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;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 


clinical Research - 1958 

Clinical Director's Reporfc-— — — — Shy 

Services Given by the Clinical Investigations Unit — Shy 

Blectroeacephalography and Clinical Neurophysiology 

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 

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 

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 


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 


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 


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 


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 


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 

39(c) Studies on Central Nervous System 

Control of Intraocular Pressure,, 

(Anatomy of Posterior Ciliary 

Nerves)-- ~ — - — ---» von Sallmann 


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—- — „-_. 


von Sallmann 

van Alphen 




van Alphen 
van Alphen 







Tans ley 

Serial No. Title Principal lavestigator 

58(c) Functional Studies in Retinal 

Anomalies and Diseases 

(Electsoretinography, Adaptometry, 

and Perimetric Light Sense Studies)-^— Copenhaver 

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 







81 (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 


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 


Laboratory of Neuroanatomical Sciences 

Summary" — — 

Individual Project Report 
Serial No. Title 



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 — ———-—-.—«—--— 












Serial No . Title Principal Investigator 

NII3DB-NA-DR-12 Development of new histcchsmical 

methods Sidman 

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 

I3INDB-NA-DR-20 Psychological effects of asphyxia 

neonatorum in rhesus monkeys-- — ----- — __=-_ Bailey 

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 , 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:.;' 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 Neurop h ysiology 

fjuimnary- —-»--—--...---- — -— — --_- — .--.__ — — Marshall 

In dividual Project R eport 

Serial No . Title Principal Investigator 

NINDB-NP-SS-la The chemistry of neural activity — Tasaki 

NINDB-NP-SS-3 Investigation of the sensory mechanism——— Tasaki 

NlNDB-NP-SS-4 Physiological studies on the nervous 

elements in tissue culture---- ■- Chang 

KINDB-NP-SC-3 Generation of impulses in spinal 

motoneurons---- — - — • — —-—--—■— — « — - Frank 

NIM>B-NP~S2-4 Effects of locally applied drugs on 

single spinal niotoneurons-— ------ — »------. Frank 


Serial i-iO. Title Principal Investigator 

NINDB-IIP-SC-5 "Direct" contralateral inhibition- — Frank 


Laboratory c f Neuroche mistry 

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 

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 $128,000 

Faibliest&as^ end Beports 79^000 $16,000 95,000 
(Xttfoneatioa Office) 

Msroct Ts^iaias 39>500 39*000 


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. 


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- 

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 

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 . 


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. 


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. 


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 


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. 


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 . 


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. 


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. 


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. 


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. 


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 ) . 


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. 


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. 


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. 


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 
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scone of its e^ratlons by tralnixig youn^^^c- scientiots in pm^tieulsar 
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eone^ts of ^id;^iiolo£^. Basis traizdsg sM. in tl;^ £i@ld 
of iitjs Bsore acute smd id.Sespresd ccBimimiea.ble dlsi^seis offers sa:^ 
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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 


CslenSm* Year, 1958 

Collaborative Heseai?da 

latiffisal Xs^titute of Isurologlcal 

Mseases end BLlMness 

^tioaaX Institutes of le&ltli 

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& rt m a. 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 




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 

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. 




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 


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, 

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 . 


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 


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 

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, 


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 

Individual Project Report 
Calendar Yeax I958 


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 


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. 


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 


Individual Project Report 

Calendar Year 1958 


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 

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 


Individtjal Project Report 

Calendar Year 1958 


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, 

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 


Individual Project Report 

Calendar Year 1958 


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. 


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, 

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 


Individual Project Report 

Calendar Year 1958 


7. Project Title: Mirror Movements and the General Phenomenon of 

Principal Investigator: Robert Cobn, U. S. Naval Hospital, Bethesda, 


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. 


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 


Individual Project Report 

Calendar Year 195^ 


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 

PART B„ Kone 

Cgp-2 HIKDB ' Spifigmiology Branch 


liidividual Project Report 

Calendar Year I958 


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_ 


Indlvid\xal Project Report 
Calendar Year I958 


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 


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 

Individiial Project Report 
Calendar Year 1958 


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. 


Individiial Project Report 

Calenaar Year 1958 


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 


Individual Project Report 

Csilendar Year 1958 


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 


Individ\ial Project Report 
Calendar Year I958 


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\ 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 


Individual Project Report 

Calendar Year 1958 


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 


Individual Project Report 

Calendar Year 1958 


1?. Project Title: Skeletal Abnoymalitias With Motor Systam 

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. 


Kurland, L. T., and Myrianthopoulos, K. C.: Skeletal Abnormalities 
With Motor System disease . Neurology 8:727-733, 1953. 

CSlp-:? NIHDB - Epidemiology Branch 


IniilLvldual Project Report 

Calendar Year 1958 


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 


Individual Project Report 

Calendar Year 1958 

P<\BT A 

19- Project Title: The Detection of the Hetero zygote in 

Cerebroretinal Degeneration (Amatarotic Family 

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 


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 

PART B. None 

EatlK&te d €bligati«as for Wt 19^9 

fetal: |l65,000 

Beljaburaefflente: 28^000 


Calsades- Ye«v- ISS"? 

