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ifiSEASES AND STROKE 






AMUAL REPORT 

OF 

PROGRAM ACTIVITIES 

MTIOML INSTITUTE OF NEUROLOGICAL DISEASES AND STROKE 

Fiscal Year I97I 

PART II 



ANNUAL REPORT 

July 1, 1970 through June 30, 1971 

Associate Director, Collaborative and Field Research 

National Institute of Neurological Diseases and Stroke 

National Institutes of Health 

Through October 31, 1970, leadership of this program area was provided 
on an acting basis by Dr. Eldon L. Eagles, Deputy Director, NINDS, On 
November 1, Dr. Warren V, Ruber was appointed Associate Director. Dr. Ruber 
was formerly Project Director of the American Neurological Association's 
Joint Conmiittee for Stroke Facilities, and had served as Chief of Neurology 
in the Veteran's Administration's Department of Medicine and Surgery. 
Dr. Ruber's appointment marked a re-emphasis of the role of collaborative, 
directed, and field- type research in the overall effort to accomplish the 
scientific objectives of this Institute. The intent of this report is not 
to encapsulate items of information found in the separate branch summaries 
but to provide an overview of the significant accomplishments and problems 
of the Institute's Collaborative and Field Research programs during Fiscal 
Year 1971, and to look briefly, though broadly, toward the future of these 
programs . 

PROGRESS AND ACCOMPLISHMENTS, FY 1971 

Increased funds provided by the President's budget and congressional 
supplements permitted orderly program expansion in epilepsy, funding for a 
pilot study of cerebral death, and sufficleut support to prevent dislocation 
in other programs. Without these increases significant program opportunities 
would have been lost. 

The Special Projects Branch continued to pursue studies of neurologic 
disorders, concentrating on controlled clinical studies for the evaluation 
of two experimental anticonvulsants and one marketed agent. These efforts 
have demonstrated the value of blood level determinations as an adjunct to 
anticonvulsant therapy and established methodologies for further anticonvul- 
sant studies. The successful transition of Epilepsy Abstracts from a fully 
supported Institute publication to a partially subsidized periodical available 
by subscription has demonstrated that periodical's scientific merit and 
value. The completion of the evaluation of the 15-year field follow-up of 
head injured veterans of the Korean campaign, and the evaluation of data from 
Phase I of the Head Injury Model Construction Program, mark the conclusion 
of two important aspects of the Institute's directed Head Injury Program. 
Funds must now be obtained and leadership recruited for continued efforts 
in this important program area. 

The process of planning and developing a collaborative study of the 
criteria of cerebral death was accomplished by staff of the Office of the 
Associate Director and the Head Injury Section of the Special Projects 
Branch. With expert guidance from an ad hoc advisory committee, an 
acceptable protocol was developed and a coordinator and collaborators were 
selected. Contracts for the study will be awarded before July 1, 1971. 



Results of controlled and limited studies using the assembled data base 
have continued to come in to the Perinatal Research Branch. Subject area 
task forces, in collaboration with the Epidemiological and Statistical 
Advisory Committee, are redirecting staff effort and attention to larger, 
more significant statistical analyses to identify major trends, developmental 
patterns, and risk factors. Interesting and significant studies of the 
influence of infectious diseases in pregnancy and childhood upon the 
development of the child's central nervous system (CNS) continue. 

A multi-institutional study to determine the feasibility of developing 
an effective sensory prosthesis for the blind has been launched. Currently, 
the study is focused on the problems of direct stimulation of visual centers 
of the brain, including the safety thereof, and the reproducibility of 
stimulation results. The future success of this project is enchanced in 
large part by the careful, expert, and conscientious project direction being 
provided by the Chief, Laboratory of Neural Control, Intramural Research, 
and his assistant. 

Epidemiologic studies of a selected group of acute and chronic nervous 
system disorders continue. Short term studies, particularly of multiple 
sclerosis, sub-acute sclerosing panencephalitis, and CNS tumors have pointed 
the way toward a better understanding of the etiology of these diseases. 

Collaboration with scientists working in a broad range of the basic 
biological sciences and in clinical medicine has improved considerably our 
understanding of the mechanisms and etiology of slow, latent and temperate 
virus infections. Successful transmission of two sub-acute and chronic 
degenerative diseases of the nervous system from man to primates has led to 
the implication of suspect viral agents — a significant step in the march 
toward understanding the etiology and thereby, hopefully, being able to 
successfully treat or prevent such types of central nervous system disease. 

PROBLEMS - FY 1971 

Despite the successes and progress of the past year there are problems 
affecting program development which remain to be solved. 

Tight controls on full-time and permanent employment restricts the 
recruitment of staff needed to implement and develop important new programs. 
This has necessitated the most careful allocation of positions as vacancies 
occur. Only through this mechanism can needed and important shifts in 
priority be implemented. 

Head and spinal cord injury has been recognized as a serious problem 
for several years. Despite generous Extramural support and a vigorous 
Intramural program, important scientific questions remain unanswered; these 
must be answered if the goal of improved treatment, coupled with improved 
prevention, is to be attained. Directed collaborative studies aimed at 
elucidating the pathophysiologic responses of brain and spinal cord tissues 
to trauma are urgently needed. Until additional funds are available and 
adequate leadership can be found — and the two needs are interrelated — studies 
in this important problem area cannot be developed. 



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The significant FY 1972 budget decrease that has been imposed on the 
Perinatal Research Branch (PRB) makes it exceedingly difficult to accomplish 
two important tasks. The first is an orderly completion of the Collaborative 
Perinatal Project. As a result of the cut, important activities in follow-up, 
sample maintenance, and quality control at the collaborating institutions 
and at the coordinating center in the PRB will be curtailed. The second task 
which must be foregone is the planning for research projects on neurological 
diseases of childhood; preliminary results of the Collaborative Perinatal 
Project indicate these would be productive. Although an orderly decrease 
in budget support and effort by the PRB was planned as the study phased out, 
the immediate and drastic decrease at this time is quite damaging. 

The Collaborative and Field Research Programs of NINDS are, by their 
nature, heavily dependent upon continuing and active biostatistical 
participation. The need for biostatisticians in these varied programs 
goes deeper than simple part-time consultation. To be effective, the 
statistical staff must be part of the research teams and relate closely to 
them. Scientific program leaders, charged with the responsibility of 
promptly producing evidence of research results to fully collaborate with 
scientists outside of NINDS, must have the explicit and constant control of 
the processing and analysis of the data upon which they rely. Without such 
control they have experienced an inability to discharge these responsi- 
bilities promptly and effectively. Therefore, it is imperative that such 
a realistic relationship with the Office of Biometry, NINDS, be established. 

LOOKING AHEAD 

Fiscal Year 1972 promises to be a difficult year in terms of avail- 
ability of resources to move forward with existing and planned programs. 
Budget cuts will effect the Epilepsy, Head Injury, and Cerebral Death 
Programs as well as the Collaborative Perinatal Project. Considerable 
resourcefulness will be required to maintain programs which have been so 
laboriously built and organized. In no way will it be possible to mount 
new programs in stroke research, in disorders of communication, or to 
pursue significant head and spinal cord injury research. 

As data collection for the Collaborative Perinatal Project moves toward 
completion, and is supplanted by less expensive, but promising, investi- 
gations of neurologic disorders of childhood, funding required by the Peri- 
natal Research Branch will diminish. It is anticipated that this will permit 
the resumption of active, directed, collaborative research in the field of 
head and spinal cord injury, and the commencement of similar efforts in 
the fields of communicative disorders and stroke, with only relatively 
minor increases in the overall funding level over the next five years . 



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ANNUAL REPORT 

July 1, 1970 through June 30, 1971 

Special Chronic Disease Studies for Collaborative and Field Research 

National Institute of Neurological Diseases and Stroke 

National Institutes of Health 

Stud y of Child Growth and Development and Disease Patterns in Primitive 
Cultur es, and Slow, Latent and Temperate Virus Infections 

This section has continued to focus its attention on long term 
studies of the human biology of the many vanishing primitive societies. Our 
laboratory research and that of our numerous collaborating investigators is 
directed to problems which have been phrased under this theme. The neuro- 
logical development and learning patterns in children in diverse cultural 
experiments in the human condition has been the major focus of attention. 
The laboratory studies on human biology, genetics and associated molecular 
biology, immunology, virology, and biochemistry have all been directed at 
solving problems which have been carefully chosen from small isolated bands 
still living in the primitive situation in which these problems may be more 
appropriately studied than in larger civilized societies. 

Our efforts to document the development and neurological patterning 
in disappearing primitive cultures has resulted in the largest archive of 
such documentation in the world. The collection and preservation of exist- 
ing cinema data from Australian aborigines, New Guinean, Oceanic and African 
aborigine groups, and American Indians, as they live as hunter-gatherers or 
primitive hoe-and-digging-stick agriculturists provides the only such docu- 
mentation of the life and behavior of man, as he most probably lived and 
evolved for over 997c, of his evolutionary history, or about one million years. 

Kuru, a new disease which we discovered and described in the New 
Guinea Highlands fourteen years ago, has been the first subacute and chronic 
degenerative disease of the nervous system of man with a transmission model 
in animals and an established virus etiology. From analogy with kuru, we 
selected Creutzfeldt-Jakob disease and other forms of spongiform encephalo- 
pathy as likely slow infections. The "hunch" proved right: Creutzfeldt- 
Jakob disease has now been transmitted to chimpanzees, independently from 
ten patients, and serial chimpanzee to chimpanzee transmission has also been 
successful through the third passage. The agent, as that of kuru, is fil- 
terable. During this year significant breakthroughs were achieved in the 
study of these two CNS diseases with the successful transmission of both 
kuru and Creutzfeldt-Jakob disease from human patients to 3 species of New 
World monkeys — the spider monkey (Ateles geof froyi) , the squirrel monkey 
(Saimiri sciureus) and the capuchin monkey (Cebus sp.) . Creutzfeldt-Jakob 
disease was transmitted from affected chimpanzee to yet another New World 
monkey, the woolly monkey (Lagothrix lagothricha) , as well as to squirrel 
monkeys. Serial passage and viral pathogenesis and characterization studies 
have been expanded utilizing these more economically purchased and con- 
veniently handled primates. These animals are also being used in isolation 
studies being conducted on other human slovj-infections suspected of being 
caused by viruses. It is significant to note that during the past year we 
have received greater than 400 specimens from patients with CNS diseases. 

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Of this number, approximately 50% (200) are brain biopsy and autopsy speci- 
mens and between 50-60 of these are from patients with Creutzfeldt-Jakob 
disease. We now believe that the two human diseases, kuru and Creutzfeldt- 
Jakob disease, and the two similar animal diseases, scrapie and mink enceph- 
alopathy, which we are also studying, form a group of diseases of similar 
pathogenesis, which we have called the spongiform viral encephalopathies. 

Our work has continued to incriminate the measles virus as the cause 
of SSPE, but we have not yet found evidence of a second virus in this disease, 
With the return of Dr. David Asher, from 17 months work in various insti- 
tutes in the Soviet Union, work has been begun on the problem of chronic 
progressive forms of encephalitis caused by tick-borne encephalitis virus. 
Several strains of the virus known to induce chronic infections have been 
inoculated into rhesus monkeys. The continuation of these studies, in 
collaboration with our colleagues in the Soviet Union, are providing working 
models for the in depth studies to demonstrate chronic virus infection as 
the cause of epilepsy partialis continua or focal epilepsy in children in 
which viral- like inclusion bodies are found in brain cells. Further, we are 
pursuing with our Soviet colleagues our studies on epidemic hemorrhagic 
fever, of the nephro-nephritis type, which occurred in epidemic proportions 
in civilians and troops during the Korean War and which still occurs at a 
somewhat lower level of incidence at the present time. 

Encouraged by our success with one of the presenile dementias and 
several cases in the borderland between Alzheimer's and Creutzfeldt-Jakob 
diseases, we are concentrating on the other presenile dementias and senility 
with particular emphasis on Alzheimer's and Pick's diseases, parkinsonism- 
dementia and senile dementia. Our work on the presenile and senile demen- 
tias, the isolation of latent viruses from the central nervous system, and 
comparisons of the many aspects of the pathological processes in kuru and 
Creutzfeldt-Jakob disease (including the presence of amyloid (senile) 
plaques and extensive glial hypertrophy with fibrillary tangles in the astro- 
cytes) has led us to be increasingly concerned with possible virus involve- 
ment in certain aspects of senile dementias and normal aging. 

In our attempts to establish infection as the etiology of other dis- 
eases of the CNS of man we have successfully isolated in tissue culture a 
strain of adenovirus from the brain of a patient with lymphosarcoma. We 
have shown the virus to be closely related to candidate adenovirus type 32. 
Furthermore, we have recently demonstrated Cowdry type A intranuclear inclu- 
sion bodies, vjhich by EM contain papovavirus-like virions, in in vitro ex- 
plant cultures of human brain from a patient with progressive multifocal 
leucoencephalopathy . 

We are trying to establish the range of cases with spongiform enceph- 
alopathy which may be transmitted; other diseases we are studying by in vitro 
cultivation of brain and other tissues, and with animal (including monkeys 
and apes) inoculations, are multiple sclerosis, amyotrophic lateral scler- 
osis (both sporadic and familial types) , parkinsonism-dementia complex, in 
both the U.S. and Guam, essential parkinsonism, myoclonic epilepsy, 
Schilder's disease, metachromatic leucodystrophy, and progressive supra- 
nuclear palsy. In collaboration with Drs ., Michael Alpers and Malcom Simons 



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of Australia investigations to seek the agent of kuru in cultivated leuco- 
cytes of patients and of experimental chimpanzees are under way, and the 
evaluation of the humoral (thymic) and delayed hypersensitivity (splenic) 
immune status in kuru in man and in the experimental disease have been made. 
Most significant has been our continued isolation of further strains 
of hidden, masked, or latent viruses from surgically sterile brain and 
other tissues of both our chimpanzees with transmitted neurological diseases 
and of human patients. The list of these virus isolates from chimpanzees is 
now over one hundred virus strains. These fall thus far into ten identified 
new virus species. Six of these have been thoroughly investigated virolo- 
gically and have been the subject of both graduate theses and papers in the 
specialty journals. 

All six viruses have been deposited in the American Type Culture 
Collection. Two of the six viruses. Pan 1 and Pan 2, have now been class- 
ified as simian foamy viruses type 6 and type 7, respectively. We have 
demonstrated that these RNA viruses have an RNA-DNA dependent polymerase 
suggesting a close relationship to the RNA oncogenic group of viruses. 

In 1969 we isolated two strains of virus from the lungs and brain, 
respectively, from sheep with progressive pneumonia in Montana. We have now 
demonstrated that both viruses are serologically indistinguishable from 
maedi, a lymphoreticular progressive pneumonia, and visna, a primary demye- 
linating disease of sheep in Iceland. Moreover, visna, maedi and the virus 
of Montana sheep progressive pneumonia are all RNA viruses associated with 
an RNA-DNA dependent polymerase. Further characterization of these viruses 
and their relationship to the RNA oncogenic group of viruses having these 
properties are under way. 

In many cases, two or more viruses have been isolated from the same 
brain. We are forced to speculate that activation of these latent agents 
may be a process responsible for some subacute or chronic neurological 
diseases. We are attempting to study the activation of measles virus to 
produce SSPE, of chickenpox virus to produce herpes zoster in later life, 
of herpes simplex virus and the simian herpes viruses to produce central 
nervous disease, and of a papova-like virus to produce progressive multifo- 
cal leucoencephalopathy. Cytomegalo virus and EB (Epstein-Barr) virus 
activation to produce the post-transplantation of "pump" syndrome in most 
subjects receiving organ transplants has been studied by Dr. Lang at Duke 
University. We are now interested in using this work as well as a model for 
the chronic and slow infections with which we work. Our current hypothesis 
is that similar processes may underlie the pathogenesis of kuru, Creutzfeldt- 
Jakob disease, and other spongiform encephalopathies and, perhaps, even be 
involved in some presenile and senile dementias. 

We therefore continue attempts at isolating virus strains from long- 
maintained sterile tissue explants and trypsinized cell suspensions, using 
the techniques of cell fusion and co-cultivation. These are being applied 
to the degenerative central nervous system disorders listed above, as well 
as to other chronic diseases. The electron microscopy, in collaboration 
with Dr. Peter Lampert, and the immunological techniques, including fluores- 
cent antibody, are being used to localize virus-like particles, gamma glob- 

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ulin, components of complement, and antigen-antibody complexes in tissue 
and in sera of patients with these diseases; and extensive serological in- 
vestigations of serum and spinal fluid from patients with these diseases for 
antibodies against a wide range of microbial antigens has been continued. 

Nutritional studies, and studies on reproduction and fertility, on 
the selective advantage and establishment of genetic polymorphisms, of un- 
usual and odd alternative ways of employing the central nervous system in 
its higher cerebral function of language learning and use, computation 
(number sense and calculation with a numbers system), psychosexual cultur- 
ally-modified behavior, and cognitive style, are providing data on alterna- 
tive forms of possible neurological functioning for man, of which we would 
remain unaware and for moral, ethical and political reasons be unable to 
produce or investigate in the clinic or laboratory, once the natural cul- 
tural experiments in primitive human population isolates had all finally 
been amalgamated into the modern civilized cultural veneer which is now 
imposed upon almost all members of the community of man. 

Growth and development studies in primitive cultures have yielded new 
evidence for incredibly delayed puberty and slow growth rates in certain 
peoples, particularly the short-of-stature populations in the Highlands of 
New Guinea. Here we find people with the mean age of menarche over eighteen 
years, and with male puberty of eighteen at twenty years. These are thus 
the slowest growing populations on earth, with the most delayed puberty; two 
thirds of their life span is spent in reaching maturity. Preliminary evi- 
dence suggests high levels of pituitary growth hormone in such populations, 
in both pituitary glands obtained at autopsy and in the serum. Growth rate 
seems to be proportional and age of puberty inversely proportional to mean 
adult stature. 

Congenital mental defect and a wide range of neurological problems 
associated with the most severe foci of endemic cretinism in the world, in 
the Highlands of West New Guinea (West Irian, Indonesia) , are under further 
investigation. Our investigation of an accumulation of cases of familial 
periodic paralysis in certain of the Pacific Islands continues. 

The discovery of new genetic factors, including haptoglobins and 
hemoglobins, with elucidation of their biochemical structure and their later 
use as markers in human population genetic studies, has been a by-product 
of these investigations. Similarly, the discovery of immunologically virgin 
populations without exposure to respiratory and enteroviruses which are 
ubiquitous in the civilized world, has permitted: 1) fundamental investiga- 
tion on the immune response in man; 2) investigations possible no where 
else on the persistence of immune response after natural measles infection 
and live attenuated measles virus immunization in the absence of circulating 
virus, yielding data important to the diagnosis of understanding of delayed 
slow measles encephalitis (subacute sclerosing panencephalitis, SSPE) , and 
3) establishing the serological identity of the agent of the 1918 influenza 
pandemic to aid in the genetic-historical elucidation of the serial mutation 
of influenza virus. By virtue of limited travel during their entire life- 
time and intensive exposure to their natural ecology, members of primitive 
groups serve as unequaled sentinel populations for revealing the focal micro- 

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bial agents that infect man in their environment. Thus, for infections 
ranging from Chagas disease and toxoplasmosis to arbovirus infections, prim- 
itive groups offer unusually fruitful subjects for investigation. In 
toxoplasmosis, filariasis, yaws and malaria, as well as in the arbovirus 
encephalitides , we have studies in progress. 

Our studies on the antigenic composition of the virus causing the 
1918-22 influenza pandemic and on the immunologic response to various close- 
ly related influenza viruses, including the new 1968 Hongkong Asian influenza 
(the latest A2 variant which might better have been designated A3) have now 
been published. However, the opportunity of supervising and participating 
in investigations of the influenza epidemic which swept through the Terri- 
tory of Papua and New Guinea, and through West New Guinea in late 1959 has 
brought additional material for further study on virus evolution of influen- 
za into our laboratory. Incidental to this work our field investigations 
also provided the opportunity to study new virgin soil measles epidemics 
and to find new isolated populations free of many nearly ubiquitous res- 
piratory viruses. 

We planned, organized and selected participants for Theme III on 
Pathogenesis of Slow Virus Diseases of the Central Nervous System at the 
Vlth International Congress of Neuropathology in Paris in September, 1970. 
This theme provided reviews and new concepts on virus-host relationships 
and significant new data on the pathogenesis of cellular injury associated 
with persistent viral infections, immunological aspects of slow infections 
and characterization and properties of subacute spongiform virus enceph- 
alopathies. New and significant findings of the neuropathology of these 
diseases were presented and reviewed. The papers presented were published 
in the Proceedings of the Vlth International Congress of Neuropathology, 
Masson and Cie, Paris. 

Our various laboratory investigations have thus evolved around the 
theme of elucidating medical problems in the cultures of primitive man, or 
man living in small, isolated traditional communities, by cautiously 
selecting such problems of immunological, virological, biochemical, genetic 
and nutritional interest which are pertinent to the long-term studies of 
growth, behavior and human biology in these groups. 



Serial No. NDS (CF)-65 OAD 1282 

1. Collaborative & Field Research 

2. Office of Associate Director 

3. Bethesda, Maryland 

PHS - NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Study of Child Growth and Development and Behavior, and 
Disease Patterns in Primitive Cultures. 

Sub-Project I: Study of the developmental patterning of the human nervous 
system (cybernetics of human development). 

a. Analysis of culturally determined methods of approach to 
symbolic representation from drawings and art forms of child- 
ren and adults in primitive societies. 

b. Analysis of the development in writing of adolescents 
in preliterate cultures. 

c. Analysis of child care and behavior patterns in primi- 
tive cultures from photographic recording (development of 
techniques and methods). 

d. Investigation of nonrecurrent phenomena (objectives and 
selectivity used in documentation of aperiodic phenomena to 
preserve maximum information) . 

e. A research archive for ethnopediatric film investigation 
of styles in the patterning of the nervous system. 

Sub-Project II: Human evolutionary studies in isolated primitive groups. 

a . Kuru . 

b. Motor neuron disease and other degenerative diseases in 
East and West New Guinea, and in other inbred Pacific Islands 
populations. 

c. Blood group genetic studies of Australasian (Melanesia 
and Micronesia), and South American indigenous groups. 

d. Red cell enzyme, serum factor and leucocyte type pleo- 
morphisms among these groups. 

e. Analysis of dermatoglyphic variance. 

f. Epidemiologic and ecologic investigations of kuru: with 
expanding, manipulatable computer records system. 

Sub-Project III: Studies of isolated Micronesian populations. 

a. Child development and behavior on Ulithi, Ifalik and 
Lamotrek atolls, and on Pais Island. 

b. Response to live measles virus vaccine in immunological 
virgin populations without circulating measles virus (special 
attention to response in susceptible adults and pregnant 
women and their offspring). Follow-up studies to determine 
persistence of antibody response and to observe these popula- 
tions for possible long-term neurological sequellae following 
use of attenuated live vaccine virus. 



