt®s'5, acteittsdly aseatal
functioiiS; are ai£ppos@dl^ I®ss tx-actiblds I@ss Inw&vt^nts
axsd perhaps l©ss s«b'Staiiti&l Cia the literal s®®®© of t,fe©
word) thsui is a parcepto Tfee fact is,. ^@ dora't y@t toow
«aoiigh to ssake s^cli a co!spmrati¥@ ^TalBation,
We do feffiowj, however^ that p©ro©ptioE is defiaitely
a lateistal act. W@ kit^ow also that in v&&t wajrs it is subject
to error ±n the seas® that B©scart@s aeaat this^ aad wpon
which h@ based his whole philosophy of universal doisbto
Furthermore s w© kisow that the seztaory messages upon whieli
perception smmt d&p®nd are theaselves also subject to
direct iaterfereac© through actios of the central iserroiss
system o A central cos^trol is eserted e^es^ o^t to the
peripheral sense orgaass^, and acting throuaghout the entire
trajectory of the ascending ''ses^ory'^ pathways » This
control 9 which was unknown until only a few years ago^
appears to be es:@rcis@d in accordance with souse kind of
internal '*valy@ systeia" which itself is reacting to previo«®
as well as coiscurrent experiences; thus^ the modulation of
incoming sensory impulses seems to be based upon
"eapectationfl" "relative significance to self," and so forth.
The valiae systesss moreover ^ is accessible to the central
mechanisias involved in "reward" aEd "puaisitoeffit , " eaiotional
experience aad expressions and the presumably more objecti-?®
and depersonalized systems of neocortex » This ccmplex set
of systems is built into the chassis^. @o to speaks and
cannot he divoroed from either the 'ascending signals coming
f r<»i the outsid® worlds, or f rem the outgoing sensory coi^trol
impulses which 'modulate the incoming messages « Evi(^ntly
the nervous systeat is continually practicing its control
over sensory pathways Just sui it has long been known, to do
in relation to motor performance. Presumably the brain can
shape o^r perceptioi^ more or less like it shapes our
ccmportments ia both cases teleological mechanisms are at
work.
At the very least ^ theaj perception is a mental actj
and the data upon which this Mental act is dependent are
themselYes acted wpon according to the cumulative patterns
of previous asei^tal actSo Th© psychologist j, from an external
view of behavior^ has long known this to be true^ and has
been trying to ccmssunicat© that to other scieffitistSo But
many scientists^, not toeing directly involved in examiEiing
the processes of -perceptioaj, have continued to preserve
- il -
a distisotion l>@tweeis what tissy Sia¥® assissied caa to©
direct IjTs clearly amd miaeqtsi vocally appreh®aded tSii*ougli
tb® s®m8&s aad what might thereafter he @ul)J®ct to s^g^tal
operations. This cai& isow h® throws:^ out on other thaz^
psychological growads.
Biomedical scieace is not aloE® in lj@isg af-£@et@d
by ©rroaeous prior assumption® o For ©saiaplej, similar
difficulties exissted ±m classical pliysics diarirag th@ last
century ^hen that discipline was cot£sid«red to be coz&cerised
offily with "irsaaimat® matter o" Majay physicists woi&dered
then whether cosacepts of force ^ eaergy^ aad sao on^ had to
do with "physically real" probieaSa or whether they were
oaly a kisid of logical isastr^meate xseeded oaly for the
time being o In the course of Biuoh theoretical and esp^ri-
mental works @i£ch primitive definitions gave way before
Bor® subtle coisc©ptiOBS which incorporated both aattar assd
eiiergy and aow flirt with a further iacorporatlosi of field
forces a ThuSp fosaerly isolated aspects of physicSs soa®
of which appeared to 11® completely outside the proper
proviBce of that discipliises have co^a to be viewed ssuch
more holistically<. fei^ eveo in a field as advanced as
physics j, there still ^©ssist great bogs of smMguities in
atomic theory -^ cosmology and other areas. The on® uni-
versally respected cosiviction is that w© must b© tentative 5
willing to tolerate ambiguitieSg acd prepared to participate
in quite' revolutionary non-cosmon-sense waye of conceiving
of nature o ^e major advances in physics have b@@n associ-
ated with greater unification of theory. I do not pretend
that brain and mlud will follow a similar history j> but 'only
that the speed of progr®@© in any disciplln® and the dimension
of contribution of , individual scientists ar© dependent upon
the tentativenes® with which ftmdsme^tal asgraaptions are
accepted.
Nowg taking the eosMSon sens© view of the ®©parat@a©ss
of mind and ®att@rj, a view which se©m© to pervade ,aost of
our cultures does that operate to our disadvantage — -as
patients s 'physicians J) professors or experimentalists? Does
it retard, our advancesient toward a uor® eoasplet© understand! ag
of life? " If ^e saw that it were disadvantageous j, would we
abandon it?
Firsts let la® be explicit. I am not atteaptiag to
make an esclusive abstraction frc^ experience j, either of
brain or of mindo I am acknowledging 9 however^ that it is
only hy conceptual artifice and cultural habit that w®
consider these separately? in effects the expression miad-
brain or brain-mind is more adequate than either mind or
brain alone. A further practical handicap of Plato ^s
». 12 ~
assttmpiioffi l3i@sid@® it© iisterfereac® with coacepti^al
advjmce^eBt lies la th® professional anta^oaisms it
cultivates =. Th© ©actreia© views are r@adilf charac-
terlseds Ttios® who to®li®v© too jssalowssly ia matter
ar@ confideist that fotological phm&om@n& can b®
"©xplaia@ci" ©sstirelj om th© basis of laws of physics
a^d chsraistrya Mud is az& lllusioiio Six^cd raised is
th® siost aathropcMorphie thiag isi ®aa» it should he
dispensed with, "Teleology j, alsOj,'" they say^ "should
have no placsj ©sceptiagg of courssj, for homeostasis
aad a f@w other sel®ct@d forms of piarposiv© . biology ^ "
It cct^es as a shock to these persons to learn that
modern physics is becoming asore aEthropoffiorphiCa ©¥@is
to adaittiag miado 1%ose who l2©Ii3v© too s©aloxasly in
mind^ .»ss the other handg csua show that laatter is in-
ferred: Th@y mistakenly coaclMd® frosii this that matter
is therefore aa illusion „ Thiak of th© impact of this
coBceptuAl isolation os a patient with a disorder of
his '^braia-ffiiffidr' This will iaevitably interfere with
the recogiation of illmessj fiiidiag professional helpj,
participatiag is the therapeutic regiffiem» seekiag reha-
bilitatioxv and eseplainirag his disorders to himself and
to society! % owis experlenc© leads wb to believe that
there is o intellectual satisfaction to fee derived from
cosjsiderijsi either of these "opposit®s" to be asa ilMsiosio
IdeaBg. act thifflgSj rwle maskiado Whea we use
concept for purposes of gisidaace ia our daily llveSg w©
laust avoid comtustng the coacept with experiejac© aM be-
lieving th© ose is a sufficient asEplanatioa of the other o
irhere caa %i^ divide th® aervows systeas to hedge off th©
limits of Blffid? Usstil we kaow enough to be able to
answer that qtsestiogi,; how cau we b® satisfied with practi-
cally coHipiet® isolatioa ija traiaiJBgs research and practice
betweeia those who ®xam±n» and treat the mindg and thos^ who
exaaiao acid treat the brain? We saimot iam®diately dispense
with such isolatiOE as exists » but hem loag should w® b®
satisfied with this as an adequate intellectual frame of
refereaca? Of course ^ until it was possible to detenaine
some of th© brain aeechaeisiffis respoEsibl© for certain mental
processes^: such isolation was perhaps isevitablej, evea
though it were acknowledged that both professional liaes
of activity ultimately relate to the same organ system.
But already there are several brain mechanisms known to b®
respOEsifol® for mental processes. Th@s® feav® h®®n discovered
through th© efforts of various mental arad sseisrologlcal
- IS ==
disciplines 3 of tea paired up with @%cli otti«i>o How much
aore effective the .collaboration ^bms@ tk@ intellectual
and coaceptual force of two ox* nor© disciplines can be
combined within single individuals o
It has now been found possible e:cp®riffi@ntallf and
intellectiialXy to cross boundaries that had so long re^
nained inviolate that their irresistabilit^ was generally
conceded, Sorae of these accomplishments are still hardier
within our grasp » At a time of such rapid technical
achiev^aent j, it is nore than usually necessary to be highly
selective of our investment of time and action into those
things idiich will lead most directly to more fundamental
understanding. It is always easy to imagine something worth
doing, but what is v-«t worthwhile is very difficult to
determine: yet^, that siore critical determination is princi«>
pally what separates \ts fr<m great achievement o Although we
are often preoccupied with dollar budgets « dollars are
seldom over most precioiss coaBSodity« We cannot dispense with
budgets 9 but we can perhaps attend more conspicuously to
more important things c 1?e can perhaps give more consider°°
ation to t^at should we do instead of simply feeling compelled
to do scaaethingT Perhaps a wastage most to be regretted OC"
curs as" a result of pursuing perfectly jxistifiable research
when a modest inv®st®@at of cresti'^e tfeiskiag might have sug-
gested s«»i@thing far better.
In every discipline relating to the nervous systems
there has occurred during the last decade considerable dis-
ciurding, or at leatst drastic compromiaing^ of some of th©
most fund8Mffieatal principles usabraced by that discipline.
There is a swiftly flowing stresMs of intellect iml mov®aa@at
in progress c This is evident all around uSj, although th®
attitude we- often reveal in describing such progr®ss
(particularly if it is our- own work) ^ould suggest that w@
are at last coming to some sort of leveling off; there is
tranquility ahead, still waters o Rather, I believe the
future of this movement irill be more swift and compelling
than in the paste I think I can hear rapids ahead » and am
looking forwasi to them^ even though they may give all of
us a particularly severe drubbiisge What are we h®Tm for?
To conquer our ignorance o
Perhaps the most reasonable way to liberate our©©lv®s
f r<m the many kinds of psychological stricture to which w©
often seem so committed is not to take otirselves too seriously.
This leavening is difficult to achieve without risking real
or imagined deterioration, especially In a self-coasciotis
environment <>
-• 14
<^KiAL commwrmi
Jtecla teas be®n said afeout "collatoorativ©"' aad
'".i33t®rdiscipliaary'" endeavors ia cositemporaxy researcfe..-,
La^t s«8i^@r Dr„ ; »> ? a;, i. ^>..--- Cl©m©iat® aad I tliouglit it ^igfet
Im of ijjterest to e^»«iiae data prepared for tfee pi-e^iotss
Aas^ial Report off eacto of thm two Iiastltistaai to ®®@
whether thmm® data r@fl©ct©d mmcfe or Xittl® eollaboratioa.
Oam of tli@ prlwB argmmnta iis@d ii2 t&@ ©stabllshm^iott of
the Matioaal Issstitiit©s of H@aXtli eispfeasis^d tli® s^spposed
advantages of brisglisg t©g©tls®r seieiitists is compl®m«fitar5'-
diseipliaes i?ho would pr@simably work together tows!J:'d tte®
soliatioja of problosss relatiag to certaisa dis®a®© catsgorieso
la tlx© esus© of tis© Basic R®s®8xcli Programs MIlDB-lIMl.,
appro^ffiat©Iy a doss©© dlscipliB@s w®r© coi&sld©red ©specially
Th© MBisal Report does isot bj sls^ m^mm r@:f.l@ct tbM
®ffit4r« ©Mount of sollateors-tlv® r®s©arca iia beiEgc. last® ad
it r®tl®cts only tbo®® projects which yielded euffiei«@at
@xp©ri»s«atai resdlts to go woU beyond the "pilot" stag©,.
Although a gives project sight to© ateandoiied lat©r witfeoMt
p^blisationj u@i2&Il]r offi® or morm &ad Bosmttm&m several
papers are published bb a coBsequeac© of a siJsgXe project .
SoiB@ projects coatiau® for y«arsj most of theffi iiWol¥© ®or®
thaia half a lea^s'^ss ©Btir® acleatific ©f forts for thm gi'e-en
y©ar^
In assembliag th® data w@ laad® no attssspt to Jw.dg®
t&© valui© of coXlaborativ® or iaterdi^cipiiisarf r©s@ax-eh;
th® facts mlMplf reflected th© sciaatists" ©stimatioMt of
how th@y had ®s[p<ind@d th©ii* r©a@areli efforts o More thaB
half of th® projects r@fl®ct®d sollafeoratioa ®^t©adii&g
b@yosid th© liisits of th© laboratory gro«p„ This Msisally
meaijs iaterdisciplinary research <, Abougt aim fourth of th®
projects ir®fl©ct@d ijster-isistitjat® collaboration throughOMt
HIHo learly »iO per c©at of th® projects wer® r@port#d fej
solitary investigators <> This Aaatial Export r®.fl©ct«s ap-
proati)Mat@ly th® amm proportioii^6 laasraiacfe as sal labor at io.r^
is aot encouraged for its ows salcag aad th®re ar© isi®vitafelf
soae handicaps to its prosacntiojSj th© remarkably high pro-^
portiOE of collaborative aeti^ity probably Jisstifi®® thm
original arg?im©ats in favor of brisgissg th® coMplism^atary
disctiplirEes tog©th®r<,
,-ii rfesoarca iii'»rol¥© r@s«arclS, putolicstioia aad e«>g«s8wiaicatioa
&t scientific wsatissgSo Tfeijs jear has ,se®a aa @xc@ll@)tit
hanrc^st of outista^diiig r@^®f&,rc.li papers fyoa tk-® ProgriMaiio
Tfe© «''atis'@ ©i2t®rpFis<@ cass r®adilf b© J^j«t.i.fi©d os th® laaslffi
of a lew of th® r©ail.f erf;SvtiiP@ icteaso T&® ®tmtM<^ of tfe®
Ps'tsgram i® fis3etla®r sseasmred bj. tfe® l^rg® avrolaer of ia¥itA~-
tioss tlmt COM® to ovat ®ci@;atists to proi^ide papers gt
Imcturmm a.Bd t© joisj oatstaiadimg isa4'!-«'@r^lty facilities «
G©&>.gid€i)S'isg til.© s,wmb©r of liigklf qualified sci©i3itig«ta In
th& Progs ^tia &Md tfe© suffiber ';*f iisport^at posts avail&bl®;,
wa as-© touMd to las® some g^^od. m®is ®¥®rf jmsn:. But is©
alsio. ®iaJof q.isit® fs^orabl© .r©cruitiB®st curreistii wfeicfe tortag
aciexstiiit® to th© Program, W® &r© still iacreasiag i®
8tr©sigtii as am intellectual aad ®xp®rim©jatal r^sourc© foa:
traisaiag aisd expmrtmnc^ of sct®istists &t a-Il l®¥elSo l@&rlf
e¥©rf gft&joy imiversitf in t'ais coautrf aad ssosm© f ift®©ffi
aaiv©rslti®s iis for®i^ coiiiat.ri®8 ar© T®pT®B®nt®4 ia ous"
Program thiB |f@aro A f@i? of oar owa p©opl© w®r® ©Bablisd
to visit Ifeteoratorl®® or attessd maetiiiga iia a do2s®a for©iga
co'aatri^s this f®aro Tfe©r# i« aow begiaaiug some ©xchwig©
witk mmn Euesia aad Polasdc W© w®y® l&osts fof tfer®e w€5®te
to Professor Jerzf loaorskij, ,l©&d of Meuroplii'SioIogf In the-
M©riCki Institut© i» Warsai?, aad for a bri®f©r timi© to fe
a-asabar of EiJUisiaaSs, ia'cl-adlfflg PToImm^oT Bfkov of hmmixig^TmA,,
PTotimworm Sarkisoi? asjd Fropi>sr-G:?a4Shcfe©akov of Moscow ^ aad
Yi£?itors from mor® thaia 22 diff^rant co»ia,tyi®3. Tli® Progx'^iis
is wjii proT@B, as sm iat®li©etua,I asd r@s^ar«h r©soarc# of
high i!.it©raatloss&i ^'@gard„
A a©w stjie of ©diicatioa®.! ®3£peri®BC© at tk« IIH
waa aaiitiated last f©ar tof Dr-o Caatosii, Em isi^it©d
ProifiBmoi' M».rtiii PoaIocIKs, &u ©SEjm© cli@mist fro® EsLgiaad,.,
to njjesd s. period ol ttm@ at tis© latioaal Ijsstitutas of
E®satfao Dr. Pollock aud b. nvmhrnr of lemdiEg 0ci@siti«3tig at
tfe© MIH ajad. n©a^bf iaboratoriC'S m&d© tfe@M3©lves available*
for as @xt®ffid©d program of isafoi'mal mmmixmrm ai»d discusssioEi®.
T&is progi-Mii was opaa to i.iet®r©st©d sci@Et lists trow, all tfe©
I;Ba:t.lt?.?t8s «>jod aearb^ labor at c-,ri®So Most of th® particlpssfi.tii
i©agag©d is* preliiaiissrf sr®adiag fend studf prior to t.fe®
n#girija,lag of t&®s® sas®tiags ais.d ma^y of th©s( @i5i,gag©d f all"
-'la© ia this sort of ^*iffit®ll®ctusi »rk.®feop" witM Dr. P®ll©c,fe
tjid ©aefe C'tfeer.., Tfe® sutcoiie memmm to fea^© li@©a vmrf S&w&m--
■-■blm aad haii ulw®mdf h^d mm import aat iiiliu@as® om h±QehmmiQ&l
■ '=*.'*'^*as'<;l!§ local If o
Two ws,joi' Sfffipci-siia «®ir© spoasored dusissg tla@ f©ar bf
th© '/Saiisie ite^^ai-els" Programc Oa@ traced tfe© d€¥®l©pm@©t of
coac'gptjs tteoxig&out tfe# first c®sst«r;f of ^'turosfea*?''^ ''■*^'* '^'»*-^
" iS -
paid ©special tribut® to J,LoWo Ttsudichuaij, tSi® f0ysd@r of
a®ur€>clie(Bii®ts'^= Projections w®?® m&d@ S^ scientist®
r@pr®s®iitiag di@ciplii&©i3 allied to a®uroch@iBi@try s^ to
th© nature of tl%® !&@@d for an understandijag of th® i&euro»
chemical ba@@s of s&^urological and psychological pif«»c@8s@s»
and appreciation w&a ^xpT®mm®^ for th® r@@©arch pot<@nti<=»
aliti®0 and oppo:s'tianiti®s of thi@ field for th® future o
Th© oth@r S^po@itm brought tog@th@r for th® first tim®,
for coneid©r®d discuss ion , ©zparts on th® raedicals, l©gal
and social problems relating to narcotic addict ion c Both
Symposia wer® w@ll att©nd©d and lively o lach l®d to a
more widespread and intensive interest in the intellectual
content of the' issues involvedj, especially mm this leads
to new r®@@axch directions <.
0»® of th® traditional ways of iaproving the
creative poiver of an organl!Bation<»-»through the vme of
expert consult ant g^-^has been even Bor© actively exploited
by th® Program this year than in the paato The national
Institutes of Health established a pattern for advice to
the Institutes through Boards of Scientific Couns^lorsSo
Boards '«9er© established for each Institute and reviewed
the independent ax^d combined ProgranSo The Boards gave
encouri^^Bent and intellectual stimulations, as well as
aiuch°=>appr@ciated advic@o A nvmb^^r of other experts
continue to advise and p&rticipat® in sore liaited aspacts
of the Prograaio
Oa@ of th© Coasuitants p -who spent a considerable
time worJls:ing in th@ Basic Eesearch PrograMg is Professor Leo
Szilard^ During ti%@ course of his period as Consultant ,
he devised a new theory for the nature of th® aging process c
This is a lusty th@ory which ''explains'* a vast ai^unt of
adventitious facts In addition to those directly concerned
with th® genetic factors in th® aging process o Ci^d.ng as it
does into a field that needs a broader theoretical basis s^
and being a theory that can be tested experimentally at a
nu»ber of points « Dr„ Szi lard's ideas will provide auch
stimulation to th^ iieveral disciplines engaged in studying
aging and will t®&& to focus attention upo^ central iinifying
concepts o Dr„ Szilard's theoretical study ha® already been
coapleted and dis^c^is^^d aaK>ng scientists at the Mil and
elsewhere and is to appear in the January s, 19509 issue of
th® Proceedings of th© national Aeadeiay of Sci^ssceo
C^ the fr±i&gm of our laore central research mission » the
Prograai particip&t®d in a nationwide television prograsij,
sponsored by th® jteisrican Associatioa for th© Advaac^saent of
: /
-Scieac® ai^d th® Hational Acadeay of S€i@Be@„ TSii© program^
C0HQB1ST» produced fey CBSg ©mpliasi^ed som@ of tli<e research
poissiMlitt®8 b^iaes ©xploit®d In laboratories all arouad
tis© world to d©t©rniis® braiis orgaalsatiou and tli© g-elatioa-
ship of tMs to afi.irologieal and psychological fu?ictions»
Th® Braiii; Story dr©w a larger aisdienc© thaa any previous
COMQOSST show aad woa the "SliogBas Al^a Edisoa Award for th©
■'best sei®nc® steow for yoisth in 1958 c'' All t^&ntj or so
of th© newspaper r@vi@ws w©r® veuy f a^orabl® aad ®v®i£
superlative aisd th® asail response^ isaisy froa studeat© ia
high schools aad smiversitiesj, iisdicated that th® program
coBveyad a good d©al of iEtellectual coateat asti®!! as
appeal o Official ratings reported that sossewhere b@tw<e©a
15 aod 30 millio?^ p®r©oxagi viewed the prograaio For th®
first time,; a sei@-ic© prograa beat "Th© Lone Rangsr, '
Oyr profoaadest concern as scientists is associated
with th® iat@ll®Gtiaal content of our diseiplineSj, aud
especially with tli® creative processes necessary to prograag
in these fields, All adiainistrativ® considerations ought
to fee directed to ©acouragep develop and @xerei®e these
essentially inteli&^ctual faculties o What can be done to
j&ake ourselves morm creative? One suggestion is that
hypotheses sho%il<^ b^ treated opposite to casual traditions
that is^ be ssore ^ ^.'soated in th©ir most initial stages
and laor© skeptieaA )^ held when th^y are better established,-.
Our guide for progress^ individually and collectively
ito paraphrase Jacques Bar^un) grows out of omr interest®^
viewed objectively ^ in the long-range and in the largest
possible sense; oust interests to improve ourselves and
our work; our interests made as self-aware as possilsl® >
It is unlikely that mankind can attain what it does not
wish to strive for and iaipossible to seek what is
conceived as unattainable.. ■.
Robert B« Livingstoaj MoD.
6..«.1J.W.5 ?' rv'- =.- ■•' -^j ~ ■iie)ii.!Q>miCv,i- SCl&SiC'SS
Will 1ft* r. Wifedl©
orgaffii.a;®d as foor S@etioas aad a Field, S%M.tt&wt.^ E@a.®a.rcfe
tn «ach @f t&@®@ M&itB h&m a distinctive Gh&r&ct&r,
alt&eugfe tte®r© is a &®a.lt&y ammmt @£ ©veriappiag aa^l
iBt@gratioffi &i isktmr^mtm wit&ia t&e wkol@a As iffiplted
by tfe© ffia®©» t&@ laboratory's iiav««^igativ@ programs,
©ispiiasia© strac:t«ir«, but feie projects" a.r@ liMxtsd t©
^*rplaol&gy , asud Ssoa® lacks a stroMg JTuffletiOffial basi@o
Various discipli&es ar® r»pr©s©ated ajs^i&g t&© swttffity-o&'e
full tiiE© sci@fiiti,ats aad four aetlv® caffi^«ilt&®ts ffiow i®
residgffle®; «®©l®gy,i, c.ftolQ.gy., ©gsbryologyj pbiys iology ^
biocia^ssistry, ©eurosiwrgery ^ 8B®dicai si©arO'l©gy j, a@ur©pat& .
pisycMelogy, asd ®v®ffi s>vfegt@trlcs ar® fields ©1 prisiary
ift.ter«at of various M#®b#r.« of t&® staffs Tfei® wide
div^irsity i^p&rt» a quality t® th@ laljoratory , fousd ia.
f©w il asy atfeer is@ux'©aaat@Mical «i«partiffi@ats iss t&is
country o
P.r«pftrati®ffi of a fertai mumm&ry o£ asys'aai r©searc&
progr©®® i® complicated lay this divarisity ®f la&tere^to
lacls Section, C&l©f feas prepared s suatsusrisslag rmport s-sd
tl&®s® aave b©#s e©abi5&@do
S©cti®a oii Meuro©yt©logy
So h., Palay, C!ii@f
Tfee work ©S tais s®ctiosi. has feees so&^idarafely
advanced, by tfe# ajEcfeaag® of oxir ©le^trom Microscop® f@r
a. a#w ffiodel i® Juise, liSSo f&e ffi®w sieroseope p©rMit«
\m to carry o® t&® work: which we lat©ffid©d t© dO' duriiag
til® previous year witu the previous laodelo
A study of turn fia© stJ'uetur® ©f asoffis lis th© ceatr&l
aervoas systeai of f i®fe®s skows that ajEopia^® coataitis
tiaree loagitudim&lly arraaged. elemeffitss sitocfeoadria,
caaaliculi, amd amirof ilaasftasats.-, Tli@ s^st pr@aslB,eiat a-f
issarlf fiii th« vo]u^@ of tfe© iargK- axosfe. t%st tfe®®«
filais^sis s.S'® riot artifact® i® deaosKtrat^ci isf pog;it.iy@
bis'^frimgejac® of tl%e »>a:offis ij& fr@sii pr*ipar®ti03®s. thlm
study urns c&xffi@4 on in collafeoratlos ^ith Dfo Bairati,;
Usai-^ersitf of Milass^ Br. CSofdoSs Ees®©*-©!! Associate,
te'o Brigtstman feas coapleted & study of tM effects-
of larg® doses of irradiation to th^ b&&® of th® toraisjs.
T&is iffii?®sti|fatioe. indicated tlsat alt^fatioas i© tlie
vascular ©lapply of aesf^ous tissis® ar® probably primary
©vests iia th© destruction of ^©rvoMS tisasti® by X-rays o
BTo BrigM-isaa im collaborstiosa «itla Dr, Albsrs
S»a« also studied the distritatiosi of tentyrylcholla-
estsrase activity i& th® CMS of @®'^€ir%l comaaon as&iualSo
He fous&d that ia tli« rat^ goldfish, aad toad, tiiis
enzyse occurs lis tis® ©sidotlaelium of ip®ss©ls, whereas
i® til® ©at aad foTsl^, thm ^nzftm resides, im tfee neurogiis-
Since iBls.ibitors of tM® esizyag® lasvf» bseia reported to
produce demyelinizatiosa iia th© fo^lj, thlm stady suggests
a relatiosss&ip b©t«ie©s» glial towtyrflf.laoli.ii®st®ras® aad
aiyeliia forisatiois by neuroglia c@lls.
DTc, Alb€8r®9 partly im coliateor&tioja with Or, Bsfady,
has studied t«o eiizyases i® tfe© ©©utrai is#rvous «y@t®»
tisat are lffi¥oi¥@d i® the HaetaboiisiB of Y-asaifflotoutyric
acid^ Both of t&ese^ giutasic decarboxylase aad 7-asiiiiso-
bwtyrate tras^^asffiiaagie ^ are fouad iia reia timely laigli
coacentratiosa® i© gray ssfttter aisd mrm virtual if atesast
from white ssattero Tlfe® role of tfees® Offljsymes iis, t&e
jaetabolisffi or ia tli« specific fuactiosa of tissu®!^ of tfc«
ceutral aervow® syat^a is sot clear o
Th® Secttoas ^ssg host to te. As^elo Bairati
CUffliversity of Milais), Dr« Eo E« Maauslldis {Yale
Ifffiiversity) ^ ^^^ ^s'« S&®lla Dojaahu© (Ctoluffibia Uai¥@r@itf1'
for periods of 6 to 9 aioathSo
Section on Sxperiffieatai Mewopatfeology
Jo Cai^erreeyer, Clii®f
The research of this Sactio® hSLS cosstinaed nitlaias
the frajse^ork of a central theise asd long raiaig© pla®„
To Stat® thlm briefly^ the alias are to reveal estra- asd
iffit.raapiaai factors involved ist tte® Rorsm®"* 3saiat©»a?®c© «f
©piaal cord structure aaid fuiaetioa,, aa*:' to ascertain th©
degrees of deviatioia txom aorssiality wteicfe will cause
EsKperisBiPs&ts haijf® b®@s& devised to diselosa r^iafiios.-
sfeips ©f ©Ktrasplsaal factors t© spiiaai cord malfua,ctioiSc.
Thes© have b©@m feeld ±a ab®yaae© for lack of syst^saatic
study of tb,© c©sst©a.ts ©f tb® epidural spae© which Burx'<aand&
th® spis&al eordo To provide bom® of tfe® m&dh a©@d©d iaifar-
fiiatioffii, a EBorpfaological litudy of tli© epidural fat, it®
r®lati©aSs variation with ag« aad species diff@r®®c®s feas
teseia carried out by Dr„ Helen Easssey aad is sow r©ady f©r
publicatioffio Th@ iaiportaac® ©f epidural fat is that it
p©r88its changes to occur aloag the vertebral colusm without
tearing tfe® comteiitSo However, tte® deposition of fat is
found to b@ of a coanplex nature tu thm growing agid adult
cats aiad otfeer species ai&d varies individually to soia©
extent o
laitiatioffi of &n eseperiaieiital approach to tfee
i®y@l©patfeies has been feasapered, furtherisor©, by lack of
iffifongatioa about tfe® volume of spisal c&rdo Exact is:a©w-
ledge has been ujaavailabl© regardisstg alterations in aoraeal
spinal cord histology and cytology occurriag in consequeaics
of reiBoviag the spinal cordo Finally, tfe© adequacies of
coiiiionly used histological techniques had to b© studied
prior to going forward with a well -controlled investigatioa
of experimentally produced myelopathies c.
Siss© of the liorssal spinal cord aad of its several
regions has been established in several species c The
size of the spinal cord aad of its regions can be estiisated
from the correlation which exists between spinal cord
volume and size ©f other anatomical structures asore
accessible for aeasureaient n
Effects of cheiiieal and physical agents us@d for
preparing histological aaterial have been investigatedo
Every step in fixatioiSj repoval and subsequent histological
treatment of the spinal cord has been surveyedo On the
basis of the iaforiaatioffl obtained through ai®asur®BS@nt of
the spinal cord and of it© component cell structures, a
procedure has been developed whereby errors in the
histological preparations are now reduced to a lilniMUM.;
aad artlfactual chfmgess avoided., We ar© bow prepared
te iavestigat© @xperiia®a tally produced voluisetric aad
nor pho logical cte.a.age@ of th@ spinal cordo
A pr@li»li£ary study toas h@®n carried out && th©
size of neuroglial nueleio These structures increase
in diaaieter significantly with the size of the spinal
cordo ThuS; when it is desired to compare an @%p@riaental
animal with a noraal one for the purpose of determining
whether or not an experinental agent has had an effect
on the nervous systea^ w@ now know that the question can
be answered only with animals of identical sixe^
Eff^cls on spinal cord voluiae and nerve cell
structure following adninistration of urea in various
dosages and for various lengths of time are being inves-
tigated In collaboration with Dr„ 2^i@snowlcZo This study
is one of a series on dehydrating agents o Other studies^
on the spitsal. cord are being conducted Jointly with
Drc Mignon Mala, using isotopes in an attempt to visual is&e
the routes of transpoxtation of tagged metabolites o
Finally, the responses of neurons and neuroglia cells to
Hotor activity and to pharmacological agents are projected
ExperiMents of simple design, it is hoped, will provide a
better understanding of the aechanisw in spinal cord
reactions and an explanation for differences in reaction
between spinal cord regions aad between the spinal cord
and bra in o
The Section has been host to Dro and MrSo Stanislaw
Zieanowicz CJa.g@>llon University^ Poland) and Dr^ Mignon
Mala CHalversity of Stoclchola, Sweden) during 1958 „
Section on Functional Neuroanatomy
Go Lo Raeaussen., Chief
At present the work of this Section is concerned
priiiarily %ith nervous pathways and connections of the
brain &md spii^al cordj, with emphasis on the neural «@ch~
aaisas of auditory aad vestibular fxmctiono Practically
all th@ studies have engaged the Joint attention of the
Section Chief and Dr, Oacek, Research Associate o
An etf^reut ii©rvous coMpoaesit of the v®«stitoulax
aerve has hme& revealed wtiiefe is cossparable ia asaaiy
anatomical respects to tli® @£f®r@sBt cochlear bundle,.. The
vestibular ©ffereats feav® been trac@d froiSi tb.©lr ©rigla
iffi tfe® lateral vestibular aucleusi througfeo-ut tfee vestibular
Hi©rv® And its braiscfe®s as far as tfe@ receptor organs ol tk«'
vestibular labyriatfeo Tfe© possible 2=@latioffl®ls.ip ot themm
effereats to t&© hair cell receptors tfe®s8S®lv@s is currasiscl.y
UMder study o
In ord®r to b®tt®r uaderstaad tke aeurai mecfeasntsffi
of h@arliagj, studi®® of ti&e auditory affereut syst©ffli, so
long neglected, feav© received partisular atteatioiSo
Poiist-to-poi!Bt iBt<trisaur©aal r©latioasbips exist lag b©twe«sft
th® orgaa of Corti aM tfee cocfeiear aucleus aisd th© !g»im#,i:
of projections froB tlie latter to feighfr auditory if.uel©ar
groups ara being r®studl@d in aior® detail tbaja h,@T0tQt&trj
by th® i3xperise®ffital aaatoMical appr©aclio
MTo Boord, a ©diversity ©f lai"ylaiad stud@ffit, com-
pl®t®d a study of tli© iaaervatiois ®f the vestibular aad
auditory apparatus of the chlachillSo This worls s@rv«d
as thesis !»».t@rial for his MoSo d©gr@@o He is coatissulRg
studies of tfe@ auditory systasi to©r® UBd@r a MS R©s®arcfe
Fellowship,, Tfe©'proJ©ct eoncersis th© ©stablishaeat of ai-.
@f f ©reat . coffipoiieist of th© cochlea iffi swbisaiiBaiiaffl v®rt©bi'
possessing a less developed heariag iBechaais® thM.m pr®s©s3V.:
in BasBBalSc The results of this ifflvestigatioim will b@
iRGorporated is&to a thesis for the PhoSo d@gr@® from th®
Uffiiversity of Maryland c
Mto Mor@st, senior isedical mtudmmt of Yal® 0&iv«r'i:i.£.;> ^
carried out aB iadepesidesat research study ©a the fiber
coBiaestions of th@ area postre»a of tfe® ^®dulla obloagata
during th® Sixmm^T under th© COSTSP program „ H© was abl®
to ©stabiisfe certain ai&atoiaical cosmactioas with issidbral®
structur®^ and to correlate the results with previous
!il©ctrophysiol©gical observatioa® at Yal^c
Bro J. Bo Waith®r of th® Majc-Piaack Gesellshaft,
Bad Nauh^im^ G@rgiaiJiy, is ©xp@ct©d to joiia this Sectioa
befor® th© ©ad of this year^ He has b©€?a study lag physit;
of s@ffl!S© r ©emptors amd ceatral semsory ■gmQh&m.im's^ ioi
several jm&rm aisd will tursi his attasatio® to the asj&tc-i.,
and physiology of th® eff©r@:at auditory and visual
Gosmmcti^omB, a.«. , Waltfear's e^.j., ,■ ...i..: is being ..,...,..^&^...,-..,.
by the Matioaal Academy of Seieac® uad@r tfe© Visit lag
Drc Leo Massopust l©ft tte® Soatioa during tfe®
cisxreut y@ar t® ace@pt asi academic app«»iatM®Bt @ls©wfe,©r©.-
Secti.osa oa B#v@lopffieffit aad E®g@ffi©ratioffl
Wo Fc, Wisscll@p Cfei®f
Tb© res®arefe aeti¥iti©s ©f ti&i® S®cti®ft i^^l i..^,.-.;^
fowr principal categories;, Ca) seureg^mssis, Cb) reg#Ki~
®rativ® potest ialit.i®s of e@atral sad p@ripfe®ral ummr&mB,
(c) @xp@ri®,@iitaily imdiaced structural alteratioait iei tk®
ceatral ^©rvo^is mjmtmm^ aad ((d) tecfesiea..! d®v®lopmtist..,
Cs) Drso SidMaa amd Miale Ixav® ©xsuiia@d tfe®
te©&a-^ior ©f cells ±m tfe® a®ural tute® off is©us® ©seforfos wltte
t®ctotqtt©s of tissue culture aad awtoradlograpfesrs, usiissg
tritiMffi-labeled tfeyMidia©o This auclsotid© i@ imc-orporated
into DMA of cells about to dlvid® and r^ssaias as a p&x-
maia@ffit Barker o Wfeol® ^mhrjos cttltured la li!^'® Burwlvmi
for a w@®k or more aM mttotie activity was vT^— -.?• ,
Mn»cl@i of cells lyirag iia tk© ®aatle layer s;o»- ^ -ce
away from t&e liaii&g ©f the umsTSt.! t-atee i®ysutfe,«(s«.t.&ssv- «®i&
DMA and tteea Migrated to tfee @p@ftdyma..l layerc, Siabseqweatij!
they isigratedi lat®rad amd prepai-ed for dtff@r®stiatloiic As
growth cartiffiued^ subsidiary psttersis ©f aitosits an,d
ftigratioB appeared « Whole orgaa |©r ®istoryo) culture has
pr®ifid©d access tcs> a ttm© in dB%ml<8>pms^nt tfeat Ixas r^siste-d
investigative efforts iii th<B pasto F®rMux@ffit labtisliffig ©f
■e®lliilar cospoiaeiits aad traciag the® dif fereatially by
autoradiography iaav® op©a©d, asa ®mtlr®lj !s@w av@mi® of
r®s©arcte ia developmental pr©c©®a®So
listogeaesis has b©@B studied la tfe® r®tlaa® ©f
BorMal mie© aa^d ia t&©s® ©f a geaetie strata wltfe dy^tropav
of tfeisorgais by BrSo SidMaa and .f@d@ro lo feistoiogicai
diff©reac@s ajjpeared uatil ttee alstli day of gestatioa..
te©tl& Koraal aad dystrophic ejmm develop iag equally uatil
tfeat tis@„ Coia® c©lls appeared a littl® earlier tbam re^-d®,,
aad tto.® r©d-feip©lsr synapses w@r® s@®i® on. tii.® ®lglstte day,
After ttee aiatfe day ta® rods aad cos-ms d©g®a©rat@d aad
sca,rc®ly a trace of tb@ pfe©torec®ptor lay@r resAiaed at
the fift®@atfe dayc ffe@ dystrophic coaditloB r®s«a8bl®s
r®t la.it is pigs^j^iatosa, a feusaa disorder of g@a©tic ©rigte
wltliilSriai' histopathology o
Cto) Invest Igatioffl of tb® pfeeaeiieiioffi of c®atrai
servous systeig r@g®n@rat ion has bean cositii&ued; Dr., Ca^pbe:^!
of Colimbla Uffiiv®rsity collaboratL^go The Koskey's spiral
cord was severed surgical ly,, le&vii&g a gap of £ to 6 msi^
Th® cord stumps and tb.is gap w@r@ th@ssi @iiclosi@d i& a snail
sfe©et of Millipore filter sterilized by radiation. Witfeovt
the ©aclosiag fiitair, tfe© eord stuasps b©eaffie capped by mmur
&&d com&@ctiv@ tissu# and a raadosi ^car t&rmsid In the gapo
With Millipore, th© tissues ia the gap were organized
longitudimally; the gap was bridged by splndle>shap@d cells
and blood vessels , asd a few regeaeratii&g ii&traspifial
neurons fe»llow@d thi@ oriented tis@u@ in crossing the gapo
Snail fascicles of regenerated neurons were present thsre
3 to 4 is»>nths after operation^ They reseasbled peripheral
nerve roots by virtue of the presence &m»m§ their fibers
of Schwannlike eel Is c Mo functional restitution was
obtained c
Dr„ Guth has continued investigations of heterogeaow.*.
nerve rege-aerat ioii and functional restitutiono In collabora-
tion with DTo Frank, It was found that function could b@
restored t© the paralyzed heffiidiaphraggt of the rat after
phrenic seurotoay by directing central vagus nerve fibers
to regenerate into the peripheral phrenic stu^po The
recurrent laryngeal branch of the vagus nerve displays
bursts of activity which are synchronissed with those of
the phrenic to the diapferagaij, wjfeich accounts for their
ability to take over phrenic function after regeaeratioiSc
These experissents have b®®a extended to the sonkeys,
Drso Campbell of Colusibia and Soutter of Boston University
collaborating^ If successful in the leoakey;, the latter
investigators plan to apply the knowledge to human patients
with diaphragnatlc paralyses o
Other studies by DrSo Guth and Co Jo Bailey certain
to pupillary function after degeneration of preganglionic
pupillodilator fibers in catSo These experiments are
expected to shed light on the question of plasticity of
central nervous connections and have theoretical isipl icat ions
of considerable interest o
alit^ssi' ffi^ui-'sai iMi': -ai^ss pursued
la coliv v Qy^ Camm-y ^ projects
aierit r«vife«,.