Bio»«tries Bresneh ^ Coliaborativa Hsseareh 

Natioaal lostitQta of Ns^r^iogiesl 

Disttasee and Bliadnaiss 

KiBtionsl Inotitistn-'^ o£ H«alth 


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 

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 

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 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.-, 


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«&lth 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 ou tside 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 


(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 

(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^rolog y 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 

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 


(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 

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. 


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 „ 


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» 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^be r®^ @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> distribution s - 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 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^y elo pB@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, Cba jp^ea os^ teproveEaggts in progra m 

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 o f applife^ati&na 
(arct including Field 


















T» Sfcafg asgleeaaents 

DTe Sdwin Hove (since Jfareh 13, 1958) 



of the 

l^tloeal Institute of neurological 

Diseases end BUsSzidss 


1. Bft fflreaB 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 

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 i a^ 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 app i lleatioas 

Ifeetlng Ko. Amoujit R 6.. -^ 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 mi l ». n il . II n iii ii i»i uM ii I i n i I n H i m —withi n H I M I i m m l y i n m «ni « ii 1 1> > n i T i-i« r i.i» n .fii ij . «j h j,i i n «j.»a 

TOTAL I1.5 2,8lj,220 25 55-6 1,^9^,092 53.1 

6» Stsjff as slgaments 

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-<^^ 


(Fiscal year ba^is) 




"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 


Ltiple Sclerosis & Other 
Oemylisiating t)i®easfe8 

arlng & Glance 


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 









318,810 T«^- 

(188,514) (kj^y 
(53,964) flea) 





























































Awarded t© d&te 

1958 .animal B«port 
of the 
Katiosal lB8tit«te of Neiaroiogiejal Diseases and Blindneas 

III. Graduate frainiag Sr^xts 

■^ • ,^^-?,^<^Q^ Mgagolog y 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 





Ho. itesiint 




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®— 1 2/31/^ I^599.,@5 


































1 6^,163 


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 


^ear of traisie^ 







eiiaical ^iimslo^ 
Sto. of Si?ais4§«® 









Average Stiii^esas 









?9€iatrie Hiaurology 
So. of l^e^tsees 









Avegaa^ Stijpas^ 



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»gti culax 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 

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. 




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 

[—- ^■— — -— ~-~"— — — ' 


fo Approvea 




Affioant ^ 


, AmoiintJ Ho . J 

Ifercfe 1958 







Jtoe 1958 







Hoveober 1958 














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 


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. ggograaala g 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&ltmt 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. @£ao gea aEd I siiprovem^gDts ig prograi as 

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 Objaetive e 

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 




^ Appsov&l 







March ]9:?8 
J?sne 1958 
Hovffifflber 1956 



^227, 3S3 



5?. 5^ 

11 M 









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 tst s a^ yaaftieu lar 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« 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 

I tfcb 

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_ 

5 T 

9w or R€vis®i 


^ AKt.., ApfSo 






sw or Revtsad 


)» Aatc Appo 




^ Koo Appo 40 


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.; ■^S,oji t.« t© be a'^' 

■:*■ 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® 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'/® 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 


New Off- Revlffisd 

^ Anrto App« 26 

Qtolagyiagology A :^ plieaticaaig 

Anto Rfeq<. 




NOo R«qo Noo Aj^ ^ 

^ No. 








^ Aato Appo Mk 

1 991^ 



$ te660 




^ No, Apfpo jO 


^ No» App» 2 



New or Revised 




f^ Aast. Appo 


5& Atato A|Jpo 28 

■■ l^tBiD ■ 



,.i.^« ^^.,50 


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 

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 


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 

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 assifflan eatB 

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. 


1958 AanuAl Report 
of the 

Nation&I Institute of MfsttrolofifLcal taeeae*© and BUadn««B 


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&lth 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^ 


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} QrmA 

CoEssitt«@ for fefplicsffite ia the laeuyslogical fJ^I-.^- 

Iir. Jmr&sim = Eac«cutit«6 S«cy©t®ry, Otolsr^ 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 



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 

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 

Sixth Maual Beport of Clinical Isi¥@stigatioES of the 
national Issstitute of leurological Diseases amd Blincimess 

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 •: 


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 

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, 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 


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 ^ 


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 


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. 


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. 


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 


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 

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 

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. 


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 » 


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 

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. 


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 


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 


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:^ 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« . 
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 - 


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 


"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 


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 

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. 


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 

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 


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- 

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 


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 


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 

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 

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 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. 