Serial No. NDS (CF)-65 OAD 1282 

c. Influenza A2 virgin soil epidemics (epidemiological, 
clinical and immunological response and discovery of popula- 
tions without previous experience with Type A or Type B 
influenza. Follow-up studies of these populations as sentinel 
populations for detection of epidemics due to new strains of 
influenza virus. 

d. Studies of infectious disease patterns in remote indivi- 
dual populations. 

e. Genetic characterization of the population of the Western 
Caroline Islands. 

f. The antigenic identification of the 1918-29 influenza 
virus from residual antibody in aged persons in isolated popu- 
lations free of influenza since the pandemic. 

Sub-Project IV: Studies of isolated New Guinea populatioiB. 

a. Slow growth, delayed puberty and early aging in Melane- 
sians of short stature. 

b. CNS defects in areas of intense endemic goitrous creti- 
nism in Highland populations. 

c. Child development and behavior patterns in the Asmat, 
Tjitak, Auyu, Kayagar, Western Dani, Kaure, Toure groups of 
West New Guinea; the Kukukuku (Anga) , Eastern Highlands 
peoples, the Biami, Etoro and Waragu people of the Great 
Papuan Plateau of Papua and New Guinea; and, the West Nakanai, 
Mangsing and Mamusi of New Britain. 

d. Infectious disease — including encephalitis--studies in 
diverse, ecologically isolated New Guinea populations. 

e. Study of the pattern of the A2 Hongkong influenza epide- 
mic in East and West New Guinea in 1969-70. 

f . Genetic and demographic characterization of New Guinea 
populations. 

g. Hereditary and genetic disease patterns in New Guinea, 
h. The Anga (Kukukuku): an intensive longitudinal study of 

growth and development, behavior, disease patterns, human 
genetics, communication, in an archaic mountain population of 
New Guinea. 

i. Research cinema films of behavior patterns of children 
in New Guinea. 

j. Study of the hazards of lowland resettlement of Highland 
groups in New Guinea. 

Sub-Project V: Studies of isolated New Hebrides and Solomon Islands popu- 
lations, 

a. Child development and behavior in Tongariki, the Banks 
and Torres Islands and Espiritu Santo. 

b. Tongariki: an intensive study of human evolution in the 
Shepherd Islands. 

c. Human genetics and disease patterns survey in the Banks 
and Torres Islands. 

d. Seroepidemiology of infectious disease in New Hebrides. 



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Serial No. NDS (CF)-65 OAD 1282 



Sub-Project VI: Studies of Australian Aborigines. 

a. Arbovirus seroepidemiological studies of Aboriginal 
groups in Cape York. 

b. Survey patterns of infectious disease in Aboriginal 
groups in the Haast Bluff, Cape York, the Kimberley and 
Bentnik-Mornington Islands groups. 

Sub-Project VII: Studies of Central and South American Indians. 

a. Human genetics and disease patterns of Guayaki and Chaco 
Indian tribes, and the Mennonite colonists in the Chaco of 
Paraguay . 

b. Child growth and development of Guayaki and Ai'yore 
Indians of Paraguay. 

c. Child growth and development of Aroyo (Moro) Indians of 
Bolivia and Paraguay. 

d. Population genetics, disease patterns and child growth 
and development of the Waunan Indians of Columbia and 
Ecuador. 

Sub-Project VIII: Developmental, genetic and disease patterns in primitive 
populations of Asia, Africa and in Polynesia. 

Sub-Project IX: Experimental developmental neuropediatrics in infantile 

programming: an empirical approach to the language of infor- 
mation input into the nervous system. 

Sub-Project X: Ciphers and notation for the coding of sensory data for 
neurological information processing: 

a. Notational systems for human movement. 

b. Ciphers and notation for human form (physiognomy, 
physique, palm printing, ear form, hair). 

c. Theoretical studies in notational problems in mathema- 
tics, linguistics, music, dance. 

d. Alphabets: a theoretical investigation into their rela- 
tion to linguistics, and the application to non-linguistic 
information. 

e. Form recognition: neuroanatomic and genetic determina- 
tion of preferential recognition, and interrelationships of 
the problems in computer programming and in the arts. 

f. The ciphering and coding of visual data as solved in the 
visual arts (drawing, painting and sculpture). 

Sub-Project XI: Racial distribution and neuroanatomic variations in the 
structure of the human brain. 



Principal Investigator: D. Carleton Gajdusek, M.D. 



9.. 



Serial No. NDS (CF)-65 OAD 1282 

Other Investigators: Clarence J. Gibbs, Jr., Ph.D. ^ Paul W. Broira, M.D. , 
Raymond Rods, M.D. , David M. Asher, M.D. , Michael Alpers, 
M.D., Vincent Zigas, M.D. , Francoise Cathala, M.D. , Richard 
Marsh and Robert Cornelius, D.V.M, , John Hooks, Ph.D., 
E. Richard Sorenson, Nancy Rogers, Mint Basnight, Helena 
Gilbert, Judith Meyer and Richard Benfante. 



Project Description: 

The Section for the Study of Child Growth and Development, and Behavior, 
and Disease Patterns in Primitive Cultures has continued all projects listed 
in the previous Annual Reports, with expansion of collaborating investigators 
as reflected in the authorship and studies of the publications listed. The 
titles of sub-projects and their subdivisions are sufficiently explicit to 
constitute the project description. 



Sub-Project I. Study of the developmental patterning of the human nervous 

system (cybernetics of human development) . 

Principal Investigator: D. Carleton Gajdusek, M.D. 

Other Investigators: Michael Alpers, M.D. , Paul Bro^<m, M.D. , Vincent Zigas, 
M.D. , E. Richard Sorenson, Judith Meyer, Peter Fetchko and 
Donald Rubinstein. 

Cooperating Investigators: Dr. Margaret Mead, American Museum of Natural 
History, New York; Dr. Ted Schwartz, Univ. California, Los 
Angeles; Alan Lomax, Columbia Univ., New York; Mr. and Mrs. 
Mark Jablonko, Columbia Univ.; Dr. Paul Ekman and W.V. 
Friesen, Langley Porter Neuropsychiatric Inst. , San Franciscq 
Dr. Peter Kundstadter, Univ. Washington, Seattle; Kal Muller, 
Univ. Arizona, Tucson; Thomas Kiefer, Dr. Edwin Cook, Univ. 
California, Davis; Dr. Gordon Gibson, Smithsonian Inst.; Dr. 
Robert MacLennan, Inter. Agency for Cancer Res., France; Dr. 
Maurice Godelier, Sorbonne; William H. Bloxam, Wayne Dye, 
John MacGregor, Father David Gallus, Father F. Trenkenshuh, 
0. Kooyers, Dr. C.K. Dresser, West New Guinea; Timothy Asch, 
Brandeis Univ.; N. Chagnon, Univ. Michigan; James Bruce, 
Pasadena, Calif.; Elizabeth and Perry Kennedy, Univ. Buffalo. 

Sub-Project II. Human evolutionary studies in isolated primitive groups . 

Principal Investigator: D. Carleton Gajdusek, -M.D. 

Other Investigators: Paul Brown, M.D. , C.J. Gibbs, Jr., Ph.D., M. Alpers, 
M.D., F. Cathala, M.D. , D. Asher, M.D. , N. Rogers, M. Bas- 
night, J. Hooks, Ph.D. 



10, 



Serial No. NDS (CF)-65 OAD 1282 



Cooperating Investigators: Dr. Stephen Fazekas, CSIRO, Sydney; Eric French, 
and Cyril Curtain, M.D. , CSIRO, Melbourne; Dr. Malcolm Simons, 
Royal Children's Hospital, Melbourne; Dr. R.T. Simmons, John 
J. Graydon, Alan Duxbury and Frank Warbuton, Commonwealth 
Serum Laboratories, Melbourne; and Dr. Robert Kirk, John Cur- 
tain School of Medicine, Canberra, Australia; Dr. Vincent 
Zigas, Dr. Richard Hornabrook and Dr. Adolph Suweri, Public 
Health Department, Territory of New Guinea. 



Sub-Project III: Studies of isolated Micronesian populations . 

Principal Investigator: D. Carleton Gajdusek, M.D. 

Other Investigators: Paul Brown, M.D. , C.J. Gibbs, Jr. and J. Anthony 

Morris, Ph.D., David Asher, M.D., Francoise Cathala, M.D. , 
MJchael Alpers, M.D., Vincent Zigas, M.D. , John Hooks, Ph.D., 
Nancy Rogers, Mint Basnight, Richard Benfante. 

Cooperating Investigators: Dr. Leon Rosen, Gordon W. Wallace, NIAID , Honolu- 
lu, Hawaii; Dr. Jacob Brody, NINDS; Chris Plato, NICHD; Dr. 
Kwang Ming Chen, National Taiwan Univ. , Tapei; Newton Morton, 
Population Genet. Laboratory, Honolulu; Dr. Stephen Fazekas, 
Sydney; Dr. Antonio Golbuu, Jose Torres, Eddie Iderug, NINDS 
Research Center, Guam. 

Sub-Project IV: Studies of isolated New Guinea populations . 

Principal Investigator: D. Carleton Gajdusek, M.D. 

Other Investigators: Drs. Paul Brown, Rajmiond Roos, Michael Alpers, Vincent 
Zigas, C.J. Gibbs, Jr., David Asher, Francoise Cathala and 
John Hooks; Judith Meyer, Richard Benfante, Mint Basnight 
and Helene Gilbert. 

Cooperating Investigators: Dr. C.K. Dresser, West New Guinea; William Bloxam, 
John MacGregor, Richard Hornabrook, John Mathews, Territory 
of New Guinea; Edwin Cook, Univ. California, Davis; Ted 
Schwartz, Los Angeles; Dr. R. MacLennon, France; Dr. John 
Hotchin, Department of Health, New York; Paul Ekman, San 
Francisco; R.T. Simmons, J.J. Craydon, C.C. Curtain, Aust- 
ralia; David Kitchin, M.D. , Columbia Univ.; Alexander Beam, 
M.D., Rockefeller Inst., New York; Maurice Godelier, Ph.D., 
Paris; E. Beck, P. Daniel, Inst. Psychiatry, London; Chris 
Plato, NICHD; Roger Rodrigue, M.D. , Temple Univ.; and J. van 
Delden, New Guinea. 



lis 



Serial No. NDS (CF)-65 OAD 1282 



Sub-Project V: Studies in Isolated New Hebrides and Solomon Islands popu - 
lations . 

Principal Investigator: D. Carleton Gaidusek, M.D. 

Other Investigators: Paul Brown, M.D., David Asher, M.D., C.J. Gibbs, Jr., 
Ph.D., Nancy Rogers, Mint Basnight and Helene Gilbert. 

Cooperating Investigators: Dr. H. Lehraann, Univ. Cambridge; Dr. Robert Kirk, 
Australian National Univ., Canberra; Dr. Jean Guiart, Sor- 
bonne, France; Dr. James MacGregor, Honiara; Dr. Roger Green- 
ough. Dr. William Rees, New Hebrides; Chris Plato, NICHD. 

Sub-Project VI: Studies of Australian Aborigines . 

Principal Investigator: D. Carleton Gajdusek, M.D. 

Other Investigators: Paul Brown, M.D. , Michael Alpers, M.D., C.J. Gibbs, Jr., 
Ph.D., Nancy Rogers, Mint Basnight and Helene Gilbert. 

Cooperating Investigators: Drs. R.T, Simmons, J.J. Graydon, A. Duxbury and 
F. Warbuton, Commonwealth Serum Laboratories, Melbourne; 
C. Curtain, Sydney; E. Beck, London; and W.C. Leyshon, NIDR. 



Sub-Project VII: Studies of Central and South Araerican Indians . 

Principal Investigator: D. Carleton Gajdusek, M.D. 

Other Investigators: C.J. Gibbs, Jr., Michael Alpers, M.D., Mint Basnigh t, 
Nancy Rogers and Helene Gilbert. 

Cooperating Investigators: W.C. Leyshon, NIDS; J.L. Sever, NINDS; K. Walls, 
CDC, Atlanta; R.L. Anderson, WRAIR; J. A. Morris, DBS, Arthur 
Steinberg, Western Reserve Univ., Cleveland; C.C. Curtain, 
CSIRO; Elizabeth and Perry Kennedy, Buffalo. 

Sub-Project VIII: Developmental, genetic, and disease patterns in primitive 
populations of Asia, Africa and Polynesia . 

Principal Investigator: D. Carleton Gajdusek, M D. 

Other Investigators: Frank D. Schofield, M.D.,. Nairobi; Peter Kundstadter, 
Seattle; Thomas Kiefer, Univ. California; L. Vogel, M.D. , 
Nairobi; Kok Ann Lira, M.D. , Singapore; Chris Plato, NICHD. 



^2l 



Serial No. NDS (CF)-65 OAD 1282 

Sub-Project IX: Experimental developmental neuropediatrics in infantile 

programming: an empirical approach to the language of inform - 
ation input into the nervous system. 

Principal Investigator: D. Carleton Gajdusek, M.D. 

Other Investigators: Michael Alpers, M.D., E. Richard Sorenson, Judith Meyer; 
Don Rubinstein. 

Cooperating Investigators: Paul MacLean, M.D. , NINDS; Paul Ekman, San 
Francisco 



Sub-Project X: Ciphers and notation for the coding of sensory data for 
neurological information processing . 

Principal Investigator: D. Carleton Gajdusek, M.D. 

Other Investigators: E. Richard Sorenson, Michael Alpers, M.D., Judith Meyer 

Cooperating Investigators: Dr. R.L. Kirk, Canberra; Patricia Hunt, Univ. 

California, Berkeley; Paul MacLean, M.D. , NINDS; Chris Plato, 
NICHD; Alan Lomax, New York. 



Sub-Project XI: Racial distribution and neuroanatomic variations in the 
structure of the human brain . 

Principal Investigator: D, Carleton Gajdusek, M.D. 

Cooperating Investigators: Elisabeth Beck, Peter Daniel, M.D. , Institute of 
Psychiatry, London; Paul Yakovlev, M.D. , Richard Sidman, M.D., 
Dr. Kemper, Dr. H. Hamlin, Harvard Univ., Boston; Dr. Peter 
Lampert, Univ. California, La Jolla; Dr. K. Earle, AFIP; 
Dr. P. MacLean, NINDS; Drs, R. Hassler and H. Stephan, Max- 
Plank Inst., Frankfurt; Dr. G. Inke, Stony Brook, New York; 
Dr. A, Hopf, Neustadt, Germany. 



13c 



Publications: 

Gajdusek, D.C., and Bro^m, P. (Eds.): Isolated and Migratory Population 
Groups. Health Problems and Epidemiologic Studies. In Amer. J. Trop. Med . 
Hyg . 19: 127-175, 1970. 

Plato, C.C.. and Gajdusek, D.C.: Dermatoglyphics of the natives of Tonga- 
riki in the New Hebrides. J. Archaeol . Phys . Anthrop. Oceania , in press. 

Gajdusek, D.C., Gibbs, C.J., Jr., and Lim, K.A. : Prespects for the control 
of chronic degenerative disease with vaccines. Proc. Inter. Conf. Applica - 
tion of Vaccines Against Viral, Rickettsial and Bacterial Diseases of Man . 
Pan American Health Organization, World Health Organization, in press. 

Fedson, D.S., Gibbs, C. J. , Jr., and Brown, P.: Problem of antibody response 
in man following initial exposure to A2 influenza neuraminidase. Lancet , 
in press. 

Zigas, v., and Benfante, R.J.: Recent knowledge of human toxoplasmosis. 
Trop ■ Geogr. Med . , in press. 

Zigas, v., and Gajdusek, D.C.: The role of serological survey: with special 
reference to New Guinea. Papua and New Guinea Med. J ., in press. 

Gajdusek, D.C., and Alpers, M. : Genetic studies in relation to kuru. 
I. Cultural and historical background. Amer. J. Hum. Genet ., in press. 

Simmons, R.T., Graydon, J.J., Gajdusek, D.C., Alpers, M.P., and Hornabrook, 
R.W.: Genetic studies in relation to kuru. II. Blood group genetic patterns 
in kuru patients and populations of the Eastern Highlands of New Guinea. 
Amer. J. Hum. Genet ., in press. 

Kitchin, F.D., Beam, A.G., Alpers, M. , and Gajdusek, D.C.: Genetic studies 
in relation to kuru. III. The distribution of the inherited serum group 
specific protein (Gc) phenotypes in New Guineans: an association of kuru and 
the GcAb phenotype. Amer. J. Hum. Genet ., in press. 

Plato, C.C, and Gajdusek, D.C.: Genetic studies in relation to kuru. 
IV. Dermatoglyphics of the Fore and Anga populations of the Eastern High- 
lands of New Guinea. Amer. J. Hum. Genet ., in press. 

Hamlin, H. , Gajdusek, D.C., Kemper, T.L., and Yakovlev, P.I.: How differ- 
ent might be the brain of Stone Age man? Amer. J. Phys. Anthrop ., in press. 



14s 



Serial No, NDS (CF) -62 OAD 969 

1. Collaborative-Field Research 

2. Office of Associate Director 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 



Project Title: Slow, Latent, and Temperate Virus Infections of the Central 
Nervous System of Man and Animals 

Sub-Project I: Attempts to isolate transmissible agents from sub- 
acute and chronic diseases of the nervous system 

Sub-Project II: Characterization and pathogenesis of kuru virus 

Sub-Project III: Characterization and pathogenesis of Creutzfeldt- 
Jakob disease virus 

Sub-Project IV: Studies on the characterization and nature of scrapie, 
mink encephalopathy and visna virus and the relationship 
of visna virus to the virus of progressive pneumonia of 
sheep in Montana 

Sub-Project V: Studies on Australian antigen and antibody in chimpan- 
zees, monkeys and primate handlers 

Sub-Project VI: Fluorescent antibody studies on the intracellular lo- 
calization and identification of viral antigens in vivo 
and _in vitro in tissues from patients with subacute 
diseases of the CNS 

Sub-Project VII: Tissue and cell culture Jji vitro studies of viral 
induced slow infections of man and animals 

Sub-Project VIII: Characterization and distribution of two new foamy 

viruses isolated from chimpanzees tissues grown in. vitro 
and their relationship to the RNA oncogenic group of 
viruses 

Sub-Project IX: Characterization of newly identified adenoviruses 
isolated from chimpanzee tissues grown j^ v itro 

Sub-Project X: Attempts to demonstrate a viral etiology for chronic 
encephalitis with focal epilepsy 

Sub-Project XI: Isolation, epidemiology and pathogenesis of mourning 
dove pox 



15s 



Serial No. NDS (CF) -62 OAD 969 

Sub-Project XII: Infectious and contagious disease management in a 
primate colony 

Sub-Project XIII: Slow Virus Symposia, Vlth International Congress of 
Neuropathology, Paris, France 

Sub-Project XIV: Studies on the ecology, epidemiology and pathogenesis 
of arbovirus infections of man and animals 

Principal Investigators: D. Carleton Gajdusek, M.D., and 
Clarence J. Gibbs, Jr., Ph.D. 

Other Investigators: Paul Brown, M.D. , John Hooks, Ph.D., Nancy Rogers, M.S., 
Mint Basnight, M.S., Robert Cornelius, D.V.M. , 
Raymond Roos , M.D., Francoise Cathala, M.D. , Vincent 
Zigas, M.D., Ronald DiGiacomo, D.V.M. , Larry Fry, Ph.D. 

Technical Assistants: Michael Sulima, Alfred Bacote, Helena Gilbert, 

Richard Thomas, Paul Martin, Lloyd Horst, Paul Horst, 
James Webster, Judith Meyer, Sarah Andersen, Monica Lewis, 
Galen Miller, Amos Fox, James Noll and Albert Bontrager 

Students: By special arrangement this department participates in the co- 
operative college programs of the University of South 
Florida, Tampa and Antioch College, Ohio. During the 
past year the following students have spent 3-6 months in 
the laboratory: Michael Proctor, Henry Theiss, 
Jerome Kurent and Mark Littlewood 

Administrative: Marion Poms, Juliette Harvey and Sharon Williams 

Project Description: 

The studies reported in this section are being conducted in the NINDS 
Laboratory of Slow, Latent and Temperate Virus Infections on the NIH campus 
and at the Patuxent Wildlife Research Center, Laurel, Maryland. They are in 
part being conducted in collaboration with the Bureau of Wildlife and Sports 
Fisheries, U.S. Department of Interior. Additional associated collaborators 
are the Departments of Neurology and Neurovirology, Johns Hopkins University 
School of Medicine and the Departments of Epidemiology and Pathobiology, 
Johns Hopkins School of Public Health and Hygiene. In house collaborators 
are Jacob Brody, M.D. , Epidemiology Branch, John Sever, M.D. , Perinatal 
Research Branch, NINDS; K. Takemoto, S. Baron, H. Levy and W. Hadlow, NIAID; 
L. Barker, M.D. , DBS. Contractural phases of this work are being conducted 
at Gulf South Research Institute, New Iberia, Louisiana; National Center for 
Primate Biology, University of California, Davis,. California; and Public 
Health Research Institute of New York, Otisville, New York, This project is 
part of the Study of Child Growth and Development and Disease Patterns in 
Primitive Cultures and is under the director of that study. (see Project 
Report Serial No. NDS (CF)-1282 (I- IX.) 



16 s 



Serial No. NDS (CF)-62 OAD 969 

SUB-PROJECT I: Attempts to isola te transmissible agents from subacute 
and chronic diseas es of the nervous system 

Principal Investigators: D. Carleton Gajdusek, M.D. , and 
Clarence J. Gibbs , Jr., Ph.D. 

Other Investigators: Raymond Roos , M.D., Paul Brown, M.D. , David Asher, M.D. , 
John Hooks, Ph.D., Mint Basnight, M.S., Larry Fry, Ph.D., 
Nancy Rogers, M.S., Vincent Zigas, M.D. , 
Robert Cornelius, D.V.M. , Francoise Cathala, M.D. , 
Ronald DiGiacomo, D.V.M. , and Richard Sorenson, Ph.D. 