Drs. ToMas of th© Ual'^eysitf of Caliiorsia assd
van WageaeB of Yale provided tfe© Sectioia i»itfe perfus«d-
fi3!:®d braii»s of four adult asonkefs iis %»hlcbi tlie pituitarj^
stalk &&d glAnd had t>@@ffi irradiated in tli« Calif oriaia
betatron msLTlj In lif@. The i»europatho logical naterial
i»as prepared bere«. and DTo, Caanserssefer reported the obser-
vations o The path of the horizontally directed bea» m&&
seen in the temporal lobes of one aniisal only ° The other
three @ho«ed no d&m&sS9 there « and resiarkably little effect
vas encountered even in the infiindibular region adjacent
to the pituitari" stalk. This technique «as used very
effectively to produce a sauill circtmscribed lesion in the
hypophysis alone «
Our project on anatoaiical correlates of the functioffiitl
chasiges resembling huaian paralysis agitans that appeared
in chronically reserpinissed a^nkeys and cats «as ter-
ainated ^aith publication of the results CVindle and
Caasaeraeyer '58))= After daily adisinistraition of this
drug C0o2 - 0.@ ag/kg) for as long as 18 saonths^ no gross
abnoriBalities ^ere found in the braiBo Microscopically
there i»ere no henorrhages; infarcts^ softening^ deayelination^
neuroglial reactions nor phagocytosis. However ^ conspicuous
cytological changes appeared in the cerebral cortex, basal
ganglia and bs'ain ste»<. Cell nuclei asad nucleoli appeared
pale asd enlarged | the karyoplasm often 8hoi»ed a "hole"
due to removal of so»e substance during the process of
histological preparation of the tissues- These vere
interpreted as ^»npat ho logical^ probably reversible
changes o Distribution of altered cells did not conform
to pathological foci of hwean paralysis agitans o This
study is the first in<»hich a specific aorphological
effect of adi&inistering reserpine has been reported »
The pattern of neuropathology of asphyxia neonatorxus
in the aonkey is being investigated by Drc Ranck^ using
material obtained from the Puerto Rico Field Station
studies c The first thorough study of the effect at 10
days of life^ after asphyxiation for 17 Minutes during
birth and subsequent resuscitation <> has been completed o
Histology of the braia of a aoiaaspliysiated monkmf of si»ilar
age was studied for control = GetosB pathology was aot s«ea»
Microscopical cl&aisges were exteasive^ alisost perfectly
bilaterally sf»«®tricalj, and localized by cytoarchitecturally
defined nucleic Cerebral and cerebellar eortex shotsed ^©ry
little daaiag@o B^t in all ot!2@r r^gioras amd in thm ispii^al
cord neuronal logs aisouigted to 20 to 100 per cento An earlf
Bwcrophage reaction occurred and astrocytic hyperplasia was
seen in saost daiaaged regions. There were no heieorrhages nor
thronboseso Hie iaferior colliculuSj, soae tbalamic nucleic
the subthalamic nucleus^ globus pallidus, reticular forsatiosi,
eye-auscle nuclei, vestibular » trigeiainal^ cochlear nuclei »
gracile and cimeate^, cerebellar^ and superior olivary nuclei
and the gray Matter of the spinal cord were asost severely
damaged o Among undamaged regions were the amygdala ^
olfactory nuclei^ aK>st of the hypothalarau^^^ lateral
geniculate bodies, pontile and inferior olivary nuclei^ &md
most of the structures along the floor of the fourth ventricle.
Primary motor and sensory elements were less affected than
Interneurons c The neurb pathological picture in the infant
monkey resembled that reported in humas infants with
"kernicterus"; and was unlike that found in asphyxiated
human beings o This is the first experimental study of the
kind in any neonatal primate.
Cd) Other studies under way in this Section were
largely concerned with developing new techniques. Drs..
Feder and Sidman d«g:.igned and put to use a freeze-substitutioa
method which has giv^n significantly improved fixation of
small tissues sssd pfeaervatios of a variety of chemical
substances in theso They have investigated structure and
function of photoreceptor cells and obtained the first
convincing evidence that con® cells are present in the
s'etinas of rats and siiceo
DTc Wolfs, Research Associate^ has explored the
s.gnificsace of acridine orange for staining neurons in
virg and ig vivo., comparing the fluorescent image of
living cells with that of fixed or injtared elements «
Acriline orange was found to be of remarkably low toxicity
and to provide an excellent tool for studying chemical
reactivity of living cells. The data suggest that healthy
cells have so free polyaaions that can bind the dye
metacHvomatically and that DMA becomes metachromatic only
after cellular injury .
Field Statioffi of Perisatsl P&fsiology
CSubeidisry of Laboratory of Meus^oaffiatoggic^l Scl®ifflc©s)
W» Fc Wiiidl©, Chief
Sxparlaseffital ija¥®stlgatios» during tli@ fiifst full
year of acti"«'itf at th® labofatories is Puerto Rico h9,ve
b®©ffi conceraed «itl£ advex-s© factors in the perinatal period
of rh@sus soiik@ys rssultimg is neurological a^d psychological
deficits is th© offspring. Th© first advers® factor tested
Has asphyxia ffisonatoruiso
InasBsuch as littl® «as krao^is al»out si^eo»atal m>m^®fm
»md nothing at all about f@tal pliysiologf of a@y sublitmas&
priaat©, it has hemm necessary to b«gi© to collect data
pertiaest to ©asperiBBestal ®furoM>||i«ftl «ork i»itfc Mosifeey
fetuses aad iafaatSc In do lag' so ^, opportuaiti^is liav®
presented thes^elves to collect aaclilary ds.tffl ia respect
to gestatioa^ behavior^ growth amd d^velopneiat. The free
rasge coloay of 300 rhesus monkefs on Cayo Santiago a^d a
caged breeding coloagy aoii auraberisg 75 large feaale aais^ls
of the sane species have sespved these purposes o
Ab observatioaal study of behavior aad social
orgaaizatiofi was started ia Juae^ 1956 ^ by UTo Altmaaii, to
obtaia iafor^atios o@ aorraal subjects o lafaats have beea
followed after birth aad their groisth aad developii^at
recorded. There is sa aaaual cycle of reproductive activity
ia this free raage coloay, births falliag ia the sioaths
from February to Jua©o Oa the other haad, th© caged mo&kmjs.
uader laboratory coaditioas ia Puerto Sico eoaceiv® aad giv®
birth to iafaats ia all aoathSo This has aot beea th®
eatperieac© ia aorthera latitudes <> Th© meaaiag of the
differeace betweea free aad caged amlsals is uakaowa. The
two-year study of social behavior has aad® possible aa
estiiaatloa of the asasisum populatioa the islaad caa supports
This should be 1^000 to 1,200 aaiaals, which will provide
aa iffiportaat reservoir for ©xperii^satal studies. Dr« Kofordj,
froiB th© IMiversity of Califoraia^ will coatiau® the iaves-
tigatioss oa Cayo Saatiago^ siac® !&>. Altmaaa has returaed
to graduate stuaies at Harvard <>
Aathropoaetric studies oa free raagiag rhesus asoakeys
have beea uader way for two years? these were receatly tmUem
over by DTo Gavaa^ of the Medical College of South Csroliaaj
who is about to add the techalque of radiography to morm
coaveatioaal measuresseats of skeletal growth c Aa attempt is
beiuig m&d'^ to astssteliah iSitamdard, phj'sic&i ^^&miX'®tm&.t& fef
whicfe accxir^.ts ©stiseat^,® of a*g® of »o.iakgf@ caa b<e aad®.
Sisac© feiiftls records ar© mvaiiabls for isor® thaj® half tte®
affiiBMaiis ois tb© islaads it s&ould b« posslbl® to collect
the ffiec^ssas-f dtts lis a short ti®^<. Cag®-r®ar®d Moiilk®y»
1*111 fee compared witfe tkos® r©ay®d bf tls©if aiot^ers isi ss
natural Mbitato
Bstablisb««!»at of criteria for si^txro logical ®sa»iaa-
tloss, especially for isfaiats was ai®®d«dc Alt&ougli th®
momkmy h&« h&®si ®^t®nsi.vmlf used for neurological @3ep@rii»@is&t®
im ma.uf institutiosas; wo satisfactorf aetaiadafds for
aetirologieal «xaHd^atioiii h&ve h^mm publislsed, Drs. Ea^ck
and *teri@a Ra»ir@£ d@ Arellano Itav® ra»d<i cositsid#ral»l@
progr@i@S'^h sj^eurological exaiaiaifitios of th® infant
moakmff aad th® program is btis^ extended hf Or»o 0oi8%»ai«
McCroskey asd JacobeoKS. A protocol has prcQ^seed through
several ©dltiomii s.md ®l&ould b@ completed im asaother yearo
Data o^ ffi@iistruatioii of rhesus asoi^efs uader staiidard
co^ditio3£S are beimg collected o Thm i@es3i®trual cycles cf
isidiiridual akonkeys are subject to vide variations «Mch
appear to be equally spread throughout tli@ year^ Dro
Jacobsoffi fouad so relatioss betweea regularity of the cycl®®
&nd fertility^ but a change is the tiiae of smtis&g. vithiig
the cycle ((day 11 ii&stead of day 14) temded to alter the
tine of "iKplasgtatio^ bleediss.g"c Dr« Hertz^ of the MaUomi
Cascer Institute^ isj collafeoratiag liith a atudy of the
efficacy of &b early pr@gma](@cy te^t. The caged breediiag
colony ha® beea ressarkably f^rtil®^ 50 pregaaacies reaultisag
froai satiags last f®i&T.
Ife>o Jacobiiioffi collfiboratiffig «ith DTo Pel^griiaaj,
Professor of OfestetricaB at the University of Puerto Rico,
is iffi¥e@tigati@g the sierve supply of the esdoiaetriusi is a
ffiusiteer of species of amiaials imcludifflg th© !8o?sk@y sd ®aiSc
The effidoasetrium of the cat was fousid to b® deia«ely suppli«id
by fiae termisal laerves arousad the esadoa^trial glas&ds and
usader the surface ©pitheliiuia as Hf^ll as arouad blood vesseiSc
The possibility of iseuroffiai participation Im se^struatiosi
is beifflg considered <>
Data. ar« b®lag collected om maturatioia i© iafaat rhm^-^m
sosBkeys aad o® car® required for rearisg them. A isursery,,
resea»bii®g iaa Masay details tteo@e iss u®e for car® of hu».^JB
iafaists, has hmm^ ®stabl islied <, E«cor«S@ ax-® k®pt of d&ilj
weighty, food iffitak©., body t@ap€ratur«p rmBpirmtorf sr-at©,
dental eruptioffi^, heart rate (WSLQ) , grasp feflex^ «tc«
Zffifaffit Monkeys Mv® sisoi»a a regular' patter® of growtfe aad
d©v@lopweffit^ but wltli wid® raage^o LittI® inf orasat io® for
iffifast asoak©!'® feas b©@js availabl®* tat t&s cag®d aad fre®
raagiffig coloai@s «itli a coisbijied bis-th s'at© of 100 ot aor®
a year «ill sooffi pi°ovid€ it.
Neurological deficits of ©xp@ri»i®2itally Induced
asphyxia have b@@ia iiavestigated by all mmteSbeTS of th@
Field Station 'with collaboi-atioa of several sciestists o®
the faculty of the Btoiversity of Ptierto Bico Medical School «
lioiakeys of knomu isating dates ner® delivered by Caeeareais
sectiois ffiear full ternio Fetuses vm^m assphyifiated fey
r@a»viag the uteria® coaiteists intact and waitiaig until
iBtra-aamiotic respiritory efforts ceased or were about to
cease before freeing th® infant fro® the fetal aeaibraaesc
Others w©r@ delivered at once to serve a® controls o Asphyx-
iation ti«®s uere varied; soae infants were able to breathe
spontaneously whil® otSiers had to b® resuscitated by
isiflatiffig their lumgs with oxygeso Asphyxiated and control
infants were reared in the laboratory and required the B»sm
constant nursing care as healthy and sick newborn huaan
infants « Motion pictt*rss t9@re taken during the experiments
and at intervals thereafter? neurological exaainations were
perforaed regularly, and a great variety of physiological
data vas recorded for later study ^ review and cossparison.
Infants which seeded u&l ike ly to survive^ as well as some
healthy infants, were killed by perfusion-fixation for
histological studies o
Infants asphyxiated for 11 o 5 ssinutes or less and
which breathed spoatan®o«slf seldom showed netirological
deficits for aor© than a day or twoo A few had deficit®
in sucking, righting and SKJtor dexterity for longer periods
—one of them for 10 days. Infants asphyxiated 7 to 17
minutes and ret^uiring resuscitation^ all had sucking
deficits and abnormalities of voluntary ^>tlon for as long
&m 3 weeks o Other defects observed w«r© retinal hei»>rrhagss *
absorsal postural reactions, failure to localize sounds,
hypotonic or hypertonic .aMsculature, a ill-nerve palsy j.
a 3"per-second treaor^ status epilepticus^ papilledema^ and
loss of t®aperatis?® coatrol« At pr®s®nt there are 3
lafasts (3 weeks to 3 stosths of ase} ^hich seeat likely
to survive with peraeaeat neurological desage. All
others aow la the aursery are overtly aora»l.
OTo C. J. Bailey has coastructed a battery of
psychological tests for surviviag asphyxiated iafaat
laonkeys aad their aoaasphysiated coatrols. Beceatly
Miss Sassoa, from Dr. Earloi^'s laboratory at the Uaiversity
of Wiscoasia^ has takea over this aspect of the project.
Sight pairs of soakey iafaats have beea started oa these
tests; these are the aalaals surviviag the asphyxiatloa
at birth aad overtly aoraal la appearaace at preseat.
So far, Bo coaslsteat differeace betweea these asphyxiated
aad aoaasphyxiated iafaats have beea revealed by the tests c
la other test situatioas the 3 iafaats retalalag obvious
asurologic&l deficits have clearly failed la learaiag test
situatioasc
The pilot study of aeurological aad psychological
deficits of asphyxia aeoaatorua ia gulaea pigs, begua by
Drs. C. J. Bailey aad Marisa Raadrez da Arellaao, is
aeariag coapletioBo This supplesaeats earlier studies la
this species by Wladle aad Becker « exteadlag it to older
ages aad exasaialag effects of lesser degrees of asphyxia.
Sigalf icaat differeaces betweea coatrols aad experlmeatal
aaiiaals la the Becker aaze at 8 to 13 w^eks of age were
eacouatered la respect to ruaaiag tiiaec With a closed
field water aaze, asphyxiated gulaea pigs 12 to 19 asoaths
old nade sigiaif Icaatly oore errors thaa their coatrols,
but there was ao coasisteat differeace la reteatioa. All
the aalmals were killed; the braias sectioaed serially and
prepared for histological study^ which is beiag carried
out at present la the Sectioa oa Developseat aad
Regeaeratioac
£^s. CoMlss aad McCroskey have begua exper iiseats la
the cerebellum, isiplaating electrodes la nuclei aad
pathways to record, acutely aad chroaically, electrical
activity dtsriag iaduced cerebellar seizures. Slace
defects of posture, lategratioa aad co-ordlaatioa ar@
prevaleat la iafaat aoakeys after asphyxia aeoaatoruie.o
it is laportaat to explore the role of the cerebelluao
other Activities,
Laboratory of Neuroa&atomical Sciences
The senior scieiitists of the laboratory of
Heuroasatoaical Sciences have been called upon to partici-
pate in a nuaber of activities which are not directly
related to conduction of laboratory experiaentationc
Several are serving on study sect ions ^ fellowship coa-
aittees and research advisory panels a
DTo Palay has served on the National Research
Council Specialty Board; Anatoay and Physiology Review, BRG;
and has been Secretary -Treasurer of the Washington Society
of Electron Microscopy during the past yearo
DFo Rasmissen is a seMser of the Traineeship Review
Board, NIMI©; and the Coeasitte® on Hearing and Bioacoustics^
National Acadeisy of Sciences -National Research Councils
The Chief of the Laboratory is serving on the following;
Husan Eabryology and Development Study Section^ DRG;
Foreign Fellowship Coonittee; DRG Anatoaical Sciences
Training Coaaittee^ DGMS; Executive Coaait tee, American
Association of Anatomists; Heabership Cossaittee, Aaerican
Acadeay of Neurology; Coaaittee on International Collab-
oration ^ American Acadeay of Neurology; Research Advisory
Panel, National Multiple Sclerosis Society; Research Advisory
Board; United Cerebral Palsy; and Coamittee on PriaateS;
National Acadeny of Sciences -National Research Council c
Sditorial tasks have engaged soae of the investigators'
ti^ during the yearo Two monographs, "Neurological and
Psychological Deficits of Asphyxia Neonatorua" and "The
Process of Aging in the Nervous System", resulted fro®
NXNDB-supported conferences and are being published in
1968 by Charles C Thoaaso Dro Palay edited "Frontiers of
Cytolo^", published by Yale University Press o A translation
of Eaaon y Cajal 's little-known book on neurogenesis, by
Dro Outh, is in press o Organization of a conference on
"Neural Mechanisas of Auditory and Vestibular Function" has
been coapleted by Bro Rasausseno A new scientific Journal ^
"£zperiaental Neurology", has been launched by Acadeaic
Press, Inco, under editorship of the Chief of this laboratory,
Dr„ Palay also serving on its editorial board.
Williaa Fo Windle
NATIONAL INSTITUTE OF NEUROLOGICAL DISEASES AW BLINDNESS
Basic Research Frosr&n
Laboratory of NeMroamatossdcal Sciences
Puerto Eico Project
Sstiiaated Obligations for FY 1959
Totals $646,900
Direct : 531 , 500
Eeiffifetirseaeat ; 115^^1©©
Individwal Projects DR 1 througls 22
NC 1 through 7
EP 1 through 3
FN 1 through 6
2. Ssctlos o® Dev©!:-
3. Bethesda, Maryland
'i- Saffie_R@ HIHDB~M~"
PHS-NIH
Iradividuall Project Report
Calendar Year 1958
JBi^'t.A.
Project Title; Developmcat of Iiatriaaic Structwres of tfec*
Human l^an.
Principal Investigators: Wo F. Wiadle and L. Guth
Other Invest igatofs ; Moa@
Cooperating Units j ISone
Man Years Icalenday year 19 '8):
Total; 0.7
ProfessioBial: 0,6
Other ; 0 > 1
Project DescriptioK :
Objectives; To stuiy ijsaesis apd ©ubseqweat embrfologitp.al
developmei&t of ae^v© fib^r t^&cts and aerve cells In tiie iiuic-ii.'si.
ceiatral a@r¥ous mjmteur.
Methods employed;^ Stud^ ^itls tfae llgtst slcroscope of
serial section© of tesam esjfeieyos stained hf a®urological silTe--'
methods to bring out 'h© fiiso details. of aeuroa growth o A coll-
ection of hui^as ^mhrfis whlc'2 hag been assembled gradual l;f cv^a-
th© last 20 fear® coTStitutea, the saaterial for th® present
m.iow findings g bargelf as a mp&v® time and out-of~fe<mr#.
endeavor^ DTc Guth fee.' traneiated the books Etudes sur la M©uro-
geadse de Quelques Vi'tibres" by So Raa^n f Cajale This rel-
atively laaccesibie :ao!sogra;i>!x is of fumdaisi^ffital i^portaace to
©11 studies is aeu./jembs'yolog^c The maffiuscript of this trass-
latiom fea® beea ac--€ipt@d for publicatioa.
Little progf^BB fmB b3©B asad© isitfe the siicroscopical
study of the ham a ensbryo seri©®^ because of priority of otk&r
projects 0 The U'-^in ste® and spiaal cord of oae specimen hav?--
bees istudied ra\i©r thorot'ighly. Literature has been re¥i©ifed
pertijsest artic"3® traaslsitedo
Serial
A Project Des^criptioa (cont'd))
Significance ', Knowledge of intriasic developaeat of the^
braxn is scanty- ibs^re has beets no ©fsieffia.tic hiisiaii study..
Most descrip't:ioy»s. of braia developmeat have tje©ii based on
Slater ia.l not s« aimed specifically for aeural styuctui^es .
Uatil we have detailed knowledge of Imtriisslc ©structural
development of ih© huxeiiaii brain,, vi® "will act liave as adequate
basis for uaderstaadirag the Ho-raial proces<5s of agiag^ adveat of
pathological coffldliloBS and sigslficaace of fxmctioisal
aberratloBs
Proposed course of the project? To continue the project
as time permits..
irt B included? Yes
. 3 ".
Serial Ho. NINDB-NA~DR~1
PH3~NXH
Individual Project Report
Calendar Year 1958
Part B? Honors s Awards j, and Pubiicatioas
Publications other thais abstracts from tiais project :
Gutb; hot Tra:::jslation: "Studies oa Vertebrate Neurogeaesis'
("Etudes sur la Neurogeu^se de Quelques Vert^br^s")
Pf So Haiadn y Cajal; Springfield, Illinois^, Charles
C Thomas Cin press) .
Honors and Awards relating to this project:
„ 4 -
Serial HOo KlKDB-.!fA~DR-2
1 « Neuroanetomical ScleRoas
2. Section on Developiae»t
and Regeneration
3 . Betbesda , Maryland
4 c Ne«
PHS^NIH
Individual Project Report
Calendar Year 1958
Part A.
Project Title; Histogenesis of norjaal and dystrophic
retinaa in micec
Principal Investigator; Eicliard Lo Sidn^in
Other Invest igatorss Ned Feder
Cooperating Units; None
Man Years (calendar year 1958) :
Total: 1.0
Professional: 0<,7
Other: 0o3
Project Description:
Objectives; To find the primary defect leading to retinal
dystrophy in mice, in the hope of clarifying the coiaparaM©
human disease^ retinitis pigmentosa ,
Methods employed; A comparison was made of the histocheia-
ical properties of the developing retina in the normal ' C" and
"C X C3H'' strains of mice and the pure "C3H" strain ^hich has
a retinal dystrophy o Carefully inbred colonies of these mice
with timed pregnancies were maintainedo Eyes were fised by our
modified freeze-substitutioa method^ embedded in polyester wax and
sectioned serially at 5 or 3|i,o Specimens were taken daily from
birth to 24 days of age in normal and dystrophic groups. A
method was developed for maintaining the whole iiK>use eye in
vitro in organ culture o
Itejor findings: Several new observations were mad© on
the normal development of the retina. The interstitial sub-
stance was found to be present at birth and increased in stain-
ing intensity during the first week after birth o The outer
segments of rods and cones appeared as slender fibrils on the
eighth day of life and subsequently thickened and elongated.
Px-o.jmct 0®scriptiois Cco®t d)
tlmxT iwi®ptlon tfeef were PAS pos»itive asd staiued for
rod®. T^® rod-bipol&f efaapsB® «a® d#aKsastra'ol® oia th*® ©iglstb
day, a©d fi« ffismfeer increasied o¥®r tls@ saintli to ®i®v@fflth dafs.
Dev®lop»^ijtal ®v®mt^ «@r© tlie sane is t^@ di'stx'opliic Mc®«
After tt'« Biffitfe or testh d&f thm rods ^md coii®8 fail@d to
d®v®ior and^ iffld@©dif fapidly dsgeaeratad^ so t&at fey tli©
fift@oatb day Mrdlf a trae® remaised of tSi« ©istir© pliotor©ce-p=
tor l.aj@r^ ®ow©'^®r^ ao hi«toc&@iiical distinctioas 'smtm^mm
norm&l ftisd df®tropl&ic retina® ^@r® foasd prior to tfe® ?k±mth day,
fl!t« orgsn oiiltur® ®xp®»>ia«ats lik@wiss® failed to d@fia@
t^« disease j Isut did elistiaate sons iurtheT possible causes,
'■jpfee retina diffes-emtiated 1@ yltro asd tormssd its 3 lay@rs of
c@II@o lod and con® Giit^r segaeMai did @ot diff@r<@iitiat€j,
«7®^ tlkougis tl&® retisia survived for manf dafs «ft@r r@acbiiisg
the stage wher@ th@&m outer »@ga@Kt@ sl&ould iiav@ foimsd. TIae
ffior»&l and dfstropi&ic r^tiisaa li@liav@d aliks—tls®^ di£f@r@^tiat®d
to tl£@ sane extent aisd slio^#d so siga^ of dsgesssration of pliotG-
receptor colls « The additioB of excels vitaeia A to the culture
Bedit» did not cause fornsatioig of outer sep&eiatSc Tiaus tlie
disease probably does not arise tecataae of a systesiic isossious
iffifliaessc© appearii^ o© the isiM-li day of lifej oi? of a«a intriiasic
d@f@ct which 3ia^if@st@ itself o^ that d%f . lather it &pp@»-r»
that the retina mtst reach a give^ stage of differeBtiatioE
before the degeiaeratioa sets ±n, but thm it^ture of the astlasa^lus
to degeneration res^ias unknown..
Signif leasee ; The ®igsificaace arises fro» the asialogr
with the hmmfn disease^ retinitis pigaeatosa , a disease of
siMlar genetic origin aisd histopathologf » Thiet hmmu di^e^^e
is cowaoulf cos^idered nm abiatroph^^ a degenerative disease
of wBkMno'^n cause in a Biature c®ll» The nouse disease is clearif
aot an abiatrop'hf; it 1® a developaei^tal disease,, i^volviag
degeseratios of iss^tiar® celiac In m&m th@ photoreceptor cells «
especially the ro4mf m^f likewise fail to i^ture In the ^jm-
trophic subjects &md indeed, there is moms electroretinogrmphio.
data to support this idea.
Proposed course of project; The work described is bei^g
prepared for publication atm.d the project will be termisiatedo
Part B isscludeds Sfo
Serial No« NINDB-NA- DR-3
1 Neuroanatomical Sciences
2c Section on Development
and Regeneration
3. Bethesdaj, Maryland
4,
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Project Title; Histogenesis in the embryonic mammalian
nervous system.
Principal Investigator: Richard L« Sidman
Other Investigators: Irene Miale
Cooperating Units: None
Man Years (calendar year 1958) :
Total: lol
Professional: 0=7
Other I 0 „ 4
Project Description:
Objectives ; To analyze the behavior of cells in the
immature mammalian nervous system in ord r to clarify mech-
anisms of normal and pathological development.
Methods employed; (1) The intact 8 or 9 day mouse embryo
was cultured in vitro, to separate the developing embryo from
its normal environment.
(2> Autoradiography using tritium labelled thymidine
(thymidine-H ) was employed. This labelled nucleotide is incor-
porated into deoxyriboaucleic acid CDNA} of ceils about to enter
cell division and thereafter remains as a permanent marker in
those cells. Labelled embryonic tissues were fixed in Benin's
fluids embedded in wax. sectioned^ and prepared for autoradio-
graphic study by Lebland's dipped liquid emulsion technique..
Major findings; (1) A method was developed which allows
culture of whole mouse embryos. Embryos explanted at stages
during closure of the neural tube and formation of somites
differentiated at a slower rate j^ vitro than in vivo, but
formed a complete neuraxis and inost somites. Limb buds did
aot form. The embryos survived up to a ".seek in vitro, with
maintenance of body form. The heart initiated its contractions
in vitro and continued to beat for up to 3 weeks 3, even though
- 7 ~
Page 2 Serial Eo. KIKPB-HA- DR--3
Part A. Project Description (cont'd)
other organs had becosae iysed. Mitotic activity ia the brain
remained vigorous for 4 to 7 days in vitro , but little dif-
ferentiation occurred c
(2) By autoradiography with thymidine-Br the basic pattern
of ceil proliferation in the iujanature neural tube, composed of
a pseudo-stratif ied colusmar epithelium, could be studied.
Nuclei lying at a distance from the ventricular surface syn-
thesized new DMA and then migrated to the ventricular surface
to divide.. Subsequently these cells again migrated laterally
and contributed to the thickening of the neural tube which
accompanied differentiation of neural cells. Almost the entire
population of cells in the young neural tube were either
migrating to the ventricular surface in preparation for cell
division or migrating away froia it ia preparation for differentia-
tion. As the various regions of the brain assumed their spec-
ialized character, a number of subsidiary patterns of cell
division and migration appeared.
Significance ; (a) One major deterrent to the study of
mammalian embryology is the inaccessibility of the embryo. The
development of a method for culturing whole embryos allows
experimentation on nutrition and oxygen requirements^ and
allows experimental surgical intervention. Such studies have
proved most beneficial with amphibian and chick embryos, and
should be extended to the mammal. On the other hand^ these
methods for culturing the ssouse embryo are less refined than
methods available for lower vertebrates, and the curled
shape of the mammalian embryo is less favorable for experimental
manipulation,
(b) Autoradiography with thymidine-H^ is a powerful new
tool not heretofore used for the study of embryological processes,
It allows labelling of cells at a well-defined period of their
life cycle, and allows these cells to be followed through their
subsequent migrations and differentiation. It should allow a
detailed analysis of when and how the various parts of the brain
form in embryonic life. This is of intrinsic value, and also
will serve as a basis for analysis of developmental defects of
the nervous system. A fair analogy can be drawn with congenital
heart disease, which has been .clarified so well by relating the
detailed embryonic development of the heart to the time during
pregnancy when the laother was ill or injured.
Proposed course of project; The results described above
are being prepared for publication. Analysis of regional
development in the brain is in progress and will continue for
at least the first 8 months of 1959,
Part B included J No
Serial Mo, NINDB-NA-DR-4
l.~ NeuroanatOGiical Sciences
2o Section on Development
and Regeneration
3o Betkesda^ Maryland
4o Same as NIHDB-MA-2-1957
FHS-NIH
Individual Project Report
Calendar Year 1958
Part A,
Project Title: Regeneration in tfee central nervous systeaio
Principal Investigator: W. F. Windle
Other Investigators: James B= Campbell
Cooperating Units: Department of Neurosurgery
College of Physicians and Surgeons
Columbia University
Man Years (calendar year 1958) :
Total: 0,8
Professional: 0c5
Other : 0,3
Project Description:
Objectives: To study factors regulating regeneration in
the central nervous system o To learn tfee role played by the
neuroglia cells in degeneration and regenerationo To attempt
to alter the normal response of the central nervous system to
injiiry in such a way that a milieu favorable to functional
regeneration will develops To learn why regeneration occurs
so readily in the central nervous system of the lower verte-
brates and is so difficult to achieve in mammals and man^
Methods employed; The spinal cord of cynomolgous monkeys
was transected in the midthoracic regioa^ leaving gaps of
2 to 6 mm. The cord ends and gap were enclosed in a wrapping
of Millipore filter; in other monkeys the Millipore was omitted
for control. Pudendal nerves were severed to facilitate
bladder and bowel emptyings Neurological examinations were
made at weekly intervals and recorded cinematographicallyc
Monkeys were killed after 3 weeks to 4 months and histological
preparations of the cord lesions studied and compared o
Major findings; Without Millipore^, the cord stumps soon
became capped by pial connective tissue and a randomly
oriented scar formed in the gap^ as was reported previously
Page 2 Serial No. Nim®-HA-DR~4
Part A Project Descriptiou (coat'd)
in cats and monkeys = In the Millipore-wrapped lesions, tissues
were oriented longitudinally and the gaps bridged by spindle-
shaped cells and blood vessels, A few intraspinal neurons from
the rostral stump regenerated into this oriented tissue and
crossed the gap, but could not be followed down the caudal stump c
By 3 to 4 months the regenerated nerve fibers had formed small
fascicles, resembling peripheral nerve rootlets by virtue of
Schwannlike cells along their fibers o it seemed that regenerat-
ing central neurons ;, reaching the gap, had acquired character-
istics of peripheral neurons c If they descended into the
caudal stump, a point as yet not proved j they lost this
characteristic below the lesion.
Ho functional restitution was observed by 4 months
Significance: Importance of studying the phenoment>;i of
regeneration in the central nervous system Is self evident
Results of this study may be applicable to almost any traumatic
injury of the brain or spinal cord and may throw light on any
one of a number of degenerative diseases
Proposed cotirse of project : To continue this collaborative
study, extending observations to longer tiroes, and younger
monkeys Reports emanating from USSR Indicate that very young
mammals show remarkable powers of regeneration and restitution
of structures surgically severed or rea^ved from the brain
Part 8 included: ¥es
- 10 -
Serial Eo. HIHDB-HA-DR-4
PHS~HIH
Individual Project Report
Calendar Year 1958
Part B» Honors J, Awards, and Publications
Publications otfeer than abstracts froE this project:
l/indle;, Wc Fo : Kegeneration in Relation to the Process
of Aging in the Nervous Systeiao Gfeapo 4 in ^T^
Process of Aging in tis© Nervous System^, " James
Sir r en, Menry ImiS~and Williaia W indie. Editors „
Springfield,, Illinois ^ Charles C Thomas, 1958.
Wiadlei, Wo F,,, Jo Oo Smart and Jan© Jo Beers: Residual
Function after Siibtotal Spinal Cord Transection
in Adult Cats„ Neurology, 8:518-521, 1958 o
Honors and Awards relating to this project: None
11 "
Serial NOo MIMDB-NA-DE-5
i. NeuroaaatoBiical Scieaces
2„ Section on Development aad
Kegeaeratioa
3« Bethesda, Maryland
4o Same as HIHDB~HA-5~1957
PHS-NIH
IndividMal Project Report
Caleiidar Year 1958
Part A,
Project Title: Functional and Structural Changes la
ReserpiKized Aniiaals
Principal Investigator: William Fo Windle
Other Investigators: Jaa Cassaermeyer
Cooperating Units: None
Man Years (calendar year 1958) :
Total: 0„7
Professional: 0«4
Other: 0.3
Project Description:
Objectives: To determine possible aaatomical correlates
of the functional changes reseabling paralysis agitans in
anima.ls under chronic administration of reserpineo
Methods eisployed: Monkeys and cats were given reserpine
CO = 2-0 „ 6 ffi^/kg) daily for varying periods of timeo After
establishing altered functional states, they were killed,
per f used-fixed, and the nervous systess studied by histological
©ethods o
Major findings: Monkeys killed after 2 years of chronic
administration of "reserpine, and cats, after a few days or
weeks, showed no gross abnormalities of the brain o Micro-
scopically there were no heiaorrhages , infarcts, softening y
demyelination, neuroglial reactions nor phagocytosis o However,
conspicuous cytoiogical changes were found in the cerebral
cortex, basal ganglia and brain steiSo These involved cell
nuclei and nucleoli, which appeared pale and enlarged and
often showed a "hole" in the karyoplasm due to reraoval of some
substance during the process of histological preparation o
These were interpreted as aonpathological , probably reversible
changes, because there was no indication of cell losSo
^ 12 ~
Page 2 Serial HOc 10'MD3-MA-M~5
Part A Project Descriptism (cont'd)
The aeuronal cliang@s were distributed lis a pattern eiaite
ualik© that of pathological chaages in hmsB.n paralysis agitaiss,
Significanc©: Present studies provid© the first indiea--
tion of possible neuronal changes resulting from prolonged
administration of low doses of reserpiae^ It should give
grounds for caution in respect to prolonged use of this
"tranquilizer" drugo
Proposed course of project: The project is dorcmant at
present b©ca.use of pressure of other projects, lack of space
and personnel o
Part B included: Yes
- 13 -
Serial Ho, HMDB-NA-dE-S
PES-NIH
Individual Project Report
Caleadar Year 1958
Part B°o Honors, Awards^ and Publications
Pubiicatioas other than abstracts from this project;
Feringa, Eo Ro and W^ Fo Wiixdie: ladiictioa of Hypokiaesia,
Rigidity and Tremor ia Primates with Ess©rpia©<
Proco 1st Internat o Conge Meur .. Sc= ; Londoa^.
Pergai3K»a Press, 1958 {In press),,
Wiadle, Wc Fo and Jan Cardaermeyer : Functioaal aad
Structural Observations on Ciiroaically
Reserpinig;ed MoakeySo Seieaces 127; 1503-
1504, 1958. _—
Honors and Awards relating to this project: Eon®
Serial ?fo. IfIHDB~NA-DR~6
1. Heiiroanatoiaicai Sciences
2» Section OB Development
and RegensratioB
3o Bethesda^ Maryland
4, New
PHS-MIH
Individual Project Eeport
Calendar Year 1953
Part Ao
Project Titles Meuronal specificity in the autonomic nervous
system.
Principal Investigator: Lloyd Qath
Other Investigators! Clark Jo Bailey
Cooperating Units : None
Man Years (cElendar year 1958) :
Total: 0„8
Professional : 0 o 6
Other; 0.2
Project Description:
Objectives; To determine whether autonomic nerve fiber®
can laaiatain the function of autonomic effector organs other
than thos© which they normally innervate »
Methods employed; 1. Preganglionic vagosympathetic nerve
anastomosis o 2« Transection of preganglionic •white rami Tlj,
T2^ and T3o This procedure interrupts all pupil preganglionics
to the superior cervical sympathetic ganglion and leaves intact
only those fibers which subserve other functions (e.g. vaso-
motion^ piloerection and nictitating sseiabrane retraction) «
Pupil size as a function of light intensity will be measured to
determine Mietfeer collaterals frosa T4 to T7 ^ill restore pupil-
lary fuactioffio
Major findings: Hone as yeto
Sigcnif icaace: This experiiaent joay shed light on the
mechanism by which functional recovery so often occurs after
sympathectoBsyo It may also clarify the still-unsettled question
of the role of the sympathetic nervous system in pupillary
-. 15 -
Page 2 Serial No. HIHDB-HA-DR-.6
Part Ao Project Description (cont'd)
fuBctioa. Finally it is also designed to determine whether
alteration of specificity of autoaomic neurons can occur =
Proposed course og project ; The project is beiisg actively
pursued at the xaoiaeQt and 'Bill constitute the major poytioa of
the coming year's worko
Part B iacludeds No
-= 16 ~
Serial No, MIIBB-HA-PE-.?