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 

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 » 


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 


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- 

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 , 


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 


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 


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 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 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 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 


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© 
(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 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' 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 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^!" 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 .19 59 
Totals $96,500 

- Direct! $84,000 

aeifflbursments $12^500 


Serial Noo KINDB'^l(C) 
lo Electroenespnalography 
and Glino Neuropbysiol 
2o EEG 

3o Bethesdag MaxylaM 
ho Continuation of 80C, 19?6 
78c, 1957 

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 Des cription: 

Objectives s Outlined in the title? described in previous 
reports and in the Methodo 

Me tn<?ds_emplo yB 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) 


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 

S ignificance to the program of the Institute s 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 proj ect; 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. 


Serial NOo NIKDB-l(C) 
lo Electroencepnalbgraphy 

and Clino Weurophyaiolo; 
2o EEG 

3o Bethesda^ Maryland 
ko Continuation of 80C, 19" 

78c, 1907 

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 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© /^ 


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 

Significa iEe 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 

P z'oposed course of the p roject; 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 


Serial Noo NIMPB^-aCC) 

lo Eleetrcf>enceptialography " 

and Cliiio Neurophysiology 
2o EEa 

3o Betiiesda, Maryland 
ho Continuation of 8lC^ 19^6 j 
79C, 1957. 
Individual Project Report 
Calendar Year 19$8 

Part Ao 

Proja pt Title s 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 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 Institut es Saise as in previoiis 

^'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- 

Serial Noo NINDB 2(0 

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, 


Serial MOo KIlTOB^lCe ) 
lo Eleetroenceplialography 

and Glino Nextrophysiologj- 
2o EEG 

3o Befchesdaj Tlaiyland 
ijo Continuation of 79C-1956s 
Individual Project Report 
Calendar Year 1958 

Part Ao 

Project T itles Elactrocorticographic studias in temporal lobe 
epilepsy and in focal cerebral seiztareso 

Principal Investigator; Cosimo Ajmone Marsan 

Oth er 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). 
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: 



Serial Noo JlJIB22sM5l— ..— — 

lo EleetroenciptSography 

and Glirio Naxirophysiology 
2o EEG 

3o Be the s da if Marylarsi 
lio New 
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 Invest igators Takayiiki F« Enomoto 

Other Ing-eatigatorss Cosimo Ajiaoaise Marsan =• Paul Gerin 

Cooperating Units s Nons 

Man Years (calendar year 1958); 
Total; 1 

Professional; 1 

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 ftrriiriga t 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 

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 

S ignifieanc © to the program of ths In stitute; 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 

P roposed course of the project s 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!© 

Individual Projeet Report 
Calendar Yeaif 195B 

Part B s 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 


Serial Noc NIMDB<=='g(G) 
lo Electro eneepnalographv 

and dirso Netiroph^siology 
2o EEQ 

3o Bethesda^ Maryland 
llo New 
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 

Cooperat ing 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) > 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 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 


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 Investigator s Ro Gordon Long 

Other Iiwestigatora s None 

Cooper ating Units ; Nojkj 

Man Years (calendar year 1958) 
Total?" 1 

Prof es sional : 1 
Other: - 

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 fi ndings; 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 

Significa nce to the pro gram 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 projetst s 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) 

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 


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 Title s EEG changes induced ; 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 » 

Significanc e to the program of t bs 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 manifest clinical tits and^ 
therefore^ caution should be exerted in such cases toward the s^on---- 
tinuation of a given steroid treataiento 

Prop osed 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 


Serial Moo_ _ _ NIKDBJC G) 

lo Electroenciphalograi)ny" 

and CliBo Neuropi^ysiology 

3o Bettesd0.j, Marfjlarid 
ho New 
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 

P roject Descri ptions 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 /^ 


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 

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 C Rq 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 i E,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 

Significa nee^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 

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 Descriptio n: 

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 Institute s 
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 


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 

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' 

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 


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 


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 


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 

Estiaated Cb ligatioae 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 

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 

Principal Investigators! Darwin Prockop 

Bushnell Smith 

Other Investigators s Andrew Engsl 

Mark Lane 

Cooperating Units s Kristof Abraham Electroencephalography 


John Oatesp Hational Heart Instituts 

Man Years (calendar year 1958): Patient Days (calendar 

Totals 1 - year 1958)? 40 

Professionals 1 

Project Descriptions 


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 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©,> 

Signific anoe to Neti rological. 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 A o 

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. 


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 » 


One patient 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 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 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 


The brain 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 „___________ 


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 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- 


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 


To study additional patients with this disorder and^ if possible^ 
obtain additional material for pathological stu<tyo 

Part B included No 


1 , Med icalTleuro logy Braacli 

2, Sectioa on Neurological 

Disorders Service 

3, Betfeesda, Maryland 

4„ CoEtiauatioa of l?IHDB-23Cc) 

ladividual Project Eeport 
Calendar Year 1958 

Fart A, 

Han Years: 

Pattest Days 



Prof ess iosial: 




Project Title: Tfe© Histopattiological aad Claemical 

Investigations of Meuroauscular Disorders 

Principal Itovestigators: G, Milton Sfey and 

Theodor Wanko 


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 

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 


' 1^'0„ 


TtjTotoKtc Myopathy 


Sscoplithalssic Ophthalsiopl@gia 




Familial Periodic Paralf'sis 




Central Core Disease 

Glycogeo-Storag© CTfpe SIo) 

IcArdle's Disease 

Late Spontaneous Ufop&thj 79 

B^ial Acidosis 


Addisoii'^s Disease 
Cushimg's Disease 
Amyloid Disease 
Salt Lcsisg Sepbritis 
Paroxysmal Ijoglofeiauria 
Weraer's Sy;adroKe 




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 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 

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 

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 

Individual Project Report 
Calendar Year 1958 

Part A. 