Cooperating Investigators: J.D. Mathews and R.W. Hornabrook, Kuru Research 
Office, Okapa, New Guinea; J.C. Steele, M.D. , University 
of Toronto, Canada; David Poskanzer, M.D., Massachusetts 
General Hospital, Boston; Richard T. Johnson, M.D. , Johns 
Hopkins University Hospital, Baltimore; J.T. Sever, M.D. , 
NINDS; A.M. Gardashyan, M.D. , Mayo Clinic, Rochester, 
Minnesota; J. A. Morris, Ph.D., and Hope E. Hopps, DBS; 
B.H. Dessel, M.D. , Veterans Hospital, Wood, Wisconsin; 
Elisabeth Beck and Professor P.M. Daniel, Department of 
Neuropathology, The Maudsley Institute, London; 
P. A. Palsson, D.V.M., and M. Gudnadottir, M.D. , Institute 
for Experimental Pathology, Keldur, Iceland; W. Hadlow, 
D.V.M., Rocky Mountain Laboratory, NIAID; J. Hourrigan, 
D.V.M., and H.A. McDaniel, D.V.M., Animal Research Section, 
USDA; E. Dustman, Ph.D., and CM. Herman, Sc.D., Patuxent 
Wildlife Research Center, Department of Interior; King 
Engel, M.D. , Medical Neurology, NINDS; J. A. Brody, M.D. , 
Epidemiology, NINDS; C. Colson, M.D. , Columbia Presby- 
terian Hospital, New York; R. Katzman, M.D., Yeshiva 
University, New York; M. Schaeffer, M.D. , City of New York 
Department of Health, and Dr. Ward, Otisville, New York; 
G. McKhann, M.D. , R.T. Johnson, M.D. , Leslie Weiner, M.D.,. 
Robert Herndon, M.D., Neurology-Neurovirology, Johns 
Hopkins University School of Medicine, Baltimore; Dr. 
Marino, Grady Memorial Hospital, Emory University, Atlanta; 
I.H. Pattison, D.Haig, D.V.M., and D. Hunter, Ph.D., ARS , 
CoiTipton, England; J.T. Stamp, D.V.M., Moredun Institute, 
Edinburgh; E.J. Field, Medical Research Council, Newcastle- 
upon-Tyne; F. Dixon, M.D. , and M. Oldstone, M.D. , Scripps 
Clinic and Research Foundation, La Jolla, California; 
N. Nathanson, M.D. , and F. Bang, M.D., School of Public 
Health, Johns Hopkins University Hospital, Baltimore; 
W. Zeman, M.D., Indiana University Medical Center, Indiana- 
polis; G. ZuRhein, M.D., University of Wisconsin, Madison; 
H. Koprowski, M.D., and M. Katz, M.D. , Wistar Institute, 
Philadelphia; K. Earle, M.D. , and R. Heffner, Armed Forces 
Institute of Pathology; P. Lampert, M.D. , University of 



17s 



Serial No, NDS (CF)-62 OAD 969 

California, La Jolla; P. Van Nuls, M.D. , Grand Rapids; 

T. Rasmussen, M.Do , University of Montreal, Canada; 

W. Greer, D.V.M. , Gulf South Research Institute, Louisiana; 

C. Espana, Ph.D., and R.E. Stowell, M.D. , National Center 

for Primate Biology, University of California, Davis: 

A. Lowenthal, M.D. , Foundation Born-Bunge for Research, 

Antwerp; F. Cathala, M.D., and C. Chany, M.D. , Hopital de 

la Salpetriere, Paris; H. Thormar, Ph.D., and R. Carp, Ph.D., 

Institute for Basic Research in Mental Retardation, Staten 

Island 

Technical Assistants: Michael Sulima, Alfred Bacote, Helena Gilbert, 
Paul Martin, Lloyd Horst, Paul Horst, Judith Meyer, 
Galen Miller, Amos Fox, James Noll, and Albert Bontrager 

Student Assistants and Part-time Temporary Employees: Robert Rhorer, 
Wesley Russell, Henry Theiss, Michael Proctor, 
Mark Littlewood, and Jerome Kurent 

Project Description: 

Objectives : The objectives of these long-term studies were established 
in 1957 with the discovery of kuru in the Eastern Highlands of New Guinea. 
They were fully promulgated in 1962 with the activation of the Laboratory of 
Slow, Latent and Temperate Viruses of the Nervous System. A.s established the 
objectives remain unchanged as follows: (1) to establish infection as the 
etiology of chronic and subacute progressive degenerative diseases primarily 
of the nervous system of man and animals; (2) to characterize and determine 
the nature of viruses isolated during these studies by eliciting physical, 
chemical, biological and morphological properties; (3) to study the etio- 
logical role of viruses in presenile and senile dementias as well as in nor- 
mal aging processes; (4) to study the ecology of disease, disease processes, 
and the nature of their causative agents in primitive and frequently virgin 
populations; (5) to determine the presence and interactions of "helper" or 
"hinderer" viruses in the pathogenesis of diseases under study; (6) to de- 
termine the nature of viral masking, latency, temperateness , incompleteness, 
persistence, interference, eclipse and immunopathological processes as they 
contribute to subacute degenerative processes of the CNS. The major diseases 
under study are kuru, Creutzfeldt-Jakob disease, multiple sclerosis, Alper's 
disease, Alzheimer's disease, Schilder's disease, subacute sclerosing panen- 
cephalitis, amyotrophic lateral sclerosis, parkinsonism-dementia, Parkinson's 
disease, systemic lupus erythematosus, dermatomyositis , epilepsy partialis con- 
tinua, ataxia-telangiectasia, progressive supranuclear palsy, polymyositis, 
Behcet's disease, post-infectious encephalitis, chronic tick-borne hemorrhagic 
fever, nephro-nephritis , hemorrhagic fever, Guillain-Barre syndrome, scrapie, 
mink encephalopathy, visna, progressive pneumoni.a of sheep in Montana, and 
diabetes mellitus. 

Methods Employed : Basic approaches established in 1962 and which have 
thusfar proved successful are being pursued. Essentially they consist of 
standard and classical techniques, supplemented by developmental techniques 
for the isolation of viruses with certain notable exceptions: (1) the 
demonstration that the genetic mechanism of the host influences the host 

183 



Serial No. NDS (CF) -62 OAD 969 

resistance or susceptibility to disease requires inoculation of specimens in- 
to an extensive array of animals, including higher apes and many species of 
old and new-world monkeys, birds and cell and tissue culture lines in vitro ; 
(2) exceedingly long asymptomatic incubation periods require experiments to 
be held, in isolated facilities to prevent cross infections, for periods of 
from 5 to 10 years before negative conclusions can be determined; (3) estab- 
lishment and utilization of cell culture lines iji vitro prepared by explanta- 
tion and trypsinization of biopsy and autopsy tissues from humans and animals 
with CNS diseases - these lines are maintained under rigid requirements of 
temperature and C02 for unmasking latent and endosymbiotic viruses, with 
considerable emphasis being placed on viral genome rescue employing co-acti- 
vation, fusion, Sendai virus medicated fusion; (4) utilization of fluores- 
cent, phase and electron microscopy to elicit intracellular viral genome; and 
(5) histochemical and clinical chemistry procedures to detect significant 
changes in humans and animals affected with natural and experimental diseases. 
An elaborate system for optimally obtaining specimens has been established 
with scientific collaborators in the United States and throughout the World. 
In addition to studying diseases in search of viruses this laboratory is 
investigating the nature of over 100 viruses either known or suspected of 
inducing fatal human diseases particularly those affecting the brain of man 
and animals. In these studies attempts are made to develop animal or tissue 
culture models for determining physical, chemical and biological properties 
and pathogenesis of the virus. 

Major Findings : During this year our studies on neurological diseases of 
man and animals provided data for the classification of a new group of infec- 
tious agents which we have called the Subacute Spongiform Virus Encephalo- 
pathies: kuru, Creutzfeldt-Jakob disease, scrapie and mink encephalopathy. 
Over 200 chimpanzees, 1600 smaller monkeys of 27 species or sub-species, more 
than 200,000 small laboratory animals including over 25 selectively inbred 
lines of mice, a variety of pure bred and randomly bred domestic animals and 
avian hosts as well as more than 50 established tissue and cell culture lines 
of human, animal and avian origins have, since the beginning of these studies, 
been inoculated with suspensions of tissues obtained at biopsy or early 
autopsy from humans and animals affected with neurological and systemic dis- 
eases. During the last 6 months alone, more than 450 human tissue specimens 
for virus isolation have been received into the laboratory; of these approxi- 
mately 260 are brain specimens and 507o of those submitted were brain biopsy 
specimens. Significant findings include the continued isolation, characteri- 
zation and passage of kuru through 4 serial passages in chimpanzees, the 
isolation and serial passage of the virus of Creutzfeldt-Jakob disease from 
10 patients to chimpanzees, successful primary and serial transmission of 
kuru and Creutzfeldt-Jakob disease to 4 species of new-world monkeys: spider, 
squirrel, capuchin and woolly monkeys; isolation of adenovirus type 32 from 
the brain of a patient with subacute encephalitis; isolation of virus from 
brain and lung of sheep with fatal progressive pneumonia in Montana and 
demonstration that both these viral isolates are antigenically closely re- 
lated or identical to visna and maedi viruses of Iceland; thus for the first 
tim.e establishing these viruses in the United States, and the demonstration 
that two viruses (Pan 1 and Pan 2) isolated from the brains of many chimpan- 
zees in our colony are RNA viruses associated with an RNA dependent DNA 

19s 



Serial No. NDS (CF)-62 OAD 969 

polymerase, not serologically related to known simian foamy or monkey 
mammary tumor viruses and which morphologically resemble viruses belonging 
to the RM oncogenic group. 

Kuru; Kuru virus continues to serve as the prototype virus causing fatal 
subacute degenerative disease of the CNS of man and is a dramatic model for 
the study of other spongiform encephalopathies particularly those referred to 
as the presenile dementias. A major breakthrough occurred during this report- 
ing year with the successful primary transmission of the disease to spider 
monkeys, squirrel monkeys and a capuchin monkey inoculated with suspensions 
of brain from fatal human cases. Although the incubation periods were some- 
what longer on primary passage in these animals as compared to serial passage 
in the chimpanzee it is anticipated that serial passage of the virus in the 
more readily available, easier housed and more economically purchased new- 
world monkeys will be associated with a reduction in the incubation period. 
Further, we now have conclusive evidence that neuropathological lesions are 
present in pre-clinical experimentally inoculated animals even as early as 5 
months following inoculation. For the first time we have evidence that by 
millipore filtration the virus of kuru passes through a membrane with an 
average pore diameter of lOOnm. Details on the characterization of kuru 
virus are presented in Sub-Project II. 

Creutzfeldt-Jakob disease : This disease has now been transmitted from 10 
humans to 12 chimpanzees and, like kuru, from humans to two species of new- 
world monkeys: spider monkeys and squirrel monkeys, and from Creutzfeldt- 
Jakob disease affected chimpanzees to squirrel monkeys and a brown woolly 
monkey. Successful transmissions have been obtained with brain tissues that 
have varied greatly in the degree and intensity of status spongiosis, a find- 
ing which lends support to the unitarian hypothesis of the disease. Further- 
more, the transmissible cases have been worldwide in their distribution 
coming from the United States, England, Canada, and Belgium. Although pre- 
viously thought to be a rare disease, since our first report on the trans- 
missibility we have received human specimens from over 60 cases. A critical 
review of the case histories revealed that 13 of 39 patients had liver 
abnormalities and at least two cases, as yet not transmitted, had onset of 
their neurological diseases during immunosuppressive therapy following organ 
transplantations. During the past year employing the complement- fixation and 
he'magglutination-inhibition techniques sera from human patients and experi- 
mentally infected chimpanzees were tested for antibodies against 10 major 
groups of viruses and 58 arboviruses representing Group A, Group B, Bun- 
yamwera group and a number of ungrouped arboviruses to determine whether or 
not any one or more of the viruses tested were involved in the disease. No 
etiological relationship was elicited. Details on the characterization of 
the Creutzfeldt-Jakob disease virus are presented in Sub-Project III. 

Other CNS diseases : Although we have been successful in (1) trans- 
mitting kuru and Creutzfeldt-Jakob disease to several species of primates, 
(2) isolating adenovirus type 32 from the brain of a patient with subacute 
encephalitis and (3) isolating measles virus from the brain of a patient with 
SSPE we are also accumulating negative data on animals inoculated with the 
following numbers of cases of specified disease: Alper's disease (4), amyo- 
trophic lateral sclerosis of the sporadic and familial types (15), Alzheimer's 
disease (15) , SSPE (65) , chronic focal epilepsy or epilepsia partialis 

20 s 



Serial No. NDS (CF)-62 OAD 969 

continua (16), progressive supranuclear palsy (10), Schilder's disease (7), 
multiple sclerosis (14), Alzheimer's disease of the sporadic and familial 
types (15), Parkinson's disease (6), parkinsonism-dementia (4), progressive 
multifocal leucoencephalopathy (5) , Reyes syndrome (4) , endomyocardial 
fibrosis (22), hemorrhagic fever nephro-nephritis syndrome (9), Werdnig- 
Hoffmann disease (1), rheumatoid arthritis (7), dermatomyositis (3), poly- 
myositis (3), leukemia (3), and 134 cases of subacute neurological diseases in 
which final diagnosis have not yet been established. These are in addition 
to the 56 cases of kuru and 68 cases of Creutzfeldt-Jakob disease currently 
under study. 

Significance : The results thusfar obtained in these long-term studies 
have firmly established a new group of infectious filterable viruses - which 
we have classified as subacute spongiform virus encephalopathies. The success 
of these transmissions warrant continued and increased efforts to demonstrate 
infection as the etiology of other spongiform encephalopathies of the gray 
matter of the brain in human diseases. Isolation of the kuru and Creutzfeldt- 
Jakob disease viruses now provides for the ultimate development of iinmuno- 
prophylactic measures for humans at risk, chemotherapeutic measures for 
treatment of affected humans and definition of the pathogenesis of these dis- 
eases in the human host. These studies further are providing the optimal 
techniques for the successful elucidation of the infectious nature of mul- 
tiple sclerosis, ALS , ALS-PD, Parkinson's disease, Alzheimer's disease, pro- 
gressive multifocal leucoencephalopathy, the presenile and senile dementias as 
well as the role of infectious viruses in aging processes. 

Proposed Course: 1) Identification and characterization of the agent 
causing disease in chimpanzees; 2) continued long-term observation of 
inoculated animals. Continued serial studies on the fractionation of serum 
specimens from these animals for the determination of shifts in the electro- 
phoretic patterns, as well as their antibody status which may be indicative 
of sub-clinical infections; 3) continued effort to develop suitable antigen 
antibody system for the study of established strains of 'slow' viruses; 
application of these new techniques to the study of human diseases; 4) in- 
tensification of the development and application of fluorescent antibody 
techniques with the model virus and other chronic viruses, such as LCM and 
rabies, which may remain latent for many years before clinically apparent 
disease becomes manifest; 5) greater emphasis on growth, cultivation and 
establishment of cell culture lines of "target organ", nervous tissue, from 
humans and animals with degenerative diseases of the nervous system, as well 
as from cases of "auto-immune" diseases in an effort to isolate an etio- 
logical agent in a controlled in vitro environment, detection of abnormal 
antigenic fractions giving indirect evidence of disease and possible associa- 
tion with known viruses and establishment of new cell lines for the study of 
viral growth, maturation, and measurement of interferon or interferon- like 
substances; 6) increased efforts to adapt strains of 'slow' viruses to 
growth, serial propagation and characterization in tissue and cell culture 
systems employing viral genome rescue techniques; 7) continued efforts 
toward the development of procedures for the successful isolation of etio- 
logical agents responsible for degenerative diseases of the CNS, such pro- 
cedures to include cell culture blocking techniques, detection of endosym- 
biotic relationship of masked, latent, or temperate viruses in the intact host 

21 s 



Serial No. NDS (CF)-62 OAD 969 

and host cells grown jjj vitro and chemotherapeutic and immunosuppressive 
regimens to lower animal resistance to infection; 8) continued efforts to 
broaden experimental host range for kuru, Creutzfeldt- Jakob, SSPE, visna, 
and, at the same time, seek out those experimental hosts whose genetic mech- 
anisms render them susceptible to other human diseases under study. 9) fol- 
low-up studies on Pan 1 and Pan 2 viruses (simian foamy virus 6 and 7, 
respectively) from chimpanzees to determine their distribution in man, pri- 
mates, and other mammalian species; to further elicit oncogenic and tumori- 
genic properties they may have; and, to further characterize what role they 
may have in the pathogenesis of experimental disease. 



22 



Serial No. NDS (CF)-62 OAD 969 

SUB -PROJECT II: Characterization and pathogenesis of kuru virus 

Principal Investigators: D. Carleton Gajdusek, M.D. , and 
Clarence J. Gibbs, Jr., Ph.D. 

Other Investigators: John Hooks, Ph.D., Nancy Rogers, M.S., Paul Brown, M.D. , 
Mint Basnight, M„S., Robert Cornelius, D.V.M. , 
Raymond Roos, M.D. , Ronald DiGiacomo, D.V.M. , and 
Larry Fry, Ph.D. 

Cooperating Investigators: William Greer, D.V.M., New Iberia, Louisiana; 
Carlos Espana, Ph.D., University of California, Davis; 
Peter Lampert, M„D. , University of California, La Jolla; 
Morris Schaeffer, M.D., and Gerald Ward, D.V.M., Public 
Health Research Institute, New York; Peter Daniels, M.D. , 
and Elisabeth Beck, The Maudsley Institute, London; 
K. Earle, M.D. and R. Heffner, M.D., Armed Forces Insti- 
tute of Pathology, Washington, D.C. ; Byron Kakulas, M.D. , 
Perth, Australia; Michael Alpers, M.D. , Perth, Australia; 
Charles Chany, M.D., and Francoise Cathala, M.D., Paris, 
France; Vincent Zigas, M.D., New Guinea; Sam Baron, M.D., 
and Hilton Levy, Ph.D., NIAID 

Technical Assistants: Michael Sulima, Helena Gilbert, Monica Lewis, 

Alfred Bacote, Judith Meyer, Paul Martin, Lloyd Horst, 
Richard Thomas, Paul Horst, Amos Fox, Galen Miller, 
James Webster, James Noll, and Albert Bontrager 

Project Description: 

Ob iectives : (1) To elucidate the morphological, biological, chemical, 
immunological and physical properties of kuru virus in experimentally in- 
fected animals; (2) to broaden experimental host range of the virus; 
(3) to determine the pathogenesis of the disease in the experimental host; 
and, (4) to develop techniques for the elucidation of the nature of kuru 
virus. 

Methods Employed : (1) Inoculation of chimpanzee adapted kuru virus into 
chimpanzees, lesser primates, small laboratory animals, embryonated hens' 
eggs and a variety of commercially available tissue and cell cultures; 
(2) experiments to determine the size of kuru virus by filtration through 
millipore filters; (3) effects of temperature, lyophilization procedures 
and organic solvents on kuru virus; (4) attempts to demonstrate neutralizing 
antibody to kuru virus in human and animal sera; (5) in vitro growth of 
explanted and trypsinized tissues from kuru affected chimpanzees to demon- 
strate viral antigens and virions through FA, EM and techniques to unmask 
latent agents. 

Major Findings : Kuru virus has now undergone 4 serial passages in chim- 
panzees. Of the 96 animals inoculated in these studies kuru has developed 
in 26 of 46 inoculated on primary passage with human tissues, and on serial 
passages in chimpanzees the disease has occurred in 11 of 15 on second pass- 
age, 9 of 13 on third passage and 8 of 19 on fourth passage. The virus can 



23£ 



Serial No. NDS (CF)-&2 OAD 969 

be directly isolated in squirrel, spider and capuchin monkeys inoculated with 
human brain. In addition to serial passage in the chimpanzee the virus is 
serially transmissible from chimpanzees to spider monkey, spider monkey to 
spider and squirrel monkeys and back to chimpanzees. Asymptomatic incubation 
periods on primary passage to chimpanzees have ranged from 14-39 months 
whereas in capuchin, squirrel and spider monkeys it has been 45 months, 25 
months, and 23 months, respectively. In chimpanzees serial passage is asso- 
ciated with a reduction in incubation period to 10-18 months. A similar 
reduction was noted on passage in spider monkeys when on second passage it 
was 16 months. In suspensions of human brain the virus has an infectivity 
titer of ^ 105 and in chimpanzee brain the virus titer is ^ lO^. In both 
preparations the virus is thermostable and remains transmissible following 
exposure to a temperature of 85°C/30 minutes. Although we earlier demon- 
strated that the virus passes through millipore filters of 450nm and 220nm 
we have only recently observed neuropathological lesions of kuru disease in 
the brain of a chimpanzee inoculated with the filtrate of a lOOnm filter. 
In addition to heat stability, kuru virus can be lyophilized, partially 
purified by high speed centrifugation and stored for several years without 
appreciable loss in infectivity. As previously reported, numerous attempts 
to demonstrate specific NT, CF, HI, or FA antibody in the sera from affected 
humans and animals have proven unsuccessful. Further, attempts to demon- 
strate a serological relationship of kuru to one or more conventional viruses 
have not yielded evidence of an etiological relationship. _In vitro growth 
of brain and visceral tissues from kuru affected animals have resulted in the 
successful isolation of over 200 strains of viruses representing 6-9 major 
virus groups. (See Sub-Project VII of this report.) 

Significance : Serial passage of kuru virus in chimpanzees confirms the 
infectious nature of this human brain disease. The successful transmission 
of the disease from human patients to chimpanzees, squirrel monkeys, spider 
monkeys and capuchin monkeys and the successful serial passage of the virus 
in these new-world monkeys provides a more economically feasable host in 
which to further elicit kuru virus properties. These later findings may well 
serve to break additional species barriers. The properties of the virus thus- 
far elicited with the exception of the size by filtration are remarkably 
similar to the properties of scrapie virus and suggest the viruses of these 
two diseases are similar. 

P roposed Course : Continued serial passage of the virus in chimpanzees, 
spider monkeys, and other new-world monkeys in an effort to elicit additional 
properties in the nature of kuru; expanded efforts to adapt the virus to more 
conventional laboratory hosts; and, attempts to purify and more completely 
identify kuru virus. 



24s 



Serial No. NDS (CF)-62 OAD 969 

SUB- PROJECT III: Characterization and pathogenesis of Creutzfeldt-Jakob 
disease virus 

Principal Investigators: Clarence J. Gibbs, Jr., Ph.D., and 
D. Carleton Gajdusek, M,D. 

Other Investigators: Raymond Roos, M<,D», John Hooks, Ph.D., Paul Brown, M.D, , 
Nancy Rogers, M,So, David Asher, M.D. , Vincent Zigas, M.D., 
Robert Cornelius, D.V.M. , Mint Basnight, M.S., 
Ronald DiGiacomo, D.V.M,, and Larry Fry, Ph.D. 

Cooperating Investigators: William Greer, D.V.M., Carlos Espana, Ph.D., 
Peter Lampert, M.D., Morris Schaeffer, M.D., 
Gerald Ward, D.V.M., Peter Daniels, M.D. , Elisabeth Beck, 
Kenneth Earle, M.D., R. Heffner, M.D. , Charles Chany, M.D. , 
Francoise Cathala, M.D., Louis Horta-Barbosa, D.V.M., 
John Sever, M.D., and Jacob Brody, M.D. 