1, Meuroanatosaical Sciences
2o Sectioa on Developsaeat
asid Regeneratioa
3o Bethesda, Maryland,
4o Ne^
PHS-NIH
Individual Project Report
Calendar Year 195S
Part Ao
Project Titles Heterogeaeous Beinaervatioa of the DiapbragUo
Priacipal Investigator; Lloyd GutSs
Other Investigators s Karl Fraak
Lamar Soutter^ Boston University
School of Medicine
James Bo Campbell^ Columfoia Uiaiversitf
College of Physicians aad Surgeons
Cooperating Units; Laboratory of Meiarophysiology
Man Years (caleadar year 1958) :
Totals 0o9
Professional : 0 « 7
Other : 0=2
Project Descriptioa:
Objectives; To detersaiae Tshetlier diaplaragmatic fraction
caa be laaiataiaed by aerves other thaa the phrenic «
Methods QMployed: The proximal vagus aad distal phrenic
segmeats were anastosKJsed ia th© rat aad the proximal rectirreat
laryageal and distal phreaic has beea aaastomosed ia both rats
aad ia moakeyso Arterial sleeves or Millipor® tubes ^ere used
for the aaastojaoses. Coaveatioaal electrophysiological record-
iag techaiques have beea employed c
Major fiadiags; 1= Diaphragmatic fuactioa is restored ia
the rat t?ithia & sjonths after vagophreaic aaastomosiSo 2o Th©
normal vagus aerve transmits efferent respiratory bursts syn-
chroBous ^ith those of the phrenic aerve. The efferent bursts
are carried by the recurrent laryngeal fibers within th© vag-us
nerve »
Page A Sex'ial MOo NINDB-M-DR-?
Part A. Project Descriptioa (coHt'd)
Si^'ttif icance ; If the iiusian recurrent laryngeal nevi^e also
transmits efferent respiratory volleys j, there is a possibility
that t&is aerve can substitute for the phrenic nerve « SBch aa
operation might eiaable patients ■with bxxibar poliomyelitis to
survive without the assistance of an artificial respirator o
Proposed course of project: Anastoasosis of recurreat
laryngeal and phrenic nerves has been performed in the sioak©y a?ad
the rato These animals will be examined for evidence of dia-
phragsiatic function as soon as sufficient time has elapsed for-
th© aerve regeneration to be completed. If diaphragmatic tunC"
tion is restored in these animals the operation ^ill be pea-foriaed
on
Part B included? Yes
- 18 -
Serial Ko, mmB~m~m=i
PHS-NIH
Individual Project Report
Calendar Year 1958
Part B; Honors, Awards ^ and Publications
Publications other than abstracts from this project ;
Guth, Lc and K. Frank: Eestoration of diaphragmatic
function following vagophrenic anastomosis in the
rato Expo Neuro 1959 (in press).
Honors and Awards relating to tSiis project:
19
io Ke'aJL'caiiatcJUical Sc
2p Section on Develop^
and Eegeaeratioa
So BethesdEj Iferyland
4 c NeT^
PHS-HIH
[adividual Project Eeport
Calendar Yeuv 1958
Project Titles Ixperimeatal analysis of the nerve fibe:;
tast© hud rslatioasSaio,
'sastigator: Lloj'd QwU
Other Isivestigators; Mone
Cooperatiffig Uaitss Mone
Ian Years (caleudar year 1958) :
Total; 0c5
^gisioisal : 0 = 3
Project BescriptioH:
Objectives: To study the so-called "troplsic iafluence'^
of nerve fibers on, taste bu.ds...
^tliods ©mpIoF®<^° TraasectioB of tfe® gustatory serves asid
reiBaervatiosi of the circuMvallate papilla ^Ith varicas .serves =
X- irradiation of tlie etrcmBvallate papillae
_^.lQg f isdJBgs ; i„ Denervation of tli© circumvallat© papilla
results ia loss of taste buds &nd thiHaing of t^e mucosal epitfe©-
litm.. 2, Eeianervatioffi of the papilla ^itla glossophairyageal or
v&^ns aer^/es results isa redifferentiatioa of taste buds vshereas
reiiiaervation -with the hypoglossal aerv© fails to initiate taste
tad format ion e 3o Mitotic counts on colchicinized^ deaer*?a ted
llBgiaal epitfesliii^ h&m failed to reveal aaj effect of dener^stiorc
on mitotic rateo B.o-^ev@T the variability o£ the metliod requires
that a large volum© of saaterial be studied » 4o Study of a sajall
suiafoer of x-irradiated toagues failed to reveal evidence of
alteratioa ia tasstes teds^
SiKnif icagjc© ; This is on© of the fe^ areas in isaich t!ie
problem of differentiation and incMction may toe stiidied In ar^
adult BiaBmal rather tli,an ia astpfeibia or emhryoSc.
- 20 ^
Pag© 2 Serial Ko, MIHPB-MA-DE-S
Part Ao Project DescriptioB (cont'd)
Proposed coMgse of project; Further viovlt on tli® project
is postponed imtil additioaal tecJaaical assistance becomes
availa&le to assist ia the preparation and study of material =
Part B iBcluded? Yes
» 21 -
Serial NOo NK^B-WA-^-DE-S
PHS-NIH
Individual i?roJect Report
Calendar Year 1958
Part Bi Hoffiors, Awards ^ and Publications
Publications otMer thmn abstracts from this project :
Guth^ L. : Th© effects of glossopliaryjageal nerve traasection
on the circusavallate papilla of the rato Anato
Eecc 128;715-732„ 1957 (but sppeariag in springy 1958)
Giithj, L. : Taste buds on the cat's circumvallate papilla
after reianervation by glossopharyngeal , vagus ^ and
hypoglossal nerves = Anato Reco 130; 25-37 „ 1958,
Honors and Awards relating to this project ;
- 22 -
Serial NOo MIMBB-HA-DIl-9
Ic. Meuroanatomical Sciences
2, Section on Development
and Regeneration
3o Betfe©sda, Maryland
4o Mew
PHS-NIH
Individual Project Report
Calendar Year 1958
Part Ac
Project Title; Nervous Systeia Pathology in Macaca Mulatta
after Asphyxia Neonatorvua
Principal Investigators: J. B= Raack and Wo F. Windle
Other Investigators: J„ CasMierEieyer
Cooperative Units: University of Puerto Rico School of
Medicine and U. So Public Health
Service Clinic, San Juan^ Puerto Eicoc
Han Years (calendar year 1958) :
Total: 1,2
Professional t o .. 9
Other: 0^.3
Project Description:
Objectives; To determine the pattern of neuropathology
in the monkey brain after asphyxiation at birth.
Methods employed: Fetuses of known gestational age were
obtained from ssionkeys bred under controlled conditions by
cesarean section close to teriffio Half the fetuses were
asphyxiated by removal of the uterine contents without opening
the chorionic saco Other fetuses were delivered as controls.
At varying times after delivery the infants were killed by
the perfus ion-fixation technique «
Serial sections of the experimental brains and brain stems
and other sections of representative levels of the spinal cord
were prepared. Every tenth slide was stained by the buffered
thionin Hissl stain and alternate tenths by the Woelcke myelin
staino Selected sections were stained by Holtzer^ Ptah and
Bodian methods and by other techniques for iron and fato
Experimental aniiaals were compared with control aniEials
of comparable age for gross and laicroscopic pathology » Cinesa-
tographic and other records of experiments were used for
- 23 -
Page 2 Serial NOo HIMDB~MA~BR~9
Part A Project Description (coat "d)
reviewing functional observations o Two were selected for
initial study c
Major findings: One ©xperiaieatal assijaal, which sho'K?ed
extensive neurological deficits during life was killed after
nin@ days and paired witls a control of similar ageo Tfe®
brain daiaag© was almost perfectly symmetrical in the experi-
mental aaiHal and was principally in tli© grey matter ^ Tliere
were changes In 'White matter and destruction of myelin
Cmyelinatioii is quite incoiaplete) ^ but these were probably
secondary to neuronal damage o There was a striking localisa-
tion by cytoarchitecturally defined nuclei ^ with daijage
usually conforming closely to the anatomical boundaries, sosie
nuclei feeing spared even though surrounded by dasaageo
The cerebral isocortes shoi?@d diffuse subtle changes of
th@ neurons with less cossplete^ staining than in the control,
yet with no clearly abnoraial neurons nor neuroglial changes o
In a few folia of the vermis of the cerebellum there was
loss of Purkinje cells and a slight neuroglial reaction=
Other ar©as of the brain, brain stem^ and spinal cord
had 2u to 100 per cent of the neurons damaged, usually to the
stage of ghost cells « There was a reaction of early
saacrophages and an astrocytic hyperplasia in most daimged
areas o Mo heiEorrhages , nor thrombosis were seeno There was
an intense "neuronophagia" in th© thalamus „
The inferior colliculus showed the most severe dagiage.
Other areas of ©xtensiv© damage were most of the thalamic
nuclei, the subthalamic nucleus^ interstitial nucleus of
Cajal, globus pallidus, the whole reticular foriaations th©
superior colliculus, oculomotor j, trochlear f, and abducens
nuclei y Kost of th© trigeminal nuclei, superior and ©edi&I
vestibular nuclei^, cochlear nuclei^, superior olive, nucleus
gracilis and cuneatus, ssost of the grey scatter of the cord
(except in thoracic segnsents) and th© roof nuclei of the
cerebeiluHSn
AiSiong the undamaged regions were the amygdala^ olfactory
nuclei^, ssost of the hypothalaisuSs the lateral geniculate^,
pontine nuclei, inferior olive, most of the nuclei on th©
floor of the fourth ventricle j, and th© stratu® aonal© and
substantia gelatinosa of the spinal nucleus of the trigeminal
nerve and the dorsal horn of the spinal cord.
Significance; This is th© first experimental aeuropa-tho-
logical study' in stu infrahuamn neonatal priaaateo Histopatfeology
Page 3
IMDB-MA.
ript
of the iafaat monkey braia is significantly different from
that of adult laaa after asphysiatioa aad is different from
that reported ia hmsaa infaatSj wiier© hesiorrliages , cortical
atrophy^ feydroceplaalus &nd vascular infarcts are prosiiaeEto
Lcti-re ia th© raoalsey after asphyxia aeosatortua
,t reported iis fexauan iiafauts with "keraiGter'as",
Proposed course of project: To coatiaii© as qwickly as
time aad preparation of ffiaterial permit =
» 25 -.
Serial Mo. MIMpBnM:^^:^
lo HeuroaaatoMical Scieaces
2o Section on De^elopstsmt
and E®g©B@ratlosi
3, Bethesds, Maryland
4o Sam® as MI»B»MA""10".195f
FHS-MIH
Ixadiiridisal Project jReport
Cal©adar Year 1958
Part A<
Project Title I The sigaificaac® of the acridia® oraag©
staiaiag of nenronm In vitro and In jMo.
Principal Imrestigator ; 1. M„ Wolf
Other Iffivestigatorss ^one
Coop®ratiiBg Units; Noise
mn Years <cal<gadar year 1058} :
Totals 0„8
£>ro£es@io]!ial ; 0 » 6
Other; ©o2
Project Description;
Object ive@; a) To establieli the fluorescest image ^hlGh
ter©athlisg aeiaroa© will display ^h®n @taiss®d ^-itls acridia®
orange (AO) s to compare it with the iaiag© displayed foy f iK©d
aeuroQs or Ib^ injured^ teased o5.i®s stained supravitally: asjd
to establish the histocheaical significance of thes© iiaages.
h) To mtudy the interact ions of AO aad the sufestrates to
which it biisds by spectroscopic methods.
,Bfethod@ .. ®ps>lo.Y®d ; a) Cultures of fibroblasits aad pig-
meat epithelial cells mer© grown In Paul perfusion cMsabers.
Cultures of chick ©Kbrjo spinal ganglia have been gro^n in
plasma clots by the Maxi^ji? doisbl© coverslip method = These
cult-ores.,, and supravitaliy traced leaterials, have been stained
«itfe AO aad eaeaaiised hy fluoresceace microscopy.
b) Solutions of AO in coiabisiatioB ^ith various poly-
aucleotides have beea studied by absorptiou spectroscopy
aad the spectra have been ^athesaticallf axialfised.
» 26 -
Pag© 2 Serial Mo, MI^SrMMm^l^
Part A, ProJ®ct Description '(cont'd)
Ma^ior fiadiags; a) Cell sonolayer cultur©s groisu,
cos^tiauously in th® pTesence of one part per milliois of
AOs, stained adequately for fluor®sc©ac© asicroscopyo Lining
cells stained orthoctoomatically Cgre©n tluoremcence') c
Reveg'siblg' isajusmd c©lls acquired metacbrosjatic ((r©d
fluorescent) granules in the cytoplaamo Witia fus^ther
injury, th© iiucleolus and tfe© ©atire cytoplasai became
metacbroKatiCo This degree of iajury was irreversible 5,
aisd tbe cells defensrated. Spinal ganglion cultures were
harder to staiia and to observe, possibly because o£ the
dense growth of satellite cells necessary for their healths
However, the aaai® sequeace of staining eveats was observed
iu the cultured ueuroiio Th© metachromaay of the irreyersitolf
injured cells ^as like that sees ±n fisted cells aad was
probably due to EHAo Th© aetachrossatic granules Been in
revereibly injured cells probably v?®re not RMA; la the sjeiwoa
their shape and distribution ^er® not tfeos© of th® Nissl
substance c la supravitally staiijed, teased retinal rods^
jBetachroBsasy was confined to th© ellipsoid sogaeat^ which
contained all the mitochondria but little if any SNA,
b) Spectroscopic studies showed that laaay polyelecti'olvtr-"
including natural and synthetic polynucleotides, can bind
AO in such a manner as to cause bound AQ i^lecules to
associate 3 or "stack" 0 Stacked AO had metachroaatic fluo-
rescence« When the dye-binding sit®s of the polyelectrolyte
were in great excess of the AO sjolecules, they distributed
theiaselves a^ong the available sites amd u^^atackfido Th@
amount of stacking was a function of th© bindiEg ©it©/AO
ratios and of the intrinsic tendency of th® polyelectrolyte to
produce stacking of bound AO. This tendency can be espK'®ss©d
as a stacking coefficient » which varied according to the
chemical structure of the polyel®otrolyt®o Calculations are
underway to test an equation which relates the stacking
coefficient and th® binding site/AO ratio to th© oaolar
extinction of bound AO at any gives ■wavelengths
Significance s AO, because of its low toxicity and
isetachromasy, is a rare tool with which to study th© chesaicsl
reactivity to living^ intact cells. Tissue cultures provide
growings aiiechanically undisturbed cells whose staining can be
observed in details Spectroscopic studies are elucidating
Page 3 S«^rial Ho, MMM:M':-
Part Ac Project Description (cont'd)
th© physico-c&eaiical basis of AO staifelag, sad ^ill Iselp
to int®r-prwt t-fe© f luoresceiace observed ia lining cells In
terijss of specific ch©iaical st^'uctures .
Proposed course of pro.lec-t t Tb,@ otesrvatioas oe
cultisres in Paiil chajabers ^@r© mad© ia eollalsoration 'isitte
Dr. Sasauel AroasoE^ Opthalsjology Branch,, Hatios^al Institute
of Sfewrological Diseases and Bliadiiess. Tfa©f ar© coKplete
&nd will to© prepared for pufelicatioss-o
TM© ofos©P¥atioas oa cultured newroas ijiill be contimied
aiffid er.t@nded.
Tij© spectroscopic studies on a quantitative t&eory of
AO MsdiBg are a collaborative ©ffort ^itfe Dr^ Daa F», Bradley?,.
Laboratorf of Physical Cliesaistrf , National lastitute of Msiitsl
Health. A prelissinary report is in press and furtfeer report-K
isill b® subsitt@d for piablicatioR am t&© calculations are
completed «
Part B iacludeds Yes
J'V'O^ *5 T5 1 'tf'^f': 'i'^ "^ '^'^V-'i-'r^' "^'^P T*'V^ ';; j^";-
Individual Frojeet Eepos,--*;.
Cs.l©ndar ifeai" 1S58
S±_Mi EoHors, Av^ards^ and FisMicatiosas
licatioas otJier than abstracts froa this project:
Bi'adley, ©. F. and M, 1, Wolf: Tfee le-oroclsejsis--:
of PoljnueleotldeB f in: Sygiposiism on tlae
Hey.roc!»es>istr5' of .W^cleoticles, : He'circlogy .
ms^plem^xvt&vf VQlmm, 1958 (in sress) /
- 29 -
Serial HOo FIMDB-NA -pR-.il
1 a Neuroanatoiffiical ScTeaces
2o Section on Development
and Regeneration
3o Betliesda, Maryland
4o Sam© as MIHDB-S*a-o
PHS-NIE
Individual Project Export
Calendar Year 1953
Part Ao
Project Title: Structure and dsesiistry of photo-
receptor c@lls
Principal Investigator: Eicfeard Lo Sidsian
Other Investigators: Ned Feder
Cooperatiag Units: Hone
Man Years (calendar year 1958) :
Total : 1,0
Professional: 0,7
Otfeer: 0^3
Project Description:
Objectives; To define the structural and chemical
properties of photoreceptor eel Is ^ in relation to retiaal
functioso
Methods employed: Histocheiaical and spectrophotoraetric
isethoHs were usedc- Most studies were carried out with mouse
eyes fixed by a new freeze-substitution method of fixation
and stained by various histoclsesBical methods o Suspensions of
frogj, cattle., and monkey retiaal rods were prepared by differ-
ential centrifugation in sucrose and their absorption spectra
were recorded with a double beam recording spectrophotometer.
Major findings : A series of new structural features ia
rodeni' retinas were described in a Symposium on Photoreception
at the Hew York Academy of Sciences o
Ca) A description was given of the "interstitial zon©"
i^ich relates the retinal pigment epithelium to the rods and.
cones o The apical parts of the pigiaent epithelial cells
contain periodic acid-Schiff (PAS) positive granules,, Th©
apical border of these cells is siicrovillous and PAS-positive =
The space between this apical border and the outer segB©B.ts of
- 30 -
Page 2 Serial MOo Mim)B-HA-i?B-ll
Part A Project Descriptioa (coat'd)
rods aad coiaes is occupied by an feomogesaous "iaterstitial
substaac®" contaia&ing acid mucopolysaccliarideo These
morphological features imply a secretory function for tfe©
pigmeat epitheliussj, with the secretion directed toward the
photoreceptor cells „
Cb) The first conviaciiig evidence was obtained for
the presence of cone cells in retinas of rats and xaic@o
Their structural and histocfeemical properties are cosparabl©
to those of cones ia other vertebrate species „ Cones ware
detected also in the guinea pig retina, in confirKmtion of
a few reports in the older literature o
Cc) Internal structure was detected with the light
raicroscope in the rod-bipolar synapse, as described with the
electron microscope by several investigators in recent years.
After fixation by freese-substitution, the synapse appeared
as a central J dense, round or oval sphere surrounded in
turn by an almost clear zone and a dense outer Margin o TIae
central sphere contained lipid and a PAS-positive substance,
while the surrounding clear zone and laarginal zone had
different histocheaical properties o
Cd) A difference between rods and cones was detected
during study of the distribution of dehydrogenase enzymes
in photoreceptor cells c DPH diaphorase, and presumably the
associated dehydrogenases^ were found in the ellipsoids of
rods and in both ellipsoids and byoids of cones c Succinic
dehydrogenase j, on the other hand, was found only in
ellipsoids in both types of cells c Studies are in progress
on a wider range of vertebrate retinas^ to determine if
this difference between rods and cones is generals
Ce) The absorption spectrum of rhodopsin shifted to
higher wavelengths when this visual pigment was studied in
intact rods than when extracted into solutiono Rhodopsin
probably exists in a different form or Molecular shape
in vivo than when extracted »
Significance; It is self-evident that each advance in
our understanding of retinal structure and chemistry will
aid our understanding of visual functiono
Proposed course of project; This project will to©
- 31 -
Page 3 Serial lOo MIMPB-MA-DR-ll
Part A Project Rescript ioa (coat'd)
terminated ©arly in Jaisuary^ 1959 c
Part B iHcludeds Yes
- 32 -
Serial Mo„ IIIBB-NA-DR-Il
PHS-HIH
Individual Project Report
Caleadar Year 1958
Part B: HosaorSy Awards .„ and Publicatioias
Publications otfe©r tlian abstracts fro® this project:
Sidamap Ro Lo^ Hietocheiaical studies on photoreceptor
cellSo Anao McYoAcadoScio, 1958 (In press) c
Honors and Awards relating to this project;
First Richard Stearns Meiaorial Lecturer^ Albert
Einstein College of Medicine, New York, May, 1958.
-33-
Serial HOo HI1DB-HA-DR--12
lo Kern'oaaatomicai Sciences
2o Section on Deveiopaent and
Eegeneration
3c Bethesda^ Maryland
4o Same as MINBB-NA-9
FHS-MIH
Ijidividual Project Keport
Caleiidap Year i95S
Part A,
Project Title: Development of new Iiistocheiaical methods „
Principal Investigators: Med Feder, Richard Lc Sidman
Other Investigators: Noa®
Cooperating Uaits: Hose
Man Years (caleEdar year 1®58) :
Total: Oo9
Professional: 0=7
Otiier: 0,2
Project Description:
Objectives: To devise new methods for tlie localisation
and cfearacterization of chemical substances in cells o
Metlaods employed: A ne-m aetlsod of fixation by freess-
substitutioa' was developed o Tissue samples were f rosea rapidly
at -175®C and then placed in a "substituting fluid" which
simultaneously dissolved ice and fixed the tissue at -70^0.
Dehydration and fi^.atioa were thus achieved with little dis-
tortion of tissue structure c The most useful substituting
fluids have been osmitia tetroxide in acetone and laercuric
chloride or picric acid in ethanol^ all at "70®Gc A variety
of nervouSj, special sensory, and other tissues were studiedo
Major findings: A nusber of new structural features ia
retina were described in another reports Skeletal isuscle was
fi^sed regularly without the contraction and distortion which
usually accosapaay fisatiOQo During study of brain meninges
infected escperisea tally with the fungus Gryptococcus neof onaaas ^
a nucleus was demonstrated in the organism for the first tlHeT"
The nucleus had an eccentric position^ a diaiaeter of less than
Ip. , and contained BHA like other nucleic The method served
for deiaoastration of nerve fibers in central and peripheral
tissues by both silver stains and supravital methylene blueo
The method also allowed preservation of all lipids in tissue j,
-34 -
Page 2 Serial Noo HIMDB--NA-BR~12
Part A Project Descriptlosi (cont'd)
as well as other aaolecular species difficult to presence, such
as acid mucopolysaccharides o
Sigaificaace: Th© method is profeably the best available
for the accurate jSfixatioB, of sssall tissues asad tfee preserva-
tiom in them o£ a wid© variety of chemical substances „ It is
already being used ia a auaber of other laboratories eagaged
±n histochemical studies „
Proposed course of project: A further analysis of the
structure aad chemical characterisation of sense organs aad
developing braia will be usadertakea when the iavestigators
establish their new laboratory at Harvard Medical Schools
Part B included: Yes
- 35 ~
Serial Mo„ HIHDB--MA-DE-12
PHS-HIH
Individual Project Eeport
Calendar Year IS5S
Part B: Honors , Awards, and Pul>lieatioas
Publications other thaa abstracts fros this project:
Feder, Ho and R„ Le Sidaaa: Methods aad principles of
fiKatioa by freese-substitutioao Jo BiopkysiCo
aad Biochem. Cytol o , 5:593-602, WSBo
Honors aad Awards relating to this project: Hoae
Serial Mo, I-TIIBB-M-BR-IS
io lleiiroaiiatomieal Sciesceg
2, Section oa B©ve3-opmsat
and Eegsaeratioii
3« Betlisscla^ IM„ aad SaiDi Jh
4o Sans as NIHDB~HA~13
ladivid'aal Project Ssport
CaleBdar Year 195S
Project Titl©: EafeairioE'' aad social os^ganizatioa of rhesus
HSOEkeys oa Cajo Saatiago, Puerto Eico
Prisscipal Investigator: Stuart A. Altmaan
Other Sja'vestigators : Hone
Cooperatiag Units
m Years Ccalessdar
Total; Oo€
Prof ess iomai s OoS
Project Descriptioa:
Objectives ; To observe and record befaavios' aiad social
organisation of Macaca laplatta jaoskeys ob Cayo Santiago ■a'itk
tlie vi®w of oMaiaiag basic control data for esperiaisats iE
aeurological aad psjclaological deficits of adverss factors ii?.
tls© periBstal psriod of tiais species. To collect aormal re-
productive and gestational data. To observe aad studj tas
Ms t feods_emj;ilo.Y®d ; Direct observatioas and 2=©cordiags In
pictures aad souad taps ^ers Had©» Th® great cojaplesit]
of the social foeliavior of primates is a result of their Qstea-
si¥e repertoire of toeliavior aad of the mB.nj ^s^'s is TAich tfeej
cojabine the elemeats of tb.is repertoire iu.to a ^ride variety of
distiBct sequences of behavior ^ or "coiarses of actio»."o Tlaiis,
±u order to Easjiisiae tla@ accuracy of predict ioES of social
Iseliavior^ it is essential tlaat ^e obtain estimates of the
probabilities of sequences of events. These ©stimates are
Msed on the relative freqweziey of eacli possible course of
action o Tlae frequeiacies sr© ofetaiaed from rEssdoia saiaples of
social iatoractioaSo Tlie sampling tliat v&s hegnn In 1957 ?fas
continued iiatil Jiia©, lf>5So
Page 2 Serial No« KIMDB-lfA-DE-13
Part Ao Ps*oject Description (coat'd)
te.jQg findings; Of prijaiary sigaif icaace, tke orgaaiEstioa
of a prinat© society can he expressed as a set of rules that
are iadepaadeat of aay particular saeabsr of the society and
which depend upoa the fact that each member of the society
passes through assent ially the sasae life history patters »
These rules can he givea as a set of probabilities of co-urses
of act ion o
Analysis of sequential depeadeasies of social l>ehavior
requires a foraml §iodel of social orgaaisatios. A stoichio-
metrical aodel, based on the aatfeejiiaticai theory of coEuauniea-
tion, has bseia developed.
rf analysing the esteasive data that ha¥©
bees collected oo the life history pattera asd on the organisa-
tion of the society has not been completed^ certain outstandiag
characteristics are noted.
Individual recognitios is highly developed in isoakeys and
is of considerable importance in determiaiag the status of the
individual in the society » For esample, a monkey must toe afola
to distinguish bet'ween those ^ho® he caa doaissate in food cosi-
petition and those ^ho doainat© hia and are therefore da^gero-as
to challenge, la addition^ he aust foe able to m&lze temporary
shifts in these distinctions to take into accouBt the changes ir=
snood of his companions 0 For e^aiaple^ a playful adult sjale can
be approached safely^ Khereas the saae male, r^hea angry, canaot
b®o Beyond that, the sK>nkey must continuously revise these
predictions as fee and his companions ajaturs.
Thus the ontogeny of recognition is of paraaount import aace
in understanding the social behavior of sK>nk©ySo The iafaat
ssoakey's purview expands a® a result of his o^n curiosity and
the strong attraction that the infant has for other xaeiabers of
the society. His ability to distinguish dangerous situations
and individuals ±b further facilitated by his saother's inter-
ventions in his ovB behavior and in that of individuals "sho are
associated isith hiia.
One of the most outstanding characteristics of the groups
of Konkeys noi? on Cayo Santiago is their reiEarkabls social
stability. In this respect^ they ar© ia sharp contrast to the
highly unstable conditions that ^ere reported shortly after th©
creation of the colony in 1938, A nussber of factors seesa to b©
responsible for this change » The consistency of th© life his-
tory pattern has already been sjentioaedo The displacesient of
aggression onto subordinates has considerably replaced the
- 38 -
Pag© 3 Serial MOo MZEDB-MA^Il-13
Part Ao Project Descriptiosa (cout'd)
coBtiiSXJOUs contests for status that seeia to have characterised
the colony shortly after its forsnation; the status of doaiaaat
meiabers of the group is ao^ "wery rarely coatested. In addition^
aggressioa no'w geaerally iavolves oisly threats; overt physical
aggression is rare. Beyoad that^ suppression by dominant Esales
of aggressiOB asK>Eig subordinates j, and the stroag attraction of
the @nti?e group to these domiaasit males' have tended to reduce
disjunctive teadencies. The cohesivemess of the group has teeea
further eahaaced by the strong boBd betiseea otother and infant
asd its persistence during the juveaile period^, by the tendency
to restrict int ex-act ions to saeHibers of th© peer group, and by
the Eiarlsed ability of the laonkeys — ail except sis of vihom feave
spent their entire lives on the island — to recogsaize individuals
and the boundaries of their group.
The annual cycle of reproductive activity that vsls reported
for 1957 ^a® again observed.. Once again^ extensive sesual
activity between adults occurred during the four isonths begin-
ning in Slid- Sept ember. Parturition correspondingly occurred
fros February through May.
Significance ; The opportunity to study a colony of rhesus
ssonkeys under free ranging conditions is unique in the ae^
^orld.
The basic research ^ith the colony of Cayo Santiago has
transformed it into one of the best-kno^n groups of free ranging
fflammals. Despite th© fact that rhesus siacaques are used more
extensively in bioiaedical research than all other primates
coijsbined^ laany fundamental biological questions about this
Hjonkey have never been adequately studied. A continuation of
research ^ith the rhesus colony on Cayo Santiago say ^ell pro-
vide answers to many of these questions.
Proposed course of project; To complete analysis of data
collected.
A short break in observations was occasioned by transfer
of the Principal Investigator to Harvard University, A n@^
phase of the ^ork ^ill begin in November 1958, ^hen another
investigator comss on duty.
Part B included; Ho
~ 30 -
Serial No,, MIM3'B-MA-DR-14
lo WeuroaaatoEiicai Sciences
2c Section oa De¥©lopm©Bt
and Eegeneratioss
3o San J-uan aad Cayo Saiatiago
4. Sam© as KI!fDB-ifA-14
PHS-MIH
ladividual Project Eeport
Calendar Year 1958
Part A.
Project Title: Physical msasiarestents of rhesus laoakeys
froia birth to old age
Priacipal Investigator: So Ac Altisana
Other Investigators: J,, Gavan and K„ Chandler
Cooperating Units: Medical College of Soutli Carolina
Man Years (calendar year 1958) :
Total: 0„8
Professional : 0 o 5
Other: 0„3
Project Description:
Objectives: To arrive at age ©stiiaates of Esoufceys by
estabrisEIng~srandard physical laeasiiresieats o
Methods employed: The techniques of anthroposietry ar©
beiag used to obtain data on: sitting height ^ head lengthy
head breadth^ foot length., tail lengthj, dentition and weigfet.
The definitions of Ac H. Schislts were usedo To these^ tlie
technique of radiographic recording of skeletal parts is
about to be added o
Major findings; It is too early to report results,,
Data are being coirected and will be analysed o
Significance; Mensuration data on th© monkeys of
Santiago Island are of prissary value as a sjeans of estisss.tiag
ageso Age determination is essential for study of growth
and development j, behavioral ontogeny and other studies c
Proposed course of project; To continue the project o To
- 40 »
Page 2 Serial Bo.
Part A Project Descriptioa iaoufd)
correlate data with tliat obtaiaad from the cagad colonic?g
ia Cliarleston, ScCo, by Dr, Gavaa who has received extra-
Mural support for five years o
Part B iiicluded: Ho
- 41 -
Serial Mo. MZHDB-HA~0R-15
lo Heuroaaatoiaical Sciences
2, Section on Bevelopsjsnt
and Regeaeratioa
So Saa Juan^ Puerto Rico
4, Same as lIIISB-lA-17
PHS-MIH
Individual Project Report
Calendar Year 1958
Part A,
Project Title: Tecliaiqu© of aeurological esaiaination of
the laoakey (Macaca saulatta) o
Principal Investigator: Jo Eaack
Other Investigators: Marisa loRo Eamire:s de Arellano,
Co M. Coafes^ Do Lo McCrosky,,
H. Ho Jacobson
Cooperating Units: Uaiversity of Puerto Rico Medical
School.. Saa Juaa
Man Years (calendar year 1958) :
Total: lo3
Profess ional : 1 „ 0
Otfeer : o 3
Project Description:
Objectives: To design asd perfect tecfeaiques for carryi-Ci;
out neiirological exaainations of Konlsey© froa the first day of
life to maturity 0
Methods employed; Adaptation of standard methodm used in
the hwBjan neiirologlcal ©xaainatiOHj, when possible.;, recordiag
photograpliically each step in motion pictures o The E^jnkeys 052,
Cayo Santiago and in tli.© San Juan caged colony are used as
subjects o
Major findings: Considerable progress has been mad© In
tbe infant ssonkeyo A protocol with "cfeeck sheets" has goa©
through several revisions and is no^ Sseiag used in comparing
asphyxiated with control aaiaalSo It is too early to analyse
the data completely o Horiaal raaturat ional patterns have b©©a
established o Deviations have heen observed after asphyxia
neonatorum^ some of ^ich occur with regularity,.
Significance: Although the sionE<©y is being tised for
~ 42 ™
Page 2 Serial Mo^ MIIDB-MA-DH-IS
Part A Project Description (comt'd)
exp©riia@atal neurological investigations ia many last itut loss ,
no satisfactory standards for the neurological exaajiaatioa
have beesi published o
Proposed course of project: To complete the study, SMd
publ isS~r'esul ts " 'iUustrat e3~toy ®ot ion picture,,
Part B included: No
- 43 ™
Serial IlOo MBIBB~m-BE-I6
lo NeisroaaatGiaical Sciences
2a Sectioa oa Developmeat
aad Regeaeratioa
3o Bsthesdaj, Mdc aad Saa Juan ^ F.I
4o Hew
Individual Project Eeport
Calendar Year 1958
Part Ac
Project Title: Horssal reprod'acti'^s fuactioa In tfee
rhesus sionkey
Priacipal Invest igators: Howard Ho Jaeofoson
©tli©r Investigators: Hone
Cooperating Units: Isaiversity of Ftierto Eico
Medical Sckool aad USPSS
Clinic^ Saa Juaao
Maia Years CcaleacJar year 1958) :
Total: 0o5
Professional : o o 1
Other: 0,4
Project Description:
Objectives: To obtain data on meastr^atioa In asoakeys
under staadard coaditioias., T© ofeserv© wketlser or aot
seasoaal variatioas ©cciaro To stuidy spoatameoMS fliict^ation
of ttos© cycles assd attempt to alter them experimentally »
To obtain data oa conceptions isa tlae sioakey asd deteraiine
ifhetlaer or not seasosaal changes in coacsptioia rate occur o To
study factors iaflueisciag the relation between coaceptioa
Cpresuasably together witti oviilation) and tlie lEsastrual rhythm
of individual soakeySo
Methods employed: The toreediag coloay in Puerto Eico
pro^i^es an esceileat source of monkeys for study „ The
animals are fed a standard optinma diet a.nd ar© iiaspested
daily for evideac© of laeastrual bleeding o Ab. atteaist ^iil be
made to alter the usual rfeytlm by meaas of impIaBted iadactios
coils wlaos© cKrreat is lead to tlie serves supplying the uterus
to iadiice toleediago Simultaaeons ©xaaisiatioa of the
esadoffietriBsi will be perforasdo
- 44 "
Page 2 Serial Ho . S? IMB3 ~gA -Bi!.-! 6._
Part A Project Descriptiosa (eoat'd)
Matiags are preseatiy. coaf iaed to what is coasidered
the aost optiisal portioEs of the menstrn&l cycle o -To determiae
th© relationship of concept ioa to the measts-aal rhythm chosen
couples will to© isated outside of the usiial period asd the
results will fe© obtained toy visual iaspectioa of the geaitai
tract o
Major fiffidiags: Th© menstrual cycle of iadivid^ai
monkeys is subject"!© wide variations » Tlaese variatioas seeia
to to© equally spread throiigho^it the year. There is no
systematic relation between regularity of cycles aad fertility „
A change ±n the time of mating withia the cycle Ci<,eo on day II
ratlier tlsasa day 14) tends to alter the time to the so eallsd
iaplaatatios bleeding o It seeias from this data that implaata-
tioa toleediag is more a fuactioa of tlie animals owa sieiistriial
rIsytSm than it is of length of residence of tlie eiaferyo In tlie
uterine cavity o If this can foe coafiraed it will require
different explanatioas for liow the toleediag is prodyjcedo
Matings in tlais saae colony show tliat coaceptioia occurs
throBghout the yearo Tkis is ia contrast to sost piiblisked
reports o The colony as a whole is resmrkably fertile, part
of whicla is teatatively attributed to the care ?^itli ¥/hiGl3, tfe®
asatiags are performed o
Lgaj
reproductioffi ia the human teve tlaeir origia iu observations
oa primates c Our data iadicate tliat some of tlse coaclusioas
dra^a from earlier work, particularly about tfe,e autoaossy and
th© fis:©d character of tlae Moi'^oiaal regulation^ require
additioaal restrictions asid laay feav© to be revamped o
Proposed course of project: To study tfee relatioaship
between the aervous systea aad reproduction by sjeaas of
electrical stisuilatioa of tlae uterus , by continued study of
the nerve distribution in the titerus^ aad by study iag factors
liaitiag the period within a givea meastrua-l cycle during
'^ich ovulation and conception will occur o
Part B included: Mo
™ 45 »
Serial NOo lTMDB-lA~DR~i7
i„ Meuroasiatomicai ^Sciences
2o Section on BeveiopBient
asd Regensration
3o Betljesda^, Maryland^ Saa J«an
4o New
PHS-Nie
ladividual Project Report
Calendar Year 1958
Part A.
Project Title: Maturation in infant rfeesus monkeys;
and care required for rearing tfeemo
Principal Investigator: Howard No Jacobson
Other Investigators: Hone
Cooperating Uai-fe University of Puerto Rico Medical
School and USPHS Clinic, San J«.a.B,.
Man Years (calendar year 1958) :
Total: Oo5
Professional: 0o2
Other: 0o3
Project Description:
Objectives: To collect data on normal cage reared infant
rhesus monkeys in order to describe growth and developaieat
and the kinds of variations encounteredo
Methods ©SBployed: A nursery, resembling in m&ny details
those" in use for the care of human infants, is staintainedc.
Records are kept of daily weights^ daily sailk intake^, tempera-
ture, respiration rateo Dentition is recorded „ Heart rates
are recorded electrically , Grasp reflex is measured routinely e
Maturation of the ability to self-feed is assesaedo
Major findings: More than 50 monkeys were born in cages
and an equal auMber ia the free-range colony o Infant monkeys
show a regular pattern of growth and developaent ^ but with
wide ranges of norjsm.lSo Any study of infant monkeys must first
start with a knowledge of what is normal o Such standards fcjr
infant monkeys are not plentiful o
Significance; The significance of this study lies siostly
in its usefulness to others establishing priiaat® colonies o It
~ 46 »
Page 2 Serial No. MINBB-HA-BE-l?