Project Titl e; Pathological Study of Intramuscular 
Motor and Sensory Nerve Endings in 
the Normal and in Neurtnnuscular 

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 < 

Pa tient 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 

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 


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 

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 

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 

Significance to Reurolof^ca l 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 

P rpposad Cpu rsa of Pro.leat s 

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^ 

3, BethgsdSj Maryland 
4o WINDB=-24(c) 


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®yed g 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 M afegrial s 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 

Part B includsd Y®» [__| K© [x_J 

Serial lo„ NXKDB-I8<c) 

1= Medical Mexirology Branch 
2, Sectioa oa Biophysical 

3= Bethesda, Marylaad 
4. Old Ko. irilDB~20Cc)) 


Individual Project Report 
Calendar fear 1958 

Part A. 

Project Title: lavestigatioms in Localization of 
Cerebral Neoplasia by Isotopic De- 

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]) 

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 

Eatimated ObliRafciona for FY 19 59 
Tofcal; $20 » 500 

Directs $19,900 

Reinibur6«a®nts $500 

2o Neuroradiology 
3= BethssdcB Maryls 
4 o New 

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 Em ployed 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 

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 Neurolo^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 

Part B included? Yes Txl No 

Serial No, _JiiiiNDB5-Jl9(c^= 

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 

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. 


Cooperatisag Baits: Noaa 

Man Years Ccalend&r year 1958): Patient Day® {calendar 
Total: o66 j®&t 1958} 

Professional : . 33 
Other: .33 

Project Description: 

O B JKiTIVEg; 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. 

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- 

Part B included: Ho 





Issdividiia.! Project Eeport 

Calendar Year 1958 



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-, 


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 

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. 

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 


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 

Individual Project .-..,...-■, 
Cai®adar ¥©ar 1958 

Serial .Wo.liroB-12Cc> 
la Medical Meisrologi- 
2, Ciissically Applied 

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 

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 


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- 

™ 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" 

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 

Principfai Investigator i Eicliard L. Irwia 

Other la v®@ti gators: Jay B, Wells &nd EeuTj J» 


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 .,„ 

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 

Taifoxiiat'iba "coac@iaing""iSen^ological' activity of 
substances which occur ±n asiiaal tissue greatly 
esihancas our knowleciga of i&orsial and pathological 

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 

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) 


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 ; 

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 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: ; 

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?-:; 

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 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'j ja Sodiiam N oa°Chlorid @ S paee 

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 

Solids, NoBfc€hlo rid.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:P3 S&® o| . , Pr gjeg.!, 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 , Ccm rs 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 


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 lle MgocIsOT to 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^eiM siuK ^ 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 Phara a^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 


Major Fin dinggg 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 

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, 

fat iest Ma terial ; Frois NM2B patients adsaitt®d for other 
purposes o 


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®. 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 


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, 

3o Towers 0; Bo 

Di8@assion [Cliaisal snd Pathological Aspects 

of -fosieitF. ^^^^ "Ageniaed" Proteisss aad Metfeioai!!® 

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 

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 
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 

igthod j,; See 5?'=IIIM}B-4 and 5CC)o 

Fatiea t latgrial; Admissions to NIM)5 wards specifically for 
this projects Pli^s patients adsaitted for otheKpi^oseSo 

Maler.FJBd laqs; 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^^ 


Caieadar Y@ar 1958 

fubli eatioMS; 
lo TeweSo Do Bo 

Pyridsxine aad Cerebral Activity 

2o Toiifer„ D„ B, 

The Nearoehmistry ©f CootuIsIvs States 
ia Folclio Jo CEdo) Ch eijejl. Pa.tholoav 
Si t|e W^.MMS. Sistm (^ IgterMl ioM-i 
Neuroohepicil S grjBog JM) 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 

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 

Projeet Descriptions 

Obl^eeti'gesj To eiaberate the role of pyridoxine CVitaffll: 
ia' eertaia seisare states hj both ia ^im and ,|j.xli?J, 


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 

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 

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 v itro 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 Prol Mtj 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, viv o and in vitro and C3) if available, with si®ila:c 
'"py^idoxine^dependeEt" patientSo 


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 



•■'-" ■'• - "'\, ■■-; :« 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 
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^OL^— — 