Project Description: 

Objectives : To elucidate the morphological, biological, chemical, 
immunological and physical properties of the virus causing Creutzfeldt-Jakob 
disease by serial transmissions to chimpanzees; to broaden the experimental 
host range of the virus; to determine the nature of the virus and the patho- 
genesis of the disease. 

Methods Employed : As described for kuru. (See Sub-Project II of this 
report.) 

Major Findings : Employing the techniques we have developed in our studies 
on kuru we have successfully transmitted a second subacute degenerative dis- 
ease of the human brain, Creutzfeldt-Jakob disease, from 10 patients to 12 
chimpanzees on primary passage. More recently we have been successful in 
transmitting this disease from two patients to two species of new-world mon- 
keys, the spider monkey and the squirrel monkey. Further, successful serial 
passages of the virus from humans to chimpanzees and from chimpanzees to 
squirrel monkeys and the woolly monkey have been accomplished. Although the 
asymptomatic incubation periods observed in new-world monkeys (9-25 months) 
are somewhat more variable and longer than those observed in chimpanzees 
(11-14 months) , these less expensive, more readily available and more easily 
housed and handled animals are providing greater facility for expanded studies 
of this disease. During the past year independent confirmation of trans- 
missibility of this disease was obtained with the development of a fatal dis- 
ease in a chimpanzee inoculated in the laboratories of Drs. Charles Chany and 
Francoise Cathala in Paris, France. The virus is stable at —70 C and is de- 
tectable in the supernatant fluids of brain suspensions that have undergone a 
centrifugation of 5000rpm/30 minutes. Complement- fixation and hemagglutina- 
tion-inhibition techniques were employed to test sera from humans dying with 
the disease as well as to test pre- inoculation and serially collected post 
inoculation sera from chimpanzees in an attempt to elicit serological evidence 
of relationship of antibody in Creutzfeldt-Jakob disease infected hosts with 
known viral antigens. No specific patterns suggestive of an etiological re- 
lationship were observed when these sera were tested with the following 

25s 



Serial No. NDS (CF)-62 OAD 969 

antigens: influenza types A,B,C; para- influenza types 1, 2, and 3; measles; 
mumps; respiratory syncytial virus; Newcastle disease virus; echo types 6, 
9, 11, 16, 20; coxsackie types A2, A4, A8, A9, and Bl through B6; polio- 
virus types 1 through 3; Herpes simplex; cytomegalovirus; varicella-zoster; 
group adenovirus; group reovirus; vaccinia; rabies; lymphocytic choriomenin- 
gitis; rubella; SV40; psittacosis; K virus; polyoma; SV5; and, 58 strains of 
arboviruses representing groups A, B, Bunyamwera super group, and minor and 
ungrouped isolates from Europe and the North American continent. 

Significance : These studies have demonstrated that a second subacute pro- 
gressive degenerative disease of the CNS of man is caused by a virus = This 
strongly supports the continued investigations to attempt to demonstrate a 
virus etiology for other presenile and senile dementias of man. These 
studies are providing new and significant data on our understanding of sub- 
acute spongiform encephalopathies of man and are opening new vistas in neuro- 
virology. The collection of specimens from 68 cases of Creutzfeldt-Jakob 
disease within a 2-3 year period suggest the not so rare, as previously 
thought, occurrence of this disease on a world-wide distribution basis. 
Successful transmissions have been accomplished with specimens from patients 
residing in the United States, Canada, England and Belgium. 

Proposed Course : Emphasis will continue on the characterization, purifi- 
cation and identification of the virus. Continued efforts are being made to 
broaden the host range. Human tissues and tissues from affected animals are 
being extensively studied in an effort to rescue the viral genome and "helper" 
and/or "hinderer" virus interactions in Creutzfeldt-Jakob disease. Additional 
characterization of this virus as a member of the group of subacute spongiform 
encephalopathies will be continued. The epidemiology of the disease is being 
pursued. 



26 



Serial No. NDS (CF)-62 OAD 969 

SUB -PROJECT IV: Studies on the characterization and nature of scrapie , 
mink encephalopathy and visna viruses 

Principal Investigators: Clarence J. Gibbs, Jr., Ph.D., and 
D. Carleton Gajdusek, M.D. 

Other Investigators: John Hooks, Ph.D., Raymond Roos , M.D. , Paul Brown, M.D. , 
Nancy Rogers, M.S., Mint Basnight, M.S., and 
David Asher, M.D. 

Cooperating Investigators: J. A. Morris, Ph.D., DBS; Jacob Brody, M.D., NINDS; 
Paul Albrecht, M.D., DBS; Michael Stewart, M.D., 
Hilton Levy, M.D., Larry Sturman, M.D. , William Hadlow, 
D.V.M. , NIAID; Leslie Weiner, M.D,, Richard Johnson, M.D. , 
and Robert Herndon, M. D., School of Medicine, Johns Hopkins 
University; Neal Nathanson, M.D. , Gerald Cole, Ph.D., and 
Don Gilden, M.D. , Johns Hopkins School of Public Health and 
Hygiene; James Hourrigan, D.V.M. , and A. Klingsporn, D.V.M., 
USDA; John Hotchin, M.D. , Albany; C. Chany, M.D., and 
F. Cathala, M.D. , Paris; H. Koprowski, Wistar Institute; 
D. Porter, M.D., University of California; Peter Lampert, 
M.D. , University of California; M. Oldstone, M.D. , 
Scripps Clinic and Research Foundation 

Project Description: 

Objectives : To study scrapie, and mink encephalopathy as animal models 
of subacute spongiform virus encephalopathies; to determine the pathogenesis 
of these diseases in laboratory animals in an effort to elicit the mechanisms 
of infection, virus-cell-relationships, nature of the virus per se and the 
role of these viruses in inducing an aging process in infected animals. To 
study visna, a demyelinating disease of sheep, as a model in the study of 
demyelinating diseases of man, particularly, multiple sclerosis. 

Methods : As previously described in earlier Annual Reports. 

Major Findings: 

Scrapie and mink encephalopathy : The recent results of our studies on 
these tw/o diseases places them in the group of viruses which we now call 
subacute spongiform virus encephalopathies primarily affecting the gray 
matter of the brain: two of man: kuru and Creutzfeldt-Jakob disease; and, two 
of animals: scrapie and mink encephalopathy. This classification is based 
on (1) each is transmissible to an animal host though not necessarily the 
same host, (2) each is associated with a long incubation period of from 
several months to several years; (3) each induces a clinical syndrome con- 
sisting of progressive cerebellar ataxia, tremors and postural instability 
which is always fatal in termination; and, (4) each manifest histopatho- 
logical lesions only in the gray matter of the brain consisting of severe 
intraneuronal vacuolation and dropout, astrocytosis , fibrous gliosis and 
moderate to marked status spongiosis without signs of acute inflammatory 
response. 

During the period covered by this report we have continued our studies 
on the characterization of scrapie and mink encephalopathy. Wa have con- 

27s 



Serial No. NDS (CF)-62 OAD 969 

tinued to study the effects of altering the immune mechanisms of the host by- 
surgical spleenectomy, thymectomy and through the use of anti-lyraphocytic 
serum, cyclophosphamide, and exposure to X-irradiation. None of the pro- 
cedures employed had an effect on incubation periods, development or duration 
of clinical disease or fatal termination of the process. In a series of ex- 
tensive experiments we were again unable to demonstrate antigen-antibody 
reactions by complement-fixation procedures utilizing crude or sucrose ex- 
tracted suspensions of brain, kidney and spleen taken from mice at 30 days 
intervals after IC inoculation of scrapie. Sera employed in these studies 
were mouse, hyperim.munized rabbits and roosters, and naturally infected 
sheep. During the course of these studies we observed that following inocu- 
lation of scrapie IC into mice, virus remains detectable in the brain, spreads 
rapidly to the spleen and to a lesser concentration is detectable in the 
kidney. Virus in the kidney is associated with a pre-zoning effect. Whether 
this is due to the presence of specific neutralizing gamma globulin deposits 
on the basement membrane of glomeruli or whether the effect is due to non- 
specific interference is under study. Our studies suggest that the reaction 
is probably not due to the production of interferon since no increased levels 
of interferon have been detected in scrapie infected animals. Further, pro- 
phylactic treatment of mice with high concentrations of potent mouse inter- 
feron given before and at intervals after exposure to scrapie have shown no 
effect on the subsequent disease. Our studies of scrapie and mink encephalo- 
pathy have further confirmed the unusual properties of thermostability, 
resistance to ultraviolet irradiation, formalin and proteolytic enzymes. We 
have successfully repeated our studies on the size of scrapie as determined 
by filtration and our findings of 20nm-30nm has been independently confirmed 
by scientists in NIAID and at Corapton, England. Administration of poly I- 
poly C given in large doses at a variety of times had no effect on incubation 
period, clinical signs or fatal termination of the disease. 

Visna : During the period covered by this report we have developed direct 
and indirect methods of demonstrating visna virus in vitro and antibody in 
v ivo by fluorescent microscopy. We have developed highly specific techniques 
for demonstrating neutralizing antibody to visna virus in sheep serum in 
tests conducted in sheep choroid plexus, lamb kidney cells and sheep testis 
cell cultures. Consistently negative results have been obtained in studies 
to adapt visna virus to mice, rats, rabbits, guinea pigs and hamsters by 
inoculation of high concentrations of virus followed by serial blind passages. 
The use of immunosuppressants did not alter the non-reactivity of animals to 
the virus. Multiple inoculations of high concentrations into mice and 
rabbits have failed to induce detectable levels of NT or FA antibodies. 

It is of significance that disease has not occurred in chimpanzees, 
rhesus monkey, cynomolgus monkeys or African green monkeys in the more than 
5 years since they were inoculated intracerebrally only or by multiple routes 
peripherally with scrapie, mink encephalopathy and visna virus. 

Progressive pneumonia of sheep (Montana sheep disease) : We had earlier 
postulated that progressive pneumonia of sheep in Montana was caused by a 
virus closely related to maedi and visna viruses [ In 1967 we obtained brain 
tissue from a sheep and lung tissue from a second sheep both of which had 
clinically diagnosed progressive pneumonia. Viruses were isolated from both 
specimens submitted. We have now demonstrated chat both strains of virus are 



28 1 



Serial No, NDS (CF)-62 OAD 969 

antigenically related to each other as well as to visna virus by cross 
neutralization and cross fluorescent antibody tests. This work has been 
confirmed by investigators in NIAID, NIH and at RML, in Montana. These same 
investigators have extended these findings to show that the viruses are BINA 
viruses associated with an RNA.-DNA dependent polymerase. This plus their 
morphological similarity to the RNA oncogenic group of viruses as determined 
by EM warrants investigation into their potential oncogenic and tumorigenic 
capacities . 

Significance : Studies on scrapie, mink encephalopathy, visna and the two 
strains of viruses isolated from sheep with progressive pneumonia provide 
optimal models for the ±n vivo and ±n vitro mechanisms that are involved in 
slow infections of the nervous system. Data can be extrapolated to assist in 
the design of experiments and interpretation of results of studies of human 
neurological diseases processes. The self fusing properties of visna virus, 
even when inactivated, offer a possible mechanism for interpreting the in 
vivo process of demyelination, and the relationship of the visna-Montana 
sheep disease virus complex to the oncogenic group of RNA viruses. 

Proposed Course : Continued studies into the mechanism of infection, the 
pathogenesis of disease and the immune mechanisms that may influence slow 
infections due to scrapie, mink encephalopathy, visna virus, and the virus of 
progressive pneumonia. To expand these latter studies to include a search for 
naturally occurring tumors in progressive pneumonia of sheep as well as 
alveolar carcinoma of man. 



29s 



Serial No. NDS (CF) -62 OAD 969 

SUB-PROJECT V: Studies o n Australian antigen and anti b ody in ch im pan - 
zees , monkeys and primate handlers 

Principal Investigators: Robert Cornelius, D.V.M. , D. Carleton Gajdusek, 
M.D, , and Clarence J. Gibbs, Jr., Ph.D. 

Other Investigators: Lewellyn Barker, M.D. , and Michael Peterson, M.D. , DBS; 
John Hooks, Ph.D., and Nancy Rogers, M.S. 

Project Description: 

Objectives : Australian antigen was first associated with hepatitis in 
1964, and it was first found in the chimpanzee in 1969. In an attempt to de- 
fine the possible role of the nonhuman primate as a source of human infection, 
and to explore the carrier state, if any, in the nonhuman primate, this study 
was undertaken. 

Methods : The NIH laboratory at the Patuxent Wildlife Research Center has 
approximately 80 chimpanzees and 350 smaller nonhuman primates. These animals 
are housed in two buildings, both of which contain new and old-world monkeys, 
and several species of each. 

In 1969 serum samples from 95 chimpanzees and approximately 50 other pri- 
mates were tested for Australian antigen and antibody. Serum samples from 59 
chimpanzees and 6 other primates were also tested in 1970. Serum was tested 
for Australian antigen by the Ochterlony micro technique for ppt. and by 
complement-fixation, and for antibody using the Ochterlony micro technique. 

Those animals possessing antigen were then bled at monthly intervals for 
6 months and that serum tested for antigen and antibody, and blood chemis- 
tries for liver function. Cagemates of any positive animals were subjected 
to the same schedule. To attempt to ascertain when the infection occurred, 
serum stored at — 70°C from previous serial bleedings, coTimencing soon after 
arrival, were also pooled and tested. 

All laboratory personnel who came in contact with the animals, their 
tissues or blood, were placed on a bimonthly bleeding schedule. This serum 
was screened for antigen and antibody and liver functions. 

Major Findings : Ninety-five chimpanzees were sampled in the first series. 
Two animals showed 4+ antigen by CF or agar gel ppt. These two animals A28 
and A62 were still in the colony when the current study was undertaken. 

Eighty-eight serum samples from 59 chimpanzees, and 7 serum samples from 
6 other primates, were tested more than a year later. Two chimpanzees ex- 
hibited antigen titers by agar gel ppt. or CF, and one chimpanzee serum con- 
tained antibody. In addition, one stumptail monkey showed antibody titer of 
1:20 by CF. 

The two chimpanzees that had antigen in 1970, were the two animals that 
previously had shown antigen in 1969, and were still in the colony (A28 and 
A62) . Chimpanzee A51 that showed antibody, was shown to have had antigen of 
1:20 by ppt. in 1968, by the testing of stored serum„ 

The testing of stored serum from A28 and A62 . taken soon after arrival in 
the colony showed that they had carried the antigen for some time. While the 
titer was decreasing, A28 had been infected for at least 4 years, and A62 for 
at least 3.5 years. 

While A28 was housed singly, A62 was caged with A51. While this is of 

30 s 



Serial No. NDS (CF)-62 OAD 969 

interest, it could not be established how long these two animals had been in 
close contact. 

Eleven laboratory personnel were bled 5 times in 7 months. Antigen was 
not detected by agar gel ppt. or CF in any sample, nor were there any liver 
enzyme elevations of significance. 

Signif icance : Two findings were considered to be of value in this study. 
The first was the demonstration that chimpanzees can show infection with 
Australian antigen for at least 4 years, and still not have detectable anti- 
body. And the second that transmission between cagemates appears to be 
possible, or has occurred. 

Proposed Course ; It is planned to continue with the monitoring procedure 
to detect infection, titer, persistence and carrier/shedding state. Immuni- 
zation of primates for the production of antibody will be attempted. The 
purpose of these procedures will be to study the feasability of the large 
scale production of antibody for use in screening of donors and blood products 
as well as to prepare purified gamma globulin for immunoprophylactic pro- 
cedures. 



31 



Serial No. NDS (CF)-62 OAD 969 

SUB- PROJECT VI: Fluorescent antibody studies on the intracellular 

localization and identification of viral antigens in vivo 
and in vit ro in tissues from patients with subacute dis- 
ease"s~ot the CNS 

Principal Investigators: D. Carleton Gajdusek, M.D. , and 
Clarence J. Gibbs, Jr., Ph.D. 

Other Investigators: John Hooks, Ph.D., Raymond Roos, M.D. , Paul Brown, M.D. , 
Nancy Rogers, M.S., Mint Basnight, M.S., and 
David Asher, M.D. 

Project Description: 

All phases of this project remain essentially as reported for FY1969. 
Notable exceptions are the preparation of several new conjugates, adaptation 
of FA technique to the in vivo and in vitro study of viral strains isolated 
from explant cultures and utilization of this technique in studies to detect 
viral antigens and antibodies in patients with multiple sclerosis, kuru, 
Creutzfeldt-Jakob disease and other degenerative diseases of man primarily 
those which cause spongiform changes in the gray matter of the brain. 



32 s 



Serial No. NDS (CF)-62 OAD 969 

SUB-PROJECT VII: Tissue and cell culture ^^ XiE££ ^'^'^^^^^ °^ viral 
induced slow infections of man and animals 

Principal Investigators: D. Carleton Gajdusek, M.D., 

Clarence J. Gibbs , Jr., Ph.D., and Nancy Rogers, M.S. 

Other Investigators: John Hooks, Ph.D., Raymond Roos , M.D., Paul Brown, M.D., 
Mint Basnight, M.S., Helena Gilbert, David Asher, M.D. , 
and Robert Cornelius, D.V.M. 

Cooperating Investigators: Harish Chopra, M.D. , NCI; S. Chou, M.D. , Univer- 
sity of West Virginia; Peter Lampert, M.D., University of 
California; V. ter Muelen, M.D,, Gottengen, Germany; 
C. Chany, M.D. and F. Cathala, M.D,, Paris, France; 
S. Baron, M.D. , NIAID 

Project Description: 

Objectives : To grow and maintain in vitro tissue and cell culture lines 
prepared with specimens of brain, visceral, lymphatic, muscle and blood 
tissues from human patients and animals succumbing to persistent, chronic, 
subacute progressive degenerative diseases particularly of the central ner- 
vous system, for the purpose of isolating viruses and demonstrating an 
etiological relationship to the diseases under study. 

Methods : Brain, visceral, lymphatic, muscle and blood tissues obtained 
at surgical biopsy or early autopsy are grown in vitro as explanted organ 
type cultures, trypsinized dispersed monolayer cultures, and as polykaryons 
following cocultivation and Sendai virus medicated fused cultures with per- 
missive cell lines for the rescue of viral genome from donor tissues. All 
cultures are serially sub-cultured and maintained _in vitro over long periods 
during which time they are (1) observed for spontaneously developing CPE, 
(2) transformation, (3) detection of viral antigens by FA, hemadsorption, 
CF, and, HA tests, (4) examined by EM for morphology and viral particles. 
They are blind passaged into a wide variety of permissive cell lines and 
tested for detectable viral antigens. Attempts to isolate viruses are also 
being made by classical techniques of inoculating suspensions of various 
tissues and organ suspensions from humans and animals into a wide variety of 
primary and stable cell lines. Finally, attempts are being made to incorpor- 
ate kuru and scrapie into mouse tissue culture cells during the process of 
cell fusion. Two strains of cell lines are currently being employed: 
C 11 D(TK) and A9(HGPRT-) which were chosen because they allow through their 
enzyme deficiencies the selection of hybrid cells which grow in the presence 
of amniopterin. Hybrid cells are also being investigated for the appearance 
of new antigens after treatment with virus. Such treated cells combined with 
Freund's complete adjuvant are used to immunize C3H/Hen mice which are sub- 
sequently tested for antibody by means of immune hemadsorption test. 

Major Findings : To date, over 1500 explants of brain, visceral, Ijrmphatic 
and muscle tissues from humans and animals dying with subacute degenerative 
CNS diseases have been grown and maintained in vitro as primary explants and 
serial cultures. Over 225 viral isolates have been recovered, the vast major- 
ity coming from explant cultures of chimpanzee tissues. Of these isolates 

33 s 



Serial No. NDS (CF)-62 OAD 969 

approximately 827o are strains of new foamy viruses Pan 1 and Pan 2, described 
in Sub-Project VIII. TL are strains of adenoviruses Pan 5, Pan 6, Pan 7 and 
Pan 9 described in Sub-Project IX; two have been strains of reovirus and 
three remain unidentified viruses from chimpanzees. One of these has been 
designated Pan 8. The very slow appearance of CPE is necessitating a search 
for a more satisfactory cell line to pursue characterization and identifica- 
tion studies. It is significant that virus isolations have now been made 
from the brains of spider monkeys (foamy- like virus) , squirrel monkeys 
(herpes-like virus) and in one instance a squirrel monkey had both herpes-like 
and foamy- like virus in his brains. Human tissues have yielded fewer isolates. 
We have demonstrated measles virus specific antigen intracellularly in explant 
cultures of brain from a case of SSPE; a single, as yet unidentified virus- 
like agent has been recovered from brain explant cultures prepared from a 
patient dying with ALS. An adenovirus, closely related to type 32 by neutral- 
ization tests and to type 27 by hemagglutination-inhibition tests, has been 
isolated from the brain of a human that died of lymphosarcoma with a subacute 
encephalitis. Virions morphologically resembling adenovirus were observed by 
EM in the patient's brain as well as in infected human embryo kidney cell 
cultures. This is only the second reported instance that adenovirus has been 
isolated from brain and the first reported instance that adenovirus can cause 
a subacute encephalitis. 

Proposed Course : Continued emphasis will be placed on in^ vitro growth of 
human and animal tissues in attempts to isolate latent, persistent, chronic, 
masked viruses and viruses which share an endosjTnbiotic relationship with the 
donor hosts. Studies are being continued to elicit the etiological signifi- 
cance of viral isolates to human and animal diseases. The effects of latent 
viruses as they relate to presenile and senile dementias and to aging pro- 
cesses in cells is being further studied. 



34: 



Serial No. NDS (CF)-62 OAD 969 

SUB-PROJECT VIII: Characterization and distribution of two new foamy 

viruses isolated from chimpanzees t issues grown in vitro 
and their relationship to the RNA oncogenic group~oT 
viruses 

Principal Investigators: John Hooks, Ph.D., Nancy Rogers, M.S., D. Carleton 
Gajdusek, M.D. , and Clarence J. Gibbs, Jr., Ph.D. 

Other Investigators: Mint Basnight, M.S., Robert Cornelius, D.V.M., and 
Ronald DiGiacomo, D.V.M. 

Cooperating Investigators: Peter Lampert, M.D. , University of California; 
Harish Chopra, M.D., NCI; H. Thormar, Ph.D., and F.H. 
Lin, M.D. , Institute for Basic Research in Mental Re- 
tardation, Staten Island 

Project Description: 

Ob iectives : The purpose of this study is to characterize two new foamy 
viruses, simian foamy 6 (Pan 1) and simian foamy 7 (Pan 2), isolated from 
chimpanzees; to elicit their experimental host range, and their physical, 
chemical and biological properties; to determine their etiological signifi- 
cance in human and animal diseases; and, to determine what etiological role 
they may have in experimental kuru and Creutzfeldt-Jakob disease in chimpan- 
zees and new- world monkeys. To determine the relationship of Pan 1 and Pan 2 
to prototype strains of simian foamy viruses as well as to monkey mammary 
tumor virus. 