Part A Project Description (coat'd)
is also essential in assessing the standards of care of a
nursery to b© able to compare it to others »
Proposed course of project: To contiaue collection
of datao
Part B included: Ho
Serial Mo. lIHDB~m-Pl™l@
io HewroaaatoBieal S©i©iae@®
aM R©g®a@ratioa
3o B®tla©©diip Mdo asd Saa Juaa,,PoBc
4c lOT
MS-MIS
ladividual Project E©p©rt
Cal@ad&r Year I©5S
Part Ao
ProJ©ct Titl®; Tfe® iatrisa^ic B@r¥@ supply to th® ©sdoaetriua
Priacipal Investigator: Howard Ifo Jaeobso®
Otfe.@r Sa¥®®tigatorss I^ai® Pelegrina Sari@go
Coop@rati©g Ussits: Bai^@r^ity ©f Pi2i®rto Eic©
CliaiCj, Sail Ju&n
Mas Years Ccaleadar y©ar 105@) :
Total; Oo®
Professional: 0c3
Otfeer; 0o3
Project D@®eriptiOE&:
Ofo^®ptiv®®; To steady tfe@ eoisrs® aad distributioia of aas-v®^
ia til®"" '@^doiffi@t"rluM of several species of aaimalSo To s©©
wfe®tfe®r t!i® si@rv@© ar© dlBtT±bnt®d t© tla® vascular ©leisaist® ©r
supply tla@ glasad^lar ®l©^(®at® as w@llo
M®tkods @mpIoy@.d: Tls® ©ndom^triim of adult cats, guia«^a
pigs akci'" rliisui 'aoBl£©ys i© staia®d by tti® methods of Bastar a,sd
of ¥@dd®ll using a®tlayl©a@ tolu© dy©o Tia© dy® ±m iiaj@ct@d iatra-
art@rially aad loyally into tls® uteria^ of ligMiy aia®®tli@ti£s@d
asaimalSo Wliol®-®oimts of tis®«© ar© prepared for @s:a®iaationo
Cotfliater ©taiaiBg; is uBm& t© show sstrwctural r®iatioiisfeip^ of tlii®
ffl©rv@ distribution c H^aa© Mt®ri ar© obtaia@d at @l©ctiv©
layst@r®©tosffiyo
Major findings ; TSi® eadomstri^aa of tia© cat is cl©ia©@ly
supplied witSi' iK'itt©"t©r®iaal aervaSo T&©®® aerv®® pass tferotagte
tSi® ®adom®trlwm and ar® foiaad approacsfeiag tls® surfae® ©pitla®liiiJi.
Ia certaia ar@as th® aerv©® s©©a to Is® artoori^iag aro^iad t!s@
Page 2 S@rial llOo HIMDB-IA-BR-IS
part A Project Description {cont'd)
bases o£ th® @ndoia@triaI glands o
Sigaificance: This is an unequivocal @s;a^l@ o£ tli@
participation of tb® nervous syst@B in an ar@a ^laicSi is
usually consid®r@d to be exclusively controlled ■ by hormones <
Tla® further demonstration of th® n@r¥© supply smy fe@lp
elucidate i^@cliani@@s of uterine bl@@dingc
Proposed course of project; To continue th® kisito--
logicai studies and compare results in th© several sp@ci@So
Part B included: Ho
io Me^roaaatomieal Sciences
2, Section on Davelopraent
and Eegeaeratioa
3o B©t^©sda, Mdo as-d Sar* M&n
4o Sasi® as 1IMBB-MA-.12<~1©S7
PHS-HIH
Individual Project Eepc^rt
Calendar Year 1958
Part Ac
Project Title ! Meuroiogic&l d®;ficits of asphyssia
i%@0£%atoru!s In macaca K.«alattai.
Principal lairestigators: W„ Fc Windl®, Jo Eo E&uck^
Co lo Comtos aad Ho Mo Jacoteoa
Other Investigators: Co Jo Bail@y, Marisa Eaiairas d®
Arellano^ Wo Stiehl, Jo Go Frontera.
Do ^Crosk®y., So ¥« Sas:©».
Coop©ratiKig Uaits: Uaiversity of Pii©rto Eico Medical
Man Years ic&lendar y®ar 1958) :
Total; lc5
Professional : 1 o 1
Otte®r: 0e4
Project Description:
Objectives : To d®t®rMia© seiit© aad sSsrosic asiarologies-l
efeasag®® cau®@d by a^pfeyxia a@c»aatorism la th@ m&caca sixslattao
M®tliQd@ yploy@d; Fetus©® of Imo^ci gestational s-ge w©r@
delivered at Caesar iaa s@ctioia shortly b©.for® full terra from
m>th®rB sated axader coatrolled coaditioaSo Fetwses were
aspliyxiatad by ressoval of th© uterine content© without ©pee-
ing th® clsorioaie ®aCo Aaimals? v^t® aapfeyxiated for -raryiiig
lasgtte® of tiM®o Others served as controls o Tliosa asphyKia--
t©d infants aot atol® to breathe spoataaeously ware resusci-
tated by ©sygea ia^uf flat ion of tli® luags toy iEteriBittent
posit iv@ pressure until tli@y ventilated t!i®BJS@lves „ E^peri-
is®ffital and control aaissals were raised with idsatical
8iaittt@aanc© aad t®stiffl$ procedures, except wfe.@K illa®ss or
B®v©rB daamg® iat©rf©r®do Sstaasiv® las® -w&b made of
«»tioa pictur© pliotograpfey aad other recordiag devices provi4'5.i©-
Page 2 Serial Mo. NIMDB.>HA.~DR^19
ipsrt A. Project Description Ccoat'd)
opportuaity for r©¥i@^ and coiaparisoa of exp®riBi©ats, Neiiro-
logical examinatioiKS «ere perfora©^ regularly.
Aaiiaals ^hose survival seeiaed msllkely and a f©w others
were killed for pathological study. Oa® infant ^as asphyxiated
by uniateatioESl r®teatioa of th@ li®ad ipithia the vagiaal canal
duriag a spoataa@ou@ br®@ch delivery o
Ma.ior fiadia^s; Of eight lafaats asphyxiated for 11.5
miautes or less aad -^hich thea breathed spoataaeously, 5 showed
ao abaor®aliti@s after recovery froa th© acut© period. Three
anisaals had deficits ia suckiag^ oa@ of which also had deficit©
for 10 days ia rightiag^ motor dexterity aad toaicity.
Of 13 iafaatSj asphyxiated for 7 to 17 saiautes aad requiring
resuscitatioa, all had deficits ia suckiag aad th@ quality aad
ezteat of voluatary motioa was affected. However » ao differ®ac@s
from the coatrols i»@r@ aoted ia voluatary laotioa after 3 ^e@k@
ia the 3 iafaats surviviag that loago
Miae resuscitated iafaats had deficits ia righting. Eight
resuscitated iafaats had deficits la reaction to droppiag.
Flame-shaped petechial h©iaorrhag@@ vsere @e@a in the retiaa ia
8 iafaats. Sis iafaats had deficits ia local isiag souado Tt?o
iafaats had hypotoaic musculature for a ^eeko la oae of these
the hypotonia alternated ^ith generalized rigidity.
Other abaorsaialities ia resuscitated iafaats observed only
once included a III nerve palsy, a 3 per second treisor lasting
7 dayjg^ status epilepticus, papilledema j^ and loss of coatrol of
body temperature «
Signif icaac® g Ssperiiaeatal evidence that asphyxia aeoaatornsj
ia a primate can produce symptoms ; comparable ta certain ones
described ia human neurological disorders of iafaacy^, haa h®®n
obtained for the first tiasSo
Proposed course of project; To continue aad extend this
project. Attempts are being made to produce more surviving
animals aad to test them psychologically. Physiological and
chemical studies of cardiovascular aad respiratory functions ^ill
be related to the neurological studies =
Part B included: Ye®
- 51 -
Serial Mo= 1|MB-M~p|j|~l^
PHS-NIH
Individual Project Report
C&leadar Year 1953
Part B; Honoi-s,, Awards, and Publicatioss
Publications other than abstracts froa this project:
Wiffidl©^ W, Fo s Editor j, "Hetirological assd psjchologicai
deficits from asphysia neonatorusa" o Springfield^
Illi&oiS; Charles C Tho^as^ ld5@.
Eoffiors and A'wards relating to this project:
» 52 -
Serial Bo. HIHDB-HA~DE~20
io Heuroaaatoiaical Scieaces
2o SectioB, oa Developaeat
asd RegeiaeratioEi
3 c Bethesdaj Marylaxid
4<, Mew
PES-HIH
lEdividual Project Report
Calendar Year 1958
Part A.
Project Title: Psychological effects of asphyxia
neonatoruia In rhesus monkeys o
Principal Investigators: Co J, Bailey, Sue Y, Sa3£oa
Other Investigators: Hone
Cooperating Units: University of Puerto Eico
Medical School, Saa Juan;
UoSoPublic Health Service
Clinic, San Juan
Man Years (calendar year 1958) :
Total : 0 o 7
Prof essioaal : 0,4
Other: 0„3
Project Description:
Qtojecti\^es; To constriict a large battery of tests tiiat
can be administered to young monkeys « To detersaine tfee
effects of neonatal asphyxia on the perforiaaac© of saonkeys
in these tests » To correlate th© results of the psychological
studies with histopathological observations on the brains o
Methods eaployed: Pregnant Eionkeys were delivered at
term by aaeans of cesarean sect ion o In some cases ^ infants
were retained in th© intact aembranes in order to asphyxiate
then; in other cases they were allowed to breathe iiaaediately
and- served as controls. Asphyxiated infants were resuscitated^
if necessary, by intratracheal insufflation with oxygen o
Activity, curiosity^ emotionality, and learning ability Vf&re
studied by means of the psychological test battery c.
Major findings: Most of the tiae was spent on designing^
constructing J, and testing the battery of tests « A series of
tests was decided upon that involves training the animals
every day for the first 2 years of li£e„ So far only S pairs
™ 53 ~
Page 2 Serial Mo„ MIMDB"M-DR~20
Part A Project Descriptioa (cont'd)
of monkeys have started this test battery, the oldest having
finished about one-third of the testing program, and the
youmgest about one-tenth c Although it is still too early to
determine what the final outcoaie of this testing program will
be, so far there appears to toe ao coBSisteat differesc©
between the asphyxiated and norEsal aniBalSo
Significance: Hot enough animals have progressed far
enough in the testing program to decide whether the monkey
differs from other aaiiaals^ such as the rat, guinea pig. and
cat J in its reaction to asphyxia neonatonuSo
Proposed course of project: The psychological test
battery is still being perfecte^o Some tests will be dropped
and new ones invented o Present animals will proceed with the
battery, and others, particularly those with a more sever©
asphyxia, will be started »
Part B included: Mo
- 54 -
Serial Mo. NIMDB-NA-DR-21
lo Neuroaaatosnical Sciences
2, Section or Development
and Regeaeration
3, Bethesda, Md. and Saa Juan^,
4, Same as NiroB-NA~15-1957
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A.
Project Title; Psychological and histopathological deficits
of asphyxia neonatorum in guinea pigs.
Principal Investigator: Co J. Bailey
Other Investigators: Wo F» ?/indle
Marisa I, R„ Hamirez de Arellano
Cooperating Units: University of Puerto Rico School of
Medicine j, S&n Juan
UoSo Public Health Service Cliaic,
San Juan
Man Years (calendar year 1958) :
Total: 0c8
Professional: Oo6
Other : 0 » 2
Project Description:
Objectives; To determine the ability of guinea pigs to
learn and remember simple aiaze problems at various ages after
asphyxiation at birth « To extend earlier experiaients of Becker
and Windle to older ages and different test situations. To
correlate physiological and psychological studies isith histo-
pathological observations on the brains.
Methods employed; As previously described: (a) asphysiatlca
by intrauterine ischemia at full term, (b) resuscitation by
intratracheal insufflation -s/ith O2, (c) study of neuromuscular
and neurosensory deficits^, (d) testing learning ability in laases
and (e) histopathological correlations, Littermate controls
■were used.
- 55 -
i»age z Serial Mo. NIMDB--HA-DR--21
Part A- Project Description (cont'd)
Ma.lor f iadJBjgs : Using the Becker alternatioB m&ze^ dif-
ferences between controls and experisjental animals &t 8 to 13
weeks of age were not statistically significant in respect to
time of ruMiing aud aumber of errors; there -m&s a difference
ira respect to repetitive errors significant beyoRd the .01
level. In another group of animals started in the Becker aaze
in the first t^o weeks of life, the controls ^ere significantly
faster in running the laase than the asphyxiated litteraatea
(p ^ »05). Using a closed field ^ater xaasej, the asphyxiated
guinea pigs (no^ 1^ to 19 aK>nths old) made laore errors thmi
tfaeir littermate controls (p « ,01) « In order to test retentioa,,
the animals relearned each mazeo There \i?as no consistent dif-
ference in the asphyxiated and aonasphyjtiated pigs. In order
to test the effect of stress on asphyxiated guinea pigs^ one
group relearned the Becker laase ^ith an electric shock on its
entire floor , except in the goal box. Ko difference could be
detected between the experimental and control aniaials. Altho^igh
there were significant correlations among various measures of
degree of asphyxia and degree of neonatal neurological deficits ;,
there -were no significant correlations among the measures of
degree of asphyxia and performance in the mazes »
Sigaificaa.ce: Asphyxia neonatorum In the guinea pig~-
previously shown by Windle and Becker to produce neurological
and psychological deficits correlated ^ith strtictural brain
daiaage in the first 8 to 10 iseeks of life-~no\? appears to ha\^e
produced effects on adult ability to learn siajpl© profolesas.
Proposed course of project : The acute and chronic
physiological and experimental psychological aspects have been
completed,, Though many interesting leads could be pursued
profitably^ the time of the investigators must he devoted to
primates nest year. The histopatho logical studies ■will be
continued at Bethesda and correlations dra^n where possible <=
Part B included I Ho
- 56 -=
Serial Ifo. EMM=M=M=M
lo Neiiroanatosaical Scierjces
2„ Section on. Develop?ner-t
and Regeneration
So Bethesda, Mdo aad ...
4o Hei
Jgart A.
Project Titles
PHS~HIH
Individual Project Report
Calendar Year 1958
Centers axid patlaiways. invol'ved in induced
cerebellar seizures,,
Principal Investigator; Co Mo Combs
Other Investigators: D, L=, McCroskef
Cooperating Units s
University of Puerto Rico Sclioo i
Medicine
UoS. Public Health Service Ciiwic,
San Juan
Man Years (calendar year 1958) :
Total: 0.8
Professional: 0.4
Other- 0.4
Project Description:
Objectives: To obtain a clearer understanding of the
cerebellar role in movement by determining the structures
involved in the phenomenon of the long-lasting induced cere-
bellar seizures. To then study the effects upon the cerebellar
seizures of electrical stimulation and lesions in the involved
structures =,
Methods employed; Initial experiments are being perforisied
on catSo The seizures are produced with perajanent iHiplantecJ
electrodes in the chronic animal o During the seizures recordings
are made from other deep electrodes.. Subsequently stimuli are
delivered or lesions are placed in the latter structures to
study the resultant effects upon the cerebellar seizure.
Stereotactic mapping isith deep electrodes is being
by the cathode ray oscillograph and the electroencephalogr-:.,..! ..
Seizure-producing electrical stimuli are delivered to the
exposed cerebellar cortex ^
- 57 -.
Page 2
Serial Mo, MIMDB->M-DE~22
Part Ao Project DescriptioK (ctxit'd)
B^.ior f jndiaR-s : This project feas been started "but too
recently for s report of resr.ltSo
Si^nif icaac©: Ataormallties of posture j, coor'iination and
quality of voluntary movements have been observed after asphyxia
neonatorurac, Physiological studies oja role of the cerebelluai In
tliese are aeeded for adequate uaderstaadiag of tfe^; pfeenome'nac
Proposed conigse of .,, project ; To carry out tia<j! stated plan.
To extead tlie study to normal monkeys aad those sufJfering neonatal
asphyxia .
Part B included 2 Mo
„ 58 -
Serial No» HIHDB-M-HC-^l
1= Meuroanatomical Scieaces
2o Section on Heiirocytology
3, Bethesda, Maryland
4. Same as NINDB-M-18«-1957
PHS-NIH
Individual Project Eeport
Caleadar Year 195S
Part A,
Project Title: Ultrastructure of the nervous systesjo
Principal Iisvestigator : Sanford Lo Palay
Other Investigators; Angelo Bairati
E, E. Manuel idis
Spencer Gordoa
Cooperating Units: Depto of Aaatoiiay;, Usii¥ersity of Milan
Dept. of Pathology, Yale University
laa Years (calemdar year 1958) :
Total; 1=9
Professional; lo4
Other; 0,5
Project Description:
Ofo.iectives; To study tfee fiae stz-ucture aad orgaaization
of Bervous tissue, particularly the syaapse, the protoplasra of
sserv© cells aud their processes^ amd the neuroglia^ -sith special
atteation to the correlation bet^iieen structure aad function.
Methods employed: The principal iastruaeat employed is the
electros microscope « Tissues are prepared for examination by
fixation in osaaium tetroxid©^ effibeddiag In laethacrylate, and
sectioning.
Major fiadiiags; A study of the terminals on the giaat
Mauthi^er cell in the medulla of the gold fish showed that the
boutonlike endiags on this cell resejuble those in Bjaimaals in
that they coataia ayriad -vesicles and a cluster of laitochoiidria o
The intrasynaptic cleft is approximately the saiae diameter.
The large club-eadings on the lateral dendrite differ frosa the
usual teroiiiials in the following ^ays: The mitochondria are
very small aad are aligned opposite the synaptic cleft o The
vesicular componeat is rather limited aad is clustered at the
sides of the ending instead of at the synaptic cleft. The
meaning of these differences is not iamediateiy clear.
Page 2 Serial Ifo^ HIHDB-M-NC-l
Part A, Project Description (eoat'd)
A study of asoplasE la the large sas^eliBated fibers of the
goldfish laedulla^ and In peripheral nerves of the rat aad cat
demonstrates that there are three longitudinally oriented struc-
tures in axoplasm: lo loag, eista'eiBely slender jnitocfeondria,
2<, their meiaforane- bound caiaalicull, assd S» slender threads,
termed neurofilaments , In the goldfish, the aeurof ilaments are
the BKJSt promiaent feature of the large asoaso They are ceatly
aligaed parallel ^ith oae another ^and fill alaiost the entire
axon. They are approsisiately 120A in diameter. la the smallest
anons^ the aeurofilaisQats are less Kiamerous and the caEalicula?:"
elements are the siost promiaesit striacturec Ho-vjev-er, ia the size
range between the biggest and the ssaallest, there is no re:gular
relation toeti^eea diameter of fifeer and .ratio of filaiaents to
canaliculi, Also^ ia the fish, there is eo x^egular relation fcat^si
the thickness of the layeliE sheath aad the diaaieter of the axon.
That the filaments are not artifacts of fisatioa is deaionstrated
toy a study ^ith polarised light optics of the fresh nerves aad
medullary tissue. The axoplasm sho^s a positive extrinsic
birefringence and a slight positi^'e intrinsic birefringence
^hich are not significantly changed by fixation, embedding, and
sectioning.. The neurof ilaEents in laasHiaalian peripheral nerves
are like those in the large nerve fibers of goldfish jaedullaj,
but certain differences are isaiaediately evideat= The filaments
are not so concentrated or so regularly disposed as in the^,
goldfish and their diaeieter is approsiaately half, i.e. 60A.
Significance : These studies are part of an intensive
program to explore the nervous system at the electron Ejicroscopic
level in order to establish criteria for an adequate fine
structural histology of these tissues. Until this is done a
clear concept of the interrelations bet'sieen neurons, neuroglia,
and their respective processes cannot emerge . The geography
must precede exploitation of the resources «
Proposed course of the project : Prelisjiaary reports of
the results on Mauthner cell synapses and oa aKoplasm are being
prepared for publication. The study of the Mauthaer cell will
foe continued, because there are morphological types of ssmapsss
on the Mauthner cell other than the ones we have studied so far«
Furthermore a study of dendrites will be carried out to learn
their characteristics in contrast to those of the asoQo A stu.dy
of the relationship between the dorsal root ganglion cell and
its surrounding capsular cells is to foe initiated in collatoration
^ith Dr. Rosenbluth. An investigation of the esstracellular space
in the laaEmalian central nervous systeai ^ill be carried on in
collaboration with Dr. Spencer Gordon.
Part B included: Yes
- 6© »
Serial No. NIlDB-NA-MC-1
PHS-MIH
Individual Project Report
Caieadar Year 1958
Part B; Mobofs^ Awards j, and PublicatioBS
PuMications otfeer than abstracts from this project:
Palajj, So L« : An electron Microscopical study of neuroglia
in "Biology of Neuroglia/' 1, F, Windle^ Ed.,
Springfield, Charles C Thom&Sj, pp. 24-38^ 1958 »
Palay^ S, L« : The Bjorpfeology of synapses ia the central
nervous systeis« Ssp= Cell Eesearch, SuDplement 5.
275-293^ 1S58.
Palay, So L, : Tlie aorphology of secretion^ ia ''Frontiers in
Cytology," So Lo Palay, Sd.j, Nei? Haven^ Yale
University Press^ pp. 305-342, 1958o
Honors End Awards relating to this project ;
Serial Hoo MimE:::Mh=mrA^
io MsTarosnstoiKieal Sciences
, 4. ^m as 1OTDB~M-19-1@57
PHS-NIH
ladi vidua! Project E©po^t
CEl@adar tear 195@
Par|_A<
Project Title: Easymatic reactions of gaiEma-a^inobutyrat®
Cy-ABl catalj?s©d toj' brain tissue.
Principal lavestigator : Eo W, Altjers
Other Investigator®: J, M» McKlsansi
R. Ac Salvador
Cooperatiag Uaits; Laboratory of Clinical Clissaistrj^ IIIMBB
Laboratory? of E®nTOQh®miBtvf » MIIDB
Mas Years Ccal©Mar fear 1958) ;
Total: 0o8
Prof ©ssioaal ; ©o5
Other: OoS
Project Description:
Ob.iectiVQa; To establish and characterise th® ®mzjm&tlc
traa@forsifttiosi@ ot -y-AB and related cosspo^Ms -sshieh occ^ir ^h®n
these cosipouads are iuciabated with brain tis^is® imder approp-
riate eoaditioa®.
Methods- employed s Spectrophotoiaetri'i, fluoriastf^g isotop©©.y
chroisatography &nd related techaiq^eso
^.lor .fiadin.^®; A soltifel© enzfu® ^hich o3sidiss©s succiaie
seiaiRldeh^d® to SBCCisie acid has beeis purified fro® feraia
Mitochondria 0 The reactioii catalysed hy this ®n^jm® rsqutr®®
diphosphop^ridia© aucleotid® a® hfdrogers accgptoro
Sobs© «vid@ac€ was obtained for the po@@ibl© iaportsacig of
y-AB in the energir aetabolism of the braiUo Comparisoa of
osfgea coasumptioB and oxidation of C-^^^^-AB in hojEogeaates of
teaia tissue iadicat® that a significant frsctio® of th@ res-
piration of braia tissue ig vitro ma^ h® associated ^ith th@
reaction® iavolviag Y-a^iMol3ut^rat®o
it* '--
Siisaif icaac© g The area of metabolism described »bov@ is
imdouMed importance ia cerebrsl metatolisnio Tb© eiiacida™
iojA of these reactions is a fimdamssital necessity for &m.
!9id@:rstanditig of the operation of the neural mechanism and th@
©rangei^nts of metabolism which vmj cause certain form's of
@urological disease o
l^oposed course of pro.lectg Further ^ork is planned for
iMaining information about the rate of metabolism of 7-emino<"
lutyrate in vivo <, This -siork ^ill require the deirelopment of a
technique for introducing C^^-Y-amisiohutyrate into the endogenous
1 of brain Y-aminobutyrate . This will be approached in three
®fSg
CD Continuous intravenous infusion;
(2) injection of a radioactive precursor of Y-aminobutfrate
toich will peaetirate the blood-brain barrier rapidly;
(3) the umm of very young animals in ^hich the blood- bra in
^?ri@r is not yet established <,
aft B included g Yes
- 63 ->
Serial MOo MJMm-'MA--m€^2
PHS-MSH
Individual Project R©poift
■ Caleadar Year 19 5@
Pagt B; Eoaors, Awards, aad Fiibllcation©
PublicatioQS other th&n abstracts fs>om tl£l@ project :
Albsr®, E., Wo aad lo Ao Salvador j Osidatioia of succisic
s@iaiald©feyd® fey sol-abl© dafeydrogeaas© from foraian
Sgieace 128; 359 (1958) o
Eoaors and Awards s-elatisig to tlais project:
^ m ■^■
Serial lo. mmm-m.-MC'^'S
lo H@wroaiiato£IciT"Sci@B€©s
2„ SactiQa on W.&m^oeytologf
4. ail® as MIl®e-''MA."21»i9S?
IMi^id^sal ^oject Report
Caieiiday Year 1958
1SI±_A<
]^oJ®ct Titles Q%saffltitativ© histoekeMcal dtstritatioa of
glutamic dscar^sflss® ia tb® aervoma sfEi^^-y
Principal Iffi%«'®stigatojr^ 1. W. Alters
Otli©r Isaveatigators: E. O.. Srady
Cooperating Uiaits; Laboratory of Haiaroch®iaistFy> ICIlfJB
las Y^&r& Ccslesdar |r®ar 1968);
Total ; 0o9
Prof ©ssiojssl ; 0 c S
Others 0<^4-
Project Description:
Ob.1®ctiiyeiB; To dstersiis© tfe© ia@iaroaiiat©adcal iocalizstios.
of glutamic dmc^ThoKflSiS® ia tl&© i£@r^ou@ mystmm^
liBtkod® ®i^lo¥@d.; Liquid scint illation co'isating; Lowrf
^1 trmssicroapalytical tecimiquies o
lia.iog fiadinges; The distritetiom of gluta®ic dsess^tosFlss?©
Ms b®®ii 4®t®rsiiia@d ±u girosis aspess of to central s@r¥oiss sf ? - :,
affid at til® iaistplogical level In s©^©fF*l ag-ea® of tb.® te-ai®. .: ,;■
spinal cord of tli© rhesus EOBk@y, The esiaf®® is virtually afe:i-at
iia ail sreaig of niiit® i!!att®r, aad is tls@ newtohfpophfmtn aiad
piaaal gls&ado It i® T^l&ttvelj &ig;ls, is certain liistologicai a-"" ^
of tia© g^©F asatter amd q\iit@ lo'is im otlsa3r@ =
SijjffiifieMcg ? l@uropljiaFSBaco logical studies laave iMics ' ^./sd
tli&t gaassffia-amiao butyrat® i® ^ po^erfwl iaisibitor of sysaptie
traa®iBi®sios» Siac© larg© aisoimts of gaima-aisiao Mtyrat© ar ^'
Y-AB decarboKylas© ay© fo%md uaiquely la tis© central ii?.©i"«foiss
Byst@isij! t&® fflof© precis© local isiatiosa of this dists'ltotioa :J r of
fasdasseatai importance tm mst&hl-kmhlng thm phjmlologlGzl re /
this Sfstassso
Pag© 2 . ,'S«riai Mo,. limB-lA-
Part '":, l>i-oj®bt Ofsscriptioa C<^osst*d}
.Froposed eoaa's®, of piroject :; fofk tg^ymiisated
Bcript te,is. b©@a s^teitted for publicatioa =
■jEci'iseecii;; xe®
m ^^
Sefiai Ho, HIDBrS:
FlS-lflB
Cal^siday Y®&-» 19S8
Pi£t_B5 l^noTB. Awards, aad Pulblicatioas
F«Micatioia@ otter tMii aljstracts from this project?
Al]b©rs, n. H. aad !<. O. Brady: Tto distributioa of
glutasaic d#cafbos:yias® Ir* the a^rvous ®fist@s
of til® s'&Qsus asosik©yo CJ» press)
^ &7 ^
lo MeuToaaatOBiical Sciesces
2= Section OB Metorocytologf
'I. Sam© as lK©B~.m«22-195?
ladi-^idual Project l@po&'t
Gal©Bdar Yeas? 1958
Part^<
Project Title: MiGro-a-adiometric measureiseat of deeartoosryiss©
react ioas.
PriBCipal lavestigator ; E. W. Aibea's
Other iB'^estigators : R. O. Br&d.y
Oooperatiag Uaits: Laboratory of Heiirocheju.xstr^'j NIM®
Maa Years Ccaleadar year 1958) :
Total: Oo9
Prof ®s8ioaal : 0o6
Otiser: ©o3
Project l>®sci"iptioa:
Qfo.iectives; To devise a ssthod ^hicsh ^ill permit th©
Bieasureaaeat of carboa dioxid© p^odwced hf ©nsyaes ia samples of
tissue ^@lg&isg t©© gsicrograsajs or lesso
^tlaods ®m&lo^®di Ultra-aicro asi&lytical tecteiques of tfee
Lo^ry tfp®i liquid sciatillatioss cousatisig-
M.lor giadiagB; A ^sdiosi^tric ®©tfe©«S for th© msas^re^ej^^t
of glutamic decarboxylaes© ©cti^it^ ±n teaia tisstae h&s hsen
developed aad applied to ssEtples of brain tissue ^eiglaieg as
littl® as 3 siicrograa^c
Sipaif ic&sce ; Th® Jistliod perssits qisaajatitative histoclaemisai
studies of d®carlK>xylas®s o An ©at2®@ratiois of the decarteosEylatisg
reactiosEi® aad tfaeir products s®rv©s to illustrat© th© importaaiee
of a study of decar bossy latia^ en&jmsm ^itla respect to uenirolog-ical
diseases :
Glutamic deearbosflaas produce® Y~a®iao butyric acid;
histidi®© d@carbosylas0 produces histaaiae;
5~hydrosy tryptophaB® decartoxylas® produces serotoBia;
3,5~dilaydrosyph®aflalasiisj© decarbossylas© produces a precm^sor of
non-®pinephti.n@ .
Page 2 S©rial Mo. mBBtM~MC.-4
Part A Project Deseriptios (cositM)
The miderlyiag specificity? of ^tls.© jsetfeod m^f allois its;
application to macro-analysis of ssleplos of bloody ®tc=; fliers
other aetlso'ds are difficult to apply »
Fropossad cotars© of pgo.iect; Work terminated asd
raaauscript laas b«@a s^Smitted for jhaMieationi.
Fart B included : Yes
^ G@ »
Serial Mo. Mil»:llbMb4
lEdividual Project E^isport
Part_B: Eonars^ Awards^ aad Publication®
Fublicatloisi® othea' thsku atetiracts from this project j
Aib®rs^ Eo Wo &nd R. O. Brady: Tli® disti'itatioa of
glutamic .^©cafbosylas© in the aervous systesi
of tfe® rfeesits saoakey- (Ii^ press}
S©2-i©l Ho. If|MIB-'M~HC"5
lo KeisToasatosical Sciences
2, Section on Nem'ocytolog;^^
3. BetlsesdE, ^x-j^land
4» Sam© as HIlBB~m"23-l©i7
ladi vidua 1 Project Efeport
Calendar Year 1958'
Part A<
Project Titl©: A f luorisstric siicrossstfeod for tisa detei'Siiua"
• ■ tion of ©ucciaic ^©saialdefeyde.
Principal Investigator: E. W. Albers
Other Investigators: Eo Salvador
Ooop@ratiag Uaits: LalKsratory of !?eiiroclaes!istry- MIKSB
^^ Years Ccal®adar year 1958) :
Total: i»0
Project Baser iptioa:
Object i'g'es ; To develop s sensitive siattjod to pes-jrit the
aasasi^sreasat of swccioic s®Mald©&fde CSSA) ins tissue sssiple-s of
10 ^icrograi^ or less.
M.I or itndltmm: SSA sa^ b© ^assured Isi^ aeaias of the
f luor@se®sac© of & derivative formed hf r-sactiois ^itls diaiaino-
I^BZoic acid. fh@ s^nBitlvitf of the i^thod is adequate aad
the set&od ca© b© applied to the stsasiaressnit of the gamma-aiaiao^
i© traasaisiaas© activity of Israia tias'sa©.
Sig^if icaiace ; T&© d©-^9lops®3at of tl»is s^tl:^od ^as a
r©qtiir©^iit for aa adequate st?ad^ of th© sietsboliss of gasaaa-
aaiffiofoutyrate, Fisrtfeer ds^elopffisiit of tlie «ltras»icro inethoci
has permitted aa iss'^estigatiosi of the possiMs association of
ga^aa-aiaiaob^tfrat® trausamiiia©© ^itla c©rtaia neuTo&n&tGmlc&l
BtruGtnreSs ami thus to a h^jsotliesis for tlse playsioiogical rois
of gamas-SMiBolsutsrrat®..
- ?i -
Page 2 Serial No. MM^:M:zi!£z§^
Part A. Project Description CcoBt'd)
Paroposed course of project; Work terminated SEd
aanuscript has beeja siabaiitted for publicatioao
Part B i^eltsd^dj Yes
Serial Eo. MUTDB^MA-HC-S
Individual Ps?oject Seporf ,
Calendar Yeas' 195S ^
Part _Bj_ Hosaos's, Awards ^ aacl FuMicatioas
PiiMiGatioES other thasi afests-asts from this project:
Salvador, E. A. mid 1. W» Allsars: The distrilnstioa of
glutaMiC'"y»ajuiEofetit^"r:le transaminase in tlae
Eer¥oi2S sj^stem ©f the rb.es sjs morilsef .. C.i''^ press)
Serial Ho. MI1BB-M-HG"@
1. HetiroaQatomieal Scieaees
2. Sectioa on M©^irocytologf
3. Betliesda, teryland
4. Sas© as MSM5B«.M-2@°1957
Iffidi vidua 1 Project Report
Calendar Year 1©5S
Project Titles S?®Bros@cr®tloa is th© s*od@Et»
Priiacipal Investigator; Milton Wo BrigMsaaa
Otl3©r lBi?©stigators§ lone
Cooperating Usaits: Hatioaal Casaear to^titiite
Maa Years (caleadar yeas* 1058) :
Total: 0.9
P^of ©©3ioaal : ©o@
Othes" s 0.3
Project D@3criptioa:
Objectives; To det©:^MB© the aios'plaologieal ©ff©ct© os tkfi
a@tiros©cff©to?y system oti C®) p^F^iologieal stismli^ i<,®o
suckliisg^ and (b) s-igradiatio^So
Methods ©apio¥®d; (a) LactatiEg female aiic© ^©re allowed to
BIAS'S© littsffs fo^ ^a2"ioMS Intetv^lB, and cMsges ia a@Bros©ci'©t©s'y
coateat of tl2@i2r hypotbalamo-hi^popljiysial sy^tesus ^@^© noted »
(b) Larg© do©©g of 220 EW s.s&fm ^feieh •^@r© direct©^ to th©
hypothalamus J T9©re gi^ea to adalt rats. Alterations iB th® bs'aia^
spl©©® aM peffiplses-al blood ises-© folloT^ed,
,fejog fiiadiaga; C^) Th©r© is a S'sleas® of aetsrosecs'etorf
material, ^hich issf he q1qb®1j associated ^itfa os^tocisij from
th@ n®uTohfpophfmlB of a significant aualse? of f@sial©eo
Cb) Th© astarosseretorj Bucl®i ar© no more susceptible to
z~ irradiation thaa ar© adjacsat rauGl©i<. TIa© earliest ckassges ±m
the brain ar© &n increas® in vascular p®ria@al2ility to trfpaa bin©
aad tfe@ forssatioa, in the irradiated aoa©, of p©t®sliia© aisd of
parivascular globules coMistiag of a glycoprotsia. Isuroglial
Pag© 2 S@a-ial lOo l|ipB:nM=l&^
Part Ao Project Descriptiosi (coat'd)
asad n©us?osial ©lemeats are relati-^ely refractorj^ A teriaisial
leucocytosis de^jiastrates timt tli® h@5iiopoi©tic tissue is ©till
able to respond (e.go to Bfstemlc InteQtlon) ^ Ten® stmct^r©
of t&© spIe©iJ3 isas msaltered.
Si^aaif igsac© % Insight into the factors ?;laich iafiu©gsc@ th©
fMS&ctiou of tia© ls^pothalaKi>-sieT!ar©aypopliys5ial systeiSo
^o^m±cowBe_of._tM.-BEQJSStt Fart Cb) lias l5@@Ei sufomitted
for puMicstio^o Fart (a) is to &« sutoittsd fojp ptiMieatioao
Part B iaclisded; Ho
-. 75 «
Serial Hoo lIISlsMrIfc2_
1. Newroanatoffiical Sciesices
2. Sectiosa on Meiia'ocytology
3. BetfeQsda, Maryland
4. Saaie as MIIiBB-M"-29-»185?
ladividisal Project Seport
Caleadai' Y©ar 1958
^rt_A,
Project Title: Sat^aaem'-oaal cfeoiiiiesterase of the ^/ertetorate
ceatrsl n^T^ous systesa.
Priacipal latest iga tor: M. W. Briglitiaaii
Otiies" Ia'?©stigatos"s: E. W* Albers
Cooperating Units : Hoe©
mn Years (■cal©Edsr f^&r 1958) :
Totals lol
Professional: 0»S
Other: 0.3 '
Rroject Dsscs-iptioa:
OMQCtive®; To Esake a systematie mtudf of t^e sites assd
probafol© fiaaction of ©strasewroaal cMoliuesteras©,
Methods ©laployed; Tfe© application of the Soelle
kiBtochemlcai g^thod &as te©s ®st®isd©«l to © species, Th®
d@v@lopM3nt of |»seudocholiae®teras© C^<^B} activity In the ehie&
embryo v&b q^aatified by a sp@ctropfe©to^tri€ isstliod.
Ma.io^ ilndinm®: iA} Th@ estraaeiironal clsiolis]t®st®ras©.s
occur ©itfeer in ths esdotlsslium <e.g., rat, goldfisia, and toad)
or isa tfee glia i®.g.f cat and rooster), TliQ^^e is so ofevioiis
pliyletic pattera of distrilmtion.