[Tidual Projeet Eepost 
ZsUndzr Year 1958 

Part B„ 


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 


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 

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© 

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) 


iBsdi^idaal f¥ojeet ieport 

Caleadasr Year 1958 

Part A„ 

Proj©st_T itles Physico^eheaiical studies @f Hiraaa C©rebi.'9spiaa.\ 

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 

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 M aterial ; 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 

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 
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 

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— ia tjgglai. 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©- 

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 

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 


Iradii^idual Project Report 

Calendar Year 1958 


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 % 

Total; 0,4 
Professional; 0^4 
Other s 

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 

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 Firadlg gs; 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, 

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 ; 

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 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.)_ 
Individtial Pi'ojeet Sepoi?t 
Calendar Year 1938 


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) 


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 ; 

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 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 


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 


MHIM, REPOK Calendar Year 1958 



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 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 


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 pat holOi| y of the iatgaoc^lar preasMge a 
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) gveitis o 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 


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. 


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: 


(^thaliBolo^ Bratjch 


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) 

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, p hy siology 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^ 


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&ltaSc 

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 


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„ 


§«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) 

IMividual Fi=ojeet Report 
Calendar Year 19^ 

Payt Ao 

Pi=®ject Titles Studies ©a Diet and Drag Ihdueed Ejcperimental 

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.ieetiveai s 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;.± 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 Einpleyed s 

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 
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 


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) 


Individual Pmjeet Report 
Calendar Year 19^ 


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 

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 . E inplqyisd 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 Sistitute s 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 

Pro.ieet g 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 

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) 

IndividmL PyoJe@t Report 
Calmdar Year I958 


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: 

O b.1eetives 8 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 

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 

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?ojeet g 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) 

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 


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 al o 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 Emplcrsred s 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 

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 

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» 

Proposed Course of Project s 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 

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®s 3 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 ? 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 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) 

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 

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" 


% Betfcssdap Marsrland 

4o New Plmjeet 

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 


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 
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;s g 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 

I¥ep@g@d Csfurae of PSi'o.lect s 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 

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 


Cooperating Units s 


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 

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) 

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) 


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 Studi es 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; 


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' 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 Pr ogram 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, 


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 , 


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 

Part B included Yes ^ No £J 

Serial Koo NIim=A5(el 

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 

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 

3e Bsthesdas -''asylard 
ho Same as RIT'1DB"66 (c) 

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^£ @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> 

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 

Hsisr© is seed f os= fte'ther 


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) 

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 Findinga s 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 Institute s 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 Cto u rse of Project s 

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 

3. Bethesda^ Maxylatsd 
^o Net? Project 

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 Bnplojyed s 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 

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 

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 

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 

Signi ficance to F^o^aa @f Ihstitute g 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 

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 

«. 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 

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 


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 

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 ?!• 


Individual Project Repoi^t 
Calendar Year 19^ 

Part A p 

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 Brnplgyed s 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 

Mitoehagidria s 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 retigolrm g 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® 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©a nee 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 

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 

E xposed Ceurge of P ro,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 Bnployed i 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 

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@ Prog iiram 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 o f 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^es g T© deteritsine \^ means ©f the electron 
microscope detectable changes in ^arionxs human c^strephi© 
Biuscle diseases as cess^ared with normal tissuso 

Methods E roplqyeds Ifeiterial obtained from traisele biopsies 
on htiman sabjects wjth muscle dystrophies ®f ■various kinds are 
iinmersed imsjediately after esccisiess in V^ soditam 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= 

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 

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 Institute s 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 Project s 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-?^ (©) 

Iisdiiridual Fi^jeet RepoH; 
Calendap Year 1958 


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(yed s 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 Ihstitute g 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 Project s 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) 

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^a ee 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«) 

Individual Pro jeet Heport 

Calaidsr I©&2? 1953 


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 

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» 

- 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- 

Si^nifiean ee to f^ograro.of Institute s 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= 

» it 

Proposed .Cotars® of .Py ^jogt s 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- 

. 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 


£'xp po ^< $ 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) 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 


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 

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 Empl oyed: 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 

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 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 se nse testings The absolute light 
threshold is determined for red and blue stinroli from 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) 

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 FladxT ga; 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 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 Prc r ^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) 

Individual Project Report 
Calendar Year 1953 

Part B o 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) 


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 

Sipalfic ance to Progg aia 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 

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&<° 

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 


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), 

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> 


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; 


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 

f . Selective isolation of the lesion at operation. 
go Fhotogr^hic and sound recording. 

h. Histological and chemical ezmBination. 


MajQg Fiad ings ; 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 


Major Fi ffidiags 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. 


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. 


Major Findings (cont'd); 

tiiporaL lot 

In teaiiporal lobe spllepsy, there is an ill-defined dis- 
turbance of body image 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.) 


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 


Mai or Findiags (c oat'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 Project s 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) 


Individual Project Report 

Cslandas Ysas 1958 

Part B s 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 


3. BeChesda, Msrylaad 

4. NIN]IB-26(e) 


I&dividual Project Raport 

Calendar Year 1953 


Project Title s 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© 


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. 