Methods : Pan 1 and Pan 2 viruses were isolated from brain, visceral 
tissues and lymphatic tissues from several chimpanzees affected with kuru, 
Creutzfeldt-Jakob disease and from chimpanzees dying with intercurrent infec- 
tions. The viruses were isolated and maintained in primary HEK cultures 
which were incubated at 35°C - 37°C. Characterization studies of Pan 1 and 
Pan 2 were conducted using HEK cultures as the biological system of choice. 
Virus purification and assay for the presence of RNA dependent DNA polymerase 
were done using the procedure described by Lin and Thormar (Journal of 
Virology, 6: 1970) • Cross neutralization tests employing antibody specific 
to simian foamy viruses 1 through 7 and mouse mammary tumor virus in HEK cul- 
tures were performed to determine antigenic relationships between these 
viruses . 

Major Findings : These studies have shown that Pan 1 and Pan 2 viruses 
isolated from chimpanzees are myxo-like RNA viruses which induce a foamy, 
syncytia without inclusion bodies in HEK cultures. The viruses do not share 
common neutralizing or fluorescent antibody antigens and neither is related 
tc any of the known simian foamy viruses or to monkey mammary tumor virus. 
They are morphologically indistinguishable from each other by EM and are 
125nm in diameter. The virions consist of a central core 45nm, surrounded by 
an inner capsule of 80nm. Virions are observed only in the cytoplasm and at 
maturation obtain their outer envelope by budding into intracellular vacuoles 
or at the plasma membrane. The morphology of these viruses is strikingly 
similar to prototype simian foamy viruses and those viruses belonging to the 
oncogenic RNA viruses e.g. mouse mammary tumor, murine leukemia, bovine syn- 

35s 



Serial No. NDS (CF)-62 OAD 969 

cytia and monkey mammary tumor virus. Both viruses are ether, chloroform 
and pH-sensitive and are inactivated by exposure to 56°C/30 minutes. They 
pass through 220nm millipore filters but not through lOOnm. Homologous 
antibody occurs in the serum of donor chimpanzees. Antibodies to one or both 
viruses are also found in chimpanzees not inoculated with experimental kuru 
or Creutzfeldt- Jakob disease. Following purification procedures two main 
areas of protein and nucleic acid were demonstrable by absorbance of gradient 
fraction at 260nm and 280nm. Infected materials show a larger peak at frac- 
tions 5 through 15. The density of the area ranges from 1.08g/ml to 1.12g/ml. 
Enzyme activity, determined by uptake of H thjonidine, was observed in frac- 
tions 5-15 and to a lesser extent in other samples tested. Preliminary 
studies on the acid-insoluble product show reaction at 37°C and resistance to 
RNase. Cross neutralization studies showed that Pan 1 and Pan 2 viruses were 
not antigenically related to each other nor is either one related to simian 
foamy viruses types 1-5; monkey mammary tumor virus, visna virus, bovine syn- 
cytial virus or the virus which causes progressive pneumonia in sheep in 
Montana. Finally, although Pan 1 and Pan 2 share the property of RNA-DNA 
dependent polymerase with monkey mammary tumor virus they differ from this 
virus in their morphology. 

Significance : Identification and characterization of two new foamy 
viruses from multiple tissues of a high percentage of chimpanzees tested 
suggest wide distribution of these viruses in nature. The CPE they induce in 
HEK cultures is strikingly similar to the intracytoplasmic vacuolation of 
neurons and astroglia observed in the brains of humans and animals affected 
with kuru and Creutzfeldt-Jakob diseases. 

Proposed Course : (1) Continuation of seroepidemiological survey to de- 
termine the distribution of these viruses in man and animals; (2) increased 
emphasis on the pathogenesis of these two viruses in chimpanzees to determine 
their etiological significance in experimentally induced diseases of the 
brain, (3) increased emphasis to determine the oncogenic and tumor inducing 
properties of these viruses and (4) expanded studies to determine the re- 
lationships between these viruses and other viruses now classified as onco- 
genic RNA viruses. 



36 s 



Serial No. NDS (CF)-62 OAD 969 

SUB- PROJECT IX: Characterization of newly identified adenoviruses 
isolated from chimpanzee tissues grown in vitro 

Principal Investigators: Mint Basnight, M.S., Nancy Rogers, M.S., and 
D. Carleton Gajdusek, M.D. 

Other Investigators: Clarence J. Gibbs, Jr., Ph.D., Raymond Roos , M.D. , 
John Hooks, Ph.D., and Paul Brown, M.D. 

Cooperating Investigator: Wallace Rowe, M.D. , NIAID 

Project Description: 

Objectives : Identification and characterization of heretofore unidenti- 
fied adenoviruses isolated from chimpanzee tissues explants grown in vitro . 

Methods : Standard classical techniques for the isolation and identifi- 
cation of viruses utilizing tissue and cell culture systems. 

Major Findings : Four antigenically distinct viruses designated Pan 5, 
Pan 6, Pan 7 and Pan 9 were isolated in primary HEK inoculated with fluids 
from explant cultures of lymphoid tissues obtained from three chimpanzees. 
In HEK and primary African green monkey kidney cells each of the viruses 
induces a CPE consisting of rounding-up of cells and the formation of grape- 
like clusters characteristic of adenoviruses. All four are DNA, chloroform 
resistant, heat sensitive viruses which are stable at pH 3.0. Each passes 
through a lOOnra millipore filter. None of the four viruses is antigenically 
related to known human or simian adenoviruses by neutralization or complement- 
fixation procedures. Homologous antibody occurs in the serum of each chim- 
panzee from whose tissue the virus has been isolated. 

Proposed Course : Seroepidemiological studies to determine the distribu- 
tion of these four newly identified viruses in man and animals. Studies are 
being conducted to determine the significance of these viruses in animals 
developing experimentally induced kuru and Creutzfeldt-Jakob disease. 



37: 



Serial No. NDS (CF)-62 OAD 969 

SUB- PROJECT X: Attempts to demonstrate a vira l etiology for chronic 
encephalitis with focal epilepsy 

Principal Investigators: D. Carleton Gajdusek, M.D, , Clarence J. Gibbs , Jr., 
Ph.D., and David Asher, M.D. 

Other Investigators: John Hooks, Ph.D., Nancy Rogers, M.S., Paul Brown, M.D. , 
Ra^TTiond Roos, M.D. , Larry Fry, Ph.D., Mint Basnight, M.S., 
Robert Cornelius, D.V.M., and Ronald DiGiacomo, D.V.M. 

Cooperating Investigators: T. Rasmussen, M.D. and S. Carpenter, M.D., 

Montreal Neurological Institute, Canada; A. A. Smorodintsev, 
M.D, and V.I. Il'yenko, M.D., Leningrad, U.S.S.R. 

Project Description: 

Occasional patients with chronic focal epilepsy are found at craniotomy 
to have active inflammation of the brain months or years after the onset of 
illness. In the USSR this condition, called Kozhevnikov 's epilepsy, seems to 
follow acute tick-borne encephalitis (TBE) . There has been one published re- 
port of the isolation of TBE virus from the brain tissues of two such patients, 
but this has never been confirmed. Il'yenko and colleagues in Leningrad have 
discovered a promising animal model for this condition; several strains of TBE 
when inoculated into rhesus monkeys by the intracerebral route produce only 
mild encephalitis, followed by a month or so later by the appearance of focal 
choreoathetotic tremors, along with histological evidence of chronic active 
inflammation of the brain, from which virus has been recovered up to a year 
after inoculation. We have attempted to confirm Dr. Il'yenko's finding using 
one of her strains of TBE virus. Four monkeys have been observed for three 
months without evidence of movement disorder, and eight more have been inocu- 
lated more recently. If the Soviet findings are confirmed we will investigate 
the mechanisms of virus persistence by immunosuppression, and by Jji vitro 
cultures of brain cells. Dr. Il'yenko has been invited to participate in our 
studies through the U.S. -U.S.S.R. Health Exchange Agreement, 

Rasmussen and co-workers have observed focal epilepsy with chronic enceph- 
alitis in 23 patients, most of them from the United States and Canada. None 
had preceding acute encephalitis, although many had a variety of other febrile 
illnesses. In our laboratories there have been several attempts to isolate 
viruses from the brains of these patients by inoculation of mice, primates and 
tissue cultures, without success. Brain tissue from a recent patient has been 
grown in vitro by trypsinization and explant; tissue is now in serial passage 
and will be fused with several indicator cell lines in an attempt to detect 
latent viruses. 



38 



Series No. NDS (CF) -62 OAD 969 

SUB-PROJECT XI: Isolation, epidemiology and pathogenesis of mourniag 
dove pox 

Principal Investigators: David Asher, M.D, , D. Carleton Gajdusek, M.D. , and 
Clarence J. Gibbs, Jr., Ph.D. 

Cooperating Investigators: E. Dustman, Ph.D., C. Herman, Sc.D., and 
L. N. Locke, D.V.M. , Patuxent Wildlife Research 
Center, Department of Interior, Laurel, Maryland 

Project Description: 

Troublesome outbreaks of pox disease have occurred among wild mourning 
doves (Z enaidura macroura) in many areas of the United States, including 
Maryland. The agent of this disease was isolated in the chorioallantoic 
membranes of hens' eggs. Of five orders of birds tested only doves and 
pigeons were susceptible. The domestic pigeon ( Columba livia ) developed 
only a mild form of disease. Dove-pox virus protected domestic pigeons 
but not mourning doves against challenge with pigeon pox virus. A serologi- 
cal study of the antigenic relationship of dove-pox virus to pigeon-pox virus 
is in progress. 



39 s 



Serial No. NDS (CF)-62 OA.D 969 

SUB-PROJECT XIJ. : lafectious and contagious disease management in a 
primate colony 

Principal Investigators: Robert Cornelius, D.V.M. , Ronald DiGiacomo, D.V.M. , 

D, Carleton Gajdusek, M.D, , and Clarence J. Gibbs, Jr., Ph.D. 

Other Investigators: David Asher, M.D., Paul Brown, M.D,, Raymond Roos, M.D. , 
John Hooks, Ph.D., Nancy Rogers, M.S., Mint Basnight, M.S., 
Luis Melendez, D.V.M., and Keerti Shah, M.D. 

Technical Assistants: Alfred Bacote, Michael Sulima, Monica Lewis, Paul 
Horst, Lloyd Horst, Paul Martin, Amos Fox, James Noll, 
Galen Miller, James Webster, and Albert Bontrager 

Cooperating Institutions: Animal Resources Branch, NIH; Armed Forces Insti- 
tute of Pathology, Departments of Neuropathology and 
Veterinary Pathology; Department of Neuropathology, Uni- 
versity of California; Department of Neuropathology, The 
Maudsley Institute; Department of Pathology, University 
of Western Australia School of Medicine; Gulf South 
Research Institute, National Center for Primate Biology; 
Otisville Laboratories of the Public Health Research 
Institute of the City of New York; Department of Patho- 
biology, Johns Hopkins University School of Public Health 
and Hygiene; New England Regional Primate Center 

Project Description: 

Objectives : To systematically study normal behavior, breeding habits, 
clinical chemistry and hematological values, clinical and sub-clinical inter- 
current infections in program and control animals employed in the study of 
slow, latent and temperate viruses of the nervous system; to establish a 
registry of normal and abnormal values observed in a well conditioned and 
well maintained colony. 

Major Findings: 1) Herpes simiae : during the later part of 1970 over 
907o of the rhesus monkey population in this program was bled and examined for 
NT antibody to Herpes simiae. A total of 15 of 97 rhesus tested had NT 
antibody with titers of 1:100 or greater. 87 of the monkeys were imported 
from India and 19% of these had antibody. There was no apparent correlation 
of Ab with age, sex or length of stay in the colon}'^. 8 of 13 monkeys that 
had Ab were or still are cagemates. 16 of the 97 Ab positive monkeys were 
colony reared. 

2) Spontaneous di se ase in program animals : None of the program disease 
problems have been observed in a variety of animal species being employed 
as experimental hosts. In primates pulmonary vascular sclerosis, pregnancy 
toxemia, mycotic pneumonitis, gastric ulcer, polynephritis , meningitis and 
Herpesvirus-T infection have been a problem. During the year newly imported 
mink developed clinical signs of Aleutian mink disease within 5 days after 
being received into the quarantine unit. This was later confirmed by histo- 
logical study. In the ferret colony we have observed spontaneous occurrence 
of liver tumors of vascular origin and bronchial asthma due to filarids. 



40 s 



Serial No. NDS (CF)-62 OAD 969 

3) Collection, tabulation and program analysis of data on normative or- 
gan weights and hematological parameters in the chimpanzee is currently under 
study. 

S ignificance : 1) Herpes simiae infection in a primate colony is of 
importance because of the high mortality rate this virus causes in man. The 
ongoing study we have initiated will serve to assess the degree of communica- 
bility and exposure hazard for animal caretakers. 2) A general diagnostic 
pathological workup is being performed on each animal (control and inoculated) 
in our research program that dies with an intercurrent infection. Such a 
surveillance provides information on diseases occurring within the colony and 
provides information for successful management of a large colony which is 
made up of old world and new-world monkeys. 3) Normative data on organ 
weights, hematological, CSF, clinical chemistries and serological parameters 
in the chimpanzee and smaller primates will assist in the rapid diagnosis of 
disease. 

Proposed Course : Continuation of this long-term study and publication of 
a compendium on normal and diseased animal values. 



418 



Serial No. NDS (CF)-62 OAD 969 

SUB- PROJECT XIII: Slow Virus S^nrnposia, Vlth International Congress of 
Neuropathology, Paris, France 

Principal Investigators: Clarence J. Gibbs, Jr., Ph.D., and 
E. Osetowska, M.D. 

Other Investigators: D. Carleton Gajdusek, M.D. 

Cooperating Investigators: Hilary Koprowski, M.D., Wistar Institute, Phila- 
delphia; Jean Lapresle, M.D., and Francoise Cathala, M.D., 
Hopital de la Salpetrifere, Paris; Richard Johnson, M.D. , 
Johns Hopkins University, Baltimore 

Project Description: 

The development, organization and promulgation of a major theme on 
"Pathogenesis of Slow Virus Diseases of the Central Nervous System" as part 
of the Vlth International Congress of Neuropathology, Paris, France, August 
31 through September 4, 1970. 

Results : Publication of Proceedings of Vlth International Congress of 
Neuropathology, Masson & Cie, Paris, 1970. 

E. Osetowska and C.J. Gibbs, Jr., Theme IV, "Pathogenesis of Slow Virus 
Diseases of the Central Nervous System", Proceedings of Vlth International 
Congress of Neuropathology, Masson & Cie, Paris, 1970: 

Johnson, R.T. : Virus-Host relationships in acute and chronic encephal- 
opathies, pp. 761-778. 

Gibbs, C.J.jJr. and Gajdusek, D.C.: Characterization and nature of 
viruses causing subacute spongiform encephalopathies, pp. 779-801. 

Hunter, G.D. : The biochemical properties and nature of the scrapie 
agent, pp. 802-817. 

Oldstone, M.B.A.: Pathogenesis of cellular injury associated with per- 
sistent viral infection, pp. 818-824. 

Simons, M.J., Fitzgerald, M.G. , and Alpers, M.P. : Lymphocytic function 
in kuru. pp. 825-826. 

Brody, J. A, and Nemo, G.J.: Response of peripheral blood lymphocytes 
from multiple sclerosis patients to guinea pig basic protein, and multi- 
ple sclerosis patients' cerebrospinal fluid and brain, pp. 827-828. 

Hotchin, J.: Immunological aspects of slow virus disease: LCM virus- 
induced anti-brain antibodies as a cause of neurological damage, 
pp. 829-830. 

Petursson, G.: Studies on viral antibodies in visna. pp. 831-832. 



42 s 



Serial No. NDS (CF)-62 OAD 969 

Albrecht, P.: Immune status of the organism during experimental scrapie 
infection, pp. 833-834. 

ter Meulen, V., Muller, D„, and Katz, M. : Immunohistological and histo- 
chemical studies in subacute sclerosing panencephalitis: An example of an 
analysis of a slow virus infection of the CNS. pp. 835-836. 

Koestner, A., Long, J.F. , Jacoby, R.O. , Olsen, R.G. , and Shadduck, JoA. : 
Canine distemper as a model of a parainfectious demyelinating encephal- 
opathy, pp. 837-838. 

Parry, H.B,, and Vince, A, A.: Scrapie disease of sheep: the roles of 
gene and "slow virus" in pathogenesis, pp. 839-840. 

Dickinson, A.G. : Classification of scrapie agents based on histological 
and incubation period criteria in mice. pp. 841-842. 

Barlow, R.M. , and Rennie, J.C«: Experience with mink encephalopathy in 
various experimental animals, pp. 843-844. 

Zeman, W.: Subacute sclerosing panencephalitis, a slow measles virus 
infection, pp. 845-849. 

Parker, J.C., Jr., Klintworth, G.Ko, and Graham, D.G.: Myxovirus -para- 
myxovirus within lesions of the CNS of two cases following vaccination 
with attenuated measles virus, pp. 850-851. 

Baringer, J.R. , and Griffith, J.F.: Experimental measles virus infec- 
tions of the nervous system, pp. 852-853. 

Rorke, L.B., Katz, M. , Masland, W.S„ , and Koprowski, H. : Experimental 
subacute sclerosing panencephalitis in ferrets. An animal model system 
for study of the human disease, pp. 854-855. 

Haig, D.A. : Propagation of the scrapie agent in cell culture, pp. 856-857. 

Beck, E., Daniel, P.M., Gajdusek, D.C. , and Gibbs, C.J. , Jr.: Subacute 
degenerations of the brain transmissible to experimental animals: a 
neuropathological evaluation, pp. 858-873. 

Anderson, R. McD.: Histopathology of the brain in kuru. pp. 874-875. 

Nathanson, N. , Cole, GoA. , Weiner, L.P., Gilden, D.H. , and Johnson, R.T.: 
Diversity of pathological lesions produced by acute virus infections of 
the nervous system, pp. 876-891. 

Hirano, A.: Further studies on amyotrophic lateral sclerosis and parkin- 
sonism dementia complex on Guam. pp. 892-893. 



43' 



Serial No. NDS (CF)-62 OA.D 969 

van Bogaert, L. , and Osetowska, E.: Etude comparfie de la maladie de 
Carre et des encephalites de la rougeole. pp. 894-896. 

Fraser, H. : Comparative morphology of ageing and scrapie, pp. 897-898. 

Pattison, I.H. : Detection of the scrapie agent in the tissues of normal 
mice. pp. 899-900. 

Zlotnik, I.: The pathogenesis of scrapie, pp. 901-915. 

Lampert, P.W. , Earie, K.M. , Gibbs, C.J., Jr., and Gajdusek, D.C.: Elec- 
tron microscopic studies on experimental spongiform encephalopathies 
(kuru and Creutzfeldt- Jakob disease) in chimpanzees, pp. 916-930. 

Sever, J.L., Horta-Barbosa, L., Vernon, M.L., Fuccillo, D.A. , Plum, F., 
and Baringer, J.R. : Creutzfeldt- Jakob disease: Virus-like particles in 
brain biopsies and tissue cultures, pp. 931-932. 

Bignami, A., and Parry, H.Bo: Electron microscopic observations in slow 
virus infections of the CNS. pp. 933-934. 

Field, E.J.: Slow virus infection of the nervous system, pp. 935-936. 

Ovary, E. , Benko, Ch., and Gombi, R. : So-called Gonatas-particles. 
pp. 937-938. 

Zu Rhein, G.M. , and Eckroade, R.J. : Experimental transmissible mink 
encephalopathy (TME) . An ultrastructural study, pp. 939-940. 

Raine, C.S., and Sheppard, R.D. : Ultrastructural observations of measles 
virus in nervous tissue, pp. 941-942. 

Masters, C.L. , Kakulas, B.Ao , Gajdusek, D,C. , and Gibbs, C.J., Jr.: The 
significance of the recent experimental observations on slow virus infec- 
tions to neuropathology, pp. 943-951. 

Kurtzke, J.F, : Multiple sclerosis as a latent infection of the nervous 
system, pp. 952-957. 

Sever, J.L., Kurtzke, J.F. , Alter, M. , Schumacher, G.A. , Gilkeson, M.R. , 
Ellenberg, J„H. , and Brody, J„A.: Virus antibodies and multiple scler- 
osis, pp. 958-959. 

Benko, Ch, , Ovary, E. , and Gombi, R. : Morphological methods in the 
cerebrospinal fluid (CSF) testing of subacute sclerosing panencephalitis 
(SSPE). pp. 960-961. 



44 3 



Serial No. NDS (CF)-62 OAD 969 

SUB- PROJECT XIV: Studies on the ecology, epidemiology, and pathogenesis 
of arbovirus infections of man and animals 

Principal Investigators: D. Carleton Gajdusek, M.D„, and 
Clarence J. Gibbs, Jr., Ph.D. 

Other Investigators: Nancy Rogers, M.S., John Hooks, Ph.D., Paul Brown, M,D„ , 
Mint Basnight, M.S., and David Asher, M.D„ 

Cooperating Investigators: Jacob Brody, NINDS ; K. Shah, M.D. , and 

F. Bang, M.D,, Johns Hopkins School of Public Health, 

Baltimore; A. Smorodintsev, M.D,, Institute of Influenza, 

Leningrad, USSR; A. Shubladze and V. Zhdanov, Ivanovsky 

Institute, USSR; B. Gavrilyuk, Institute of Biophysics, 

Serpukhovsky Region, USSR; V. Il'yenko, Research Institute 

of Influenza, Leningrad, USSR; J. Casals, M,D. , 

R. Shope, M,D. , and W. Downs, M,D, , Yale University, New 

Haven; C. Wisseman, M,D. , University of Maryland; 

R. Hornabrook, M,D„, Kuru Research Office, Okapa, New 

Guinea; F. Schofield, M.D. , Nairobi; J. Sever, M,D., NINDS 

Project Description: 

Studies are continuing on the following areas: 

1. Epidemic hemorrhagic fevers - chronic and persistent infections 

2. Seroepidemiology of arbovirus infections in ecologically 
isolated primitive indigenous populations: 

a) Seroepidemiology of Alaskan populations 

b) Seroepidemiology of the populations of the Caribbean and 

Central and South American countries with particular 
reference to Puerto Rico, Bolivia and Paraguay 

c) Seroepidemiology of Australasian populations 

3. Japanese B encephalitis studies on Guam 

4. Persistence of arbovirus infections in man and animals 

5. Isolation and characterization of the virus causing nephro-nephritis 
hemorrhagic fever syndrome in Southeast Asia 

Significance and Course : These are long-term (5 years and more) studies 
being conducted on a continuing basis. Seroepidemiological and viral ecology 
studies are being conducted in primitive cultures and specific patterns of 
disease determined. These studies will continue until statistically sound 
data are collected for publication. 