CB) As ©srlf as tis© fiftla. da^ ot iacisMtioa^ t&ar© is
appreciable FChE activity -^feich is tlsiis attributable to tfea
spo^ioblast. Tfe© ©^teyo l3©co®®s ©tssceptibl® to a n^aalser of
selective FCliS isilaibitors at tfe© tis© of greatest iacrease In
mmb®T of aewroglia cell®,
Sigaificaacg : Fiudiiigs of sp©ci©s dif foresees in tli@
localiaitioa oi estrsaeisroasl cbolinestefases afford &n &pprose&
to til® elucidation of tise fiiiactioa of these esaymss. Tfeer©
appears, to S^ a relatios feet^een tk© adveat of IC&l acti^'ity and
s»yelog©rjes4s ia t&@ esibryoc
- VG -
Pag© 2 Serial HOo MI^B~HA-MC»7
Part Ao Project ©©script ioa (cosst'd)
Proposed Coutb® of project; Tla© isork osi sp@ci®s
differeaees ia locaiizatioa of tli©s© eassfm©^ is baisig prosested
for publics tioao
Part B inclydedj T©s
Serial Ifo. SIM^JIh^Mtl.^^
Iiidi¥iduffil Proj@ct Eapos't
Cal®fficlar y@&r 1958
Part B; Honoris^ A'sjardSj and Publications
Publicffitiou® otfe©r than abstracts iron thlm project:
Brightsaafflj Mo 1. aad So ¥, Alb©yss A major coapoaeat of
aeuroSijpophyaial tissu® associated isitSi satldiaretie
activityo Jc l©uroeh@a<, (in press) »
Hosioi'® aad Awards s-elatiag to this projects
Serial lOo Iimiito-~gii~iaP-.l
lo M®^r©aaat©mical Sci@sic@s
2o Sectioa on Essperiia^atal
leuropatliology
So B@tfe©sda, Maryland
4o Saa© as HHDB~1A -SO-iSSI"
SBdividwal Project Eeport
Calesadar Y©ar 1058
Part Ao
Project Titl®: Fatlog©®@tical factors ia th® d©¥®lopE©nt
of ®y©lopathi®s o
Priacipal Investigator: Jaa, Cassmeraeyer
Otfe®r Investigators: H©l®2a Easasey, Migsioa Mala asaci
Staaislaw Ziemnowics
Years Cealeadar year 1958):
Total: lc8
Professional : 1^4
Otfeer: 0^4
Project Description:
Objectives: {&) To ototaia detailed isforsatioss alsout aormal
feistoT^Ieal"""aad physical characteristic© of th® spiiaal cord to
dissclos© factors involved is siaiiit@iiaBC@ of tlmsu® t^UBl&n aad
tolood eirculatioao Cb) To assess histological characteristics ©f
varied fmactioaal conditioa© of aeuroim assd ae^arogiiao Cc) To
correlat® cytologieal characteristic© .of th@ spiaal cord with
diff@reat d®gr@®@ of fuactioisal abnormal iti©Sc Cd) To clarify
the raechaaism of c@ll daiaag® aaid progr@@JSiv@ aatur@ of tiss^®
chaiag@s within th© spiral cordo
Methods^^ @iaploy@d: ¥olua@tric aad karyo®®tri© sa©asur®K@ats
w®r@ pevioinmhoik aorssal spiaal cords of s®v@ral sp@ci@©o
Techaiqu©® w®r® staadardis©d which ws.d® it posslfol® to pr©par@
microscopical 3@ctiOE&s of uniform app@araQc@o
Major fiadiiagB: Thr©@ series of invest igatioja® hav© be©a
conducted and their r@@ult@ will b® adapted to ©sp©riEj@ats oa tfee
spinal cord:
Ca) On th® basis of n®w raethods developed h@r© it wag
possible to iseasure rapidly th© volim© ©f th® ©atir® cord, its
regions and segments; repeated ei^aminatioBS w@r@ sad© on tli®
intact epeciaseaSc Th© percentage ©is® of each region wass foimd
Pag© 2 Serial MOo lUmm-W A-^W-^l
Part A Project D®©sriptioa (cosat'd)
to vary witfe species, ©„gc., sioakoy, dog^ cat, rabbit aad
oposssufflo CoBSiderabl® individual variations w@r© found
witfeia ©acli sp®ci@So
A furtlssr us© of tiiis voIuim©trie ®©tho,d led to th@
observation that tli@ volim® of tli© spinal cord caa be
ostiaated accordijsg to as ©quatioa based oa a correlatios
b©tw®®B th© volussfi® of th@ spiaal cord aad either body vmiight
or leagtls of leg boseSo This as®tfeod s&b tri@d with equal
accuracy in moakeys, cats aad rabbits «
Cb) Tfe© combia®d volissi@tric aad morphological control
of tfe© various ©taps ia th® preparation of histological
sectioQS l©d to a r©visios& of th© ©atir® procedure used
for a®uropatl&o logical iav®stigations„ Th® liistological
material is no'w obtaiaed fr©© of ®©v®rai of the uawasited
changes ^icls usually occur in th@ spiaal cord^ With tfe©
precautiosss adopted, particularly tiaose concerned ^itk fissa-
tioa and r®sioval of th© spitial cord^ a mmb©r of aoaEp®eific
o©ll cfeaag@s w®r@ avoid@do As part of tfeis project,
Dro Mignoa Mala^ is proc®@diBg witfe staadardizatioa of auto-
radiograpliic t®cfeBiqu@®c Prop@r Isaadling of tissue through
f isEatioa aad @iab®ddins has given a basis for detenaialBg tfe®
aoriaality of spiaal cord architecture, c®ll appearaae© and
auclear sis® ia ®oaSs®ySj cats, rabbits ^ cfeiachillas^ rats
aad B»ic®o The shape aad &±ze of neuroglial aucl©i In th@
cat, vsmrm fouad to b© influ@ac©d by tfe® histological
t®chai«|u@s us@d; this iaflueac® was greater oa c®lls of tfe©
thoracic regioia thaa oa thos® of th® cervical regioac Wh@s
a poor fisativ® was @japloy@d C®ogo, th® widely r®ooii®©ad<gd
fizatioffi by subia@rsioa of th@ sp©ci®®a with 10% formal ia)
th® ill @ff©ct was diff®r@at through various r©gloas of th©
spiaal cord as Judged hy th@ mmb®T of dark asurons aad
pykaotic aeuroglia ©@lls through the seggmats of th© spiaal
cordc
((c) Two karyojaetric surveys w@r© p@rform©d oa two types
of material, th@ oa® aiai®d at th® fuactioaal sigaificaac© of
neuroaal auclei aad th® other coac©ra@d with iadividu&l
variatioas i® the sis® of a@uraglial aucleio
M®uTonn ia the braia of chronically r@s#rpiaia@d Esoak^ys
were fouad to coataia ©alarg©d ausl@i aad aucl®olio This
cytological reaction was regarded as evideac© of a fuaetioaal
abaorsaality of tessporary aattar@o Ho periM-aeat pathological
Pag® 3 g©?ial llOc IfII?BB-im«.SF~l
Part A Project Bsscriptioa ioont'd)
(shaag@© were saotisd i® tlie €®Btral aervows ®y^t@m iBcl^adiag tli©
spiaal Gordo
Tfe© otfe©r kary©is@tric studj of ss@^l^ogliaI auclei In tlie
cat d@^2i@trat@«l tkat th®j w®r® of the ©aa© sise thro^gla t!a©
oes-Fical aad thoraelc rsgioas of tk© spinal eord^ How@'<f@£>,
tla© siE@ of astroeyt© asael@i ctsaaaged a® a liaear £uacti©Ei to
tSa® volisn© of tte© spiaal cordo
Sigaifieaffie©: The ©valwatioa of th® sigaif icasice of tfes
research resialts already ©b hasad isfill b© divided iat© tto©©
divisions whicla igadead form &n integral part of th® overall
project o
Ca) ¥©l?^i3©tric studies of tli© spiaal eos't! combined witli
ia@asiar©©©Bts ©f loag boa©s of th@ a:2t realties ©aabl© las aox?
to d@t@raiis© wfe@tla©r th© Kpieal cord sis© ia a gi^ea cas©
aig^t half© ehaaged as th© eoiBS©qu@ase of an @sp©risi®ato This
has not h®@n possitol@ to estafeligfe taatil tli© pr©s©at tackaigu©
was d©v@lop®<l fe©r@o Asaotlier asset to thi© teckaiq^© is that
w® ar© ia a positioa to select asaiiaals of equally ^is©d spiaal
eord after haviag ©easwred tli® I@agtl& of tke aaiaal's
estreiaiti©®, tfeis ®ay prov® Sa@lpf?al tor t&i© plaaaiag ©f ©ssperi-
M@nts witia phasiaacological age^t© ia ©rd@r to stsidy tlsseir effect
©sa e©atral aervotss ©y@t®®s of similar eis©c
Cb) B@fof® aay ®xp@ri^®sts could h® p@r formed a ssascta ae@d©d
coatrol ©f ©tar teistologieal a®tl&ods iaad to to© coBpI®t®do Th®
careful formalatioa of a standard preparation of l^istological
tisstses has ali»@t cosapI@t©Xy ©limisaated th@ pven^uc® of
art©f actual cell changes from ous* microscopical sections o ThnSf
w© ar© mow bos'® coaf idaat sbo^t th® results of microscopical
iffivestigatiosas In partietalas? afeout tfeos® of th® ©piaal cordo
Ps'©'^iou£5ly, tSi® study of this ©rgaa saor© tSiaa asy othej? was
haffip®r@d by th© occurreac© of UMBeroBS art©f actual c@ll cfeaagQSo
Since a larg® wmls®r of ia'yestigators hav® interpreted th®s©
cell claamges as represent iag asx intra vitas process^ asaay '
©rroiaeouB eoncl^sioE® Isav© been reached about result© ©f esp®T±~
a@QtSo Tlx© ©tasdardiaatioa of histological is®thods forced aa
iaportaiat part of autoradiograplaic iav@stigati©a© In ord@r to
idoatify properly tSa© sit® of tagged ©ub©tase®s and tl-&@ asaotiat
of depositioSo Faramouat to all aorpl&ological st^dy is to
preserv© as mada as possible tS&© sis® ©f tissw© stTUQt\ir®Bp
their relative di®tassc@ sad distritatioB ©f particles c
Pag© 4 Serial MOo MISDB-MA-SP-l
Part A Protest Description Ccosit'd)
Cc) Tk© ©b®@s?vatioffl that tli® si^© of glial m.ie,le± vai'ied
wit^ tla@ t®eS5aiqu©s used asad tls@ ©is© of asiiisals poiated out
two isportaat fact®* aaaely, Cl) ^s>rpl?,ological studies aiaed
to ideatify changes ia auclssr sis® mist urn® siaterial treated
accordisag to tli® h®st t®chnlqp.®s available, aaid (2) comparison
between the sis© of c©lls in aa ©spes'imeistal aad a noraai aEimal
±B permissibl© oaly when tia® asi®als are of siiailar siss»
Til© sigaificaac© off asalysisig tls® sis® of auclei, atael©oli
was broiaght oiat ia a study witli c&rosic admiBistratiosa of
r@s®rpin© to moEksySo Aay changes ia auclear siss© is ^ssisally
tak©® a© aa m-wl&eue® of efeaag@g! ia iatracell^lar metateolisjs of
is^cl@ic acido Tku^, th© karyometric r©3ialts siiglat i^disat©
tlmt tlais nuclear fiaactioa is d®raKg©d tJ^roughout tfee aervotan
syst©® a© a eoias@qu®ac© of chroaie resQE-pia© tr®atis®ato Biff ©rest
parts of th@ toaia tod responded with diff©r©iat iateasity to tlse
tr©atsi©ffito Coacomitaatly ¥;itli th® eellislaa!' claaag©^ th® asiiaal©
©^ibited th© syadroa© characteristic of taasaa pas-lsiasoBisai.
Froa tSa®s@ studies w© hav© also Igarned t&at tli© only material
iis©ful for ©tudi@© oa cell fisfflctiosa is tSa@ ©a® ^iclsi Isas beejs
feaadled with th® grsateet ear@« Maay papers dealing ?/it&
©iiailar subjects h&v® nsed t<§qlmiqu@s wlii©li are not well rsaited
for muoh stiadi@® aad tls,@ir stateaeats h.&v® to b© reevaluated ©a
th© feasis ©f E®w ©xp@riB@atSo
Proposed coarg® of project ; Futt^r® ©^perimeiats Bill he.
coadwcted' aloag two" separa't© l'ia@® :
Ca) Estr&spijaal factors coE©©ysi©d witli spiaa.1 ©ord fuaetiosts
will be studisd ia Joiat ©sp©ri®©ats witls Ds'o H., Eamsey o& tlm
epidural space o T1i©b© @^p©ria@)at3 will Sorm a cosatisauatio® of
tiro nsarly c©apl@t®d st^idies by Dr. !» tess©yj, oae oa^ tSi©
topograpSiy of t!a® epidural fat of tls© c&t aad aaot&©r' study ©s
epidural fat ©f tk® aoak®y aad rabbit ia co^parisosa with that of
th@ cat,
Cb) latraspiaal factors ia^olv®d ia spinal cord fBastioa®
will b© studied ia several ©speriiaests, tiaes© ®sp©ri^©ats f^ill
b© directed toward© thr©© ^&±u aspects of ®sp©ris®istal neuro-
pathology o
Esp®ri!a@Qtal a®Mr©patlaol©gy fraq^esatly r@qtair©ss aa®afe®©ia^
trepaaatioa and «s® of detsydratiag agsats, tfesr@for© a prelisisary
stwdy with Dr, Si@raaowicS w&b plaa,ffi®d for tife® purpose of ©s&iaiaiag
"■ 82 ....
Pag© 5 Serial lOo lIliBB»IIA ■■SF-i
Part A Project Descriptioa Ceoat'd)
to wfeat d@gr@® tfe@ spiBal cord r®act© to ©ob® of tia@g© ©zp©ri-
iB®]atal coMitioaSo Sou® eats imd@rw©sit cs-aaiotoHy ia
aa©stfe®sia assd otl8®rs w@r@ tr@at©<d with wr@a for w&rylng tiia@
aad dosag®„ Tte® brain arad spiisal cord material was messf^red
volMiiS©trisaIIy and ttea pr®par@d lai®tologically„ Tls© nuterial
is b©iffig r®adi@d for aicroscopical ©saaiaatloao
Affiotls®r s@t of ©sp®rim©at© will b® e©at®r©d oa a
correlation !3®tw®iga d®gr©© of asrphological cfeaagss am.d fuactioriai
d@iicits of aewroaSo Aaiaials will p@rfora a c@rtaia aisomat of
Motor activity slaortly b®f©r® th®y ar® killado 0'to©rs will
receive different drug© which ©licit fwaetioisal atoos?©aIiti@s
as &ft@r iaj@©ti®a witfe aa@stfe@sias r@s©rpi2a@p acetyl pyridia®.,
curar@g aad isasulia„ A ©©ri©® of aaiaals will b® subjected to"
liv@r da®ag@ of diff@r@at typ@®^ t&@s@ ©speg-iassats ^^ill b@
coatrolled by r@p@at@d toiockeiaical ©^a^aatio^s of tli© feloodo
Through! tJi@s@ ©t^idies w® feop® to Is® ia a position to aecoisat for
Bom® of tfe@ p@culiariti@® ©f spisal cord pat&ology and diff@r-
©ac©s In ©liaiG&l maaif ©stations b@tw©©sa tfe© rostral aad caudal
regions o AltSi®wgfe ttes® studies ar@ s^aialy concerned witfe tls©
spiaal cords, tls® braia will b@ ©zaaiasd t©0|, aad t&© eoacli^-sioas
to b® r®ada@d will b@ f»pplicatol@ to tfee ©iitir® central n®TVQUB
ssystesso
Kssp®rSjs@at© '^111 fo@ dir®©t@d toward® tfei© factor® rege-
lating traasportatioja of substaae^s to tls@ B@ur©ssSo For tfeis
purpo©©' Dro Migffioa Mala lias b©©sa cnrrmutlj @agag@d iia ©t^dyisg
til® distribtatioa of isotopes tliroiL^g^ th® braia aad spiaal cordo
Furthmrsmre g a morpfeological stisdy oa th® disstribustioa of neuro-
glia aad %mss®la h&s b@®B coat©a&plat®d tlsat ^@ m^j lay t!i©
fouadatloa for a diserassioa about tk@ iiil£©r@at factos"® iaYol¥©d
ia coatrol of traa^portatioa ©f isub®taiac®s t&rougM t&© serTOisis
ti@g5u©o Histological mat©rial ka© fe©®a prepared for ml&rors-
copical ©stamiaatloa of th® spiaal cord^ ia ^nkmjBs, e&tSj,
rabbit£(^ ckiaoliillas^ rats aad mic@o
Part B iaclud@d: lo
Serial MOo lin>lIi3-M-lP'-2
io l©^aroasaatosaical Sciences
So Sectioa on l^perisseatal
Neuropathology
3o B®th©S£la, iarylsad
4c fSaisie as 2IIMDB-m-32-.19i7
Sndi^idual Project Esport
Cal©sadar ¥@ar 1958
Part A,
Project Title: Di©trifoutioa ©f fat In the epidural spae®
Priaeipal lEi'?r@stigator: Helea EaB^sey
Otfe@r lavsstigators: Mou®
Cooperating Uaits: Mono
laza Y®ars {©s-l^^a^fes? year 195S) :
Total: lo©
Profess ioaal : © , ?
Otla®y: 0«3
Projeet Beseriptioa:
Obj©ctiy®s: To study th@ aisatoaieal di^tributiosa of fat
ia tS&@' spl&sil ©pidi&ral spa©® ia several ©p©ci®s, to shew tfe©
variation T?it!a ag© aad other faetos-s, to study tla© bistologf
of @pid«a2'ai fat asid to cosipar© tfei® witli t!a© histology of
©tte@r adipose tiB@u®.
Method® ®mploj®d: Epidural fat was colos'Qd i£ yjLv© by
OK-al a^ialst ration of Siidaia III dissol"i«'©d ia oil„ llaiMals
w©r© killed Isy p@rftisi@©--fisatioa; tla® ires'tebral coIuse was
removed aad tti© boss® deealeif iedo Dissect ioas to dsaoBStrats
tla© epidiaral fat ia its proper ^-elatioa© wes*© photograplaed
for eomparativ© study o
£3aJor f iiidiaga : Complete series of sp@ciB©ESS tov© b©ea
E©CBismiat©d for adult catSj, kit teas 1 day to 24 weeks of ag@j,
ratebitss and fmnt®yBo A f®^ sp@eis@iss of ©©veral ©tla@r sspecies
hav@ b®@a ©tudl©c!o Epidural fat ia each of th© sp@Gi@s
®3£aBia®d sfeowed a differ@sit, characteristic pattern of distri-
ct ioso
Pag© 2 Serial Ko . lilggj^i^^rggril.
part A ProJ®©t Description C<s®at'd)
Pietrib^tioa of fat ia tls© ®pidaral space of tia® cat was
©kown to follow a distinct a©taa@ric patters character ia®d fey
losigitudinal lateral masses coim©ct©d by s®ga@atal dorsial
iaass@So Th® patters was laid dowa disrisag tts© first 12 w@®&©
a£t®r birtfeo listologically thi® tissue diff@r®d from, adipos©
tissB® ©ls@wh©r@ ia th© foody^ littl® fibrous coaaectiv© tissue
beiffig pr@s©2ato Its coapositioa aad its r©lati'/© freedom £roH
attaclaE@iat nak© it well adapted t© it© fuactioao
Sigaific&Bceo TSais is th© first complete d@seription of
th® di@po@it'i€»s^"'6f fat tlaroisglsoMt tk@ ©pidisral spac©o Spidaral
fat s©rv®s as a luteicaat und protect ioa for th© spiaal cord,
disra^ B@rv© roots aad blood v®ss@Is against tfeeir feoay
surrousadiagSj, particularly dwriag s»^®®@nt of th® vert@teal
eol^smao Biac® t^©s© vislaerabl© structiares &rm eoataiaed witlaia
a closed spac® a.z\d iEflu©iaced hj pr®ssu2'© ckaages la it^ all
struetures of th® ©pac© aasim© bowb playEiological sigaificaas©
as w©llo
Proposed coiar®® of project: A coBpl@t@d B^aascrlpt on th®
aaatomy asjd histology ©rTSe^idural fat ia the cat will fee
subssitted to aa anatoaical joariaalo Bat© on t&® ©pidMral fat
of tfe© rabbit asid th® Eoak®y ar® being analysed aad will be
readied for publicatioa isa th@ a@ar fmtu?®o Exp@rim@Btis iais'-olv-
iffig dasmg® to gtructmres ©f tl&@ ©pid^aral spa©® will b®©©B® part
of aaotSser project o
Fart B igs©liid@d: Wo
15 -
Serial lo 11MBB-MA-.EP-3
1 o Meusroaaatomical Scieaces
2„ Section on Ssperisiental
Meuropatlsology
So Bstfeesdaj, Karylsiud
Individual Project Eeport
Caleiidar Year 1958
Part A.
Project Title: Structure of brains of monkeys ii), -wshich the
pituitary glajid had been irradiated isith
Isigh-eiiergy deuteroas .
Principal Investigator: Jo Camaiersaeyer
Other iQvestigators: Coraeliiis A. Tobias
Gertrude ¥an Wageaea
Cooperating Units: University of California
feiversitj Medical Seliool
Man Years (calendar year 1958) :
Total: loO .
Professional : 0 <> @
Other : 0 o 3
Project Description:
Objectives; To mirvsy e^teat of sts-uetiiral claaage in
brains of amiisals previotisly subjected to foeaais of high-energy
deuterons focussed on the pituitary gland .-
Methods eHtpio;^ed: Four feaaie rhesus laoBlseys ^ere
irradiated on tlse Califoraia Betatron prior to seswal mat?arity,
Several years later they "^iere killed by the teclinique of
perfusion~f isatiojs. The braijjs ^ere reasoved and exasained for
gross pathology o Appropriate portions v^ere sectiosaed serially
and studied for microscopical defects.
Major findiags: T^o brsias ■©ere perfectly normal. Oae
brain shoised moderate chaages in hypothalamic Biiclei directly
connected v^ith tlie pituitary » One brain showed severe degen-
erative chaages in the parts of tbe braia adjacent to tlie
pituitary; these changes ^©re restricted to the patb'©ay of tfee
beam.
Page 2 Serial MOc lIBIDB~M-EP-3
Part Ao Project Description (coat'd)
Si^nif JcaHce ; The study of carefully prepared braias
following deuteron irradiation lielps towards a better mnder-
standing of tfee biological effects that tfee organisM may suffer
after expostsre to a variety of radioactive sources ia particular
at high altitudes, Th© encouraging aspect of tlie ciorpliological
results is a confirmation of the idea foriaulatecl by the Cal-
ifornia scientists that this type of radiation^ when properly
applied^, may fee used as a tool to destroy tissues -si'liicb. are not
easily accessible «
Proposed course of project; A report was giveB to the
investigators at the Uaiversity of California . They xsill
incorporate the fiadings i^ith their data aad prepare a fiaal
report «
Fart B included;
... s? ^
Serial Mo= Mil©B"JfA"-FM-.l
Hem.'oarf.a t oui ;,
4. Sggje as MIMJ-B»m-^-.34.»ZSS'
ladlFidiial Project Report
Calead&r Year i05S
Part^,
Project Titles A Btndy of the auditory affeysj-jt sRd «f fE=«T*?-.->-j'
PriEcipal issvestigator: Graat Lc Sassiiissejs
Cooperating Units;. Hoae
Total;- OJb
Professional j © o S
Oth@r;: 0.4
Project Description;
Otej actives,; To cosstiim® to explore and ^©i;@ai unlsnomi
anatOMicai a©uro0al comnectioas of the sff®'r©nt ajid ©fferest
auditory sfstsis; to leasrsi sosjetlilfsg about tia© asiatossiical ekcI
fimctiosal iat^rrelstionsisiips of th®®© two ^ymtmiBBc
CD Til© questiois of tose or fr®q«efficf coMuctiosj iocai-
i»@d -witlsiB. til© ascesjdiag pathway ie poorly wsdes'stood, Qei.®
of thm m&jor ofejecti'^@s i© to dsteraia® tfee 'poiat to poiat '
iiateramiroisal hook-up that evid©ii,tly e;Ki8ts feetiseea tls© o?g®?a
of Corti ais<S the cocfeleas' s?.uc-l@us and th© pi-oJ^ctioBS frosa
tiie Isttsr to th© superior oiivsry coispl^x aiad to th® faiglier
auditory awcl@i. '(2) Also to l@arE tmrm abo«t tlae iats'issic
aisd ©xtriasic recurreint or feed-back coaK©ctioa® of th®
cochlear nucleus. (3) To dotei'aiae wMtm^ or fiot tfee eoefelsar
aucleuis possesses €ff©r®iit a^uroas t&at isn©rvat© t&@ Mir c®ll©
of tto® cochl©ao
-. 88 »
Page 2 Serisl Mo. mmB-MA-Wll-'l
Part Ao Project Description (cont'd)
Methods ©aployed; T!i® n®'^3r aad more effect lire tecla-
niques &r@ cMefly depended lapoa'for deEKjnstratiosi of ajsonsl,
preterminal (Mauta-Gygas method) and terminal or synaptic
d®g©n@ratioa {Hasmussen metliod) . Th© employment of these
techniqu©© on ©ssperisjent.al subjects in ^hicli small isolated
lesions are properly placed makes it possible to reveal iiis-
portaut complexly arranged neuronal connections.
Ma.ior findias^s; Histological preparations of 35
©xperimeatai animals {cats and cfeinchilla^) isaife been com-
pleted but tJse series is not yet s^ifficient to complete the
picture of tb® afferent projection arrau,ge®eat ia the coclalear
nucleus .
Significance: ibre exact knowledge of the auditory
system is essential for a foundation upon which to design
intelligent physiological experimentatioa and for interpreta-
tion of the results that lead to uEderstaadieg of the nst^ro-
mechaniSBi of feeariago
Proposed eoura© of pro.ject ; To extend the study of
the afferent systesi is a similar fashion to higher levels
of the auditory system including th© auditory coi.'tex.
Part B ii&cluded: Mo
Serial lOc MIMBB-I'IA-Fl-P.
lo ll©uroas3a.tomical Sciesices
2o Section ois Fiimctior.al
Sewr®as.Et®mj
3„ B®tfe@sda^ Mai«ylan.d
4„ 33mm as HIlBB-!m-3f>^l@f57
Iiidi^idtial Project leport
Cal©adar Y®ar 1958
Part Ac
Pr©J©ct Titl©° A corralativ© feistopatlsologlc and genetic
Btudj of tfe@ he&rlmg mecfeamisia in a ©trair^
of coageBital d@a£ gaiaea pigSo
Priaeipal Iiav®atigator: Go Lo Rassms®©a
Otfeer Investigators J Moa®
Cooperating Uiaits; DFo George Jayj, ge.a®tlc±st of the
Laboratory Aids Brasaclis; is
r®spoasible for th@ br@©diffig aad
®tndf of tfe® inlieritaxic© of d@af-
gMinea pigs to this isesctioa duriag
tls© y@ar 1957 o
fca Y®ars Ccal©adar y@ar 1S5S) ;
Total; ©=,8
Prof ©g^ioaal : 0„4
Otfe®rs @c4
Project D®eicriptioa5
Objective®: (1) T© d@t®rmia® tla© factors of iRfe®ritance
r^ipoiiIlI¥"loF tli® fesariag defect o Bro Jay is tfe® principal
investigator of tfei® plsa^® of tli® project <, <2) Th^ chief ais
of tfe® investigator of thlm s®ctioii is t© find, if possible,,
tfe® iisideriyiisg ^»rpto logical «l©f®cts ^hieh lm&,<& to r@gr®ssio5S
of thM hearing saecfeasigs aad ultimat® d®afss©^So
M®tl&od@ employed; S©riss,l s©cti©a^ of th® petrous boass
aad braia ®t©i»s of aaisaals raagLag ia age froa birth to two
asad ©S8,© half y©ar„ ar® prepared by variotis appropriate
tecfeaiqus®® for tte© st^sdy of ta© diffe,r®at tlEmxms &.nd cjto--
logicml @i®»eatSo lyeliss aheatfess ©.ud asoraal d©g«ftsiera.t ioa
Pag© 2 Serial ^Q q .. _ g jggg-M -.»-2.
Part A Pr€*J©ct Bescriptioa (coat'ei)
t©©liaiq«@s are also ©laployad to deteraia© tfee stages ot
nturosial d@gea®ratloiio
Major fiadiags : A histological ©ttsdy of tte© petrous
honm&~or'^ aBlssalf ©f tfe@ IIH 'waltsing strain' is nearly
cosipl©teo Til© patteological cfeang©® noted tfeus far ar©
©sseatl&lly similar to thoB® described toy Lwrie ;(1941) from
a ©sailer ®@ri@s of Waltsers g®cur©d from B-nothmr Qolonjo
Th© Iiair cells ar@ tli® most suscaptibl© cellwlar ©lejsents
of tis© organ of Corti beiag the first to disappear ei2tir©iy
by tfe® ©ad of tSs© first ^@eko Early atrophic cfeanges ar@
noted in tli@ stria vascularis a@ a tfeianirag of th© capillar
b®d Bad Govm-Tlmg ©pitiieliiira but th© atropfey is aot complete
even is ttes older aaim&ls of ^sr® tfeaui two year® ©f ag®o
Otlier cellular ©l©is©atss of tfefs orgaa of Corti fad® away nores
slowly and tn a c®rtaiis ®®qu®Btial fashioao All r®Miaats of
tfe© organ of Corti hav® disappeared in aniiaals of fiv© i^stfes
of ag®o Tfe® cochlear s&euroiss ®lot?iy atrophy lat@r oa but a
f@w resistant aff©r©nt aewroj^ persist ±u tSi© oldest anisal
studied.
Of partienslar ister@st ±m tfe® fat® of tSi© ©ff©r@nt
eocsfelaar fibers located lis t!i@ Jw^tagaaglioaic ^spiral of tbe
coclileso Tlies© fiber© wlios® cells of origin ar® raore
r@sot©ly located is tla© CHS ar© not ©atirely r®si^taat to
tisis k©r@dit&ry disease^ Tfees© fib@rs also drop ©wt after
tte© first year aad ao trae® of th.@m i® fouisd ia thm two year
old axii^alo
Th%m far ia tis© study no irorpJiologle&l d#f@ct feas fe®«>a
foimd ^i©!i miglit acGotsat for tli® r@gr@ssiv® cfeaffig©gi is tfe©
eoelsI@ao
@ig,saificaac©c Obe®rirati©ias resulting fro® tfeis project
should b@ip^o uiad@rstamd i&) similar typ© iiili©rit©d deB.inc^s^.
occurring in feuMia aad Cb) thm resaltaat progress iv® patfe.o-
logical ckaages tfeat ®ubs@qis©atly occwr iis. all ®t.ruct^rc-?s of
tfe® eoclile&o
Proposed coiars@ of project: To cosiplete t!i® sttady during'
195S and to r@pert r®suTfSo
Part B isi©lud@d^ No
.=- Bl »
Serial WOo NIMIB-M--g!f^3 __
2o Section on Fmictional
NeuroaaatOBy
3 c, B@tii©sda, Maryland
4. Sass© as MISI>B--MA»36-»lgS?
PES -HIE
Individual Project E©port
Caleadar Tear l'®58
Part A,
Project Titles An experimental stwdy of th© s<©dial ioag-
itudiaal fascicialus of the brain st©si and
spinal cordo
Principal Xave^tigator: Leo Co Msssopust Jro
Ot.fe@r Investigator®: Ga Lc Eagiswssea
Coop®ratiag Units: Hoiae
M&n Y®ars <cal©iid&r year 195S):
Total: 0 = 9
Prof @s^ iojsal : 0-„ 6
Other: 0,3
Project Description:
Objeetiife® : To determine tfe® origia and teralRatiosi of
fibers in th@ M@dial longitudinal fascicniiu® is central
nervous syS5t©» witfe particular atteatioa to its tersiiaation
on cell® of th@ aat®rior feosra of tli© spinal cordo
M€5tS&ods ©^ployed: Tli© makiag of lesions at diffaresit
l©v®lI~orTfe© s®dia'r"loagitudinal fascici?,lu® aM in thm
various a«©l®i of origia of its fibers o Aaalysis of a®tiroa&l
comi®etloa@ by th@ us® of a:£0£%al a^d terminal degenaratiois
adtliods described by Kauta-^ygas lS54c
Major findings: A aotewortfey findisig sine® tfe© last
axmual report is smm&r±zmd in tk@ Aaatomical Record of I95So
Tfei® coaceras a co®poss®nt of tfee saodial losigittadisaal
fasciculus wfeiclJ arises fros th® saidbraia in an area oecupied
by th® iat®r@titial siuc1®mb of Cajal, Tfe© fibers cour®© ia
the dorso-sedial part of tfe® medial lo^gitudissal f asciculiijs ;
tuna sfearply veatrad at the level of tls© facial collic-alws
aad tarainate ia tli© maia inferior olivSo
- 92 -
Pag© 2 Serial NOo NIMIB--HA--FH~3
Part A Project Description Ccoat'd)
3igai£ieaiic#: This ^t«dy will turmlBh isforBatiou on
thm t©raliiatiO!i of descesdissg pathways from the brais stem
to aaterior horn cells o It should be of value to tlio-se
astiropliysiologists studying the ©lectroaie behavior of
anterior horn cells which are affesctsd by tte© tensiaatioB
of d©sc@isdijBg pathways froja tli@ braia steaio
Proposed cours® of project: Thlm project vf&s termln-
at©d ISIiSIFo* thim yearo If-fee smB^script is ia th© final
stages ©f preparation o
Part B iacluded^ Mo
Mi
- 0S -
Serial Ho. MIH
2o Section on Functional
4. B&sm as MIM)B-.M-3?~li5f
ladiiridual Project Siepos't
Calendar Year 1958
Part A.
Project Titi©: Heisroaal cona®ctioas &nd t&e fiinctioaal
sigBificance of the iate^pediancular micleMS.
Friiscipal lavestigator : Leo Co ^ssiopMStj Jr,
Otiser latest iga tors: G. L» Easia^ss.ss^i
Cooperating Units: Hose
Man Years (caleadar year 1@58) :
Total: OoS
PsotesBlQn&l : @ <> 5
Other: ©.S
Project Ssscriptioa:
|eotl^@s: Ca) To dotevmln® tfe© ae^s'osal eoaasctions
of th© iaterpe'd'jincwlar qmcIqiss, partieularly the ®ff@reat
coaaectioas aljo^it ^hich llttl® is kao-^a. {b} To deterain®^
if possiMejB tSs© fuaetional. significance of this aucletis
^hicii aaf ia^olve both viseeral processes and/or sossatic
SK>tor processes.
„^tijiodg ^emplo^ad ; (a) Ssiall ©lectrol^tic Issios© ss*©
to fes coaf iaed to tls© isatarpeduacislar aucleiss itself in
order to avoid isjjury to stsrroimding ©tr^ctisre^ fjisich •®oiild
cosplicste iat©rpr©tatio3a of the results. This veatrslly
placed aticlais® bas fe©retofor® te©®si destroyed fross a dos'sal
approach ^aicla 3a@c©ssarily destroyed siasj^ ©truetures along
t&© ©lectrod© pat!s. Cb) l^fe® esiiploy®ei5.t of physiological tasts
ia ItKstfe acute and diroaie preparation© to determin© tfe©
ftanctiosaal integrity of certain visceral ajodsaaisss hj m^&nB
of recording ©lectroaically alteratioi:^ in tempmr&tur®
regulatory ■ ability, vasosK>tor chasjgeSj fesart rate cSaaage®
aad otisersc Further,, aay abBormal feelaavior dta® to destr miction
of tlie aucleus «ill ba studied <■ {c} TSie ©Kperissatai sistsrial
» 94 »
Page 2 ^ Serial No. NINDB-NA-FK--4
Part A, Project Descriptioa {cont'd)
^iil h® prepared according to certaia axoaal and termlna.!
degeaeratioa t©c&aics in order to bring out tSae result lag
degeneratioB.
Ma.ior fiadia^g; Ihie to tlse teelmical difficulties
iavolv©d ia tlie surgical approach to tla© iaterpedunculas*
nucleus ; osly one of 12 anlEials operated ^pon possesse€l a
^ell-plac8d lesion ia the aucleisss. More successful exper-
iments s!e©d to be done.
Si^Bif Icaace : This project will provide basic aeat»
omical information on tlie efferent f ibor connections of tliis
nucleus, a subject atout ^hicla little ©spsriBeatal data asrista.
Farther, it ^ill pro'^ide soa© knowledge of the possible
functional significance of tfeis nucleus.
Proposed cours® of project ; To inactivate project
during 1959.
Part B included 5 Ho
B5
Serial Ho, NIMBB~HA-gI^5
1 . MeuroajsatoaiGal Scieaces
2. Ssctioa on Fijaetioaal
Meuroanatomy
3. Bstliesdaj Maryland
Indlvldn&l Project Seport
Part A.
Project Title: Tla@ coBsparative anatomy oi th© efferent
cochlear bundle in selected siibaianssialian
vertebrates; an ©sperisaeiital study.
Friacipal Investigator: Eobort L, Boord
Other liavestigators : Grant L. Easjaussen
Coop®rat;iag toits: Tfeis project ^as initiated 1 July,
1958 under the auspices of FsaMie Health
Service Eesearefe Felloisship #g003 under tae
sponsoj?slsip and guidance of Dr., Qordon M.
Rasm, Dspartiaent of Soologj^^ Uaiversitsf of
larylaad, and Dr. Grant L. EasM^ssen, tlais
latjoratory.
Man Years CcalQadar ^ear 1958) : ,
Fffof essional : ©»S
Other: ©,,2
Project Description ;
Objectives: {1} To establish tlae |>r©s©ace of aa efferent
cocibil@ar bundle in safemammalian vertebrates. (2) To stiidy its
central and peripfeeral anatomic relationships to ths acoustic
nerve compleis.
Metftods @aplo?Qd; The surgical aiidlia© incisions in t!i©
medulla froja the dorsal approach, at tfe® level of tSae facial
genua, ^here fibars of tls© ©ffereat cochlear biandlo Mi?© fo^sn
defiaitely ©istablished to cross in masaaalSo Follo?5iag th©
placessent of favorabl® lesions, serial SQctioas of tiae fcrsisi
stem. aM inner ears ars prepared hy degene^atioB isetlaods. Tlie
Nauta-GygaK tschaic for degeaeratiug saons is applied to iiitra-
medullary Katerial and tis® Sudan black and Marcfei m^tljods are
employed on the peripheral portion of the acoHBtic aer'sfe.
Pag© 2 Serial No. HIMBB-MA-FM-S
Fart Ao Project Description (cont'd)
^■ior ^iRdiBigs : ' Ho major results to report. Serial
seetioas of th© brain stem and inasi* oars of tlsre© pigsoss and
one Caiasaa liave teen prepared by appropriate histological
techniqiiQS lm.t observations at this time are too incoaplet©
to preseat definite aEatosaical details eoncorriiag th® presence^
origin or courts of t&e effes'eat coelslear handle in tiasse forms.
Sigaificaac©; The disclosure of an ef fersst eoaiponsiit
to tfe© cociilea laas resiaited ia ssucls apecialation eoneerning its
rol© isi th® Iseariag procsv^s a© isell as its ultimata central
aad paripheral taraiaatioas. While %h® sffer®at Isiaadle has
been sho^a to be present in esssntiallf coastant form tiirougb-
oiat th@ massaliSj its pressac© has not heen deteraiiaed in
lower fonas. A study of tlie ©fferent cochlear "miadle in lo^er
forms ^itli a simpler auditory apparatus ^ill lead to a better
uiiderstaadijQg of tha functional significaac© of tfeis feuadl©
and provide p'h'^logen®tlc e'^ideaca upon 'shicli to base f latere
physiological aM anatomical considerations.
Proposed co«rs@ of pro.lect ; This project ^ill eontinti©
tmder tlie present researck fello'ssfeip grant nntil 1 Julj, 1959 o
EovievQTi it may b® neeesssry to csntiaus study fesyond this time
ia order to cosiplete histological preparations and obser'^'atious.
The resi<® of tfeis project 'sill l^e tEsed In the preparation of
a diss©rtatioa for tlie degree of Doctor of Philosophy and
Psrt B iaclisded: No
- 9? -
1 . Ke'uroanatomical Scieaees
2o Section on Functional
4. Nes
Iisdiiridisal Project Eeport
Calendar Year 1S5S
Pa£t_A.
Project Title; A study of an eff®r©at cosaponeBt of the
vestilsulas' nerve arisiag from the laedulla
ofoloiigeta «
Priacipal Investigator: Graat L. Sasmuss©^
Other Investigators: Eiclsard Gacek
Cooperating Units: Hoiae
liaa Years (calesdar year 1958) :
Total: 0.?
iProf ®@sioB.al : 0o4
Other: ©.3
Project Descspiptioa:
Ob.iectives; To Isaara saor© afeout t&e origia and partie-
ularly th® mod® of termiaation of tSi© efferent t?©st±bi2lar
fite®rs, i.©.,, to d®t@rm±n® ^iietfeer or aot the liair receptor
cells receive efferent as «©11 as th© Tsell~kaoi?n affereat
fiber tarminsls.