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. 


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. 


gpoposed Ciourse of she Pro ject; 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. 


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 


3o Bethesda, Maryland 

4. Same as NINDB 195? 26(c) 

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 Year s (Calend ar 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 E p^£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 F indings? 

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 


3o Bethesda.. Maryland 

]+o Same as NINDB 195? 38(c) 

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 Investigator s Jo K. Van Buren, M, D» 

Other Investigators; None 

C ooperating 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 This study is now terminated.. See reference given 

Signifi cance 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 project s 

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 


Serial No. /)////li B- & ^cj 

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;.-; 


3. Beihesdaj Maryland 

Uc Same as KINDB 1957 27' 

Individual Project Report 
Calendar Year 1958 

Part A, 

Project title ; Studies of Involuntary Movements 
Principal Investigator s J. M. Van Buren, M. D. 
Other Investigators; Maitland Baldwin j M. D. 
Cooperating Units? 

Man Years (Cale ndar Yea r 1958); 

Total; .25 
Professional; =25 
Other: o25 

Patient Days (Calendar Year 

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 
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 

Part B included Yes f^ No ftj 

Ssslai No, WimB 66 ic) 

lo SurgicaTTIeOTologr Branch 


3'. Bethesda, Maryland 

ho Saroe as NINDB 1957 39(c} 

Individual FroJBCt Report 
Calendar Year 1958 

Part A. 

Project title ? Pain Mechanisms 

Principal Invest igators J. M, Van Buren, M. D, 

Other Investigatorsj^ Mildred Blevins 

Cooperating Units ; 

Man Years (Calendar Year 1958) ; Patient Days (Calendar Year 

Totals olO 211 

Professional; ,05 
Others „05 

Objectives The essential interest of this stu(^ is centered 
ijpon methods for evaliiation of pain 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 

Individual Project Report 
Calendar Year 1958 

Serial No. NINDB - 67 

1. Surgical Neurology 


2. Primate Neurology 

3. Bethesda 14; ^^^r-.i^ 

4. NINDB~43 (c 

Part A . 

Pro. iect Title s Study of Cortical Intracellular Potentials- 

Principal Investigator ? Choh-luh Li, M. D. 

Other Investigators None 

Coo perating Un it 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 Project s 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 

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 






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 

Cooperatin g Unit ; None 

Man Years (calendar year 1958) 
Totals .25 
Professionals o25 
Other? ,25 

Proj ect Description s i: 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 

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 

2. Primate Neurologv 
3» Bethesda 14, Mar • 

Individual Project Report 
Calendar Year 1958 

Part A^ 

Prelect Title s 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,; 


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- 

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 

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: 

Individual Project Report 
Calendar Year 1958 

Serial No, NINDB__-.70 
lo Surgical Neurology 

2. Primate Nsuroloi.-f/ 

3. Bethesda 14, " . 

4. NINDB-41 (c) 

Part Ac 

Project Title? Neuromuscular Transmission in Hypothermia 

Pr incipal Investigator ? Choh-luh Li, M. D„ 

Other Investigators s None 


Man Years (calendar year 1958] 
Total ! ,25 
Professionals „25 
Other* „25 

Project Descriptiong ■ 

Patient Days (calendar year 1958) 


[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 ^ 

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 


2. Primate Neurology 

3. Bethesda 14s Maryland 

Individual Project Report 
Calendar Year 1958 

Part A. 

Project Title ; Effect of Cooling on Conduction of Impulses in Cranial 
and Peripheral Nerves » 

Principal , Inv estiga tor ? Dr, A. Ortiz, M<, D, 

Other Investigator i Dr» Choh-luh Li, M«, D. 

Cooperating Units; None - 

Patient Days (calendar year 1958} g 

Man Years (calendar year 
Totals ao 
Professional » oOS 
Others .05 

Project Descripmon s 

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 Findin gs 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* 

Signific ance to Neurolo g i cal 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 Project s This study will be continued and may be extended 
froiD nerve fibers to nerve substance » 


No k1 

Individual Project Report 
Calendar Year 1958 

Serial No, NINDB - 72 (c j 
1„ Surgical Neurology 

2. Primate Neurology 

3. Bethesda 14s Maryland 

Pa rt A. 

Study of Pharmaceutic Agents Acting on Various Cortical 
and Subcortical Structures of the Brain. 

Principle Investiqator t 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' 

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> Neoro loqical Research ; to be seen 

Proposed Course of Projects This study will be continued. 

Part B included Yes [J No ^ 


Individual Project Report 

Calendar Year 1958 

Part A. 

Projact Titles Properties of C 

Principal Invest i cjators ; 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 Descri ption s 

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. 