45^ 



Serial No. NDS (CF)-62 OAD 969 

Publications: 

Asher, D.M, : Focal neurological disease with chronic encephalitis in 
children and in an experimental primate mode. Proceedings XIII Interna- 
tional Congress of Pediatrics, Vienna, 1971. 

Asher, D.M. , Gibbs, C.J., Jr., and Gajdusek, D.C.: Experimental kuru in 
the chimpanzee: physical findings and clinical laboratory studies. 
Brain, 1971. 

Asher, D.M., Locke, L.N. , and Gibbs, C.J., Jr.: Isolation of the agent of 
mourning dove pox. Bacteriological Proceedings, 71:190:vl36, 1971. 

Basnight, M. , Rogers, N. , Gibbs, C.J. , Jr., and Gajdusek, D.C.: Character- 
ization of previously undescribed adenoviruses isolated from chimpanzee 
tissue explants. Bacteriological Proceedings, 70:178:vl59, 1970. 

Basnight, M. , Rogers, N.G. , Gibbs, C.J., Jr., and Gajdusek, D.C.: Charac- 
terization of previously undescribed adenoviruses isolated from chimpan- 
zee tissue explants. Amer. J. Epidemiology, (in press), 1971. 

Beck, E., Daniel, P.M., Gajdusek, D.C., and Gibbs, C. J. , Jr.: Subacute 
degenerations of the brain transmissible to experimental animals: a 
neuropathological evaluation. Vlth International Congress of Neuropath- 
ology, Masson and Cie, Paris, pp. 858-873, 1970. 

Brown, P., Cathala, F. , Gajdusek, D.C., and Gibbs, C.J., Jr.: Measles 
antibodies in the cerebrospinal fluid of patients with multiple scler- 
osis. Proc. Soc. Exp. Biol. Med. (in press), 1971. 

Brown, P., Hooks, J., Roos , R. , Gajdusek, D<,C. , and Gibbs, C.J., Jr.: 
Creutzfeldt-Jakob disease: attempts to identify the agent by CF antibody 
relationship to known viruses. Nature (in press), 1971. 

Brown, P., Roos, R. , Hooks, J., Gajdusek, D.C., Gibbs, C.J. , Jr. 
and Dowdle, W.R.: Viral antibodies in humans and chimpanzees with 
Creutzfeldt-Jakob disease. Amer. Acad. Neurol, (in press), 1971. 

Gajdusek, D.C: Slow virus infections and activation of latent virus 
infection in aging. Advances in Gerontology, 3: 1971. 

Gajdusek, D.C: Slow viruses in autoimmune disease. Am. J. Clinical 
Path, (in press) , 1971. 

Gajdusek, D.C. and Gibbs, C.J., Jr.: Transmission of two subacute spongi- 
form encephalopathies of man (kuru and Creutzfeldt-Jakob disease) to 
new-world monkeys. Nature, 230:5296, 588-591, 1971. 



46s 



Serial No. NDS (CF)-62 OkT) 969 

Gajdusek, D.C, and Gibbs , C. J. , Jr.: Degenerative neurological diseases 
of viral etiology: scrapie, kuru and Creutzfeldt-Jakob disease. In: 
Atypical Virus Infections - Possible Relevance to Animal Models and 
Rheumatic Disease. The Arthritis Foundation (in press), 1971. 

Gajdusek, D.C, Gibbs, C.J. , Jr., and Lira, K.A. : Prospects for the con- 
trol of chronic degenerative diseases with vaccines. In: Proceedings 
of the Second International Conference on Application of Vaccines Against 
Viral, Rickettsial, and Bacterial Diseases of Man (in press), 1971. 

Gajdusek, D.C. and Gibbs, C.J. , Jr.: Persistent, defective and slow 
virus infections of the nervous system of children. Proceedings XIII 
International Congress of Pediatrics, Vienna, 1971. 

Gibbs, C.J., Jr. and Gajdusek, D.C: Characterization and nature of 
viruses causing subacute spongiform encephalopathies. In: Vlth Interna- 
tional Congress of Neuropathology, Masson and Cie, Paris, pp. 779-801,1970. 

Gibbs, C.J., Jr. and Gajdusek, DoC: Transmission and characterization of 
the agents of spongiform virus encephalopathies: kuru, Creutzfeldt-Jakob 
disease, scrapie and mink encephalopathy. In: Immunological Disorders 
of the Nervous System, Res. Publ. Assn. Nerv. Ment. Dis., 49:XXIV: 
Williams and Wilkins, Baltimore, 1971. 

Gibbs, C.J. , Jr. and Gajdusek, D.C: Virological studies of agents iso- 
lated from subacute and chronic neurological diseases. Proceedings XIII 
International Congress of Pediatrics, Vienna, 1971. 

Hooks, J.: Characterization and distribution of two new foamy viruses 
isolated from chimpanzees. Ph.D. Thesis, The Catholic University of 
America, Washington, D.C, 1970. 

Hooks, J., Gibbs, CJ., Jr., Cutchins, E.C, Rogers, N.G. , Lampert, P.W. , 
and Gajdusek, D.C: Characterization and distribution of two new foamy 
viruses isolated from chimpanzees (in press), 1971. 

Hooks, J., Rogers, N. , Lampert, P., Gibbs, CJ., Jr., and Gajdusek, D.C: 
Foaray viruses of chimpanzees: electron microscopy and fluorescent anti- 
body staining. Bacteriological Proceedings, 70: 180-181, 1970. 

Lampert, P.W. , Gibbs, CJ., Jr., and Gajdusek, D.C: Experimental spongi- 
form encephalopathy (Creutzfeldt-Jakob disease) in chimpanzees. Journal 
of Neuropath, and Exp. Neurol. (in press), 1971. 

Lampert, P.W. , Earle, K.M. , Gibbs, CJ., Jr., and Gajdusek, D.C: Elec- 
tron microscopic studies on experimental spongiform encephalopathies 
(kuru and Creutzfeldt-Jakob disease) in chimpanzees. In: Vlth Interna- 
tional Congress of Neuropathology, Masson and Cie, Paris, pp. 916-930,1970. 



47 



Serial No. NDS (CF)-62 OAD 969 

Masters, C.L., Kakulas, B.A. , Gajdusek, D.C., and Gibbs, C.J., Jr.: The 
significance of the recent experimental observations on slow virus infec- 
tions to neuropathology. In: Vlth International Congress of Neuropath- 
ology, Masson and Cie, Paris, pp. 843-951. 

Rogers, N.G. , Gibbs, C.J„, Jr., Gajdusek, D.C., Andersen, S.W., and 
Basnight, M. : Visna-like agents from brain and lung of sheep with Montana 
sheep disease. Bacteriological Proceedings, 71: 180: v72, 1971. 

Roos, R. , Rogers, N.G. , Basnight, M. , Chou, S. , and Gajdusek, D.C.: Isola- 
tion of adenovirus 32 from human brain in a case of chronic encephalitis. 
Bacteriological Proceedings, 71: 194: vl55, 1971. 

Roos, R. , Gajdusek, D.C., and Gibbs, C.J. , Jr.: Liver disease in Creutz- 
feldt-Jakob disease (subacute spongiform encephalopathy). Amer. Acad. 
Neurol. 1971. 



48 s 



AMUAL REPORT 

JULY 1, 1970 THROUGH JUNE 30, 1971 

EPIDEMIOLOGY BRANCH 

COLLABORATIVE AND FIELD RESEARCH 

NATIONAL INSTITUTE OF NEUROLOGICAL 

DISEASES AND STROKE 



Introduction 



The major areas of activity of the Epidemiology Branch during the 

period of this report have been: 

I. epidemiologic studies of neurologic diseases; 

II. laboratory studies related to the epidemiology and immunology of 
neurologic and perinatal diseases; 

III. the continued activities on Guam and the Trust Territories; 

IV. genetic studies of neurologic diseases; 



The following professional personnel have joined the Epidemiology 
Branch: Dr. Charles E. Morris, Associate Professor of Neurology at the 
University of North Carolina, has been our Of f icer-in-Charge on Guam. 
Dr. John Stanhope, an epidemiologist who was with my staff here in Bethesda 
was transferred to Guam for the current fiscal year. Dr. Roger Bobowick, who 
completed his residency in neurology, joined my staff in Bethesda. Dr. Jeffrey 
Allen, who completed his internship in Seattle at King County, is in the 
Genetics Section. Kathy O'Meara, Biologist, is also in the Genetics Section, 
and Marjorie Matthews,' R. N. , is working with the Epidemiology Branch. Mr. Otis 
Turner, Statistician, joined our staff for 6 months in Epidemiology, but 
subsequently accepted a promotion to Associate Director, Division of Maternal & 
Child Health Services, and Gary Cooper, Medical Technician, is currently 
working in the Laboratory. Dr. A.V. Bird, a neurologist from South Africa 
has joined the Branch for 6 months as a Visiting Scientist to pursue studies 
in the epidemiology of multiple sclerosis. 

The following people have left the Branch: Dr. Roger Detels left to 
accept an Associate Professorship in Epidemiology at U.C.L.A. , Renee Owens, 
Biologist, left to join her husband in Texas, Jane McNew, R.N., left to get 
anM.P.H. degree at the Johns Hopkins School of Hygiene and Public Health. 

In terms of organization and planning for next year, we are seriously 
hampered because the position of Deputy Branch Chief is vacant. Dr. E. 
Michael Holden, who is finishing his second year of neurology residency at 
the Boston City Hospital, will go to Guam as Of f icer-in-Charge for two years. 
Dr. Robert Stern, who is finishing his internship at Mt. Sinai, New York 
City, will be joining the Branch in Bethesda. 



It 



We have pursued the mission of the Epidemiology Branch in studying the 
epidemiologic aspects of neurologic disease in which, through our approaches 
we can contribute significantly to the understanding of specific aspects of 
neurologic disease, which will be of immediate value to physicians and patients, 
We believe that through the technique of specific hypothesis testing in 
defined populations rather than through large descriptive population studies 
we can perform this function most effectively. Our major contributions 
during this year have been: As an offshoot of our Guam studies we have 
demonstrated that CNS dopamine metabolism is diminished in amyotrophic 
lateral sclerosis patients in the United States. This is the first demon- 
strated chemical abnormality in the CNS of ALS patients. Through studies of 
SSPE we have shown that this disease apparently results from the combination 
of an abnormal infection with measles and a subsequent zoonotic trigger. 
Epidemiologic studies of MS in California and Washington State have produced 
provocative information suggesting that a protective factor exists among 
people born in low risk areas and further that the risk of developing MS can 
be altered by migration after puberty. We also found that rates for Japanese 
and Chinese for MS are consistently low in California and Washington State. 
Serologic studies of MS continue to implicate the measles virus and a 
possible familial factor. Several difficult studies have been established 
and are proceeding well. These include studies of the natural history of 
Parkinson^ disease and the effect of L-Dopa, twin studies on multiple 
sclerosis, studies of ALS among veterans and a case/control study of Jakob- 
Creutzfeldt disease. We have made significant advances in diagnosis and 
treatment of acoustic neuroma and studies relating to I.Q and retinoblastoma 
and torsion dystonia continue to bear promise. 

I . Epidemiologic studies of neurologic disease . 

Multiple Sclerosis 

We have completed the first phase of our studies of multiple sclerosis 
in California and Washington State and have made the following observations. 
Orientals who have an apparently low rate of multiple sclerosis relative to 
their latitude of birth have low rates of multiple sclerosis after migration 
to California and Washington. Further, second generation Orientals persist 
in having a low rate of MS suggesting a possible genetic factor. People born 
in Washington State had higher rates of MS than those born in California 
which would be expected according to the north-south differential of this 
disease. People migrating from northern states to the West Coast had higher 
rates when they migrated to Washington than to California, implying that they 
either acquired protection in California or the causative factors are less 
common in California than in Washington. The implication of this is that the 
causative factors persist after puberty, which is contrary to current belief. 
Those who migrate from low risk southern areas did not develop a high rate of 
MS either in California or Washington (small numbers). This implies that 
by the time of migration they have protection from multiple sclerosis. If 
protection can be acquired as these data would seem to imply, there is some 
hope of developing a preventive mechanism for MS. 



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In collaboration with Dr. John Sever, Head, Section on Infectious 
Diseases, Perinatal Research Branch, C&FR, NINDS, we have pursued our studies 
of measles serology in patients and family members. Recent data suggest 
that measles antibody is slightly but consistently elevated in MS patients 
when compared to controls. This elevation is not consistently encountered 
among siblings of the same sex of the MS patients, suggesting a possible 
familial factor in this disease. We are still hoping to extend our studies 
to the Shetland and Orkney Islands where an ideal population for study exists. 
A brief trip to these islands confirmed the observation that the rates of MS 
in these small populations are three times higher than the highest rate 
reported anjnA/here else. 

We have developed several new studies of the epidemiology of multiple 
sclerosis. We are investigating some 15 or 20 twins from the Veterans Twin 
Registry in which one of the twins has MS, in order to determine if there are 
distinct variations in exposure or disease history prior to onset of MS among 
the twins. We are also conducting patient, sibling studies of a small group 
of MS patients in which the mother is alive and available. We hope to 
detect possible differences in early infections such as we encountered with 
SSPE or in other factors which have been suggested as having possible 
relevance in the etiology of MS. Further, through the contract mechanism we 
are involved in a study of 6,000 veterans with MS in order to further 
elucidate migratory patterns and predisposing factors in this population and 
selected controls. 

Amyotrophic Lateral Sclerosis (stateside studies) 

Following our observation on Guam that ALS patients had diminished CNS 
dopamine metabolism, we extended our studies to stateside ALS patients and 
again encountered diminished CNS dopamine metabolism. This finding has now 
been confirmed by others at NIH, at Columbia, at Harvard and at Montreal. 
Initial treatment trials of ALS patients with L-Dopa have not yielded 
impressive results. The observation of a chemical abnormality in the CNS of 
ALS patients is to our knowledge the first substantial chemical abnormality 
in the CNS of these patients, and hopefully will lead to greater insights in 
abnormal neurometabolite patterns among patients with motor neuron disease. 

We continue to follow 12,000 statesiders who worked for more than one 
year on Guam between 1945 and 1955. The final information on approximately 
207o of the 2500 people in this group who have died continue to trickle in. 
We have now encountered two patients who died of ALS which does not seem 
excessive for a population of adult males. 

We are pursuing our epidemiologic study to test the hypothesis that 
pancreatic insufficiency is causally related to ALS which has been suggested 
by several investigators. We continue to follow some 3,000 veterans who had 
duodenal ulcer between 1948 and 1958. In one group only vagotomy and 
pyloroplasty was performed, while in another group gastrectomy with Billroth 
II anastomosis was conducted and hence the patients had induced pancreatic 
insufficiency. The clinical phase of this study has ended with the suggestion 
that an excess of non-specific neurologic disease occurred in those receiving 
the Billroth II procedure as opposed to those receiving vagotomy and 
pyloroplasty. We have no evidence of ALS in these groups. We are now 

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attempting to follow-up through the Veterans Administration on causes of 
death in this unique population. We are having difficulty in accumulating 
the final data but there is a suggestion of an excess of several neurologic 
diseases including Parkinson's disease and multiple sclerosis. 

We have developed a protocol with Dr. Gilbert Beebe, Director, Follow- 
Up Agency, National Research Council of the National Academy of Sciences and 
Dr. John Kurtzke, Chief, of the Neurology Service, Vererans Administration, 
Washington, D.C., to conduct studies of ALS veterans patterned generally on 
the initial MS study among veterans. We have also developed and field 
tested a detailed questionnaire to be administered to 200 ALS patients and 
200 patients with brain tumor in VA hospitals in order to detect possible 
predisposing factors to ALS. We will also analyze exposure to Guam as a 
possible predisposing factor to this disease. 

In conjunction with our studies of multiple sclerosis in California and 
Washington State we have analyzed patterns of ALS among patients bom in 
California and Washington and among migrants. In striking contrast to our 
findings with MS we have detected no geographic patterns in the incidence 
of ALS in these populations. 

Parkinson's Disease . 

In collaboration with the National Parkinson's Foundation, Inc., in 
Miami, Florida, we are studying the effects on long term administration of 
L-Dopa and the influence of L-Dopa on the natural history of Parkinson's 
disease. We are following a cohort of up to 500 Parkinson's disease patients 
who developed their disease from 1962 to 1965 (pre- L-Dopa) and will match 
these individuals with a similar number who developed their disease after 
1968 and are receiving L-Dopa. Our initial findings suggested a possible 
cohort phenomenon with patients in the L~Dopa treatment group being 5 to 6 
years older than those in the pre-L-Dopa group. We have reviewed several 
other series of patients and have not been able to confirm this impression. 
It is important however that an adequate understanding of this observation is 
achieved because if there is a cohort phenomenon, it would imply the gradual 
disappearance of Parkinson's disease as has been suggested by some investiga- 
tors. 

We are continuing our study of identical schizophrenic twins receiving 
phenothiazines in order to determine if phenothiazine- induced extrapyramidal 
disease is the result of a genetic inability to handle these drugs. Analysis 
of our first six sets of twins suggests that there is no genetic control over 
this phenomenon. Further, we have been studying the appearance of Parkinson's 
disease among blacks and whites and have the impression that the disease 
occurs less frequently among blacks. In our studies of drug- induced Parkin- 
son's disease however, the rate of disease is similar in blacks and whites. 
In collaboration with Dr. Thomas Chase, Chief, Unit on Neurology, Laboratory 
of Clinical Sciences, NIMH, we have shown that schizophrenics who receive 
phenothiazine derivatives and who do not develop extrapyramidal signs have 
significantly elevated dopamine and seratonin end products in their CSF. 
Those who do develop extrapyramidal signs have significantly depressed levels, 
particularly of dopamine end products in the CSF. Thus, a possible mechanism 

4t 



can be postulated that over- stimulation of dopaminergic cells from pheno- 
thiazine utilization resulted in exhaustion of these cells in a certain number 
of cases and subsequent development of parkinsonian features. 

Subacute Sclerosing Panencephalitis . 

It is extablished that measles in some way is causally related to SSPE. 
As a result of our case/control studies we have determined that SSPE develops 
in individuals with very unusual measles histories. In up to 2/3 of the 
patients (P=.001) there was either no history of measles, measles under age 
one, or measles approximately 2 months after exposure to chicken pox. 
Further, we have confirmed the observation that SSPE occurs with a strong male 
preponderance and essentially in rural areas. Since measles even under age 
one does not follow this male, rural pattern, we believe some form of zoonotic 
triggering mechanism, perhaps an animal virus, is also involved in the 
pathogenesis of SSPE. Our patients had a significantly higher rate of 
exposure to sick animals including dogs and fowl, but not cats, than our 
controls. 

Serum samples collected from patients and their families and controls 
and their families are now being tested for measles antibody and will be 
available for studies of other appropriate agents. 

Stroke 



As the result of a field trip to Panama, a potentially valuable study 
of stroke among blacks has been developed and awaits funding. We wish to 
explore the relative effect of genetic factors and environmental factors on 
the extraordinarily high rate of essential hypertension among blacks. 

Jakob-Creutzfeldt Disease 

We are conducting a case/control study of 40 patients with J-C disease. 
Approximately 15 patients and control have been examined and interviewed. 
Analysis of these data are not yet available. 

II. Studies on Cellular Immunity in MS 

Our studies on lymphocyte function in MS have continued in an attempt 
to further define the role of cellular immunity in the pathogenesis of 
demyelinating diseases. These have consistently failed to show any 
significant differences between MS and normal persons. However, our results 
demonstrate that suspensions of brain material (especially white matter) 
are definitely stimulatory to lymphocytes in culture. This has been true 
regardless of the health of the donor. The nature of this effect is uncertain 
but must be considered in all investigations on the response of lymphocyte 
from patients with neurologic illnesses to CNS antigens. 

During the year we continued to investigate the effects on circulating 
lymphocytes following measles vaccine. Results have been inconsistent; in 
some experiments a depression in lymphocyte transformation was seen while 
In others no effect was apparent. Attempts to demonstrate a lymphocyte- 



5t 



depressing serum factor following vaccination have been equally negative with 
multiple sclerosis patients and normal control persons. These findings, 
although involving limited numbers of patients, are in contradistinction to 
reports by other investigators of an ant i- lymphocyte factor in the serum of 
patients with MS. 

Studies on Cellular Immunity in Neurological Diseases other Than 
Multiple Sclerosis 

In order to investigate the possible role of lymphocyte-mediated 
immunity in the Guam ALS and the PD syndromes, we initiated a cooperative 
effort with our staff on Guam. The lymphocyte transformation technique 
was carried out with the lymphocytes from selected patients on Guam; at 
the end of the tissue culture period, the lymphocytes were "harvested" 
and sent to our Bethesda laboratory where the remainder of the DNA extraction 
procedure was done. The results of these experiments indicate that such 
studies are feasible despite the limited facilities and laboratory personnel 
on Guam. Definitive studies are now underway. 

Basic Studies on the Lymphocyte Transformation Phenomenon and 
Related Tests of Cell-Mediated Immunity 

Liquid scintillation quantitation of tritium incorporation by dividing 
cells has been adopted as the best measure of lymphocyte transformation. 
Using this technique we have completed a series of experiments on the effects 
of a synthetic double-stranded RNA (poly I-C) on vaccinia virus induced 
transformation. The results of these experiments have substantiated and 
extended the observations described in last year's report. Complementary 
studies using a double-stranded RM. virus (Reo-1) have not shown any effect 
on lymphocyte transformation. Other laboratory studies carried out by 
Dr. Jeffrey Allen have extended our knowledge on the effects of storage of 
lymphocytes for hours or days prior to initiation of culture; we anticipate 
that these will prove useful in future investigations using cells from patients 
with diseases such as subacute sclerosing panencephalitis (SSPE) and Jakob- 
Creutzfeldt disease. In addition. Dr. Allen has initiated a series of 
experiments to study the effect of the measles virus on lymphocyte transforma- 
tion induced by non-specific mitogens and specific antigenic stimulants. 

Because of reports that cellular immunity to CNS antigens in MS patients 
can be demonstrated by a different technique (human macrophage migration 
inhibition), we are now initiating studies using this method to complement 
our lymphocyte transformation investigations. To learn this technique. 
Dr. Nemo spent one week in Dr. Bartfeld's laboratory at New York University 
and is currently setting up the method in our laboratory. 

Studies on Chronic and Congenital Viral Infections in 
Experimental Animals 

Approximately 50 mice whose mothers received injections of the minute 
virus of mice (MVM) late in gestation have been observed for one year. The 
only abnormalities observed to date has been the occurrence of solid tumors 
in two animals. The first such animal was lost; the second was found to 

6t 



have a large tumor in the area of the right uterine horn with metastasis to 
the liver. Preliminary results suggest the presence of MVM in both the 
tumor and the liver and raise important questions as to the role of chronic, 
congenital MVM infection in the etiology of solid tumors of the mouse. 
Further studies are in progress to evaluate the oncogenicity of MVM and cell- 
free extracts of the uterine tumor. 