Methods employQgl; T^s Sisdaa felack techaiqia© prs'^ioiasly
described "by Easmiisseii (1333) for study of Walleriaa degssei'atiort
of n©rv@® of tSi© p©trows toa©s is iaploy©d siafeseqweat to pro-
d^ctioa of lesions is the 'Vestibular nuclei and other regioas of
the hiadtoaiii. Serial s@ctioai@ of the vestilsalar root and all
its peripheral brandies ar© steadied in © series of 31 ©speri-
a©Htai animals (24 cats and 7 chinchillas) . Silver &nd
©sthylea© blu® vital stains '^ill be employed for deiaosastratiEg
tfe© ultimate tersinatioa of the efferent©.
Major findiaggi; Dwriag th© past year a layeliaated
nervous componeat of the vestibular aerve has I>©©b revealed
^hich is eoffiparabl© in isaay s^rphological respects to the
Page 2 Serial So. MI^.^A:zSM.rS___
Part A. l^oj@ct Description tconi: ' d)
©ffereat coclaleftjf tjus-dle.: For the first time vsstibuslar
©fferests h&r® ls<Bea traced froB a central origiu. aad tferougb-
otst tSs© Testilwlsr nerv© sad all its teaaehes as far as |:h@
receptor organs of tlie vestitelar labyrinth. Th® esseatial
findings ha^e been reported by abstract in tlie Matomical
Record, 130 s ^61-362^ 1958.
Slgralfi c asce J As yet ao pfojsio logical atusSf has teosa
attempted to elucidate t!s® fuECtlon of th& vestilmlar eff©r£=i».t©..
Ho^e-'rerj, oise m&f sp©c"alat® o?g their fianctioaai sigsif icsKss© or
th© Msis of f?ljat ia too^U- ateowt tMe CGchl®&ic &itev&nt bundle
^feieh .has heen dem>nstrmed to eaert a regalatorf or iEhite?. 'o-;--
iaf iueacs oa the auditory aenrous iKp«t (Gaiasabos) .
gyo.gS.gM. 9.9MM® oi Pro.i ®gi s To contiawe the stwdy is,
order to ®©ttl«i the import asst question coacerniag the ultiMiate
terssiBatioa of tis® @ffer#iits iB the vestibular receptor organs •
Fsrt B iaciuded? Mo
k^jsmar-y Eeport of Lsabo^ratory of Biop&ysies
Calendar Year 105S
Keanetla S. Cole, CMef
l^e ceiTferal objeetiire of the Laboi^atos'y of Biophysies
is to ^inde^stand tlje aature asd tiae inplicatioas of -^e ion
jaovemeats fiiadsuaaatal to tlie initiation aad propagation of a
nerve impulse » T^e staffs 3o1m. Wo Mooreg Eicliaz'd FitsEiigha
Robert So TaylOA% Jolm .H« Gebhart^ sad Eraest E» WlaiteoKb, have
progressed towards this objective^ in part 'syith the eollafo-
oration of Jose del Castillo^ eiiaical Hetarophysiology^, ISHBB^
Seymour Lo Friess^ Haval Medical Research Sastitute^ sad the
Computatiosj. Labos»atory,j national B^aresu of Staa^ards,
The characteristics of iadividaal ionic saovemeats as
first determiaed from meas^srements of the sqisid ason saesibraae
eurreat esd potential iiader eontrolled electrical, geosietrical
aad ionic coEditious have led to fas? reaching coaciiisionso Bat
ststoseq'aesst work has made it necessary to imdertake bm. esami-
aation of the exteat to 'which the: sasasured meabraae c^asTS'esits
de^^s.^. tspoa the adequacy of these controls.
The tentative conclusions from the aeasuresieats^ analysesa
amd cosputations ao^w available for the squid assos are that the
iatemal ctsrreat electrodes of 20 ohm cmo" resistance sb drivesi
froffi the 0c2 ohin aiaplifier outputs in routine use have allo7/ed
a reasonably adequate coatrol of the laeabr^ie potential over a
5 laso length c The effects of the external electrode geometry
OS the control do siot seem to be serious but these and the
effects of sm i:snide3itif ied meisbrase resistance have not been
satisfactorily resolved,
Aa iadicatioa of the effect of snob, aa iatemal electrode
is that it caa reduce the impulse tr^isit tiiae froai the nos^sl
I sisec, for a cb, to less than | {isec.
In the course of this work the need to measure mesabraiie
current^ without iatrodueiag bm appreciable poteatial differ™
ence^ has bees met hy the invention and use of a ne^s feedback
circuit aad the stabilised wide baad electrometer preamplifier
for faithful measuresieat of saicroelectrode potentials has bees
improved v^ith about a 10 fold reduction in noise and ripple »
Frajikenhaeuser's method for measurement of the potential
inside a node of Ranvier from an adjacent node has been success-
fully adapted to use the improved electroaeter presjsiplif iers
and operational amplifier techniques available heroo A prelim-
inary examination of the characteristics of the node storied that
■when the dixf icialties of coatroliiag the mexabraae potential
thrcagh the high iBternodai resistance vsere overcome^ ioaic
current characteristics similar to those of the squid a^coa
were foiazid and wei-e is agreement with independent results <>
Although sosae progress has been made, there is still the
need for a careful study of several laethods^ including those
originally used by Hodgkin ajad Huxley^ for the separation and
empirical representation of the sodium and potassium com-
ponents of the squid mesabrane ion current* It has, however^
been possible to show, by variation of the initial conditions,
that the original representation of the potassium current
transient is only satisfactory under liiaited conditions while
the assumption of the independence of the two components has
received further support o
The finding that in 0o5 M external KCl the squid aiem-
brane has the predicted negative resistance characteristics to
permit and all-or-none action potential suggests that the normal
distinction between sodium and potassium is only relative o Yet
the survival of a high peals sodium conductance to the end of a
long experiment showed that this membrane mechanism can be very
rugged. Preliminary investigations with ethanol, ether^ chloro-
fossn^ acetone^ di02£ane^ eserine^, and cevadiae indicate that the
potassium conductances were unaffected while the sodium conduct-
ances v;ere markedly decreased hy some and increased hy none,,
In contrast to eserine^ Holothurin, from the sea cucumber ,
blocked the frog sciatic nerve without change of velocity and
on a single node also^ irreversibly and independently of pHo
New derivatives of the ethylene dianine type showed that^
although a cis form blocked when the trans form did nct^ morels
diamine units were no more effectives
Further analog computations of Hodgkin-Husley equations
with modified time constants have given action potentials with
recovery plateaus that are in better agreement than before with
those produced by some membranes «
The error in the earlier SSAC digital computer code has
been found and the recomputations on the IBM 704 compl«3ted„
Computations of the entire uniformily propagating action poten-
tials at 6,3°Co and 1S,5°C, have been completed^ and the cal-
culated net ionic fluxes per impulse found to have approximate
agreement with eisperimental results. A digital code has been
constructed to compute the more difficult problems of the initi-
ation of an impulse in a medullated axon with modified Hodgkin-
Husley membranes at the nodes^ Preliminary computations show
both the origin of an impulse at an electrode and its propagation
with a constant velocity along the ason.
- 2 -
NATIOIAJ^ mSTITOfE OF ISrUROIXiGICAL BISiSASBS AMD EUlSHMEBi
Basis Resoarcii Progrsa
Bstiiaated QbligatioBS for F¥
Totals $150,500
Direct: 130^000
ReimbBS'semeat ; 26 . 500
ladi^iduai Projects 1 through 5
Serial Ko.lllDB^B°.l _ ____
1. Laborato:cy of liopSysic;
3, Bstliesclaj, liSiTjl&nd
4. CostiBuatioa of ISSf
PHS^HIH project
lndxvldu^.1 Project Eeport
Calendar Year 1958
Pa^t A,
Fs-oject Title: loaic Permeabilities of tiae Sqtaid Giaat Asson
Principal Investigators: loore^ J. ¥,. Cole^^ K. S.
Other Investigators: Taylor, S. S., del Castillo^ J. (Cliii.
1
Cooperating Uaits: Section on Cliaical Nsuroplaysiologyj, BIHDB.
Marine Biological Laboratory ^ Woods Hole.
Massachusetts
Maa Years:
Total: 4.6
Professional : 3.8
Other: O.S
^TojBQC iJescripxxoa.:
Objectives; The interpret at ion of nerve f^Bction in terias
of~fait' ionic transports aad t^e eliacidation of tb.e
striictures aad mecSiasaisms hj which these transports are
coatx'olled.
Me tliods _EBgj.oyed ; The iJsstr«imentatioB reiaaiaSj, fsiada-
mesataHyj tiae saaie as taat first used at Woods Hole in
1947 to measure directly tlie 'ion carreiit crossisag tla©
squid assos. meaibrans foetv^eea a long aasial electrode
witiixB. tlie aso32 and an essternal coEcoatric eJ^eetrode
after a controlled chaage of potential difference across
tlis Hesibraae, Mieropipette-calojael and e5:-ternal=-ea,loael
electrodes- have been used for jaeastiremsBt of tbe seabrane
potential asid control of tlie membrane cBx^rent to miniaise
tlie effect ox isiternal and external electrolytic resis-
tances; and to siBiplify tiae axial electrode design^
constr^5ction and operatioa. Tlie Laboratory eliopper™
stabilissd broad-band electi'OBotsr has isade t&e drift
of meiabrane poteBtial raeasurisig equipment during a.a
esperisxojit negligible. A new eisrreat Eieas"arijag circuity
waich iiatrodiicss a jaegligible potential drop across its
inpitt, has feeen developed In the Laboratory^ effecti"/ely
Bsod at Woods Hole d'aring the 1958 sq^sid soasoaj, and
s'abatitted fox' '0isblicatio2i.
Part A, Project Bescriptioa (eoBt'd'l
Major Fiacliags: The p:c- ...;.„:;._ ;-::^ xavestigatioas witli
"~™s^iiiolj etfeer^. ealoroform^ acetone^ dio^aae^ eseriae^
and ce'^adine liatre shotA'a that tke steady state potassiiisi
GOnductaBces v^qtq relatively ■una.f feeted v&ereas the
peak sodium coiiductaaces were saarkediy decreased b^r
some bxat isaereased by aoia©. In apparent contrast is
tlie f lading of asi Bneliaaged peak sodiiam cGiidnetaiace at
the end of a loag eKpes'iaieat . Tae procaine results
■pTevlously reported iiave been submitted for publieatiojiv
Personnel have not been a^vailable to laalse satis-
factory progress on the general problem of tlie routine
separatiOB and empirical repressntatioas of the sodJ.tiia
and potassiissa aosapcsaeBts of the meiabrane ioa currents.
Howei^er^ coasidera.ble evidence has been obtained Is.
support of the original asstsmption tliat these eoapoKeiats
are iadepeadeat of each otiaer. The potassi'toa current
traasient caai be expressed in tlie Eodgkisi--IMsley fora
only if an ejeposieat of at least tw'enty-f ive is issed ±n
place of the original valwe of four. Tliis in turn im-^
plies asa evea smaller reverse potassiiia conductance th&n
previously sisspe.cted aad leads to a laeiBbras,© laobility
of 10--"^ that In vi&ter casts doiabt upon tlie utility of
the formtslatioa.. Th.e findiag of a negative resistance
cliaracteristic v/ith tfee asoa la 0.5 M KCl has b©ea
submitted for -pBblicatioa and is not oalj predictable
by ttee original equations B,tkd a rational fesisis for tlie
observed a,ll-or~aGsie actios potesitials but also siaggests
that tl3.e tssual distiiactiOii he'meen sodium aiad potassium
as well as lithiBsi saay be HOi'e cfaaatitative than qiia-li-
tatixTe .
Some years agOj^ wixeii it becasie possible to obtais
and aiaiata-ia mtons liaving up to 6 ma/c-ra'^ of peak sodiiim,
tlie first of several complications appea,red in the ioaic
ciirrent pattera tliat liad aot bees seen ±n the original
esperirseats v;itli peak sodium ctarrents of about 1 Ea/cBi"^.
Duriag tSie 1958 sqtiid season^ tlie effects of resistaiiee
in the output circtait of the eoatrol amplifier and of
tiae surface iiepedaace of the axial electrode upon tlie
potential difference across tlie Hembrane at tlie control
poiat and at ctfeer points u'itlaia the aieasuria^ dhm&ber
vere investigated vjitn. various arraagesients of additioasl
essternal aad internal asoa electrodes.
Asaislifier resistances well beloi^ one ohm. em. and
asial electrodes of 20 ohm. em.^ effectii^ea gave a coBtroI
of tlse membraae potential to v;ltliiB a tevi millivolts.
Ajaplif ier and/or electrode resistances of abo^lt one ohm
cm. aEd 75 dbs& csi. respectively allowed some anomalies
wfeile for sc3me^ha,t hi.gh.eT ^e^iH.^^^i^^iceB m® messbE'aae
potential erroj!;' coisid be teas of nailliTi'Olts u'ithia
sailllsieters of the cositrol sicroelectrcde. Tfc,ese
espei'imestal aEd aBalytical approaclxes &-£@ not yet
complete but it is escpected tkat tb.© iShenomena. are
to be rather ■©ell espiaiaed in physical tesss. Tli©
effects Qf tlie eistsmal electrode geometry do not,
B®em to be serious fo^t. they asd the effect of aa
"tinidsatif led siembrane resistasxce &ave aot heen satis-
factorily s^esol'^ed. In spite of these resorvQ-tlo^jB ^
it seeias probable that tfee meBsbraaes of strong a2r.ons
laave been ro^tiiaely measured imder eonclitioEis not
dasgeroiisly far froa those assumed aad sought and
that tiaey Ixave claas'acteristics similar to those of
the asosis tiiat were easier to control.
Sigaif icggcg_ to MXSTDB Beseareh; The receatly developed
coa5^¥~iS3~'Se~^aiure^5"€s of the socli-om and
potassium ioB. aovesiesits across tiae squid &Mon sEembrsBe
offer a ne^^ Iiigliiy specific^,- and general approacla to
the factors uaderlyisg aoraiai a-iad patliologieal aervoiss
processes wlsica casmot as yet be iBvestigated directly
In hlghoT aaisials aad aaaa.
Proposed Cotxrse ®£_^^ject: Tlie detailed arad esteasif/e
aaalyiis oaT^ve p^'esent s-ecords will be cGntintied \i'it}.i
■e^e objectives of
1. a reasGaably ecssaplete aad critical essaaiination of
tlse origisal Hodgkia-Sessley f ormBlations ,
2. more satisfactory and objective tecliniques for t&e
empi2?ical forsiulatiosss of tl?.e ssensbrase cliaracterist.
3. the resol^tica of the eoaveatioaal playsiologj/^
patliology aad phajjaiacology of iae3?ve isito tli© ioa
persieability variables,
4. establisliiag of practical e^psrimeatal roiitines ano.
geseral plaaaiag of f'jrtfeer expefiaeatal workj
5. providing bases for t&eoretical iBveBtigatiosas of
the Hec!iaaiss»s of ioa persieabilitiesc
icl'-ided - yes
Part E;; Honors^ iiV.'ardSj, aad F-uoilcations :
Publicatiojtis other than abstracts frosa this project:
Schwauj H. P. and Colo^ K, B,.^ "Electrical physiology:
/alternating; c-arroat adnittaace of coils and tissues"..
Accepted for publicatioa in Medical Physics ¥ol. III.
Chicago^« Otto QlBsser., Ed,^ 'fe&.r~Eo6E'~PnSllsEers .
project
■•di'/idaal Project Eeport
loaic Permeabilities of Nerve Membranes
Theoretical . Investigations
Priacipal ijivestig"
Other lavestigatorf/.
Al;.r^OSAS-'.^iC£i J is..
-; rating CoaputatiOB. Laboi HatiOE.&-
of Staadards
Man Years:
Total: 1.5
Profess ioaal ; i . 2
Other t .3
Project Descriptioj,?.: ^
Objectives: To investigate the bases,, coaseqwesices and
essteiGSions of iosic permeability concepts and iafonaatio:.;
by matheE: aaalysi
Methods Eapioyed: Matiaematieai aaalysiSj, usiag th
of JioElimear differential <5qw.atioiis said digital
analog compntevs. The electrosaic analog compBtej.- Ill
tlais. laboratory has becosae a:a indispensable tool for
this project^ although many problesis remaia which require
the use of the digital eompiiter of the Hatioaal Eiire&.-a
of Staadards.
Llajor Fiadiags: Tke error in the earlier SEAC cok;
~ oF°^e""locigkin-IIuxley equs.tioas for the sqisid r.
brane has been found <def i^-itely to -.-result from
error peraittiag overflow ia the computation oi
traii^scsndeBtal fuactioss near tlieir siagBlairities. 'Ai:
resulted ia tbe appearance of t\vo spurious singula^' pc
ia the phase space for the cyrreat claisip case. One of
these points^ a tluresliold saddle poiat^ produced wrc:."';:'v;;
diverging phase plane tr-ajectories. Eecomisutationr;
tlie H'ationa.l Bureau, of Staadards IBM 704 of the mer:
a.ctioa potential have Qhown tlant iateraiediate sised
isaptilses eaa foe obtained fcy a suff ivciently s-cciirate
a result w'silch had ox-igiaalij' bee» expected from the
forsi of t^e eqiiatiOBS. The accuracy a?equi3?ed (foe iter
than 1/10") iSj hou'ever^, experi2ien.t,a,lly tiiiattaisia,ble3
and the Hodgkin-Huxley equations adeqtiately represent
am all-or-aoae
Coffipiatatioas of the cojaplete Hodg^ia-lmsley
propagated action potential for 18.5° C. aad for 6.3®
C. were fiaislied with velocities of 18.7 is/sec. and
12.3 B/sec respectively and the net traasfers per
impulse of potassiism^ sodium, and leakage ions computed^
to slio?; the effect of temperat«are on them. The figisres
for potass iiaa showed only a rough agreeiaent (30%
difference) with tke experiaeatal results of Shanes aad
gave a Qiq of 0.36^ compared ivith Shanes' valtie of 0.52.
Coding has been completed and a fev; exploratory
computations have been made with the 704 for the case of
a noa."Space~clsmped aoded aerve fibei*^ vjith flodgkin-
Huxley membranes at th$ nodes., so saodified as to have
the capacitance aad restiiig condiietaace found ±'a frog
jierve. These shov; the initiation of an impulse at a
stimiilatijig electrode^ aiad its coasequejat propagation
v^itii constant velocity. The difference between the
two velocities .already obtained^ 13.6 m/see. for a 0.01
msec, stimulating current piilse^ aad 11.9 m/sec. for aa
infinite disration ptilse, siaggest^ hov/ever, that a truly
coastaat velocity has not yet been reached, aad f--— !•'— -
comptitatioas are aeeded.
A large nujaber of computations of the cosir
Hodgkia-Huxley equations unvder vario^is coaditioEs i;.i:.;-i
laodif icatioHS ha-ye heen made with the Berkeley analog
computer. The analysis ia terxas of the interaction of
two siib-systems of the models p3:'eviously reported^ has
been verified and extended to e^cplain the effects of
iSiltiplyiag the time constants by various factors.
One snch modified systesSi. \5iith the time coiastiont for
the potassiusi activation 100 times normal and that for
the sodiiaia isiactivatioa 0.35 tlaies norsial^ nearly dupli-
cates experimental pla-tea'a actios potentials found Tvith
the driig TEA {Tasaki and Sagi^vara) , The saembrane
condiictaace changes are less accurately reproduced, smd
these are be lag studied fisrither.
The osciila-tioas obtained u'ith the ecfaatioas
lander a space current clsjap are the resitlt of the i;:..'.-
bility of a singular isoint ia the phase spa.ce. Thy®
the raage of coastaat current vBlzies vjhich prodiice an
inf iaita traia of iiapialses caa be predicted from ca-lca-
lations of stability. In additioa.^ a saall raage of
Part A. Project Desc3;ipti :.
cui'rei}.' J.ties results ia finite trains of
impulst-o. ...a. the mathematical basis for this phes-o-
menon will require further study. Under a resistive
clamp^ an intermediate condition between current and
voltage claanp, oscillations are obtained \?lien the
claiapiag resistance varies from infinity down to a
value betvveen 300 and 500 ohsi cm.^ while a threshold
phenomenon is still obtainable as low as 50 ohm cr-i.
Tills latter va.lue sets an aT>proxi!aate theoretical
upper limit to the permissible series resistance for
voltage-claffip measurements. Preiininary calculation,?.,.
assuming an ideal electronic control system, show
that anomalous inensbrane currents, including "notches"
and'bscillatioas'i can occur in a Hodgkin-Huxley axon
and an ideal electronic control system. With a
necessarily siaplified geometry ,, these anomalous
currents are found for a high resistance axial elec-
trode alone or in combination with noa-uniform
properties along the axon such as fiber diameter.,
condition of the aeiiiibrane or variations of the current
electrode impedance. These computations are similar
to observations of our ou'n and others on real axons
and are being continued for a more detailed comparison.
Significance to NINDB Eesearch: The concept and the
'meaiuremenis'"'15i~The""so5iii5^ potassiuns. ion axoveiaents
across the squid axon meinbrane offer a highly specific,
and general approach to the factors underlying normal
and pathological nervous processes which cannot as jQt
foe investigated directly in higher animals and man.
An analj/sis of the mathematical properties of the Hodgkin-
Huxley equations malses possible a better understanding
of their possible modifications and their liisitationSj,
than is obtainable by a purely physical interpretation.
Proposed Course of Proj^ect: The analog cozaputer will be
us°ed to^soXve" 'the~51o3gkin-Huxley equations under a
number of physiologically important conditions^, espec-
ially the changes of esr-citability near threshold. The
analysis of their mathematical propertiesj, v;hich has
already been useful in understanding experimental
results, \vill be continued, especially to investigate
further the conditions under vmich oscillations can
arise for both perfect, and imperfect space clamps. ArA
atteiapt v/ill be made to ir^odify the equations on the
basis of recent ^'Oltage-clamp experiments on the frog
node-j so as to provide a saathematical model for this
important case. The study of propagation of an impulse
in a non-space-clasaped noded nerve will be continued.
icluded "■-> yes
Proiect Resort
art B-; Honos'Sj Awards,, aad PubiicaixoBS
Publicatioas other tkan abstracts fropa this project;
Cole^ K. S,f Atttosiewics, H, h.^ aacJ Rabinowitz, P.
x'iiiitoiaatic computation of aerve Gjicitation, correction.
J., Soc. Indust, Appl. Math.. 6:196^, 1958.
Cole, K. S. Nervous system Sxcitation. aad propagatioa
of nerve. Accepted for pisbiicatioa hy Medical Physics
Vol, III, Chicago, Otto Glasser, Sd.^ Year "°5obk"?SSTisliers .
1. Laboratory of Biophysics
2. «-==.^-.«-™~
3. Betliesda;, Maryland
4. Continuation of 1957 pro;
PES-HIH
Individual Project Eeport
Calendar Year 1958
Part A,
Project Title: Correlation of Acetylcholinesterase
IsaSiibitioa With Herve Action
Principal Investigator: Whitcosab, E. R.
OtSaer Investigators: Moore, J. W. aad Friess^ S. L.
Cooperating Us^its: Kaval Medical Research Institute
Chemistry Department, Mount Sinai Hospital
Man Years:
Total : 1.0
Professioaal: .8
Project Description:
Objectives: To detexmine the inhibition spectrrai of
acetycholiaesterase drugs aad to detexiaiae their
blocking action pa the nerve impulse.
T7~Frog Sciatic~lTerve: A length of desheathed frog sci"=
atfc ''nerve is exposed to a cosipotsad of selected geo--
metric and electron distribution. The time course aad
exteat of the depression of the A fiber actios poten-
tial aad the ehajage of its ooaductiOB time are
compared vjith the In vitro anticholinesterase activity
of the agent.
^- §i^3.e Fiber; A single fiber from the toad sciatic
nerve TJi isolated and mounted across an air gap for
recording from a single node. The node is stimulated
directly at the rate of 1/sec. The time course and
amplitude of the threshold and the action current of
the node are studied as functions of concentration and
pE levels of the drug solution and unsaturated electron
distribution of
or Findings;
1. The effects of holothurin^ a neurotoscin obtained from
the sea cuciHSjber, were studied using the desheathed
bullfrog sciatic nerve and the node of a single toad
f i ber -
impulse ^-.i/.. ...:>.. .^^-..:.^',^..,....^ -^- - --
versible iBaaner, As the ilse ir
sciatic nervQ is being b:.,^.--..'.- .. .—^ ^.v. ....,rved by
decreasing height of tJje action potential, ti
GO noticeable eliaago in the velocity. On a s^....-.
nerve px*eparation, cojapounds like eserine sreduce
velocity as tbe action potential is decreased, ■
applied to the single nodo^ holotlrarin Bl)olishec
action current irreversibly Id. 1-5 in.5.nutes at a
concentration as lev/ as 0,5 win. This irrevers5.bl;2
effect was accompanied hj a fast rise in threshold.
The effectiveness of this xieurotosin is not altered
by pH level in the pH 3,7 ~ 7.6 ravi;-e , Again th:- -•
in contrast to eserine which is approxiiaately 1/
potent^ blocks in. a revorsib- • ^^^'"•■"ner ., and with a.
effectiveness that is depec the jM level.
The properties of liolothnrin are being si
in collaboration v;ith NBEtl and the Monnt, Sinai Hos-
pital,. Chemistry Departaeat. A report, '"Some Pharjaa-
cologic P3:*operties of Holothurin., an Active Neurotoxin
from the Sea. Cucuaiber" , of its properties is novi being-
prepared for publication in the ■' Journal of Pharmacc''-;-
and E;j:yeri.mental Therapeutics' ,
In connection v/itb. studies in stareocheiaical inflnencef
on the hydrolysis of cyclic acetates, the effectiveness
of the follov/ing compounds to block the conducted
impulse in the bullfrog desL ' " ' : ■ sciatic nei've T^as
determined
a) tetraaiethylstreptamine"2HCl
ramethyltetracetylstreiatamixie
ClG%0^^2^^S ^"^'^ • ^^02. 26
c) jiayoinosasiine
Cglij^^HGg jn.w. 217,14
d) scylio inosaiaine
CgH-i7N05 m.w. 211 .1 A
e) desoxystreptaiaine
CxG%2^2^3 ^'^^- SIS. 19
,/a ^fjl^O-i ,~,
' -rclohejranc
dlspla.;; ■■c-3'',, ■ this
set
.-- - /.aness -til
blGcki.£;g tise coaducced iapulst; . Tisis iuvesti-
gatioa v;as coxiducted ia col labor at ioii ivith HMB'I»
The report '-Furtlser 'Response of A^icetyldaoiiii-^
esterase aud of. Conduction In Bullfrog Sciatic
liei've to tlie Stereocliezaistry 6± Ajaiae Inhibitors
T. 1, '■■ is being jjreps^red for p>jblxcati02i in
A series of cxperisier^
arid i:a;piittjde ^of tlie actiosi Gurreat ai;.:
to deterjaine if the pli level could altei,* 'che
•-eSi'L^cnse of ffee node to the tertiary fors of
TIae vairiaiicas so far observed have obscured any qiii
gressi:. ^i© t^aresfeold increased,,
the tiae for e»eriB.e to exert its Kasiai'usi effect
■vas as le-Jifr s.s 20 mlmites i.33, sose trials,.
Complete reGOverj of tiie actioa Gur'^-ent aiupii--
i-ude f/as obtained after trr=eatiaent ia several
triais hut is ■ ^sly possible-
%ijdf icaBce to JIINDB Eesea^ii^: Tkese studies aid ±n
eTaXiIatT^~lEe~roTe~or ace€ylcliolinesterase la
nerve £iinctioji asid ia ciaai-acteE'isiEg tke coaditioa
for oae type of nerve iiapnlse blockade.
Course of Psfojeet: To coatin^ae lasing the de-
sheathed frog sciatic serve as a saeaiis for studying
tl3.s effectiveness of the propagated nerve ijapulse.
To modify the single node recording pi'ocedure ia asa
attempt to obtain a quantitative relatiOBshiL3 bet^^eeii.
the in vivo pjroperties of anticholiaesterase eompO'UE.cl<
&.nd , tEeir effect on the action curreat axid/or thrssho'.
To ccsabine the results of tfeese tn/o procedusfes tc
tfee role of acetyleholiriesterase „
■-luded ""■ no
1. Laboratoiry of Biophysics
3 . Bethesda^ Maryland
4, Coatiiiuatiosa of 1957 ps^oj
PHS-HIH
ladivldtsal Project Report
Calendar Year 19 5S
Part A.
Project Title: Membrane Potentials of a Lobster Giant .teon.
Principal Xavestiga.tor: Dal ton j J. C.
Other Investigator: Taylor, R. E.
Man Years:
Total : 0
Prof essioaai : 0
Other: 0
Project DescriiDtioa:
Objective : To test the possibilities of the lobster giant
aason as a preparation to be issed in general studies of
ion perjsieabiiitieSj to compare this axon with other
nerve preparations which have already been investigated,,
and to use the preparation in studies of the actions
of certain pharmacological agents \9ith special reference
to jaeaibrane oscillations and the productioH of
repetitive activity.
Methods Employed; Restiag potentials and spike potentials
~f rdss "the lobster giant axon are obtained by means of
an ix^tracellnlar microelectrode and recorded by con-
ventional electrophysiological instruments. A circu-
lation and cooling systezn perraits changes in ionic
composition an.d concentration, and the admission of
drugs. Electrical stimulation of the nerve may be
external or internal.
Major Findings; Any one of the three giant axons in the
lobster cireuiaoGsophogeal coiamissures iaalses a rather
simple, hardy preparation for the present investigations.
Xt appears to have good possibilities for further
experimentation on the types of ionic problems v.'h±r-h
have heen under investigation in this Laboratory.
The investigation of the effect of various
Eigents has been continued and concluded. Ethyl alcohol,,
acetone and probably dioxane cause repetitive activity^
a slowed return of the spilse potential, 'Without osci-
llaticns, tou'ards a reversibly decreasing resting
Serial. Ho, HI^iDB-B-^
Part A. Project Descriptioa (cont'd.)
potential c Tlie effects of ether aad cb.los?oforai v/e5?e
simiiar escept that an appaireat fuihibitory action
issiaall^ X5^®'^'®i^«ted repetitive activity. The actioa of
YeratridiE© was also similar biit aot completely
reifersible. The effects of procaine viQve a linear
S'iinGtlon of the smoimt of tlie tertiary f osra over the
pH I'ange 6.5 to S.7S isi^j^estigated. Tlie resting
potential decs'eased aad the action poteatiaX decreased
a.B.d lesgtlse3?.e<3 biat without chaag© of form is the late
phases .
Sigaif icaace to HIHBB Eesearcb : Tlaese iavestigatioas are
"" one aspect 'of~iSe general probieias of ionic perme^-"
bilities imioh. are tmdei' investigatioin In this Lalsoratory^
and^ as SBcfe^ are a mode of attack on the unflesclsring
q-uestiOHS of general nerire fnaetion. "Hsiey are useful
in pro¥idi3sg additional e^ideaee both of tise coamon
features and of the siagularities of ner^/e phenomena.
TIaey have a deeper siguif icaace in that they present
a possible transition betweea the iosic permeability
analysis now only available fox* the esotic squid ascos
and the neiirological problesjs of higher asaimais asj.d
man.
Proposed ConvBe of Project: The escploratory work on the
XoDS^Ser'"^OB. 'Ts largely completed aad has beea dis-
Goatiauied K?ith the return of the principal in-zesti-
gator to inaetiire duty. Soae more detailed worls on
procaine Biay be laijderttJJseu, and as nevi techaiques can
be developed or present procedtsres miiaiaty.s'ised^ the
i022.ic curreats la the lobster a^oa mosabraae will be
deteraiaed — to be tested as the coEsmon •andea.'lying
basis of the eoaveiitioaal phenomeaa novj be lag
investigated.
Part B inclisdejl
Isidividual Project Beport
Galoade.2" Ye&r 1958
Pa£t_^s Honors, A-wardSj, aad Psabllcatioss
Ptiblicatioas other than abstx'acts from tlnis projects
Saltosi^ John C, Effects of Bxtes-aal loss oe MeMJs^ase
Potsatiais of A Lobster G-iajat kKon, Accepted for
pufclieatioa la J. Gea, Physiol.
Serial Ho„ HIHD-B-B-5
lo Laboratory"©? Biophysics
2„ ^«„-^„=-
3 a Bethesda^ Maryland
4. ContinuatioB, of 1057 psfoj
PHS-HIH
Individual Project Report
Calendar fear 1958
Part A „
Project Title; loaic Permeabilities of Hodal Mesnbraae
Principal Investigators s Moore, J, ^hg del Castillo^ J.„
CClia„ Neurophysn)
Cooperatiiag Unit: Section on Clinical Heurophysiology^ JJIHBB
Man Years ^
Total; 0„2
Professional % 0 o 2
Others 0
Project Descriptions
Objectives s T&e measurement and the analysis of the ionic
components of the membrane current in the single frog
node to attempt an interpretation of the many phenomena
characteristic of this most useful present represen-
tati'y'e of medullated nerve fibers o
Methods Smployed; A technique developed by Fraaleenhaeuser
Tor "f he frog "node has been successfully adapted to nse
the improved electrosaeter preamplifiers and .operational
a^nplifier techniques developed here during the sq'uid
axon investigations o With it.p the essternal interaodal
leakage resistance is effectively increased to allow an
accurate jaeasiire of the nodal membrane potential » An
analogous technique was also developed to minimise the
ejsteraal leakage of the controlling current^ injected
into the nodal aeiabrane through one other internodOo
Major Fiadingsi A prelisainary study of the accuracy of
iSe voT^Eage claMp on the node showed the difficulty of
feedback contr-ol through the high interaodal ssseoplasBn
resistance, Vfith easily obtainable values of this
resistancec, the node potential and current showed
violent oscillations. With enough further reduction of
this resistaiice., adequate potential control and con-
tinuous current characteristics ivere found and v;ere
siiailar to tliose of the squid axon meHbranes, A manu-
script "iln Electronic Electrode" by Moore^ J» W. and
del CastillOj J. has been submitted for publication in
the Institute of Radio Engineers Convention Records
Part A„ Project Desc: :.,;.. ..-..ca (cont'd^j
Sigaificsace to KIMDB Research; Mednllated i:....-... -. ^..;_
are'sucli impo-irEaEt eojiipoaents of aeurological systems
that it is aot onij' interesting but practically
important to knovj if — and if 30,5 to what extent —
tSie concepts and inforiaatioia now available oialy for
the squid ascon also apply to the nodal jnembraneo
Proposed Course of Project; Fraaicesahaeixser aaad Dodge
Have estaFITsheci the basic similarities of the nodal
aad sqisid axon membranes » The project Bay be held in
abeyaace iiatil accumulated results on squid a.3eons
are analysed and codified »
.rt B iJselBcled ™ Mo
LABORATORY OF NEUROPHYSIOLOGY
Wade H. Marshall, Cfeief
Itie productivity of "the Laboratory of Neurophysiology
has coBtinued at a high level and either directly includes, or
touches upon loany areas of knowledge.
I^e Section on Limbic Integration and Behavior is roll-
ing in the graad manner lander the inspiring leadership of Dr.
Paul MacLean. A study of midline nuclei of the liiabic system
is under\5?ay. These systems are very important for integrated
behavior, Itiis research is particularly pertinent for the
general field of alcoholism (Korsakoff's syndorme aspect of
alcoholism) , and for certain aspects of the schizophrenic
syndrome. Dr. MacLean's laboratory studies oa this and other
projects are being largely done on the squirrel eoakey, the
use of which he has developed into a very cosaprehensive tool
for correlated observations on physiology and behavior.
Another important project is the investigation of relation
of visual apparatus to limbic system.
This Section is carrying out an. extensive series of
experiments involving biochessical differences in various
regions of the brain using ani^^ls ranging from souse to
monkey. Specific chemical poisons affect different regions
accordiag to dominance of differing saetabolic processes.
These researches contribute to basic knowledge of the brain
and have specific implications for problems of alcoholism and
drug addiction. A study on brain s^chanisios involved in se^cual
activity is underway ia the squirrel monkey. The precise
aechanisaiis i» the brain involved in sexual activity have re-
mained elusive until very recent years, and these studies
proMse to contribute real inforBm.tion on brain laechanisas of
this isaportaat function. It has long been realized that the
persistance and relative aperiodicity of the human sexual
appetite, as compared «/ith lower primates, was one of the
strong socially integrating forces in the evolution of human
society. The importance of sex ia psychiatry requires no
elaboration.
In the Section oa General Neurophysiology certain studies
are being carried out v/hich complemeat the above discussed re ■
searches oa the limbic system. .The large pyraiaidal cells in the
hippocampus are attractive ©les^nts on which to employ unitary
electrical analysis. Itiese neurons are otherwise uniquely
arranged for experisjeatal analysis in that the basilar dendrites
are arranged dorsally and laterally and long apical dendrites
are directed centrally. Work is going forward to analyze the
direct electrical excitation, and antidroaiic excitation through
basilar dendrites and orthodromic excitation through the apical
dendrites. This section has also concluded work begun in the
previous year on the action of curare on the neocortex. No
~ 2 -
specific actioQ was foiaad aad this has beesi so reported la the
Traasactioas of the Asericaa Hetiroiogical Associatioa. . lavesti -
gatioa of th© direct cortical sfespoas® is being coratiaued. The
direct cortical response, first ia^estigat©d by Adrian, li®s had
a" peculiar history. It has beea cosasidered to represaat, pri-
aiarily, electrical activation of deadritic appendages of cortical
Eeurous. It has beea ius®d ©xtetasiyely as a, criterioa of cortical
reactiveaess by investigators doisg esperii^nts involving local
perfusioK of cortical vessels or perfusion of the whole braia.
If this reaction constitutes an adeqtmte test it should be
sensitive to aaoxia. Our tests have shos?a that it, in fact.,
is very resistance to anoxia compared to the aaosic sensitivity
of known synaptic transfer reactions. Work is also proceeding
to test the widely held notion that activation of neuron so^a
or axon process is not necessarily involved in the direct cortical
response. The direct cortical response of the hippocampus is
under siailar investigation. Work is also going forward on
direct estimation of K ion transfers in the neocortex during
action of drugs (Ga^sia Amino Butyric Acid) .
During the suamer the head of this section Emde a scienti-
fic mission trip to Braail to clear up some disagreesaents about
spreading depression with Dr. Aristides Leao, the discoverer of
the phenomenon. Dr. Leao clearly showed that tetanic stimulation
of the callosal system of the rabbit can initiate SD. "Bais is an
important point which we -have failed to confirm and about which
there were no reports of confirsiation from the several other
laboratories which have worked on spreading depression. Soae of
our special technics, the triangular pore electrode assembly for
direct cortical response work and the Bak high input impedance
amplifier were introduced to Dr. Leao's laboratory. Six lectures
and semnars were given by m& in Brasil . At the end of my tour
of duty I was awarded by the Brazilians a trip to the Indians in
the Upper Xingu Basin as a sseaiber of a special medical mission .
I saade sosie curious observations which may be worked up later in
a formal publication. Aiaong these observations is the identifica-
tion of what will probably be accepted as the aost ancient trans-
quilizing agent. I was also impressed with the nany opportunities
for various kinds of research among the priasitives of Brazil where
several decades of courageous and kind treatsient of the Indians
by Brazilians has sjade the Indians friendly and cooperative .
Dr. Freygang is currently working' on ion exchange patterns
in E^sjbranes of single muscle fiber preparations. There are so®s
distinct advantages is using such preparations and he will con-
tinue this work during the coming calendar year in Cambridge
University J England in collaboration with Professor Hodgkin.