Met hods Em ployed 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 t o 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 

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 

Ee£iiaat@d Obligations for FY 1959 
Total: $119,500 

Directs $55,200 

Rsimburssment : $64, 300 

Serial No.J!f!!^LlZiiiL„ 
lo Surgical Neurology 

2. Clinical Neuropathology 

3. Bethesda^ Maryland 

4. New 


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); 
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, F indings; 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 

2» Clinical Neuropathology 


3, Bethesda, Maryland 

4. KINDB 35 (C) 


Individual Project Report 

Calendar Year 1958 

Part A. 

Pro.iect Title; The Localization of Myosin i n Human 
Striated Muscle by Fluorescent_.Antx~ 

Prin cipal Investigator; Igor Klatzo, Mo D. 

Oth er In vestigators 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 Ye ars (ca lendar 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 

Proposed course of th e project; This project is 

Part B included; Yes ^ No [^ 

Serial No, ^I^™ 75 


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, 

Honors and Awards relating to this projects 

Serial mo„ WINDS - 76 (C) 

1. Surgical Neurology 

2. Clinical Neuropathology 

3. Bethesdaj Maryland 

4. NINDB 37 (C) 

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); P atient Days (calendar 
Total; .25 year 1958); 
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 


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 

2, Clinical Neuropathology 

3o Bethesdaj Maryland 
4, New 


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); 

Professional! '25 
Others "25 

Pro.iect De scriptions 

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 

2, Clinical Neuropathology 

3, Bethesdaj Maryland 

4. Formerly NINDB 33 (C) 


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); 
Professionals .05 
Other; .05 ct 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 

2o Clinical Neuropathology 

3o Bethesda, Maryland 
4. New 


Individual Project Report 

Calendar Year 1958 

Part A. 

Prciect Title? A New Me thod 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 
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 


2. Clinical Neuropathology 

3. Bethesda, Maryland 

4. NINDB 29 (C) & 30 (C) 


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); 

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. ^^^^ ^ 


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 

2« Clinical Neuropathology 


3. Bethesda, Maryland 

4, NINDB 31 (C) 


Individual Project Report 

Calendar Year 1958 

Part A. I 

Prelect Title; Study of the Effects of Hypothermia on 
Injured and Normal Brain Tissue. 

Principal In vestigator; Edward J. Laskowski^ M. D. 

Other Investigators; Igor Klatzo, Mo D. 

Man Years (calendar year 1958); Patient Days (calendar 
Total! .25 year 1958); 
Professional; .25 
Other: .25 

Pro.iect Description: 

Obj ectives; 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 co urse 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 

Part B included! Yes /^ No /™7 

„ . , ., NINOB 81 (C) 
Serial No, 


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 Obligatio ns 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) 


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) 
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, 

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) 


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,, 

Goope ratinR Units ;^ National Naval Medical Center and Waiter 
Reed Aroiy Hospital. 

Man Years (calendar year 1958); 
Total; .25 
Professional: .25 
Other : , 25 

Objectiv es; 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. 

Signific ance 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. 

Prop osed 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) 


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 

Cooperat i ng Unit s; 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 
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) 


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 

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 


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 Investi gator; 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^ 
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 
abnormalities J this compares with only 0.48 percent of 
the abnormal children in the non-diabetic control group. 






























































S_ignificarxe to Maurologic aX 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) 


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 „ 

Princip al Investigator; Anatoie S, Dekaban, M.D. 

Other Investigators; None 

Cooperating Units; None 

Kan Years (calendar year 19 58); 
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 

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. 

Pro tect 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. 

Othe r Inv estigators ; Marie J, Kendall j B„A. 

Cpo|;egatii; g Units; None 

Man Years (calendar year 19 58); 

Total; '.05 
Professional: .01? 
Ofcher; .05 

Proj ect Description ,^ 

Ob jectiv es: 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 I Smployed; 

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 

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 

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. 

Significan c e 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 f or FY 1959 
Total; $24,500 

Directs |24,000 

Eeistbursf^ent: $500 

2<, S©cti©E oa Cliaieal 


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 l a'^estigators; Hone 

Goopegat iag Uaits; Mom© 

Man ¥@as>s Patiemt Days 

Cc&l®ada r 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@d i 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 Fi adiags; Koa®, Th® apparatus has bsea bisilti 
laboratos-y spac® i® beiagarraagsdi s^^a® preliaiBss-y 
traiaiag of moa^@y@ h^ k@gaao 

Sigaif i caae® 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 t h is 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) 

2o Section on Cllnlcml 

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@@ 

Friiaeip al liases tigat org Ho Laasdell 

Other Zu iTQStigators; 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 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 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^estigat ogs; 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 

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 

M ajor 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 » 

Signifi caBe® @f th© proggam to tiiie Za stitate; 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 

Totals $26,©®© 

Dlrecas $25,50® 

Serial MOo HIMDE-9I (c) 

2o Seeti©sa on Pais 


Iffidi-^idaal project E©p©rt 

Gal©mdar ' Year 1958 

Part Ao 

Pr@J®ot Titl®; Fl«@thais© Studies „ 

Priacipal Iza^es tigatog; 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 

"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 F iadingss 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 


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 

2o Section on Pain 
3o Bethesda^ Maryland 
4o New 


Individual Project Report 

Calendar Year 1958 

Part Ao 

Project Title ; Hypothermia in Neuroanesthesiology. 