Other experiments with MVM have suggested that maternal infection 
early in gestation may result in reabsorption of the infected embryo. This 
problem will be studied further during the next year. 

Our studies on the epidemiology of SSPE led us to hypothesize that a 
fundamental step in the etiology of that disease may be an initial exposure 
to measles virus at a time when persistent maternal antibodies might interfere 
with the normal development of cellular immunity and hence might allow for 
an unusual persistence of the viral genome in certain long-lived cells such 
as neurones. Dr. Nemo and Mr. Cooper are currently attempting to develop an 
animal model for such a system and are investigating the course of infection 
with measles virus in young mice whose mothers were inoculated with measles 
a few weeks prior to mating. No results are available at this time. 

Studies on the Minute Virus of Mice (MVM) 

This agent was selected for use in the development of a model system 
for the study of chronic and congenital viral infections. It is hoped that 
an elucidation of mechanisms involved with establishment of chronic MVM 
infections will contribute to our understanding of certain chronic neurologic 
diseases whose etiologies may also involve a persistent, latent or 
recrudescent virus infection. MVM has been reported to cause cerebellar 
hypoplasia and congenital ataxia but has been the subject of limited inves- 
tigation because of technical difficulties involved with its detection, 
quantitation and propagation in high concentration. The efforts of Dr. White, 
Mrs. Sutton and Dr. Nemo during the past year seem to have resolved these 
difficulties: they have developed methods of propagation yielding virus at 
concentration 100 to 1000 times higher than previously possible, and have 
improved their infectivity quantitation techniques from a six week rather 
unreliable test to a one week procedure with sharp endpoints. In addition, 
they have demonstrated the growth of MVM in 4 different continuous cell lines, 
although reports from other laboratories had been confined to only 2 primary 
cell types. Of particular interest is the development by Mrs. Sutton of 
chronic MVM infection of one of these cell lines. This system is proving 
useful as a continuous source of large amounts of virus and may advance our 
understanding of chronic infections in vivo . 

Experiments in two cell types have substantiated the impression that 
MVM replication requires that the infected cell divide, even though the virus 
can infect and persist in a non-dividing cell. This observation may be 
relevant to the viral latency which may be a prerequisite to the pathogenesis 
of SSPE, J-C disease, and progressive multifocal leukoencephalopathy (PML) . 
Our working h5rpothesis is that MVM infection, like certain papova virus (SV-40) , 
polyoma and perhaps the papova-like agents involved in PML is associated with 
an integration of viral DNA into chromosomal DNA., and that this relates to 



7 t 



the tendency for MVM to persist in non-dividing cells. Experiments are 
underway to test this hypothesis by defining the sequence of molecular events 
which occur in the course of MVM infections in vitro . 

III. The continued activities on Guam and the Trust Territories 

The epidemiologic patterns for ALS and PD on Guam have been updated 
to include the past five years. A slight but apparently not significant 
decline in rates was noted. The pattern of age-specific attack rates, 
however, has not changed. This suggests that we are not observing a cohort 
phenomenon which would have occurred as a result of a massive common exposure 
at some time in the past and implies that the cause of causes of ALS and PD 
on Guam are still operative. L-Dopa trials on PD patients are in their 
15th month with some observable benefit in the extrapyramidal features of 
this disease. Drug trials in ALS patients with L-Dopa, isoprinosine and 
placebo are in their second month. The studies are "blind" and no dramatic 
improvement or untoward responses have been observed. We have further 
confirmatory evidence that CNS dopamine production in PD patients is markedly 
depressed while CNS dopamine production in ALS patients of Guam and in the 
continental United States are significantly comprised but to a lesser degree 
than encountered in PD or classical paralysis agitans. 

We are not attempting to develop an assay of impaired CNS dopamine 
metabolism in which we used expired air and urine rather than CSF to conduct 
our measurements. The technique is now being used on patients and controls. 
If is is feasible we will conduct studies of CNS dopamine metabolism in 
family members of ALS and PD patients and also in geographic areas where the 
diseases seem to concentrate. 

We conducted a follow-up study of the congenital blindness among the 
Pingelapese population on the Eastern Caroline Islands. Six patients have 
been seen at NINDS and the diagnosis is now believed to be achromatopsia 
with the single reservation that the high myopia encountered in the affected 
patients is perhaps a separate entity. The unusual age distribution of 
patients observed in our previous trip report which had suggested a possible 
extrinsic factor was not encountered in our most recent survey. 

IV . Genetic studies of neurologic diseases 

The two main interests of the Genetic Section continue to be movement 
disorders, especially the torsion dystonias, and hereditary tumors of the 
nervous system. In addition the Section is embarking on a study of a highly 
inbred group of Irish Tinkers living in the Southeastern United States who 
are thought to have several unusual genetic traits. 

Our efforts regarding the dystonias are directed toward determining 
the basic neurochemical defect in the autosomal i;ecessive and autosomal 
dominant forms. Ten of our patients have participated in a study of 
catecholamine metabolism on the research ward of Dr. Thomas Chase, NIMH. 
Results to date, although not pinpointing a specific abnormality do suggest 
we are dealing with the abnormal pathway. Evaluation of various therapeutic 
regimens using L-Dopa, peripheral dopa decarboxylase inhibitors and other 



8t 



drugs is being conducted by collaborators with patients from our series. No 
single regimen seems effective for all patients so that tailoring of drug and 
dose for each patient is still necessary. Finally, we continue to see new 
patients and their families with unusual movement problems. One interesting 
group which has emerged consists of professional musicians troubled by a mild 
movement abnormality which, however, is sufficient to impair proper performance, 
This may be a distinct new entity or else may represent the heterozygous state 
of autosomal recessive dystonia. 

Acoustic neuroma remains our chief interest in the area of hereditary 
neoplasms. On the basis of our study of the large Pennsylvania kindred with 
acoustic neuroma and review of the literature we concluded that this seemingly 
rare trait might actually be fairly common if the appropriate population were 
sampled. Therefore, we looked for the presence of bilateral disease and 
positive family history in individuals with onset of sjmiptoms of acoustic 
neuroma at a young age. To date we have information on over 2100 individuals 
in 51 families and have found at least 4 with positive family history of 
acoustic neuroma. Since only 4 families have been reported in the literature 
we have been able to double the number of reported families by this approach. 
The interesting question of relationship between neurofibromatosis and 
acoustic neuroma is being investigated through clinical study, cell culture 
techniques and chromosome analysis. 

The retinoblastoma study, which deals with perception of affected by 
sighted individuals, is moving into its final stages. Preliminary results 
show no significant difference in performance between patients and unaffected 
sibs used as controls. This is at variance with the two reported studies on 
this question. Chromosome studies in familial cases show no abnormalities 
unlike sporadic atypical cases. 

Our interest in genetic factors in ALS and PD as seen on Guam continues 
through study of select families on Guam and among migrants living in 
California. 

Groundwork is being laid for a comprehensive genetic, demograph and 
medical study of the Irish Tinkers of Murphy Village, South Carolina. Since 
this group is understandably wary of strangers, great care is being paid to 
establishing rapport with elders and other informants within the community. 
We are enthusiastic about this project since it is through study of such 
isolates that many new recessively inherited traits have been described. 

A canvass is being made of isolated communities along the coast of 
Maine for similarly inbred groups. 



^ 



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CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 1971 

NATIONAL RESEARCH COUNCIL, FOLLOW-UP AGENCY (PH43-64-44 ) 

Title : New Epidemiologic Study of Multiple Sclerosis in U.S. Military 
Veteran Population 

Contractor's Project Director : Gilbert Beebe, M.D. 

John F. Kurtzke, M.D. 

Current Annual Level : $34,700 

Objectives : The contractor will perform an extensive survey of multiple 
sclerosis among veterans of the Second World War and the Korean War. 
This will be an update of an initial survey which included 600 patients 
and will include approximately 6,000 patients. Patients will be matched 
with controls to determine geographic patterns, socio-economic status, 
urban-rural localization and numerous other variables which have been 
tested for multiple sclerosis. Parallel information will be available 
for Negro MS patients and controls and white female MS patients and 
controls. In addition an extensive investigation of migrant and non- 
migrant populations to and from high and low risk areas for multiple 
sclerosis will be conducted. Emphasis will also be placed on the 
relationship between the communicable diseases experienced in childhood 
and subsequent multiple sclerosis. 

Major Findings : The project has just commenced and data is still being 
accumulated . 

Significance to NINDS Program and Biomedical Research : Multiple 
sclerosis is a major neurologic disease in the United States. Previous 
epidemiologic studies have indicated that there is in the Northern 
Hemisphere a north-south gradient in the rates of multiple sclerosis. 
From a previous study, certain interesting correlations were noted 
among military personnel who developed multiple sclerosis, such as an 
excess of people with higher I.Q.s, with urban residence, with higher 
socio-economic status and with defective eyes. The recent emphasis on 
risk of multiple sclerosis among migratory populations is of crucial 
importance since it must be determined if the causative factors of this 
disease are more prevalent in northern areas or if there are protective 
factors or mechanisms operating in southern areas . If clear patterns 
can be delineated by this study, the results could suggest in which 
populations we must concentrate in the search for the etiology and 
prevention of multiple sclerosis. 

Proposed Course of Project : This project was initiated in March 1971 
and will run for three years . 



lot 



CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 1971 

THE JOHNS HOPKINS UNIVERSITY (PH43-67-134 7) 

Title : The Epidemiology of Parkinson's Disease 

Contractor's Project Director : Abraham M. Lilienfeld, M.D. 

Irving I. Kessler, M.D. 

Current Annual Level : 

Objectives ; The contractor will: (a) Select a group of hospitals to 
provide sufficiently large groups of patients with Parkinson's disease; 
(b) Assemble groups of patients with parkinsonism, Parkinson's disease 
and similar neurological deficits who have been hospitalized for any 
reason or seen in any hospital clinic; (c) Assemble matching groups of 
patients without parkinsonism symptoms who have been hospitalized at 
the same institutions and who are similar to the cases in age, sex, 
race, and other pertinent demographic characteristics; (d) Select a 
probability sample of physicians (neurologists, general practitioners, 
general surgeons, and internal medicine practitioners) and from them 
secure a cohort of patients with parkinsonian symptoms who have not 
been hospitalized; (e) Compare the patients and their matched controls 
with regard to epidemiologic characteristics of possible relevance to 
the incidence of or morbidity from Parkinson's disease, including 
familial mortality experience and smoking histories; (f) compare, 
within the patient groups, the epidemiologic characteristics of those 
with specific parkinsonian symptom complexes; (g) Compare the hospital- 
ized and nonhospltalized parkinsonian patients in order to test whether 
there are methodologic biases in studies restricted to hospitalized 
patients; (h) Assemble a large group of patients known to have had 
Parkinson's disease, parkinsonism or similar neurological deficits as 
the basis for an analysis of the overall and cause specific mortality 
of the decedents; (i) Reach conclusions as to the adaptability of the 
case-control method proves to be inadequate, design other more suitable 
methods for the epidemiological study of Parkinson's disease. 

Major Findings ; The first phase of the analysis of this study concentrated 
on previously reported lower rates of Parkinson's disease among smokers 
than non-smokers. In the population sampled in the present study, this 
difference was not encountered and in general, patients were less likely 
to have ever smoked, and smokers tended to smoke less than a comparable 
group of patients without Parkinson's disease. Further, analysis of 
the data are currently underway. 

Significance to NINDS Program and Biomedical Research : At present, 
understanding of risk factors and natural history in parkinsonism is 
limited to a few studies primarily in clinic populations . The 
extension using a case-control matching could provide important 
information for this extremely important neurologic condition. 

lit 



Contract (PH43-67-1347) 

Proposed Course of Project : The project has run for two years. The 
second year was funded at a reduced rate. Therefore, analysis has been 
more time-consuming and the final report is not yet available. 



12t 



CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 1971 

THE UNIVERSITY OF CALIFORNIA-BERKELEY (PH43-68-36 ) 

Title : Health Survey of Stateside Guamanians 

Contractor's Project Director : Reuel Stallones, M.D. 

Dwayne M. Reed, M.D. 

Current Annual Level : 

Objectives : The contractor will: (a) Analyze mortality experience of 
Guamanian residents of California for the past ten years in comparison 
to that of California residents generally; (b) Contact adult Guamanians 
residing in California and enlist the voluntary participation in the 
survey of approximately 1,000 subjects; (c) Secure family and individual 
health histories from recruited subjects, and administer a screening 
examination comprised of: neurologic examination, psychological 
examination to test for dementia and motivational aspects, EKG, BP, 
blood tests for cholesterol, triglycerides, uric acid, and glucose; 
(d) Conduct similar examinations of a matched group of Guamanians 
residing on Rota Island, Saipan Island, and Guam; (e) Code and process 
data gathered and analyze results, demonstrating whether or not disease 
patterns vary between Guamanian residents in the U.S. and the general 
population, and between Guamanian residents in the U.S. and Guamanian 
residents in the Pacific. 

Major Findings : The investigators documented an unusually high rate of 
hyperuricemia and hyperglycemia among Chamorros living on Guam, in 
California, or on Rota. The patterns of diabetes and gout differed to 
some extent. The major conclusion was that there were no important 
differences in disease patterns as a result of westernization. 

Significance to NINDS Program and Biomedical Research : Establishing 
a baseline on Guam for metabolic disease and cardiovascular disease 
as well as some knowledge of psychological patterns and patterns of 
neurological disease will be invaluable to our studies on Guam. They 
vri.ll also provide important information concerning patterns of disease 
among migrating populations. 

Proposed Course of Project : The project as originally proposed has 
terminated and a report received. A portion of the funds were unspent 
and the investigators were authorized to pursue their study in another 
Micronesian population on Palau. The field work is complete and we 
have received the final report. 



13t 



CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 1971 

THE MASSACHUSETTS GENERAL HOSPITAL (PH43-68-982 ) 

Title : Screening of Blood and Urine for Abnormal Amino Acid Patterns 

Contractor's Project Director : Vivian E. Shih, M.D. 

Current Annual Level : 

Objectives : The contractor will perform cromatographic screening of 
approximately 1,000 samples each of blood and urine for detection of 
disorders of amino acid metabolism among residents of Guam and the 
Trust Territories. Testing will be performed by routine methods 
developed and utilized in the contractor's laboratory conducting 
repeat screening on tests of blood or urine samples as needed in 
order to confirm or nullify the significance of unusual patterns. 

Major Findings : No major abnormalities in infants, retarded children 
or ALS or PD patients were encountered in the survey of blood or 
urine of 435 patients with various diseases and 574 normal infants 
and 20 normal adults in Guam and other islands of Micronesia. One 
girl, apparently healthy but with a history of seizure disorder 
(apparently a febril convulsion), many years ago, was found to excrete 
an unknown sulphur amino acid. Other non-specific differences in 
amino acid patterns were encountered in different populations and were 
apparently the result of dietary factors in the given communities. 

Significance to NINDS Program and Biomedical Research : As the study 
continues on Gxoam we become more convinced that metabolic diseases 
play an important role in causing amyotrophic lateral sclerosis and 
parkins onism-dementia. In addition, on many islands in the Trust 
Territories we notice abnormal genetic patterns of disease which 
suggest that certain inborn errors of metabolism may exist. Should 
we identify these specific metabolic errors on Guam or in the other 
islands we may gain important insights into the cause of the neurologic 
diseases in Guam and by using population isolates gain information 
concerning metabolic pathways in a given population which would provide 
information on metabolic processes in human populations anywhere. 

Proposed Course of Project : A final report has been received and a 
joint publication has been prepared for submission to a Pediatric 
Journal. Further investigation on the family of the girl with the 
previously undescribed sulphur amino acid is underway. 



14t 



CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 1971 

THE JOHNS HOPKINS UNIVERSITY (NIH71-2026 ) 

Title : The Study of Regional Differences in Stroke Mortality 

Contractor's Project Director : Dean M. Nefzger 

Current Annual Level : $21,937 

Objectives : The contractor will: (a) Analyze death certificates of 
all veterans dying in 1967 in Georgia (high mortality area) and five 
Rocky Mountain States (low mortality area) . From the certificates 
approximately 1,000 certified CVA deaths in each area and 100 randomly 
selected controls will be chosen for further analysis (2,200 cases 
total). From these basic data, the frequency of reported CVA among 
veterans will be compared with male populations in similar areas to 
determine if the geographic variations reported for civilians occur 
among veterans; (b) By review of available hospital records and when 
necessary by physician or family interview, the validity of the diagnosis 
will be established in order to estimate the relative frequency of 
mistaken diagnosis or failure to make the diagnosis of CVA; (c) By 
review of the accumulated information on veterans dying of CVA an 
estimate of the relative frequency of specific types of CVA will be 
compiled; (d) All verified stroke deaths and all errors in death 
certification will be analyzed in terms of geography, age, race, place 
of residence, marital status, from the point of view of sources of 
information (competence of certifying individual) and other variables; 
(e) During this investigation the complete Military and Veterans 
Administration folders will be reviewed for a subgroup of 50 cases and 
controls per state in order to evaluate the usefulness of these records 
in subsequent studies. In addition, all other avenues of ascertainment 
of a valid rate of CVA among veterans will be explored in order that a 
definitive study of veterans population be conducted in the future when 
the great bulk of veterans of the Second World War arrive at the age of 
high risk for CVA. 

Major Findings : A total of approximately 1,000 stroke deaths and 2,800 
deaths from other causes have been compiled. The data on these cases 
are ready for final analysis. A preliminary review of the stroke cases 
in Georgia which has a high mortality rate from stroke versus the 
Rocky Mountain States which have a low reported mortality from strokes 
has revealed surprising and potentially Important information. It 
appears that this wide discrepancy may be purely artifactual and the 
result of different reporting habits by physicians in the two areas. 
It is uncommon for stroke to be listed as underlying cause of death in 
the Rocky Mountain States. This finding is at variance with the Johns 
Hopkins University Epidemiology of Stroke study which we supported by 
contract (PH43-66-920) . 



15t 



Contract (NIH71-2026) 

Significance to NINDS Program and Biomedical Research : The 
epidemiological patterns of stroke are poorly understood, although it 
is suspected that there are regional differences throughout the U.S. 
Any information confirming these differences and indicating a cause 
for these differences could lead to a better understanding of causation 
and prevention in this important cause of morbidity and mortality. 

Proposed Course of Project : The contract has expired, but final 
processing of data is not complete. The investigators have received 
a contract of $21,937 to complete this work. We are awaiting a final 
report . 



16t 



Project Title: 



Serial No. NDS (CF) - 55 E 201 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Studies on amyotrophic lateral sclerosis/parkinsonism- 
dementia complex of Guam (ALS-PD) 



Previous Serial Number: Same 



Principal Investigators : 



Jacob A. Brody, M.D. 
Charles E. Morris 

NINDS Research Center 
John M. Stanhope, M.D. 

NINDS Research Center 



Other Investigators; 



Jose Torres 

NINDS Research Center 
Francisco Leon Guerrero 

NINDS Research Center 
Manuel T. Cruz 

NINDS Research Center 
Olivia Cruz, M.D. 

NINDS Research Center 
Roswell Eldridge, M.D. 



Consultants : 



Kwang-Ming Chen, M.D. 

National Taiwan University, Taipei, Taiwan 
Yoshiro Yase, M.D. 

Wakayama Medical College, Japan 
Leonard T. Kurland, M.D. 

The Mayo Clinic, Rochester, Minnesota 
Donald W. Mulder, M.D. 

The Mayo Clinic, Rochester, Minnesota 
Haruo Okazaki, M.D. 

The Mayo Clinic, Rochester, Minnesota 



Cooperating Units ; 



NINDS Research Center, Agana, Guam 

Special Chronic Disease Studies, C&FR, NINDS 

Laboratory of Slow, Latent, and Temperate Viruses, 

C&FR, NINDS 
Department of Epidemiology, School of Public Health, 

University of California, Berkeley 
The Mayo Clinic, Rochester, Minnesota 
Department of Pathology, Massachusetts General Hospital, 

Bos ton 
Amino Acid Laboratory, Massachusetts General Hospital, 

Bos ton 



17t 



Serial No. NDS (CF) - 55 E 201 

Department of Neurology, Wakayama Medical College, 

Wakayama, Japan 
Trust Territory Health Office 
University of California, Los Angeles 

School of Public Health, University of Hawaii, Honolulu 
Unit on Neurology, NIMH 

Section on Neurology, University of Montreal 
Department of Neuropathology, Albert Einstein College 

of Medicine, New York 
Department of Neurology, Neurological Institute of 

Columbia University, New York 
Neuropathology Branch, Armed Forces Institute of 

Pathology, Washington, D.C. 



Man Years 



Total: 

Professional; 

Other: 



3/4 
1/4 



Project Description: 

Objectives : To determine the cause of ALS and PD, and to determine the 
epidemiological, clinical, neuropathological and physiological significance 
of these diseases and to develop therapeutic approaches to the diseases. 

Methods employed : Routine methods for epidemiological, clinical, 
neuropathological, and neurochemical and therapeutic investigations. 

Major findings : As of February 1, 1971, we were following a total of 
90 patients. Of these there were 30 confirmed ALS, 13 with sxispect ALS, 
12 patients with definite PD and 35 with suspect PD . During the calendar 
year 1970 there were 15 deaths from ALS, 3 deaths from PD, 3 deaths among 
PD suspects and no deaths among ALS suspects. Of these, autopsies were 
performed on 17. 

We have completed the 5 year updating of the NINDS ALS and PD patients. 
We have analyzed most of the data and have completed early drafts of 
manuscripts for publication. The rates of ALS and PD have been declining 
slightly but apparently not significantly. Age-specific death rates for 
1950 - 59 and 1960 - 69 are identical indicating that no cohort phenomenon 
relative to some form of early common exposure or practice is apparent from 
these data. This indicates once again that the caxises of these diseases are 
still present on Guam. ALS and PD each caused 10% of all deaths among 
Chamorros on Guam over age 25 through 1970, Thus 1 in 5 adult Chamorros 
will die of one of these diseases. 

Analysis of offspring of 100 ALS and 100 PD patients and matched 
controls will be completed shortly upon securing further information on 
a small residual of people. We hope that analysis of this data will furnish 



18t 



Serial No. NDS (CF) - 55 E 201 

familial and possibly genetic patterns of the diseases. 

Once again we have noted several microfoci of ALS and PD patients in 
various areas on Guam. Prior to ray next trip in August our staff will 
attempt to identify these foci and map them in order that we may conduct the 
appropriate field surveys during my stay. 