Further work has been done on Freygang 's hypothesis ad-
vanced last year and based on excellent laboratory work that the
dendrites and a large part of the soa^ dendritic aseaibrane in cer-
tain classes of neurons are not electrically excitable (see last
years report M-NP-GN-3) . This work has been confirmed hj further
«> 3 -
iavestigations by Freygasag aad Frank ■using Frank's tecliQic of
placing one electrode iisside a sseuroa and aaotbier electrode just
outside of the membT&nQ. These experisi^i&ts done oa the anterior
horn cells of the cats' spinal cord confirmed Freygang's conclusions
made oa the principle cells of the cats' lateral geniculate. The
major portion of the soaa sseEabraae reacts only passively during
activation and if the axon is discharged by the activation the soEia
aeabrane is not electrically esscited b.u,t remains passive . Itais is
a very important result in the current progress of knowledge. It
is now generally accepted that souse nerve cells operate in
this manner and that others do not. In the forssr type, it
turns out that the extracellular potential is an IR drop across
a passive ^mfcraae and that this current is the first derivitiv© of
the potential recorded intracellularly.
Drs. Frank and Fuortes are proceeding with use of clamp-
ing technics which show that the ^esbrane resistance does not
fall to a negligible value during the peak of the spike which is
in line with all above work of Freygang and Frank.
Drs. Frank and Sprague (University of Pennsylvania) have
found support for the idea that what is called direct inhibition
probably occurs through short process iate rne urons .
Dr. Frank and Dr. Pa ton (Royal College of Physicians and
Surgeons) devised Hsethods for admnistering saaall quantities of
drugs near neurons while recording from electrodes just outside
the cell and within it. This technic is useful for tests of
specific cheiaical transmitter agents. They fotmd no evidence that
gasma asiino butyric acid had any such specific action on anterior
horn cells of the spinal cord. Dr. Frank has been particularly
honored by an a-ppointEeat as a lE-eEiber of a scientific saission to
visit Russian laboratories.
The work of Dr. Tasski and his collaborators involving
the execution of critical experiiaents on the transjuission of
the nerve isapulse is now widely recognised as a major contribution
to science . The principle that active ©lectrocheMical forces
maintain activity in the nsT'^Fe laembrane, whi-sh is aost clearly
demonstrated in the two stable state experiaent now seejas to
have been proven. It appears that our confidence in, and support
of Dr. Tasaki has been Justified in every particular and that
his ideas are either accepted or seriously examined and considered
with great respect throughout the world.
Itiis group is also launching a general attack following
up their evidence that production of the serve action potential
is not a simple physical' phenomena but involves active biocheaical
processes. To this end they have another large scale program in
action, using intracellular glass pH electrodes, intracellular
and extracellular chemical isanipulatioa and electron parasiagnetic
resonance spectroscopy for detection of free radicals.
- 4 - . .
Aa investigator workiag as a guest in Dr. Tasaki's
laboratory at Woods Hole turaed up a very ijaportant decisive
argument iavolviag the two stable state concept of the nerve
membraae. Briefly this experiment showed that a fayperpolarising
respoase could be propagated if the external K ton was jaade
very high.
Dr. Tasaki and his collaborators are engaged in a general
attack on the processes iinderlying initiation of sensory nerve
isipulses in the retina, the cochlea and the skin, on all of
which Dr. Tasaki has niade important contributions for many years.
This section has in the past year seriously attempted
to do electrical analysis of neuron and glial cells grown in
tissue culture. The results were sufficiently premising to make
continuation of this work an urgent matter. In tissue culture
preparations the cells can be chezaically manipulated and the
electrodes can be placed under visual observation. This latter
point is very important. Dr. Chang has continued tbe observations
on glial cell excitability in the cat brain.
Bs'Bo Tasaki and Spyropoulos^, who have been invited to
write th& section on nerve excitability for the 1959 edition
of Annual Reviews of Plifslology, were also invited to participate
in the International Biochemical Congrecs in Vienna during
August of this year. Dr. Spyropoulos accepted the invitation and.
also took the opportunity to visit several important laboratories
in Europe to gather better information for the Annual Review.
His reception was most impressive and, according to Dr. Spyropoulos,
indicated the very great respect with which his chief. Dr. Tasaki,
is regarded throughout Europe.
The Section on Cortical Integration is proceeding with
methods to stoi'e on tape patterns Of electrical activity from a
25 electrode array, recodeing them into brief diphasic pulses
and feeding this pattern back into the array as a stimulus pattern.
The Section Chief has proceeded with further work on psychology
and physiology of isolation, sojae of which was done as a consultant
of the military on space travel problems.
Dr. Strumwadsser is proceedin^i with studies on CNS mechanisms
in hibernation Hibernation is an important scientific subject
providing many opportunities to make crucial observations
on temperature and time factors in the CNS as well as fundamental
metabolic studies. It also provides important information
pertinent to clinical research on hypothermia techniques for
surgery. Dr. Strusawasser is also proceeding with unitary electrical
analysis of visual and auditory integrating mechanisms. This
woi'k is being done on the frog brain, and is already yielding^
important inforsnation on fundamental problems of attention,
habituation, discrimination and e:stinction.
MATIOHAL IHSTSTOTE Of NSUROLOQICAL DISM3SS MD BLIMMSSS
Lftboratory of Neuropbfslology
Section 03£i Brai% St©8i
Eatiaatgd Oteligfetioi&s for FY 19§$
Total; $19^000
Directs IS.SOO
l®iab«8rs®a@at : §00
Individual Projects - lone
MTICMAL IHSTZT^TE OF HEllJaOLOGZCmL DIJSEASIS MB BLINMISS
Eatiigiatgd Ql>ligatioBS tog F¥ 195g
Totals $85,000
©iFect: 70,500
Ssdividffisl Projects 3^ 4^ aad S
HATIOH^L INSTITUTE OF liEUBOUSI£CAL DSSMS^ MB BLlWmEBB
. Labofatery of Nsaspop^ysiology
iestioB osi tb@ Spiaal Cosrd
Estisiat@il Ofeligatloas for Fl ISSS
Total 2 $76,S©0
.: IS,O0O
Ig&divid»&l Fs>@J@ct@ la, ' S aii«l 4
1.
Serial No, !fINDB-NP-SS-la
i„ Lab, of Neurophysiology
2, Special Senses Section
3o Marine Biological Lab<, j,
Woods Hole^ Mass^ ;
Univ., of Micho* Abh Arbor^
Mich,, and Bethesdaj, Md^
4o Combination of 1 and 2, 1957
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A„
1
Project Title: The chemistry of neural activity
Principal Investigators: I, Tasaki and Co S, Spyropoulos
Other Investigators: J, Chang, A^ Bak^ M„ Ezay and
R, Sands o
Cooperating Unit: Laboratory of Physics^
University of Michigan^ Ann Arbor
Man Years (calendar year 1958):
Total: 2„5
Professional: 1,5
Other: 1,0
Project Description:
Objectives: Experimental studies carried out in this
laboratory cast some doubt to the generally accepted
view that production of action potential in the nerve
fiber is essentially a physical pbenomenono The main
objective of this project is to find out the relation-
ship between biochemistry of the jierve and the process
of action potential productiono
Methods Employed: The method of intracellular injection
has been used to study the effects of various chemicals
upon the squid giant axon. The method of measuring
the iatra-ceilular pH with a micro-glass electrode
has been developed. The single fiber technique was
employed to investigate the mechanism of action of
various chemicals upon the vertebrate aerve fiber.
The method of "electron paramagnetic resonance spectroscopy"
was used to investJ-gate ■^hetker oa* not there is
PHS-NIH S.
ladividual Project Report
Calendar Year 1958
Part A. Proj, Desc. (cont, ) Serial No, NIHDB-2i;P- SS-la,
page 2
produetion of free radicals associated with nervous
activity.
Major iFJEdJEgs:
(1) Using an electrocheiaical model of the serve
(iron-aitric acid or cobalt-hydrochloric acid system) ^
the mechanism of action potential v;as iayestigated„
The results obtained indicated that many known
properties of the nerve fiber membrane can be
satisfactorily reproduced in the models The similarity
bet\&-een the electrical behavior of the model and that
of the nerve membrane is considered to support the
two-stable state hypothesis of action potential
production. This hypothesis postulates that there are
two chemically stable configurations in the excitable
membrane and that initiation and abolition of the action
potential represents transitions between the two states,
The membrane potential^, steady and transient^ is assumed
to be determined bj the configuration of the membrane
and the ionic environment »
(2) Usijag an extremely small glass electrode^ it
was possible to determine the pH of the a^.oplasra directly.
Both a calomel cell and a glass electrode (made from
glass capillaries sup^jlied by Beckiaan Co, of California)
were introduced into the aiioplasm of the squid giant
axon and the intracellular pH was determined by
measuring the i^otential difference between the two
intracellular electrodes. The intracellular pH was
found to be 7,4 with a probable error of 0.1 in pH
unit. The effect of various agents upon the intra-
cellular pH v;as investigated by this direct method,
(3) With a view to comparing the process of
action potential production in lower unicellular
animals with that in the well-differentiated nex've fiber ^,
the properties of the "hyper polarising response" in a
protozoa^ Koetiluca^ were investigated. With two
microelectrodes inserted into the cell sap of the
protoaoaj; xt was found that the passage of an inward
current through the cell membrane could induce an
all--or-noQe respos^se which represented a transient
lowering of the iatraceliular ootentialo This
PHS-KIH
Individual Project Re^jort
Part A^ Proj, Desc. (cont,) Serial No, HIMDB-MP-SS--]
page 3
hyper polarizing resijonse was associated with a
simultaneous reduction in the meMbrane resistaacCo
This response could not be eliminated by replacing
95% of the sodium ions in the surrounding sea water
with choline or potassium ions,
(4) The nature of the hyper polarizing response
in the squid giant a.^on (discovered by J„ Segal)
vifas investigated by the method of measuring the membrane
impedance dui-ing activity. It was shown that the
process of production of a hyper polarizing response
represented simultaneous variations in the membrane-emf „
and in the membrane resistance. The results of this
investigation were interpreted as supporting the
two-stable state hypothesis.
Significance to Neurology Research: This investigation
is expected to contribute to the understanding of the
normal function of the nervous system.
Proposed Course of Project:
So far J, the attempt to demonstrate free radicals
during neural activity was unsuccessful. It is planned^,
however^ to repeat EPR (electron-paramagnetic resonance)
spectroscopic observations under different experimental
conditions.
Using squid giant aiions^ the relationship between
the oxidative metabolism and the hyper polarizing
response in the nerve membrane will be investigated.
The investigation of the effects of various chemicals
upon the process of action potential production v/ill be
continued both on the squid giant a^-ion and on the
vertebrate medullated and non-medullated nerve fibers.
Part B iHclucled Yes<
PES-HIH i*»
Individual Project He port
Calendar Year, 1958
Part B: Honors, Awards and Serial No. NIHDB-SiP-SS-la^
Publications page 4
Publications other than abstracts from this project:
Tasaki^ I, and Bak, .A„ Discrete threshold and
repetitive responses in the squid axon under voltage
clamp. Am, J, Physiol. ^ 193: 301-303^ 1958,
Tasaki;, I. and Spyropoulos^ C, S, Non-Uniform
response in the squid axon membrane under 'voltage-
clamp', Am, J, Physiol,^ 193; 309-317, 1958„
Tasaki., I, and Spyropoulos^ C, S, Membrane conductance
and current- volt age relations in the squid a^ion under
'voltage-clamp' o Am. J, Physiol., 193: 318-327^ 195S.
Tasaki^ I. and Bak^ A. Current- volt age relations of
single node of Ranvier as e:i^amined by the voltage-ciasip
technique/ J„ Neurophysiol.^ 21: 124-137, 1958„
Eagiwara^ S, and Tasaki^ I, A study of the mechanism
of impulse transmission across the giant synapse of the
squid, J, ghysiol,^ Load,. 143: 114-137^ 1953.
Brady^ R, ^ Spyropoulos^ C, S„ and Tasaki.^ I.
Intra-a.ional injection of biologically active materials,
rsiol, , 194: 207-213. 1953,
Tasakij, I„ Conduction of the nerve impulse. Chapter
in: Handbook of Physiology^ (in press),
Tasaki^, I, Physiological properties of the myelin
sheath arad of the node of Ranvier „ Chapter in:
Progress in Kettrobiology, Korey^ S, R, and
Murnlserger_j, J, !„,, eds. (in press) „
ionors and Awards relating to thxs project:
Dr. Tasaki was invited to participate in the symposium
on "Membrane States: E^^citation and Inhibit-oa" held
at Washington University in St, Louis,, Missouri,
Drs„ Tasaki and Spyropoulos were in^^ited to participate
in the Second Int. Symposium on the Mechanism of
LiCitation held at Humboldt University in Berlin
(neither doctor was able to attend this symposium.).
PHS-NIH 5.
Individual Project Report
Calendar Year 1953
Part B: Honor s^ Awards^ Serial Mo, HINDB-NP-SS-laj
etc, Ccoiit„) page 5
Dr^ Spyropoulos was made a member of the Marine
Biological Laboratory^ Woods Hole,, Mass,
Dr„ Spyropoulos was a participant in the
International Biochemical Congress^ Vienna^ Austria
Drs„ Tasaki and Spyropoulos were invited to write
the chapter on Nerve S:fiCitation and Conduction for
the Annual Review of Physiology,, 1959,
6o
Serial Ho„ NINDB-HP-SS-3
1„ Lab„ of Neurophysiology
2, Special Sen^ses Section
3„ Bethesdap Maryland
4„ Continuation of 1957 project.
PHS-NIH
Individual Project Report
Calendar Year 1958
Part A„
Project Title: Investigation of the sensory mechanism.
Principal Investigator: I Tasaki
Other Investigators: C„ S, Spyropoulos^ M, E„ Easy
Cooperating Units: none
Man Years (calendar year 1B58) :
Total: 1,16
Professional: „66
Other: „ 50
Project Description:
Objectives: The main objective of this project is to
clarify the laechanism' whereby nerve impulses are initiated
in the sensory endings^ particularly in the cochlea^ in the
retina and in the skin,,
Methods Employed: (1) In the investigation of the cochlear
mechanism^, guinea pigs are usedo The surgical procedure
used to expose the cochlea has been developed previously in
Dr„ H„ Davis' laboratory Ccf„ e.g, Tasaki et al,
J„ AcQUst, Soc, isios l!i?54j, 26: 766), Comparing the cochlea
of so-called waltzing guinea pigs with that of normal
animals^ the source of the endolymphatic DoC„ potentisJ. has
been essplored, (2) Using excised eyes of gold fish and
carp, so-called graded retinal potential were recorded by
the technique described by Svaetichin (Acta Physiolp Scande ,
1956, 36, supply 134), It is planned to investigate the
photochemical processes in the fish retina by the us© of
the electron-paramagnetic resonance spectroscopy (in
collaboration v/ith Dr„ R, Sands of the University of
Michigan) „ (3) ' Using the technique for recording electric
responses from individual non-medullated fibers in the
cutaneous nerve (see p^ 126 in Tasaki, lervous Transmission,
1952), th© properties of the non-iaedullated nerve fibers
PHS-HIH Y„
ladiyidual Project Eeport
Calendar Year 1953
Part A„ Pro jo DesCo (coat J Serial MOo HIlDB-IJP-SS-3
page 2
arising Ijg the toad skin have been iavestigatedo
Major Findings;
(1) The histological examination of the guinea pig
cochlea used in the previous physiological investigation
was completed in Dr<, G, Rasmussen's laboratory „ It
was shown that there were no hair cells in the cochlea
of the walt2sing guinea pigs in which nearly normal endolymph-
atic DoC potential had been recordedo This finding
proves that the endolymphatic potential in the cochlea is
not generated by the sensory cells„ This is in agreejaent
with the previous finding attributing the source of the
potential to the stria vascularis„
(2) In preliminary experiments on the fish retina,^,
results were obtained indicating that the graded retinal
potential may represent an extra-cellular potential „ No
definite relationship was observed between the resting
potential (recorded with a microelectrode) and the
possibility of obtaining graded retinal potentials in
response to light stimulationo The naicroelectrode could
be moved a considerable distance without losing the recorded
potentials,
(3) Th© threshold^ chronaxie^ the duration of the
action potential and the conduction velocity were measured
on a large number of individual non-medullated cutaneous
nerve fibers^ The analysis and a statistical treatment
of the results obtained are in progresSc
Significance to Nein'ology Research; This investigation is
expected to contribute to the understanding of the process
of sensory perception.
Proposed Course of Project; anphasis will be placed on the
investigation on the retlSl and cutaneous sensory mechanism.
An attempt will fo© isadej, in collaboration with th©
physicists in th® laboratory of physics in the University
of Michigan^ Ann Arhor^ to test whether 'or not there is
prodviction of free radicals in photic stimulation of th®
retina. It is planned to study the nature of adequate
sensory stijauii in the skin for evoking action potentials
in various cutaneous non-medullat@d asoas.
PHS-MIH
Individual Project Report
Calendar Year 1953
Part Ao Projo DesCo (coEt,) Serial Ho<, MIMDB-HP-SS-3^
page 3
An attempt will foe made to correlate the nmaber aad
the siaes of tiae noa-msdullated axons in. a Remak bundle
with the electric responses recorded from the saiae btmdleo
Part B included Yesx:s:3E
PHS-NIH 9,
ladividu-al Project Report
Calendar Year 1958
Part B; Honors^ Awards^ and Serial HOo HIMDB~MP-SS-3p
Publications page 4
Publications other than abstracts froui this project:
Tasaki^ lo and Spyropoulos^ Co S,, Stria vascularis
as source of endochlear potential „ Jp Heurophysiolo j,
March (in press) ^ 1959,,
Honors aad Awards relating to this project:
Dro Tasaki was invited to attend a Symposium on
Elec trophy sio logy of the Visual System^
10,
Serial lfo„ NIHDB»HP°SS-.4
1, LatJo of Keurophysioloii^jf
2^ Special Senses Sectioa
3, Bethesda^ Maryland
amd Galvestoaj, Teisas
4, Hew
PHS-KIH
Isadividual Kroject Keport
Calendar Year 1958
Part A„
Project Title: Physiological studies on the nervous
elements in tissue culture „
Principal Investigator: J„ Jo Chang and lo Tasaki
Other Investigators: W„ Hild and M, Wolfe
Cooperating Units: Laboratory of Heuroaaatoiaical Sciences^
Tissue culture section; Tissue Culture
Laboratory^ University of Te3£a.S"
Medical Branchy, Gal vest on^ Tessas „
Man Years (calendar year 1958) :
Total: „83
Professi?3nal: o83
Project Description:
Objectives; The technique of tissue culture provides us
with a unique opportunity of investigating the function
of the nerve cells and the glial elements in the vertebrsite
central nervous system under direct visual observationo
Furthermore^ the investigation of the gradual change in
th© activity of these elements during the course of their
developsaent from th© immature stage is ejected to give us
a better understanding of the behavior of mature cells^
The main objective of this project is to study physiological
properties of the nerve and glia cells in tissue culture aad
to compare these properties with those of the cells in vivO;,
Methods Employed; The method of recording intracellular
potent! als~'wSth liyperfine glass pipette electrodes is used
in the study of the nervous elements in tissue culture as
well as in the in vivo investigations,, Phase-coatrast
microscopy combTied with the use of micromanipulators
specially designed for the present purpose enables us to
observe the position of the micro© lee t rode in the nerve
and glial cell impaledo
PHS-IIH 11,
Individual Project Report
Calendar Year 19 58
Part A, Project Des„ (cont^) Serial No, HIHDB».lP-SS-4^
page 2
Major Findings;
It was foxmd tbat the astrocytic glias in tissue
cultiire develop a slow "electric response" to direct
stimulation with extracellular electrodeSo The nerve
cells in the same cultiire media (stimulated ia a simila.r
mamier) produce short action potentials ot the order of
1 msec in duratioKo The duration of the glial respoass is
roughly 3000 times as long as that of the action potential
of the nerve cello The amplitude of the glial response
varies with the stimulus intensity; using strong stimuli
responses of 30 - 40 mv can be evoked in a reproducible
manner o
Electric stimulation of the astrocytic glia was
found to evoke a slow contraction of the cello It was
necessary to us© a time lapse camera to demonstrate this
slow contractiono The duration of the contraction phase
was lo4 to 3o4 mino^ and that of the relaxation phase
was 6-16 min/
Slow electric responses which resemble the glial
response observed in the tissue culture material were
demonstrated with a hyperf ine microelectrode pushed into
the cat cerebral cortex^ It was inferred that the glia
cells "in vivo are capable of developing slow electric
respoffises as the astrocytes in vitro are^
Significance to Heuro logy Research; This investigation
is expected to "lead us to better understanding of the
function of the brain.
Proposed Co'urse of Project; In the previous experimeats
on the tissue culture material^ a considerable difficulty
was encountered because of the presence of a strong layer
of plasma clot in which the cells to foe studied are imbedded
It is planned to repeat the previous experiments using
tissue culture material in which collagen gel is used to fix
the cells on cover slips.
The interaction between the nerve cells and the
glia ia the cerebral cortex will be investigated.
Measurements of the electric impedance of the cerebral
cortex following a sti-ong electric shock is expected to
reveal some aspects of the behavior of the glias in the
cortex,
Pa.rt B included Yes
PHS~HIH
Individual Project Report
Caleadar . Year 1958
32,
Part B; Honors, Awards^ and Serial No, NIHDE-NP-SS--4^
Publications page 3
Publications other than abstracts from this project:
Hild^ W„^ Change J„ J„ and Tasaki^, I, Electrical
responses of astrocytic glia from the mammalian
central nervous system cultivated in vitrOo
Experientia^ Basle,, 14: 220-221^ 1958 „
Tasaki^ I, and Change J, J^ Electric response of
glia cells in cat brain. Science., 128: 1209-1210^
1958 „
Changp J„ Jo and Hild^, W„ Contractile responses to
electrical stimulation of glial cells from the
mammalian central nervous system cultivated in
vitro „ J„ Cello Comp^ Physiol „ (in press) o
Honors and Awards relating to this project;
Serial Ko. NINDB-KP-SC~3
1 . Lab of Meurophysiology
2. Spii&al Cord Section
3. Betfoesdaj, Maryland
4. Hew
PES-NIH
Izxdlvidtial Project Report
Calendar Year 1958
Part A
Project Title: Generation of impulses io spinal motoneurons
Principal Investigators: K. Frank and M. G. F Fuortes
Other Investigators: Ml. Becker and P. Nelson
Cooperating Unit: This project has been carried out with
the cooperation of the Ophthalmology Branch of NINDB. Dr.
M. G. F. Fuortes has devoted considerable time to the work
of this section.
Man Years Patient Days: None
Total: 3.0
Professional : 1.3
Other : 1,7
Project Description:
Objectives; To determine which parts of the motoneuron
contribute the various components of the recorded action
potential and the quantitative aspects of the changes each
part undergoes.
Methods Employed: Concentric micropipettes have been
developed which can be advanced independently and which
are small enough to permit both to be introduced inside
a single motoneuron. By measuring the curi*ents through
the outer pipette required to maintain the potential of
the inner pipette at any desired levels inferences can be
drawn regarding the extent of firing over various cell
parts. Some of the advantages of voltage clamping tech-
niques can thus be realized in studying the behavior of
motor horn cells.
Major Findings: Only preliminary findings are available
so far . These indicate that the earlier assumption that
the membrane resistance falls to a negligible value during
the peak of the spike is false. A transient in clamping
current occurs during cell excitation which has two com-
poaeixts like the "A" and "B" parts of the undamped
Individual Project Report
Calendar Year 1938
•Jart A. Major Findings (cent.) Serial No. NI1DB-NP~SC~3
potential. The "A" current transient follows antidromic
excitation over a wide range of clamping voltages indicating
that it arises from an area only partially clat&ped with
this technique. Repetitive firing of the "A" area follows
claiaping at depolarizing potentials. A relatively long
time delay Cl msec) occurs between application of a
depolarizing clamping potential and the development of the
first "A-B" current transient.
Significance to Neurology Research; This is fundamental
neurophysiological research designed to further development
of hypotheses capable of more successful prediction of the
observed behavior of the nervous system.
Proposed Course of Project; This technique of voltage
clamping in the motoneuron will be continued and extended
to the analysis of synaptic activity. It is too early to
say whether further information can be obtained from this
method on the repetitive activity of motoneurons^ but if
it appears feasible it will be exploited. On the basis of
the results of this study it is planned to construct elec-
trical models of the motoneuron for further comparative
measurements .
Part B included Yes No X
14
Serial lo. 1IMDB~KP"SC~4
1. Lato of neurophysiology
2. Spinal Cord Section
3. Betfeesda, Maryland
4 . New
PHS-HIH
Individual Project Report
Calendar Year 1958
Part A
Project Title: Effects of locally applied drugs on siagle
spinal motoneurons.
Principal Investigators: E. Frank and W Patoa
Other Investigator: M. Becker
Cooperating Units: None
Man Years: Patient Days: Mone
Total : 1.5
Professional: 0.6
Other : © . ®
Project Etescription:
>jec
aad inhibitor actions on spinal motoneurons.
Metbods Employed: Concentric micropipettes are used to
record simultaneously from inside and just outside a spinal
motoneuron. Test drugs placed betv/een inner and outer
pipettes are applied to the surface of the cell by ionto-
phoresis. Consequent changes in Eesibrane potential,
excitability J, meiabrane resistance and spike generation are
monitored by the inner pipette. Comparisons of drug-induced
changes with those following natural synaptic excitation
may permit Identification or elimination of a test drug as
a possible chemical transmitter substance.
Major Findings: No drug has been found which mimics either
synaptic excitation or inhibition when applied with this
technique. It was found that a number of factors affect
the movement of drugs in the outer pipette: lontophoretic
field applied, hydrostatic head, capillary forces aad
electro-osmosis. Because the control of these factors
requires a separate minor study for each drug tested to
determine the form and manner of applying it, the method
15
FHS~MIH
Individual Project Report
Csleadar fear 1958
Part A. Major Findings (coat.) ■ Serial No. liIlDB-HP-SC-4
is considered too cumber sorae for use as a geEeral survey
techaique. Px'eliminary fiadimgs on gasMia amiEO butyric
acid (GAM) indicate that this drug applied outside the
cell reduces first excitatory postsynaptic potential
(EPSP) J, resting membrane potential^ then inhibitory post-
synaptic potential CIPSP) and finally changes th© cell
SBembrane so that the outer pipette falls inside and the
cell is destroyed. There is ao indication that this drug
is the physiological inhibitor substance actioig in the
spinal cord.
Significance to Neurology Research: Identification of
transiaitter~substances woal3"be a~laajor step in the eluci-
dation of basic mechanisms in the nervous system.
Proposed Course of Project: Since this technique has not
proven practical as a general survey method it has been
temporarily discontinued. Should other techniques indi-
cate that a particular drug is a likely candidate as a
chemical transmitter or inhibitor substance, this project
can be reactivated to test the drug.
Part B included Yes Ho X
16
Serial Ko. NIHDB-MP-SC-S
1 . Lafe of Neurophysiology
2. Spiaal Cord Section
3 . Bethesda^ Maryland
4 . New
PHS-NIH
Izidivldual Project Report
Calendar Year 1958
Part A.
Project Title: "Direct" contralateral inbibition.
Principal Investigators: E. Frank and J. Sprague
Otber Investigators: M. Becker and C. Terzuolo
Cooperating Unit: Department of Anatomy j, University of
Pennsylvania Medical School
Man Years: Patient Days: None
Total : 1.5
Professional: 0.7
Other: 0.8
Project Description:
Objectives: To determine if there is reasonable time for
an inhibitory interneuron to act in the so-called "direct"
inhibitory pathway.
Methods Employed: Measurements were made of the latencies
of postsynaptic potentials elicited in motor horn cells by
stimulation of ipsi lateral and contralateral dorsal roots
in the Sg-So region of the cat's spinal cord. The possi-
bility or an interneuron acting in this "direct" inhibitory
pathway can be weighed against the time available for such
action.
Major Findings; After all other times have been accounted
for, there remains an extra latency of 0.3 to 0 . 65 msec in
the inhibitory pathway over that of the excitatory pathway.
It is considered a very distinct possibility that this
extra time^ while short, is occupied by conduction of the
inhibitory volley through an inhibitory interneuron.
Significance to Neurology Research: The claim of Eccles'
group in Australia that inhibitory and excitatory synaptic
knobs cannot occur on branches of the same afferent cells
17
PHS-MIH
iQdividual Project Report
Calendar ¥®ar 1958
Part A, Significaiace to Neurology Research (conto) Serial No.
NIM)B-NP-SC-5
and that this requires an interneuron in the inhibitory
pathway is supported by the present study.
Proposed Course of Project: Ho further work on this
project is contemplated.
Part B Included Yes No X
18
LABORATORY OF KEimOCHMISTEY
1958
Robert B. Livingston^ M.D.
Acting Chief
Wntil more space becomes available :to the lab-
orator^^j, it will not be fe^ible to establish the one
or two desirable additional sections and give a more
adequate woricing space for the t^o present Sections.
Until such a ^imejr-^©~liave-fead~t©- -aband©B-ou3?~seaz!Ch^
for a Laboratory Chief. During the year, we sub-
mitted plans to the N£H which reflect our space and
budgetary considerations pertinent to the esspansion
of this Laboratory. We are hopeful that a new -^
building will be constructed here to accommodate
some of the growing meeds of the Basic Research
Program and that such construction will be avail-
able for occupancy sometime during- 1964. The
Laboratory, of Heurochemistry will: at that time
be due for. major es^pansion. It. .Is our. plan in the
meantime y.. aft^r. construction appropriations have ..
been committedj to recruit and assemble together
the .necessary, personnel to make thiis. the most
effective, and. resourceful, establishment for the
investigation of neurochemical problc
Not a moment is to be lost in this enterprise
because advances in biochemistry and related achieve-
ments in other complementary mental and neurological
disciplines a3fe ripe for imaginative- esploitation
hy a strong tea® of neurochemists. Although the
Laboratory then will be much e^^anded and as yet
only two elements of the total breadth of interest
in this field are represented^ it is clear from
what we already have within the Laboratory and
within the Prograss theefctl^js s^paasion of neurochemistry
will represent growth iroa. strength- within this
mtial- discipline.
1. The Section on Physical Chemistry. Dr.
Alexander Rich; former Chief of the Section on
Miysical- :^^slstry-j was attracted away ^ • much- to
our regret-y, to -the- post- -of- -Associate Frofessor-
at the Massachusetts Institute of- -Technology.'.
8ot& hslor® sm<a aft®? fei® iea^iagj Dr. Riefe was
eoasidera&ly torsp a^ ft®re we^ by Ms «i#p&rtur@o
Ferhaps the outst^diiig differential Ii9ti»»eei tli@
two poaitionsg fifora Dx-o Bicli'« poi^t of ¥!««»
was tliAt he haS sover Isefore p&rtici^atsd is
itniTorsity Ilf® estmpt as a ®ti&<i«ato Mow It
vould b9 possiMe for him to Join aa ®2E«;®ll@ii%t
faculty, «zt®a«l Ssis t@a€l&i^ at ^hteh b» tm
very adept, as a liosia fide tesiure PTOfessoF^,
and sorroimd himsmWwJW& hmmi^ 4tciv9 of
graduate stttdeats. Mt&otis;l& W9 regret Dr,>
Bieb'8 leairingj, we are proud of tli® eauiiple of
our quality k» I9ill represent at MITo Moreover,
tSie le»so% is ^11 tskm& t&at «e slsould do all
in our power to Juetify t!»e feelissg aaong our
senior staff that they already belo^ to a
first rate faculty; and to continue to l»iiM
up the Hia as a high ^ality teaehiiig instmtaen-
talityo for aaay of our staff, these attitudes
and functions relating the individual to his
colleagues and to the Institutes a.r@ of central
inporti
We were vmsj fortunate In Imtti^ ahle to
recruits as a i^w Chief in the Action on Physical
Ghenistrys Uto Sidney Ao l^rnhard, Dtc Bernhard
received his Sli,Do Degree fron Coli^S;»ia University
in 10911 under a mc Postdoctoral f^illowship he
received further L>al»oratory training with Frof o
Unus Pauling at the California Institute of
Technology concerning the theory of specific
Molecular inter &ctiosi» applied to Mological
systess C19&1-1@53]) and with Dro lo Qitfreund
and ^of o f»J.W. Soughton at the IMiversity of
Casteridge, Ingland, on applications of this theory
to ensyise afmtmm C1S53-.I9M) o Dto lernhard has
had a variety of teaching experiences » Es transfers
to the Mm tram the Haval lS»dical Msear^ Institigte
where he was doing research in the Division of
Physical Moch^aaietry. Dr. lernhard was already
hefore this transfer quite closely associated with
the Section on Biysical Chesistry and with the
Laboratory of Cellular fhaneacology . Se is especial«
ly interested in the physical~cli«seic&l basis of
action of ennymes, the ^^aracterisation of active
sites on the mmsifmas ^^ i^ the physical -chesiical
sanipulations which j^ay account for e^^siatic a^tioiic
Dr<, Bers^ard took a leaine of ahatej^^ during
the first six months of hia s^ppoii^'^s^nt tm ortfer to
l.B'^'estigate tfee possibilities of iswltifuitictioffial
participation. 1b. specific eatalysis,, at the Depart-
aeat of Biophysios,,/ Wei^aaim Institute o.f Science y
Rehovotbij. Israel 0 Dro Beriahard will retinrn to
begia laboratory work at the IIH early ici Februaryy
1959 o iB the aeaatiaej, working closely with other
nembers of this staffs Dr, Eernhard has plaimed a
certain amount of reorgaiiizatioi^ of the Section for
sore appropriate space utilissatiosto As a closely
collaborating group^ DrSo David Davies^ Da^^ Bradley
and Stanley Glaiieer are conti^ulBg without abatement
their individual research prograies and especially
those which will assist Dr. Rich in the completion of
studies in which he was a collaborator. At the same
time they have begun phasing in activities coajoiniag
the interests of Dro Bernhardo Actually the shift
represents less a matter of program discontinuation
and substitution but rather a shift of €»sphasis in a
continuing program <,
Dro Gary Felsenfeldp much to our regret ^ also
left the Section on Physical Chemistry in May^, 1958$
to become an Assistant Professor of Biophysics at the
University of Pittsburgh, Dro Felsenfeld during his
stay here made many important contributions the last
of which involved theoretical considerations regarding
the likelihood of the' double stranded DHA molecule and
its analogues to suffer errors in molecular replication
by omission or cc^ission or misalignment of strands o
The Section on Physical Chemistry has continued
its investigation of the important nucleic acids DMA
and RHA for which it has already received an inter-
national recognitioKo For this purpose ^ they have
manufactured a number of synthetic polynucleotides tram
an enzyme of high yield and quality o This has permitted
a thorough examination of the structure of polyadenylic
acid. Several different complexes of polynucleotides
have been re-examined and verified by the Section^ giving
increased confidence that EHA»like molecules can assume a
helical configuration similar to DHA, Further evidence
has been elicited for the mechanisms by which the DMA may
be separated into two strands which then rapidly recombine
with matching substrate materials to form new strands as
in genetic replicationo Mew concepts of the hydrodynamic
behavior of large and complex molecules have been proposed
on the basis of studies on DMA and RMA, These same
molecules have also been examined carefully fr<M the point
of view of their linking with acridine orange and other
dyeso In collaboration with the Laboratory of Meuro-
anatca&ical Sciences, this study has been extended to the
vital J, and hopefully differentials staining of the natural
nucleotides within nerve cells in tissue culture.
One of the most important aspects of biologically
active protein systems relates to the way in which
the protein partly or wholly stirrounds metallic
ions and the relationship which obtains between the
metal and its protein coordination group » This
has been a major pursisit of the Section. Thus^
the relationship of cobalt to vitamin B12 smd
iron to cytochrome C is being carefully examined.
2o The Section on ffpAd Chemistry, The Section
on Upid ChemislEry^ under the leadership of Dr, Roscoe
O. Brady, has made notable advances in understanding
the synthesis and metabolism and the requirements for
such activities in lipid constituents of the brain o
Of special not© is Dro Brady's recent discovery that
aldol condensations of the Snoevenagel type« formerly
recognized only in organic chemical systems ^ take place
in the synthesis of sphingosine and fatty acids «
Undoubtedly^ such reactions will be found to play
a key role in the lengthening of carbon chains in
a large number of important biochemical substances.
It is already suggestive that this reaction occurs
in the formation of certain steroids « The discovery
of such an important general reaction offers an
advantageous example of the fruits of basic research c
The enzymatic synthesis of sphingosine from
palmitic aldehyde and serine is now established.
Similai'lyj the stepwise condensation of 2-carbon
fragments in the biosynthesis of fatty acids has
been shown to take place with activated molecules
of malonyl coenzyme A. Dr„ Brady has demonstrated
the steps and requirements for formation and activa-
tion of the malonyl compoundc One of the lipid
constituents of the nervous system having a high
turnover rate,; the inositol phospho tides,, has
been shown to be formed through intermediary cytidine
diphosphate diglycerldeso A further basic study
relates to the investigation of the mechanisms
of formation of compounds which contain aromatic
rings c Again; a gereral key to many biochemical
problems is being sought.