Principa l Investigator; Kenneth Do Hallj, Mo Do 

Other Investigators ; Edward Laskowskig Mc D« 

Forbes H. NorriSj, M<, D. 
Williara Lee prltchard, M<, D. 

Cooperat ing 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. 

Signifi cance to neurolog ical 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 

Propose d C ourse 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 


Individual Project Report 

CaleBdar ■ Year 1©58 

Part Ao 

Project Title; Succinyl Cboline in Awake Craniotomy o 

Principal Invest igator; 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; 

Obje ctives ; 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 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 

Prop osed 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, 

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 


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 t o Ne ta gological 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® Project s 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 


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. 


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® 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- 

_ 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 - 


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® ¥© 
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 



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 
abstractness p 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 

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 

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 scaaething T 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 

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&, papers fyoa tk-® ProgriMaiio 
Tfe© «''atis'@ ©i2t®rpFis<@ cass r®adilf b© J^j«©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; «>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^ 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 o f 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,^ E@a.®a.rcfe 
tn «ach @f t&@®@ M&itB h&m a distinctive Gh&r&ct&r, 
alt&eugfe tte®r© is a &® 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,, ©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&'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 is pigs^j^iato sa, 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 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 ®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 

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 

£^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 

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~" 


Iradividuall Project Report 
Calendar Year 1958 


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 ; > 1 

Project DescriptioK : 

Objectives ; To stuiy ijsaesis apd ©ubseqweat 
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 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 


A Project Des^criptioa (cont'd)) 

Sig nifi cance ', 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 

Pro posed co u rse of the proj ect? To continue the project 

as time permits.. 

irt B included? Yes 

. 3 ". 

Serial Ho. NINDB-NA~DR~ 1 


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« 


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 , 

M ethods 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 


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 „ 4 

Project Description: 

Obj ectives ; 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. KIKP B-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 

(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 


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 

P roposed co tirse 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 


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 

3« Bethesda, Maryland 
4o Same as HIHDB~HA-5~1957 

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 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 f indings: 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 

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©< 
Proc o 1st Internat o Cong e 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 


Individual Project Eeport 

Calendar Year 1953 

Part Ao 

Project Titles Meuronal specificity in the autonomic nervous 


Principal Investigator: Lloyd Qath 

Other Investigators! Clark Jo Bailey 

Cooperating Units : None 

Man Years (cElendar year 1958) : 
Total: 0„8 
Professional : o 6 
Other; 0.2 

Project Description: 

Ob jectives; 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 

S igcnif 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^ 


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 : « 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 

Part B included? Yes 

- 18 - 

Serial Ko, mmB~m~m=i 


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: 


io Ke'aJL'caiiatcJUical Sc 
2p Section on Develop^ 
and Eegeaeratioa 
So BethesdEj Iferyland 
4 c NeT^ 


[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 : = 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 

_^.lQ g 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 i cagjc© ; 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 


Individual i?roJect Report 

Calendar Year 1958 

Part B i 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 

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 

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 



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 cour se 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 


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 ; » 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 irreyersitol f 
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" 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 

1 a Neuroanatoiffiical ScTeaces 
2o Section on Development 

and Regeneration 
3o Betliesda, Maryland 

4o Sam© as MIHDB-S*a-o 


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 

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 

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. 


Serial HOo HI1DB-HA-DR-- 12 

lo Kern'oaaatomicai Sciences 
2o Section on Deveiopaent and 

3c Bethesda^ Maryland 
4o Same as MINBB-NA-9 


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 o naaas ^ 
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 

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 BiopkysiC o 
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 repertoire 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 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-1 3 

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^ 


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 

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 


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 : 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. MZH DB-HA~0R-1 5 

lo Heuroaaatoiaical Sciences 
2, Section on Bevelopsjsnt 

and Regeaeratioa 
So Saa Juan^ Puerto Rico 
4, Same as lIIISB-lA-17 

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 „ 
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 

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 

Pro posed 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 

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 


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 


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© 

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 pro ject; 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 

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 pro ject ; 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^ 


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 

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 : 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. NIMD B-NA-DR-2 1 
lo Neuroaaatosnical Sciences 

2, Section or Development 

and Regeaeration 

3, Bethesda, Md. and Saa Juan^, 

4, Same as NiroB-NA~15-1957 


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 : » 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 

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 

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 jndia R-s : This project feas been started "but too 
recently for s report of resr.ltSo 

Si^n if 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» HIHD B-M-HC-^l 

1= Meuroanatomical Scieaces 
2o Section on Heiirocytology 

3, Bethesda, Maryland 

4. Same as NINDB-M-18«-1957 


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