Dr, Iternard W. Agranoff^ in the Section on
Lipid Chemistry y is currently being supported by
the Institute for a year's training with Professor
Peodor Lynen at the Institute for Cellular Chemistry
in iSinichp Germany. Since his training with
Professor Lynen will include work on the determination
of oxidation and reduction of lipids toy meaiis of
certain dye techniques (available only in Professor
Lynen's Institute) j, we are looking forward to Dr„
Agranoff's return and the fruitful application of
these techniques to research within the Section
on Lipid Chemistry o
tSsGtioa on Lipid €h&mi.Btvw
Eeseag^h Goapletsd in tliis seatioa dug-iag the past fear
feas rciStjlted in; tlie disGOvarj of a sew biological meehaaiSB for
the ieagtlieiU^ of carlscm chaias, T^e a®wl.^'-deserabe<S ^©aotioa
is an aldol eoadasisatioa of th© Esosvessgel t^^p© whic^ ha^ sot
been pg»©vioi3SI^ detseted ia biological s^steaso Its fundaiaeatal
i'oie tn ^hjB%olog±Gal ' p:roQ©ss©s was f i^st eloeidatsd ip th©
eoijr'se of iav©st4gati©as on the eas^sstic sjstliesis ©f sphiago-^
siae^ the feasio ooaponeat ©f matei?ials called spMsgolipids
^Isieh ar© ispoi^tast' cosstitiieuts of the sjelin slisath of newv^So
Tiae l3ios;^tlj.esis ©f spljiEgosia© is qatal^sod fey es^-mes ps-esest
is b^ais tissy© and t&e esssistiai ^eactios is tn© ©oBd@asats.QS
of palmitio aldelifde witli £11 appsfopriatelf activated . molesul© .of
tlie aaiso acid seg-iae*,, 1?iiQ s©i»iSiiO is aati¥at©d hj th© fomatioa
of a Selsif f bas©-metai coiaples featwestx se^iis©^, pfS'id©zaX phoat- '
pliats. {^itsssis B^} aad • siaiiiases© touBi Hades' tlies© oosditioas^
th© B©tfes*'l©ae earboa atom Ccarbou 2) of ses-jlae beooiaes aegati-welf
Gliarged and is in a .f airoE-abl© coaditioa f ©r -eoixdesisatiop ^itli the
cas-boafl ca2?l3©B atora of palmitic aldehyde which dairies a poBi-
tive dipole sicm®nt« The pTodvict of tl^is s?eactioa is di^fdrb-
splii^'osia<s ^ieh ■sufeseqO'Satl^f is ©sidi^d t© splaiisgosin©^ ■
It was appa:^©at f2?esa th© coati'ibytioiis of sei?@i?al ^orBjes-s
ia th©' field of " lipcgeaesis that the secliaaisaj of biological
foraatioii .of t^ttf aciis i^eaiaised ^ses»laia©d<, Bs:p©s?l®©ats
rscoatlp' csapl@t©«l i^ this seetioa isdieat© tliat th© feiosfatfeesis
of fattf aeids p^ohoblj alsoocous's via an alSol eoMeissatioa. of
tlie Sao©^©^ag©l tfp©* ' T&b sssesearF ^©aotasats fof this pi?«3©ss
sii?© aliphatic al^Qh^des e©otaiaiag sa ©"^©a su^es" of ca^oa atens
^!ii©!i coadesse witli th© . activated ^©thf laa© carfeOB of sjaloayl ,
eoeagpj© A<, Tlie pro-soct. of the r-sastioa is a p-lifis'os:^ f att^
aoid de^ivatii?© of ©oeas^^ A-^ho^e ©Ixaia leagtli has feeea
increased Ijy 2 oa^bsn atoiES^ tli© ca^*bosyl cas?i>OE of malom^l
c.oQliEfiae A feeiag «3isplaa©d ia tli© coug'se, of the eoBdessi^ f'sae-^
tiooo 'S'lie. resoltiag"' )S05BF©os^ is dQhf&%ted^ asM tlje ^asatJiss^sted
dQirivative' is s?©dtjo©!fi ■with a laoledtil© of ' triphos^osf^ldisa©
aiiGleotide, Tfe© t&iol estes" of tli© a©wlf=^prod'Oised sati3X*at©<l
fatty acid is i^adBCsd wita 'asotli©^ aolee-ale o2 teip&osF^PFyi?idis©
Galoot ide to tlie ^©spaQti¥e alipiiatie al^elayde alid is thvis
capable of yadei'goiisg fos^tbe^ ooad©asatio& i^ith aaotlie? solee^si©
of saloayl eosa^^siQ A« Tk© iralidit^ ot thim ^e^atton s&hmk©
depends ui^<m the aliilit^- of tieey^s to eatalf g© th® " foi'sjatios of
til© f'sqioii'ed aaioajl eosssps© A fro-m asB'tfl eoe^fsa^ A ss?l dai'feoa
di©2:ideo' We sair© ^©©©stif deEioastrated tla© ^usj^jatie das^o^ia^'
tioB of acetj'i. c©©a^.i3e A to f©^a fsal^afl aoeasi^© ii ia t.lie F^es-^
©so© of ssagstesima "iops mid. ads^asais© •teip&osplJ.^teo
' Mm^theT' iBpoi^t^^t "aoatPiMitioB f ^©n tSiis seetisa d^^i^
the ^ast f©ar is tii© el^cidstioa ©f- th® aecbasisia of tli© feiosfB«
tresis of istositol g&ospljatides, fliss© isat©S"ials ai^ pr©s©at ia
SBl^&ttvolf high coBcentrEtioa In brain and nevve tlBBue^ Tli©^
as'e especially impos-tast feecatise of tlseir high metabolic tisniote^
rate. ' Accordingly^ these materials hB.ve feesa implicated as
participants lu the pli^siological iirocesses carried oa fey aerv©
tissise. It was discovered that inositol pliospliatides ar© forsied
tlsrough the partieipatioa of a 2i©w class of rsacti^© istersaedia^ir
aetaboXic coBpounds called cyti4isi@ diphosphate diglycerides
(CDF-diglycerides) 0 These coaponuds are cosspoged of glycerol^ 2
ssolectsles of fatty acids^ &nd the gietabolicaIi;sr Importsiut atscleo-
t'MQs cftidia® diphosphate ,, lEositol reacts with CBP-diglfceride
ia th© preseac© of the appropriate ©nsyae system to form inositol
phcsplaatide, Cytidiae laoaophospliate is split off In the coiwsm
of the reaotioao
iBotlJer projest »3nd®r iuvestigatioa at the ps-eseut tisse is
the study of the foriaatios of the gl-aco-=cereD2rosi<ie clas© of
sphiBgoXlpids in tisstses obtained from patients with Gaucher 's
disease,, "Oiar -iaitial investigations indicate tisat spleaic tissise
obtained frosj these patients does catalyse tlie syatkesis of
cej?-el>rosideso Tliis finding thevetove ps^dbahlf represeiits a
defissitive coatsrifoistioa to-??ards the eliaoidation of tlio ©tiologf
of tfei3 disease.
There is evidssc® Miich iadicat©^ that spbiagosiae ,
essihibits the properties of aa assti-coagi3laat„ We as:e begiaaiBg
iavestigatioBS to detsraine th© Ic^'el of fre© spliiagosiae in tla©
plasaa of normal isidi'ridiaals and patients with certaia lisiabrrliagic-
diatfeeses ia ordes? to t?F "^^ detersjia® if siach coaditions ©re
related to abEtorssalities of sphipgolipid i2©taboliSE§»
dtliQi? problem^ ci^vr^ntlf undoi^ i2^©©tigstioa- la tliis
sectioa 'deal i/itfe the ^echaaisss of the f oirfijatioa of 'cliolest^roX-
txud coispoimds . which oostaia arosjatic riags* Sttsdies hB.v@
recentl-g bee-m iisitiated oa tia© possible antigenicity ©^ ^©S'taiia
sphingolipidSo Sss-tia obtained from patients with sasltipl©
sclerosis" will he exaaiuod for the ps-esaace of specific anti-
bodies agaiEst tls@s© lipid, aatigen©*
NATIOMAL IMSTITUTE OF KEirf«)LOGICAL DXBEASSS MD BLIHDITESS
Basic Eeiseaych Prograai
Laboratory of Nsuafoehemistry
Section osi Lipid Chemistrf
Estimated Obligations for FIT 1959
Totals $86^500
Directs $72^000
B® iiibtirsement % $1 4^ 500
Individual Pro j eats - l^ 6^ 7^ 13~15
Serial Ho. MI.1DB-HC--1-
2o Lipid Chemistry
3, Bethesdaj, Mdo
4.0 ContiHuatioE
PHS-HIH
Ijsdividtaal Project Report
Calendar Year 1958
Part Ac
Project Titles Biosynthesis of Sphingolipids
3E»riacipal Investigators Roseo© Brad^
Other Investigators s Sbes'hard Trams
Cooperating Units s None
Man Years
Totals 2oQ
Professional? loO
Others 1.0
Project Description
Project g lavest. _
XSpt3F"costaiaing; sphiugosin©
Projects lavestigatiou of the forsiatioa of QompleK cerebral
Objectiy<
Bphingbiipids and tc deterssia® the rat© of tursso¥®r &ud
metabolic fat® of these stibstaaceSo
Methods employed s With ths use of new ea^paatie t@chiiiqti©s
'deV'e!Lbpe<k ia %klh laboratory for the investigatioa of the
forsaatioa of sphingolipids^ ©xperiment® ha^e beeo Goadwcited
oa the mechanism of formation of sphiagosine^ the basic
coastitueat of complex cerebral lipids of th© sphingolipid
class c Discoveries in this laboratory hair© demonstrated
the pathway of formation of sphingolipide called eerebro-
sides o Investigations have been initiated on th@ format i©E oi
abffionaal glucocerebrosides which acozmuXate in the reticislo-
endothelial cells of patients with Gaucher "s diseases
Preliminarlr etisdies along thesie lines indicate that splenic
tissue obtained from these patient® does catalf'ss© th® ^fis-
thesis of glucoasrebrosideso
« 1 »
6I©I®gIiar~iiF®ati©a. of ©phingosim® has b®®a ccaaplstQda
Tb.® r©s%lt® biaf'« lii4 to th® jiipc©^@rF of a a@w r@acti©a
for th@ biological I@agtb.@siag @f carb©® ©feais^o ftoi®
r@act.i®a tm aa aidol c®md©mgiati©a ©f tfe© fe@©¥astagal tfp@^,
a irsactiOB tamlXisx t© ©rgamie €la@mi-j@t^ which hM a©!. b®@»
pr®fiiD%P.@if tl©t@cted ia biolofie&l sy^t®®®*, Th® feiosymtte-
f^i® ©f sphiagosia® is ©at.alfs^d fey ®aaym@^ iaa brain tissM®
as&d th@ p,^iacipal ^©aetio® is a e®ad@asati@s b@tw@®m a
iss®l®eiiX@ of paliaitie ald@hyd® ^±th aa &ppr@pTl&t®lf acti-
^*at@d m©l©€?.iil@ ©f th@ lamia© aeid seria©*, S®ria© i®
acti"^at@<i bf th® formation @f a Sehiff ba®®«a8®tal c&mpl®^
b&twmmu @tf ia©^ pyridoxal pho^pbat®^ aad saagam^g© i@^@o
Under th©s@ coaditioa®^ th@ fflethyl©a@ carboa at«Mi^ c&sbem
2^ ©f ^@rla@ b®c®m©® a®gativ®ly charged aad im thvm is a
favorabl© c@aditi©s for eo^dsssatio® «ith th@ cafboafl
cas'bois atoiB^ ©arboa 1^ ®f paLsitic aM@hyd@ which carfl^s
a pssitiv® dipoI@ Mom©st« ^@ ism^diat® prodwet ©f thie
C0ad@sa®ati0a r@aeti©25i i© dihydr®®pbiag©@im© whicfe 1® @iys-
®©qti@iitly ©xidizad t© sphisg©sisa®«
Sigmifieaae© t@ H^agelogieal M@@aii@@ r@a©agchg Slp.C5©
spbing'oHpIHS cosffipfmaTas-g® pa]Ff~W th© c«p©a@iat® ©f
th© »y©lia mhrnn-m «>« a^r^©®,. -re^sarcb d©aliBg with %Um
©©chasi^B ©f th® f©s'ffiati®a ©f th®@@ ce^poisad.^ igi of utmost
tMporttt&Qm f®!- th@ %iad@f tafeisig of a rati®aai approajefe
towards th@ tr©atai@at &i d®iBy<gli2aati?sg dim^uMm mtSLt@mo
F%irth®na©re^ th@ fiadimg that tis-s«@@ ©btaised frciB pati^at©
wh® ar® aff liet@d with Iip®dystr©pl»i€ c®aditi©®@ @^eh a,®
Gaisch®r*s di@®a®@ d© appar@iitlf form ^5h©s® 3Eat@iPial@ 4a mitu
mvmt lead to a r@-©valwati©a ®f th® c©ac©pts ©®?ie@raisg fSi""
etiology ©f tfe@®® dis@a^@s aad )®h©mld ultiisataly pv&wtdm a.
basis for appropriate ©©s-rscti^© i8@ss«a'@®o
■ Pgop@®@d ©®^ire@ @f pr@j@et£ It i@ py®p©s@d t® c0atisis#
ia??@®t4ga€f©ii°"OT~m® losmatiom ©f e©mpl©s lipids^ Tifsmi®®
obtaia©^ fr^ p&ti©ats with Gaiseh@i*<'s dimum®^ Hi^sass-
Pick"® m.m®m®- nad^ if p®P^ibl@^ Tay^Saefe® di@@a©@ will fe@
®apl®y@do Fiirth®raii©r©5 iav©gtigati©a ®f th@ m^t&bQli&m ©f
e©iBpl@K lipid® im is®ysial aad dis©a®@d ®tat®® will b® iMJd©r=
tak®ao la this ^©gard^ cax'b©a<-lab@X@d radi©acti^@ i}phiag@»
mi.mm im at pr@s©at bsiag pr@par#d aad th© fat© ©f thlB
QomponmA will b@ imf©$stigat@do
Past B 4melM©d ¥@§ X H©
», 2 »
sj^iivt^^w:^
PHS-ISE
la^^ividtRal Pro^met Export
Calessdajf Year wm
Part Bs HoaorSp Award® amd PTisfelleati©^^
P«bli€ftti©as ether than Abrntrwatm trms thim pr©J®€te
lo Brady 5 Ho Oo asad ^w^ly Go Ja Th@ ©azyaatle ^ysitii@@ii>?^
®f ^pMag©sia@o J<> Biolo Chmsk. ^ gggg 26-3I,, 19§8,
2o Erady^, Ho O05 WormtcAp Jo ¥»« aad Koval^, Go Jo
Th® ©aaymatic syath®siii of sphlagosia® II„ Fisyth^r
stMdi©s OSS. th@ ffleebaaisB of tli@ reaetioao Jc BJelo
Cheaao ,, ^2h X072- 11076 ^ X0£8o "" ~
3o B«rtoa,, Eo Mo.? S©dd^ m. A„ j, amd ayady^ E„ O.. Studias
OB th® biosyathsisi^ of g&laetolipldso M@Mro.lQgyg 8jp
Si^pl@a©at 1^ 84-85^ 1958o _-.=»—=. =..
lac©i'p®ratioii of gElaet©s® isto galactoiipid^o
Honor® and Award® relatiag t© this projects
- 3 «
Serial H©o NIMDB-MC-6
2o Lipid Ch@iai^try
4o CofitiiQuatioii
PHS-HIH
ladividiml Ftq^^ ®ct Report
Calendar Yaar 1958
Part A<
Project Tltl®? Bio©ymth®©i® of Ar^aatie Compowad®
Principal lavestigators Romctie Brady
Otfe@r latest igat or ss Ho Wo Sl@g@lmaii
C©©p®ratiaf Ifeitgs Plaat Physiology Laboratory^ PiOia®©rimg
Il®g©arch Groiap,, Uo Sc Departaieat of Agri€mityr©|,
Belt@i¥iH©5 Md:,
Maa Y@arSc°
Totals 0o4
Prof ©ggioaal s 0 c 2
Others 0<>2
Projeet I>@scriptions
Projects la^estlgatiosa @f th® aechaaism of fOMsatioss of
ari^atic compoimd@o
OMectivesg To discover -ta® ©asyasatie patfeis^ay of foraiatloa
OS aronatic coaapcimd® whici do not ari®© via th@ ®@doh@pt^~
l©s® diphosphate pathway.
Methods ^iployedg Th@ ®aK;/iaatic ®ynth®®i® ©f emipl@x
aroffiaiic molecules will b®, investigated issiag staadas^d
@i)%^ologieal ia®thodiii a© wall as radioactiv® tr«5t€@r
t®chaiqta®®o
Major fiadiaggg It has b®©a ascertai&^d that labeled
acetat® ±m a preferential precursor tor tfe© biological
formation ©f c©»poi!iadii coataisiag b®aa©i3i@ rimgiio Th^r©
©xists^ particularly iit plasit tissue f,, a \^i©l©gical rout®
for b@az©ij.® riag forma tioa other than ^isi sedohgptiil©®®.
quisle ^ ehikisiic acid^,: etCa
«. 4 -
Sgrial H©o MIIDB-1C-®
Fafft A,, c©iitlmM@d
■Indlmpmnmnbllitf ot thmmm sat ©rials for @l@€;tr©s transport
si2^g©o'stsR that tfe®y Bight @x®rt a e®asid@rafel© r®l@ ia tli'§
«3ossdix€t40ffl of th® ?a®y¥@ isapislss®*,
Pi'@p@gi@d c@mf®® ©f project I Attrnw tte® ps'@pa.rati©a ©fa
®atl®factorF®Iiyi©""iyife« for th® foiwatioa of th®
fe©ss©ii© fi^g ccwpo^sm^diSi, it Is p:r©p®@©d to iav@®tigat© tlii»
r®acti®a pr®c@ss paytictilarly with .regard t® th® iiklife®©d
that th© r©actio?s ale© ©c€«r® bf 'waf ©f aa a.ld@l c0Bd©a©«8i"-
tiea ©f th@ i&i©@¥'effiagal typ®^.
Part. B is&cl^M®d Y©m M© X
» 5 »
Serial Ifeo MMDB-MC»-7
2» Lipid Chmmlmtxf
3, B@tb©@ds&^ Mdo
4 o C©at inwat ioa
PHS-NIH
ladi^id^al Pr@J@ct ll©port
Caleadas" Y@ar 1958
Part Ac
Pr®J@ct Titles Ifetsibolisas ©f laositel
Priaeipal Investigators B©raard Agraaoff
Otli@r Im¥®sti gators s H©a®
Cooperating Units s M®ae
Mas Years g
Totals 2oO
Pf©f©ggi©aals ioO
Others 1*0
PK-oject D@®criptioa
PgtQjeetg Iav®^tigatioa of th® metabolisa of iaogitolo
Objective's To l®ari& th@ aatur® of tfe© biO(gyath@tic path-
way for th© foymatioa ©f i^osltol-comtaiaiag cocspoiiads i®
sterv® and braia tis@i&eo
Methods effiplQy®dg A sp€M;tr©photometric ss®th©d for the
d@t@nBiaaii©a ©f iaositol ha^s b@@a developed ia thi@ labor*
ato:ryo la additios^ the m©tab®lisai ©f radioaetiv© iao^itol
hai! b©@a iav@@tigat9d ia whole aairaals^ tlmmw^ .^lic©e^ and
appropriate 9Qzya® pr®paratioa@ prepared fr^a ti^^me
extraotSc
Major fiadiagss Th@ pathway of the ©nzyisatic iaeorp©rati®a
©f i!&o@itol lato iffionitol lipidgi h&m b@®a @stabli@h©do
This iacorporatioi^ has b®®n i&vmd to be d@p@ad©at lipoa th®
pr®m@ne® o£ cytidine aucleotidea aad maga@@iuiB iosk^c It
hati b@©a discover@d ia th@ cour@@ o£ these iav@@itigatiomi«
that inositol phosphatide^ ar@ foiled through th® partiei°-
patioa of a ja@w typ@ ©f r^^active laterraediary metabolie
sosapomid coa^i^tiag of cytidia® diphosphate diglye©rid@:io
■^©s® coropouad® ar® coaposed of glycerol p tw® Molecule® ©f
„ 6 -
Part A^ continued
fatty acid® asid th@ metabolically importast is«cl®©tider
cytldin@ dipho@phat®o Fr@© iso^lt®! i-eaeta with cytidia©
diphosphate diglye@rid@ in tfe@ pr@e©ne@ ©f th® appropriat©
®iJ2iym@ ssy®t©ja to form iaositol ph©spliatid©o Cytidia© ja®a©-
ph0@phat© is split off ia th@ coisrs® ©f the reaction »
There has h@©n no prmvloxm description ©f aa ©xaetly aaal©-
goiis bioch©Hiicai yeactioa of this aatHr@o Iav@stigatio©s
in other labor atori©® hav® recently coiafirased oiir £iadii»g
of this si@chaaism tor th@ formatioa of iaosit©! phosphatide:?*
Sigaificaoc® to Neiarologieal Diseases research s Siac@
inosltol-containiag lipids have the highest tisrpo¥@r rat®
of all braia pho@pholipidja^ a study of th© iB@tab@lic path-
ways for the foraatiosa and fat© of these material® is
particularly iasportaat^ It has b@@a postulated that tto@
turaoT^er of this Bsat@rial is related to metabolic pr©©@ss©@»
peculiar to aerve cellSo
Pro|)08ed course of project? In additios t© desBoastratiag
the pathways ©i the foraatioii of iaositol phosphatid©^ it
is apparent from the stixdies completed that inositol im
incorporated iato compounds other thaa iaositol lipids o
la other words^ inositol may be lialced with peptide® or
perhaps aucleotideSo Th© Idestificatioa ©f momm of these
aiat®rials must b© undertakea^o, e^d it ha® beea demosistrated
that the processes whereby iuogitol im iacorporat^d iato
these materials i@ distinctly digsimilar tTom the m@qu®siG®
of reactioas required for the foraatioa of the lipid
iaogitol phosphatides p The pathway of th© forioatiosB of
coBspouads other thaa lipid iaositide® will be iii¥@stigat@dc
Furthermore^ aa attempt will be mad® to isolate aaturaliy™-
occurring cytidia® diphosphate digiycerideso Th@ir idea-
tificatioa mo ta^ h&m related upon studies with labeled
coiapouadPt, Further ia^e^tigatioa iato the isetabolic
reactivity of these materials will then be «S!id©rtak©Bo
Part 6 imclMed Y®m X No
^ 7 ^
S@3PlaI i^e »■ !r-,T(r>»! _. as?-
PHS-MIH
C%l@Ma? Y@as> X938
Part Bg HoRors^ Award® p aad Pablicatioa®
Publicatlosys ©th©!" thaa abstracts froia this p3roJ@cts
la Agyanoff^ Bo Wo auci Brady ^ R, Oo Stwdies with tritii^
labeled inositol o NenTQlogyg 8g S-uippXemaat 1^ 79~S0i.
1958o — -
2c Agraaoff^ Bo Wo ^ Bradl©y^ Ro Mo ^ aad Brsdy^ Ro G^
Tb@ ©Qzymatic systhssi® oi iaositol phosphatide® o
Jo Biolo Ch@iBo^ 233: 1077-.1083-, 19S8o
Agraaoff 5 Bo Wc Low Lsi^el tritiyja eoaatiag t@chi,iqw®S;
ppo 220-222o Chapter ins Liqiiid Sciatillatioa
C©mtiag^ N@w y©rk,p PergajsioHFrHs^ 195^ o '
Hoaor^ aad Awards r@latiag to thi?« projects
=. 8 -
serial HOo MI1BB-MC-X3
2o Lipid Ch©iBi©try
3o Bethesda^ Md^
4o H@w Proj®et
iKdi'ridiaal Proj©et Report
Caleaday Year 1958
Part A.
Project Titles Th© lff®et of Sphiagosiiffie oa Blood Coagtslatioa
Priaeipal In'cestigatorg Professor Sug®si Heebt
Oth^a^ lEvestigatorss Eobqo® Brady
Cooperatii^g Unites Hone
Man ¥@are
Totals lo3
Prof @8«iioiial i 1 „ 0
Others 0o3
Pr©j©et D©®eript4oa
Prpjeets lavoiitigation of th© antl^^eoagisla^t activity of
lphing^iii@o
Obj#etive@g To lears bow sphisgosin© reacts with tfe© eoia-
ponente" ireguired for tfe@ process of blood elottiag to d®laf
this ph^siologioaJ, phenomeooiio
■ Methods fgaployedg Purified sphisgosia® will b® investigated
ill speeif ic reaetioms required for th© elottiisg of blood to
atteaapt to ©stablisJi th% lociss of th© aEti-€oag«laijt proper-
ti®« of sphis^osia©a
Major f ip^agsg Sphisgosta© has be©ia id@stif i@d ais a
Somaliy^oeewrriBg aatl-coagtslaiato Also^ it has hmmm foasM
that sphiagosia© fo^ia® iaelwsioa eompouods ^th a siui^er ©f
losig ehaia lipid d@rivativ©So . It i® eoisc®i^abl@ that th©
ra@€haiii®i!i of aetio^ of sphiegosi^® is that of a e(»ipl@%iog
ageat which r©Bsov@s from th© mmqamwe.® of reaetioae invol'^ed
ia blood elottiisg^ a i£onaallf«o€€iirrliig conposest required
for ooagulatioiEo
Sigaifieaae® to Heurological Dis@a«e» r#a®arehi The fisadisg
that brea&dowB products of the myelin shea^^lsshibit pof®s^tial
aisti«coag«iafflt properties serit® iavestigatioiio
» 9 »
Part A., coQtiEtied
Proposed courae of pro j ee t s A Eaiero^aaalytical d@t@riaiEkatloia
3?op tHi"©®tiaiaiioB""oif' tii© aKotssjt of sphiagosia® is plaiaaa of
itofmal and certaia oases of fe@®oIftic diseases asid laaaif^sta-
tioiAS of deficieaeiei! 4n th© clottiag pi'oeeeii will be ®xaa-
isi©do Til© w®e of th© Isotop© dilution t®elisiiq?3e with '
tyitiated spMsigosiae will b® @ssplo^©d fos- this d@t©rmiaati©iXc
Part B ii8clod®d Y®m Ho X
» 1© «.
Serial lo. MIEBB^MC^^^M
2o Lipid crmmXstrf
4.0 l©w Project
Caleffidar Year 1958
P£>©jeet Titles Smfmatie Sfi»tli@#ig8 of Fattf- Aeid®
Coopes-atiog Halts? Moss®
V.a.n fearis
Totals lo5
Professional s e o 5
Project D©scifipti©ii
Pgojeetg Investigation of xh^ mm€h&&%m^ of tfe® fos^matioxi ©f
Ha^ali3°"tliat th© ©siifiaatio sfntlifiSi^ of lossgefeaiia fattf
acids o©e*MP® hf an, al4©l co'^d@issat4©sj ©f tfe© feo@ir#aa,g©l
tfp@ fo@tw<s«fia sliphatic aldCiS^cles suad ssalonfl eo#^EfK« Ao Thm
pT@m@ut ©bJ©et-4'^#® as-e to rys'ify t.bi© @©s,fm® sf^t®»a :F«ii|?oii«
s4bl@ fos'tlis s©E>i®g ol if^actio^s asci to id@istlff t.ii® liite*-
®®«liarf g>@a©tftat® iiss tki® i'roe'^i^So
Methods ^ ^^ ^ploy®d g A sfstl©''!© org&aii? proe^dyr® feas 'b@aa
3w©^pMTorll@ pff@paratic£i of radioaeti-^e aialosiFl. eoenaf^©
A„ TMii siateria,! wi?^! hm y^iafi as syfestrate for Um isiregti-
gatioa of t&e eosdag-satioE reaetioiio Alipto.at4c alsi^fefaet
fi'ora twa t<J fasis?t@eii ssarbos atoes ir* l©p.g"tlj. will fee ©.xatiiaed.
tow th^lw a&ilitf f.o pafftic:.p>at@ a.® substrate® 4si tfee
eosdsEisipg r@a«5ti©ji„ Comt-est.iofsal @ii;^f»e fra;eti©;itati©iii
proe©d«f>!ir® will to© yssed to pi^epare tfee fe-q^isit© ©E^fis©
.s^®t®ffi® to b® #a£a®.ta@do
Major fiydiiig@g It w&m dmmrbMtrmtmd im this lai9©fat©s^ that
feosic^siag®! tfp@o Tfe© s@®@s.ias^f .r®a©tafflts fof tis.® pffo@@gi)
^ 11 ^
P»i?t Ao ft: oat:
ajr© allp!sati€2 aldefefdes eoatatmiisi' ais #ve^ nmsib^r ®l.
■saloEi'l e.o®ag;^pi@ A.. Tfe® ps'-sdiset ©f tte i?©a.€ti©«s is
l@agtls lias |j©@s ia«r@as@d Isf ti?o eajrtoois at©a©o Tla® e© b-r
is d^Iiyd^atsd and tte® yssat^fat^d deriirati^® i«s
#et@f of th@ s@w3,f f©i5K!i@d satufat^d f attf aetd %m r<^tl&i * '
wita a .«i®e©Sid ssoleeisl® of t:s'ipliQ®.pte©pf^idi®© mG^lt;ori4i.
fom t&© s^espseti't'® altpteatie al^iSsSifd'gSo TU® ^ld»hy4<^ Ir
tMiM iia Fositi©© t© yM^i-g® a fyr-th#r iv^-mt^sff-.v* . .^ - ^3*'>
f«.i*s9atia.?a of th®s® lipidis S'@qM!i5?©i? to a saj©:?' dmgM&m l&#
p&s,"t id patios ©i' i©Bg elia4.a latt^ acid® eonta-iiilBg l5#t-."':-*'-
a kao-wl^dg© of tte ia©©Jiai3.i@m ©f th@ foijaatios ©f fattf aeid-A
Pwo^&&@d eoniF^^j-jf Pf©j®fJ" la addition "t© thm pf@pai?i'-i
^:trat@d tfeat ti®si8i@ ©utraetp icostais ©aapaes. wMmU cat;*
aad magi^esiym i©m@o It is tli©ygfe.t that thim -reaatiosa r-**' =-■'
P3FOC©©,® ia ti@@.ii@ ©^tracts is tfey® of eossid^rafele ■
It in ps'opos©d to att^spit t® p»;riff tfec* @]s^,^i8@ ei'StfeAK ■•
^'©spom^ibl® for thim r^aetiora t© ®tydf t&e st©i©lii«5ffl@t?^
©fetaia lasigfet i»t© ffe,® ffisefcaai®^ of aetioB ©f tte ^^it;*.^
©ltH0iig& th,® ©xaet ©atiir® of its pai"tt©ipati©s is at t.h%^
Fart B iieeliided f@s I Mo
~ 12 -
Individual Ps'ojeet Rspoft
Cal©adar ¥©ar 3.S58
Part^ Bg UonoTB^ Awards^ and Fublieatioa®
Publieatloas othe^ thaia a^styaet® from this px°oj|®ctg
Brady .9 Ro Oc Tte@ _ ©usspiatie syatli@si.s of fattj acid® tof
aldol eondensatious ?^o«o Nato Aeado Seio^ 44^, 993-998.,
Honors and Award® x>©latiiig to this pi-ojeetg
« 13 =
Serial Ho, KIlIB-MC~i5
2« Lipid Ch&mimtww
3o Bethesda^ Md»
4„ K@w Project
PHS-MIH
Ijadiiridtial Frojeet Bepoft
Caleaday Y@ar 195S
Part A„
Pro j ©at Title; Biomjnth'^Btm of Cholesterol
Priacipal Investigator? Eomeoe Brady
Other Investigators s Ebe^lsard Traajs
Cooperating Uaitgg Hoa©
Man Years
Total? 0^8
Prof ess ioaal % 0 „ 3
Others 0o5
I^oject Deseription
\9et% To iav©stigat© tfe@ isechasiisiB of conversion of
Proje
S€!Val'bai€ aeid to ofe.olesterol<
Objm
i#etiv#@g To ©Iweidat© t&© aaturi* of the €oad@ij®ati©a
^'tioii' betwees nol®e-«le@ of mevalofiic aeid for tfe© forsBa-
tios of fameseiie and squal©i%e which ar@ preeursors of
cholesterol c
Methods employed; Phospborylated isit«!riffi©diat©s ia the
proposed sequei^e of reaetioss betwe©a sjevaioaie acid aad
sqt3al®ii® ^ill b© prepared^ The meehanisitt of th® cosd@^®atios
reaction will b© investigated by siiitabl<^ tracer techiiiqis@®o
Major fiiadingsg The project has jwst beea iaitiat@d^
Sigaifieanc® to le^rologieal Disea®©® r^giQarcbs The iraportaue®
of cfeoleflteroi as a ooRpo>aeiit of the Myelin sheath of mwv@®
and itss relation to arteriosclerosis are well kisovno The
l:aoi?l@dg@ of the mechiassiSHi of it® torma.tlou m&j be of coneido-
erabl© benefit for the ®twdy of appropriate' therapeutic ageats
to facilitate itB foraatioa ia certain coBclitioiis or retajrd
its accwasulatioB ia other® o
» 14 -
Serial MOo MllDB-lC.^4,5
Part Ar, ©oBtiaued
ggoposed^ poisree of project g Model s?@actioss 'will b@ is,r«?@8ti-
gated ' ia' Vitro using 'appropriately activated 4isteriE@diai'f
<soi»poiiS¥e°°3S?XV@d £rois me-valosie acido Studies will b®
MiBdertaJKeitt on th® activatiou of carbon atoms w^ich participate
in tfe© actual carbois«»carboB eondeiisatio© r®aetioa for th©
fonaatios of ©qisaleae and iiltimat®!^ cholesterol o
Part B included Yes Ho X
= 15 »
NATIONAL IH3TITUTE OF NEUROLOGICAI, DISEASES AND BLIKDKESS
Basic Reeearch Program
Laboratory of Ueurochemistry
Ssetloa OH Ggaeral Meurochemistry
fatioas for FI 1<
Total: S253«000
Direct: « 210 ^ 000
Reimbursement s $k3^ 000
ladlvtdual Projects - 12 through 13
Isadividtial Project Ee,.
Caleadar Year 1958
Part Ac
Project Title: Tlsno-Motov Coordinatioa in a Lower
Varteferate o
PriEelpal Investigator; Robert B, Livingston,, Mo D. and
Magdolaa A, Iraayif. M. Do
Other Investigators: Moae
Cooperating B'aits: The Director of tfe© Co S. National
Zoological Park
(Dr., Theodore H. E©ed) .
Man Years (calendar yeas' 1958):
Total:
Professioaals loO
Otijer: 0
Project Description;
Objectives; To deteraiae tlie anatomical pathways lying
between tlie~retina aad the eye motor nuclei ^ and to exaisiae
toeha¥ioral chaages r©siilti:ffig from istterfereEce witli differ-
eut parts of tfecse pathi?ays « Lower . vertebrates provide
some adiraatages In hmrlp.s more limited circuits relatlag to
visiso-Motor coordiaatiouj yet tke Cfeajieleoa saust depend
upoffi such circuits to catch its food^ liviag flies wfeicfe m.
captures .by throwing its tongue long distances = The ana-
tomical organization of tlie eye ^otor nuclei and the
extraocular musclem In th® CfeameleoE appear to follow tlie
usual vertebrate Bch&m.®^ Nevertheless the Chameleon
possesses ass estrame degree of independeiEac© of eye iso^emeiits
and yet is able to direct both globe® toward the same target
It has generally been supposed that the aaimal was able to
utilisse binocixlar ®b well as uniocular double visioHc This
study represents an erample of our contiBuiag iaterest ia
the Eiechaiiisss© cf sensorimotor coordiBatiOBo
and &: dege?s@yatiOffi '-
^■'^ '"■ ?1:B!^^3MB-' ' sigiaiglcaBt differeoces
'iabc'iiaf aea sy food-catehiisg ability' ia
i'. , This appcis; -a true whether oEse ey© Is ms- ■
patched or whether oae ®fe is remoTedo
Owr a^atoaiieal fiadiags fea-v© co^finaed aad exteisdisd tfeosfe
of three pre'^ious isTsstigators asid ©ur o-wsi previous figgdirags
-•alatiag to structures iiit@rm@diat@ te-®twe©E thm ©y®s aad =• ■'^-
ei'© motor apparatos ,. T''h^ 'smiav ?8,'?d ma-^'t dir^et f!,fsK!RSfini5. r^, •
are aow recogaizab.l
B igaif ieaac® ^ ta^tfaie Program of th© las t_l ty ta ; Stmdies
of this ETSl""siT'a- Fi')«;T7i1:,o &n umiSm'-'^''^r!^^htf: 'ET"&<imecir i ran Ui^^
£rogos©d Cowxs® ©f Project; Watil It will to© posmlh:-
Cteaaftleosi ^ismo-iaotos" circ«i.its thim project will b© gi^©s ;.
3o it^-fekifciS'iiSi,; ^i\i^\i.&iSt.'
4. Nei;?
ladivicl^al Project Report
Calendar Y©ar 1958
Part Ao
Project Title: Vestibular Influeiaces on Spical Kecb
Principal latrestigator: Bo S„ GerijaEdt^ M» D„
Other Investigator.<3 : MagdolRa Irsuayis 15= D.» Sid Q1 '
^, D,j aad Eobert B, Liv:^
Cooperating Halts: Moae
Man Years (calQisciar ye&r 1958) :
Total; 1*2
^ofessional; 1<,2
Other:
Project Descrip'tioa:
Objectives^ To gain a better tinderstaadlEg of
jaervonas integration wlilcli takes place bet-^eeii ye;-
asd spiaal meejiaisisms lis thm fin&l composition o;?
ton@o pOBtiire.5 &.nd locoaEot4o2i<, By this stwdy it .
be possible to les,ris certain general prirsciplee-
to t&e tvMotlou of tise ©er^ous sjst©m<, Tlie k'
an esaiapl® of our coatia-aiag interest in tb^
sensoriaotor coordinatioa.
Metltods Biaployed; Stimulation of tJae vestibv^
is fey the ieehffiiqti© of Cseraaadt and AadsrEsoB In
catSo Natural ®tiiiu?^atioa as hy acceleratlos of
columns in th© seaiicircislar canal does sot provi&i -
syncferoay of isapalssa to perait an easy traeiag of ^.
along the fisnetionally iaportasat cesstral patlswayr
have been reeord^d froax a ^as-iety of spiaal meT^'?.
roots and small fil-sseats of tli© vesitral roots »
inpwt ©tiffiwlatioa has Iseesi applied to the ceralse'-'
various peripheral nerve® and dorsal roots.-
Ma^or i; .atioii i ssadss of t!i@
aat^iSaiS"- -.ar ©'^••e __ .,>oBses recorded
asd upper spinal eord, M aaal^sis feas 5s©©u, mad© of tl$e>
gross fuffictioaal pathways followed fey -vestibular ©noised
responses tr&m their pois?.t of ©atry ©a on© si«S© of th©
brainstem to their outflow via ©piKal nevwes alosjg both,
sides of the meurasis aad of tls.@ patterned respous^s of
both gasraa and alpha veatral root fibers o Tli©r© ai*®
difference® isi tlie efearaeter of t&e spinal omtpijt of
vestibular origin a® distributed to tSi© ispper aaad lower
limbs 0 Vestibular isapulses giT@ rise to faeilitatorf and
isshibitory patterns whicii meem to be lin&ed to jaeoJiaaisms
relating to r©eiprocaI spiisal activities aad wfeieb is-^oly?
comples interactions with local spinal reflexes „ ¥©r
isspulsee crossiag to th© contralateral side^ at the feu ._
level are siafficieist but are aot necessary for tte coistia-
lateral spiaal effects. BliminatioB of both fewlbai* and
local segmental crossiage leaves evidence for interst'diatc?
fuactiOEal Gervico-tfeoraco" lumbar crossings c 3»srprlsiJiglFr
the coatralateral vestibular a^clei do jsot tfeesKselve®
respondo Moreoverj, they can b© removed ^ithowt imterferi;-.-:
with either ipsilateral or contralateral spinal respoasas
to vestitelar gtiiaolatioiao Other evldeace reinforces th.m
import aaca of retieylo-sptKal pathways and vestibtslo-
reticular lategfatiouo
Spiixal vestifeislar responses are laarkedly iuf luer* ;
neck BiMSCle proprioceptors « Ijeagtheaiag of the d.
sauscles markedly iiafeibits or abolishes such respo:i,_.: _
laspulseis elicited by vestiljular aM peripheral aerve
latiois iffite^act '5?itfeia certain limited regions of the
cerebellum, Eeaoval of the cerebellMm or application of
silver nitrate crystal® to the pertiaejst eerebell^vc" r^^.^Ur
produces aa iisaediat© am^Bntatioa of tfe@ spiral \
r©sposs®j deuEOBstratiKig a tojaic ishlMtory c©.r*^&:s'
issfliaesac©.,
Sigiaificaace to the Prograas of tirie lastitiste;
mechasais®© msderiyiRg auscle toaej. post-are and lc«.;...,.
are basic neisrophysioiogical probieKSo A»y iiscr©a®©':"i
knowledge of aiechaBisus of circisitry in th© central lii.. /.
system and especially integration between the classical a.c<:
the diffusely projecting systems In asiimals wit^^'-'rj- ^vs-s --
anesthesia is of ©special current value o
Bdii««ohi
r
• LIBRARY
NIH
" Amazing Research.
Amazing Help.
http://nihlibrary.nih.gov
DATE DUE
1 0 Center Drive
Bethesda, MD 20892-1150
301-496-1080
OAYLO.B
PR.NTEOmUS.
Mar/Ian^ 200J4
NIH LIBRARY
4 0129 1397
^j\R - 3 1?^ -
Live Music Archive
Librivox Free Audio
Metropolitan Museum
Cleveland Museum of Art
Internet Arcade
Console Living Room
Books to Borrow
Open Library
TV News
Understanding 9/11