Results of our study of patients with ALS, PD and controls to determine 
the rate of CNS synthesis of dopamine by administering probenecid and 
sampling the spinal fluid at appropriate intervals are available. A 
manuscript has been prepared and accepted for publication. 

The data clearly indicate that PD patients synthesize very little 
dopamine in the CNS while in ALS patients dopamine production is compromised 
but to a lesser degree. Similar findings among stateside ALS patients were 
encountered. This is perhaps the most significant finding emanating from 
the Guam studies. It is the first documentation of an altered CNS metabolic 
pathway in ALS and may lead to a breakthrough in the understanding of the 
pathogenesis of this disease. 

In order to explore the apparent deficit in dopamine production in the 
CNS of Guamanian PD patients, ALS patients and some controls, we are studying 
CNS dopamine synthesis by a new method developed by Dr. Chase and his group 
in which we sample urine and expired air rather than CSF. Results are not 
yet available. 

In addition to routine case finding and documentation we are continuing 
trials with L-Dopa. At present of the 8 people in the original trial, 5 are 
still receiving the drug after 15 months . One patient started on L-Dopa died 
but we were unable to conduct an autopsy. A paper analyzing our first 6 to 
8 month experience with PD patients on L-Dopa has been prepared and submitted 
for publication. In general, extrapyramidal features in most of the patients 
improved to some degree on L-Dopa while there was no clear evidence of 
amelioration of the dementia. Some heightened awareness and interest, 
however, was observed. Rigidity responded best in our series while tremor 
also appeared improved by L-Dopa. Bradykenesia was not improved to the 
degree which we expected, suggesting that part of this feature of the disease 
may be the result of destruction in non-dopaminergic areas of the brain 
(frontal lobe) . We are following all patients carefully in order to determine 
if L-Dopa actually affects the natural history and life expectancy in this 
degenerative fatal disease. 

We are planning to initiate treatment trials in other PD patients using 
a peripheral decarboxylating agent (MD 486-Merck) and much lower doses of 
L-Dopa. A protocol has been developed for this trial in collaboration with 
Dr. Thomas Chase, Chief, Section on Neurology, Laboratory of Clinical Sciences, 
NIMH, and the Merck Pharmaceutical Company. 

Significance to biomedical research and the program of the Institute : 
Almost all ALS patients are now in one of our drug trials. At present 7 



19t 



Serial No. NDS (CF) - 55 E 201 

patients have been receiving L-Dopa since late October 1970, and 12 patients 
are receiving isoprinosine since December while 8 patients are receiving 
placebo. The studies are double-blind and we analyzed our early results. 
Dr. Chen and Yase re-evaluated all patients and scored their results and 
our medical staff gave subjective evaluations of each patient's status. 
Dr. Stanhope and I have the code and we compared results. It is clearly too 
early to make any definitive statement. Patients are tolerating medication 
extremely well and are enthusiastic and cooperative in our studies. Our 
staff has worked very hard and diligently and each patient is seen at least 
once a week. Because of this heightened interest and attention and our 
insistence that patients eat at the time they take medication we have 
introduced a rather interesting potential bias. Patients appear subjectively 
improved and are eating better than before. We should be able to evaluate 
this factor over time by comparing patients on drugs and on placebo. The 
studies will remain double-blind and when Dr. Stanhope leaves, the code and 
pill dispensing will be supervised by Mrs. Hernandez. We believe our 
isoprinosine series is now sufficient and new patients will be placed in 
the L-Dopa (with Mk 486 when available) drug trial. 

Dr. Haruo Okazaki, Section of Experimental and Anatomic Pathology of 
the Mayo Clinic continues his systematic study of the occurrence of neuro- 
fibrillary changes in the brains of approximately 100 Guamanians who died 
of causes other than ALS or PD . Dr. Okazaki visited Guam for 4 days in 
December as a consultant and discussed relevant matters with our staff 
and with the pathologists of the Giiam Memorial Hospital. 

In Guam we have perhaps the highest incidence in the world of motor 
neuron disease and of a primary CNS degeneration. The documentation of the 
epidemiological, clinical, and neuropathological aspects of ALS and PD, a 
major neuromuscular disease and an important primary CNS degeneration have 
added to the world's knowledge concerning these neurologic diseases. In 
fields in which there are no known causes and no known cures, data such as 
these provide one of the most likely avenues for development of concepts and 
facts which lead to causes and cures. We are also exploiting this unique 
opportunity to test new drugs of potential benefit to patients and through 
our studies we have discovered a dopamine deficiency in ALS patients on 
Guam and in the U.S. that is the first promising lead in the understanding 
of this disease. 

Proposed Course : In addition to pursuing the above studies we are 
planning the following: 

Lymphocyte transformation: These studies have been interrupted because 
of the increased clinical activities related to the drug studies. 
Mr. George Nemo is preparing a new protocol and material v/ill be sent 
to Guam to pursue our investigations of possible immune factors in ALS 
and PD. 

Tissue culture : Some work on tissue culture from fresh brain material 
continues on Guam and in Bethesda under the supervision of Dr. C. Joseph 
Gibbs, Head, Laboratory of Slow, Latent and Temperate Virus Infections, 

20 t 



Serial No. NDS (CF) - 55 E 201 

C&FR, NINDS. We hope we will be able to pursue these studies more 
actively perhaps by future visits to Guam by Dr, Gibbs and Mr. Nemo. 

We are awaiting results of the chemical analysis of neurofibriles from 
Dr. Michael L. Shelanski, Assistant Professor of Neuropathology, Albert 
Einstein College of Medicine. He received two hemispheres of Chamorro 
PD patients ' brains in whom autopsies were performed within three hours 
after death and material was frozen in liquid nitrogen. Five brains were 
sent to Dr. Andre Barbeau, Director, Department of Neurobiology, Clinical 
Research Institute of Montreal. These include ALS and PD patients and 
controls and Dr. Barbeau is conducting his to-chemical analysis in order to 
determine the patterns of dopamine and other substances in various areas of 
the brains . Material from general autopsies conducted previously and stored 
as tissue blocks in Bethesda were returned to Dr. Loerzel and Dr. Varona for 
review. 

Honors and Awards : Official Citation of Commendation from Legislature of 
Guam to NINDS Research Center (Resolution #381 of 10th 
Guam Legislature) 

Publications: Brody, J. A. and Chen, K.M. : Changing epidemiologic patterns 
of amyotrophic lateral sclerosis and parkinsonism-dementia 
on Guam. Motor Neuron Diseases , Grune & Stratton, 1969, 
pp. 61-79. 

Chen, K.M., Brody, J. A., Kurland, L.T. and Elizan, T.S.: 
Patterns of neurologic diseases on Guam. II. Clinical 
and genetic aspects. Neurology , 20:954-964, 1970. 

Brody, J. A., Hussels , I., Brink, E. and Torres, J.M.: A 
preliminary report on hereditary blindness among the 
Pingelapese people of the Eastern Caroline Islands. 
Lancet, 1:1253-1257, 1970. 

Brody, J.A. : Phenomenal incidence of amyotrophic lateral 
sclerosis and parkinsonism-dementia on Guam. Presented at 
the 97th Annual Meeting of the American Public Health 
Association, Philadelphia, Pa., November 1969. 

Brody, J.A.: Studies of the phenomenally high incidence of 
amyotrophic lateral sclerosis and parkinsonism-dementia on 
Guam. Clinical Society and Commissioned Officers Association 
Fifth Joint Meeting - March 31 - April 3, 1970, Washington, 
D.C. 

Reed, D., LaBarthe, D. and Stallones, R.: Health effects 
of westernization and migration among Chamorros . Amer . J . 
Epid . 92:94-112, 1970. 



2it 



Serial No. NDS (CF) - 55 E 201 

Brody, J. A., Chase, T.N. and Gordon, E.K.: Depressed monamine 
catabolite levels in cerebrospinal fluid of patients with 
parkinsonism-dementia of Guam. New Eng. J. Med . 282:947-950, 
1970. 

Brody, J. A., Hirano, A. and Scott, R.M.: Recent neuropathologic 
observations in amyotrophic lateral sclerosis and parkinsonism- 
dementia of Guam. Neurology . In press. 

Chase, T.N., Schnur, J. A., Brody, J. A. and Gordon, E.K.: 
Parkinsonism-dementia and amyotrophic lateral sclerosis of 
Guam: Effect of probenecid on monamine catabolite levels 
in cerebrospinal fluid. Arch . Neurol . In press. 

Schnur, J. A., Chase, T.N. and Brody, J.A. : Parkinsonism- 
dementia of Guam: Treatment with L-Dopa. Neurology . 
In press. 



22t 



Serial No. NDS (CF) - 63 E 1103 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Neurological diseases other than ALS/PD on Guam 

Previous Serial Number: Same 

Principal Investigators: Charles E. Morris, M.D. 

NINDS Research Center 
John M. Stanhope, M.D. 
NINDS Research Center 

Other Investigators: Kwang-ming Chen, M.D. 

Mayo Clinic 
Jacob A. Brody, M.D. 
Leonard T. Kurland, M.D. 

Mayo Clinic 

Cooperating Units: NINDS Research Center, Agana, Guam 
Mayo Clinic, Rochester, Minnesota 

Man Years : 

Total: 1/4 
Professional: 3/16 
Other: 1/16 

Project Description: 

Objectives : A survey in 1954 by Donald W. Mulder, M.D. and Leonard T. 
Kurland, M.D. gave the impression that not only ALS, but also other heredo- 
familial neurologic disorders seemed unusually prevalent while multiple 
sclerosis and perhaps CNS tumors are uncommon. The objective of this study 
is to try to determine the validity of this data and to see if it is related 
to ALS and PD . 

Methods employed : Since the establishment of this Center in 1956, we 
have occupied a unique position on Guam. It is the only neurological 
consultation service available to all ethnic groups on Guam and sees most 
neurological patients at Guam Memorial Hospital and Naval Hospital. Therefore, 
it is expected that most of the significant neurological cases are eventually 
brought to our attention. Because of this unique position we hope to 
determine the frequency of various heredo-familial neurological disorders on 
the island. 

Major findings : During the year 188 patients with neurologic diseases 
other than ALS and PD were seen and 99 EEC's were performed. From 1960 

23t 



Serial No. NDS (CF) - 63 E 1103 

through 1966, in conjunction with ongoing studies of amyotrophic lateral 
sclerosis and parkinsonism-dementia on Guam, 1,028 Chamorro patients were 
referred to our neurologic clinic. In comparison with other populations and 
particularly that of Rochester, Minnesota, the residents of Guam had higher 
rates of convulsive disorders, myotonic dystrophy, peroneal muscular atrophy, 
and hereditary ataxias. There was no indication of an unusual incidence of 
central nervous system neoplasms, and no cases of progressive muscular 
dystrophy, myasthenia gravis, or indigenous multiple sclerosis were seen. 
No patient with proved classic paralysis agitans was observed in the Chamorro 
population. One sibship of 13 was followed in which 4 patients died of 
various brain tumors and 2 of acute nyelogenic leucemia. 

Significance to biomedical research and the program of the Institute ; 
This study adds to the general body of knowledge being collected by the 
Branch regarding the island of Guam and provides information on diseases 
possibly related to ALS and PD . 

Proposed course : We are expanding studies of neurologic diseases on 
Guam and the Trust Territories using the same techniques. 

Honors and Awards : None 

Publications: Chen, K.M., Brody, J. A. and Kurland, L.T.: Patterns of 

neurologic diseases on Guam, Arch. Neurol . 19:573-578, 1968. 

Chen, K.M., Brody, J.A., Kurland, L.T. and Elizan, T.S.: 
Patterns of neurologic diseases on Guam. II. Clinical and 
genetic aspects. Neurology , 20:954-964, 1970. 



24t 



Serial No. NDS (CF) - 66 E 1319 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: A search for automimmune mechanisms in the pathogenesis of 
chronic neurological diseases by the use of peripheral 
lymphocytes 

Previous Serial Number: Same 

Principal Investigators: Jacob A. Brody, M.D. 

George Nemo, Ph.D. 

Other Investigators: Minnie Toure, Biologist 
Gary Cooper, Biologist 

Cooperating Unit: None 

Man Years : 

Total : 1/4 
Professional: 1/12 
Other: 3/12 

Project Description: 

Objectives : To study the role of the small lymphocyte in the pathogenesis 
of neurologic disorders suspected to be of autoimmune etiology. 

Methods employed : Peripheral lymphocytes from patients with multiple 
sclerosis (MS) were challenged in vitro with specific antigens. The 
incorporation of tritiated thymidine into DNA during the synthetic phase 
of lymphoblast transformation is used as an indicator of lymphocyte 
responsiveness. In order to quantitate tritiated thymidine incorporation 
more accurately, our laboratory has converted from autoradiography to 
liquid scintillation spectrometry. Liquid scintillation is more sensitive, 
far less time consuming and subject to a minimal degree of hximan error. 

Major findings : Lymphocytes from normal patients and patients with 
multiple sclerosis were challenged with brain antigens and cerebrospinal 
fluid from patients with MS. Basic protein extracted from defatted guinea 
pig brains was also used as a test antigen. The results show that no 
significant lymphocyte transformation occurred in the samples tested. 

Significance to biomedical research and the program of the Institute : 
Since it is well established that the lymphocyte is the mediator of cellular 
immunity, the lack of a significant lymphocyte response as demonstrated in 
our study casts serious doubt on the hypothesis that MS is an autoimmune 
disorder . 

25t 



Serial No. NDS (CF) - 66 E 1319 

Proposed course : It may well be that only a small proportion of the 
total lymphocyte population sampled was sensitive to the antigens tested. 
The total number of reactive cells may have been too few to elicit a 
measurable response. If this supposition is indeed correct, then 
experimental manipulations designed to elevate the response to detectable 
levels might prove fruitful. 

Several reagents are currently being tested for their ability to 
enhance lymphocyte transformation. Preliminary data indicate that 
Polyriboinosinic-Polyribocytidylic acid (Poly I:C), a synthetic double- 
stranded polynucleotide, increases the lymphocyte response to vaccinia 
virus as much as 40%. 

Honors and Awards: None 

Publications: Brody, J. A., Harlem, M.M., Plank, C.R. and White, L.R.: 

Freezing human peripheral lymphocytes and a technique for 
culture in monolayers . Proc. Soc. Exp. Biol. & Med . 129 : 
968-972, 1968. 

Brody, J. A., Harlem, M.M., Kurtzke, J.F. and White, L.R.: 
Unsuccessful attempt to induce transformation by cerebrospinal 
fluid in cultured lymphocytes from multiple sclerosis patients, 
New Eng. J. Med . 279:202-204, 1968. 



26t 



Serial No. NDS (CF) - 66 E 1320 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Stateside Guamanian study 

Previous Serial Number: Same 

Principal Investigator: Jacob A. Brody, M.D. 

Other Investigator: None 

Cooperating Units: NINDS Research Center, Agana, Guam 

School of Public Health, University of California, 
Berkeley 

Man Years : 

Total : 1/6 
Professional: 1/12 
Other : 1/12 

Project Description: 

Objectives : This study was instituted in July 1966 to determine if 
ALS and PD occur with the same high frequency among Guamanians who have 
left Guam. Since the bulk of the stateside Guamanians are in California, 
efforts have been concentrated there. 

Methods employed : A household census was completed in the fall of 
1967 which included information on neurologic disease. Names of heads of 
households were obtained from relatives on Guam, the office of the Guamanian 
representative to Congress, local Guamanians, social organizations, an eight 
year old NINDS California Guamanian registry and from other Guamanians 
already living in this country. Household information was obtained by 
trained Guamanian interviewers living in California and by personnel from 
the Branch. Follow-up examination of suspect cases of ALS and PD was 
conducted by specialist physicians. 

Ma j or findings : The ALS rate in California is as high as it is on Guam. 
Two patients with presumed Parkinson's disease and/or dementia have died and 
autopsy studies in one are consistent with parkins onism-dementia although 
the patient's age is unusually advanced and in other changes were more 
compatible with paralysis agitans without dementia as seen in the United 
States. These findings are being included in a report by Brody and Hirano. 



27t 



Serial No. NDS (CF) - 66 E 1320 

Significance to biomedical research and the program of the Institute : 
The results suggest that a genetic factor and/or early exposure to an 
environmental factor is responsible for ALS and PD on Guam. The PD patient 
in California is the only PD patient ever encountered off Guam and the 
Marianas. The disease is not known to occur in non-Chamorros . Since the 
rate of PD off Guam is lower than that of ALS it suggests that these 
diseases are not a spectrum of CNS diseases with a single cause. The 
patient with paralysis agitans is the first documentation of this disease 
in a Chamorro. He lived in the United States for 8 years which may be a 
clue as to the incubation period of paralysis agitans. 

Proposed course : It is planned to maintain contact with this migrant 
population over the years because of the valuable clues we may gain regarding 
etiology of amyotrophic lateral sclerosis and parkins onism-dementia as seen 
on Guam, as well as more general medical, epidemiological and social data 
as the culturation proceeds . Thus far only Guam-born Chamorros in California 
are old enough to develop ALS or PD . As the population ages we will attempt 
to determine if these diseases occur at high rates in United States-born 
Guamanians . 

Honors and Awards : None 

Publications: Eldridge, R., Rosario, J. and Brody, J. A.: Amyotrophic 
lateral sclerosis and parkinsonism dementia in a migrant 
population from Guam. (A preliminary report.) In Trans . 
Amer. Neurol. Ass . 93:204-206, 1968. 

Eldridge, R., Ryan, E., Rosario, J. and Brody, J. A.: 
Amyotrophic lateral sclerosis and parkinsonism dementia 
in a migrant population from Guam. (A full report.) 
Neurology , 19:1029-1037, 1969. 

Reed, D., LaBarthe, D. and Stallones, R.: Health effects 
of westernization and migration among Chamorros. Amer. 
J. Epid . 92:94-112, 1970. 



28t 



Serial No. NDS (CF) - 66 E 1321 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Japanese encephalitis on Guam 

Previous Serial Number: Same 

Principal Investigators: Roger Detels, M.D. 

Jacob A. Brody, M.D. 
C. Joseph Gibbs, Ph.D. 

Laboratory of Slow, Latent and Temperate Viruses, 
NINDS 
Other Investigators: None 

Cooperating Unit: Laboratory of Slow, Latent and Temperate Viruses, NINDS 

Man Years : 

Total: 1/3 
Professional: 1/6 
Other: 1/6 

Project Description: 

Objectives : A Japanese encephalitis epidemic occurred in 1947 on Guam, 
but, according to serologic studies, involved only 20% of the population 
before apparently disappearing from the island. It is the objective of 
this study to determine if Japanese encephalitis virus (JEV) is persisting 
on Guam at a low level and to determine why it has not established an 
epidemic pattern as in Japan, Taiwan and Korea or an endemic pattern as 
in Malaysia, despite the presence of a suitable vector and reservoir hosts. 

Methods employed : Sera will be collected from Guamanians born prior 
to, during and after the occurrence of the 1947 epidemic and will be 
analyzed for antibody to JEV and other Group B and Group A arboviruses. 
Sera will also be collected for antibody screening from animals. Mosquitoes 
will be collected to determine the types of culicenes present on the island 
which might act as vectors. 

Major findings : Eighteen percent of sera from 498 Guamanians born 
since 1900 contain hemagglutination inhibition antibodies to JEV. 
Twenty-one percent born prior to 1950 and 8% born since 1950 have HI 
antibody to JEV suggesting that there has been Group B arbovirus activity 
on Guam since 1950. Nonetheless, only 1 of 100 pigs bled had HI antibody 
to JEV . Culex tritaeniorhynchus , but not Culex annulus , has been identified 
on the island. 



29t 



Serial No. NDS (CF) - 66 E 1321 

Significance to biomedical research and the program of the Institute : 
JEV was thought to have disappeared from Guam contrary to the usual pattern. 
However, the finding of HI antibodies to JEV in 8% of Guamanians born since 
1950 but not in pigs when all the known necessary ingredients are present 
for JEV to be either endemic or epidemic invite investigation to determine 
the uniqueness of Guam and thus contribute to the knowledge of factors 
important to the epidemiology of JEV. 

Proposed course : Tissue culture neutralization tests will be done on 
all positive HI sera and an aliquot of HI negative sera using several 
antigens in addition to JEV. Further sera are being collected from 
Guamanians born since 1950 for neutralization tests. The C . tritaeniorhynchus 
from Guam will be subclassif ied, since Barnet has proposed that only 
C. tritaeniorhynchus summorosus acts as a vector for JEV. 

Honors and Awards : None 

Publications: None 



30 1 



Serial No, NDS (CF) - 67 E 1325 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: One year experience of all births on Guam with special 
reference to diabetic complications 

Previous Serial Number: Same 

Principal Investigator: Jacob A. Brody, M.D. 

Other Investigators: John M. Stanhope, M.D. 

NINDS Research Center 
Anne Kantor 

Office of Biometry 

Cooperating Units: NINDS Research Center, Agana, Guam 
Office of Biometry, OD, NINDS 



Man Years : 




Total : 


1/4 


Professional: 


1/12 


Other: 


1/6 



Project Description: 

Objectives : The present project was designed to expand the scope of 
this study by analyzing all births for one year. During the year 1965 
there were 2,523 births and we compiled data on date of birth, place of 
birth, birth weight, birth order, length of gestation, birth defects, age 
and race of parents, maternal complications such as diabetes, etc. 

Methods employed : A study by Yen in 1963-64 revealed an unusually 
high incidence of abnormal carbohydrate metabolism during pregnancy among 
the native population of Guam. He also found that obesity and large babies 
appeared to be a constant finding in mothers with abnormal carbohydrate 
metabolism and were connected with an increased incidence of maternal and 
perinatal complications . 

Major findings : These data revealed an excess of low weight and high 
weight babies among diabetic mothers. Differences were not statistically 
significant and in general infant mortality on Guam is similar to that in 
the United States. 

Significance to biomedical research and the program of the Institute : 
The data will be of value for public health and anthropological studies. 

3Ib 



Serial No. NDS (CF) - 67 E 1325 

We will also conduct prospective retrospective studies on the use of birth 
weight as an indication of diabetes in families and communxties . 

Proposed course : This study has been terminated. 

Honors and Awards: None 

Publications : None 



32 t 



Serial No. NDS (CF) - 67 E 1485 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Analyses of abnormal urine and blood amino acids metabolism 
among Guamanians 

Previous Serial Number: Same 

Principal Investigators: Jacob A. Brody, M.D. 

Vivian Shih, M.D. 

Massachusetts General Hospital 

Other Investigators: Jose M. Torres 

NINDS Research Center 
Manuel T. Cruz 

NINDS Research Center 

Cooperating Units: NINDS Research Center, Agana, Guam 

Amino Acid Laboratory, Massachusetts General Hospital, 
Boston, Massachusetts 

Man Years : 

Total: 1/6 
Professional: 1/12 
Other: 1/12 

Project Description: 

Objectives : Earlier observation indicated that indigenous Guamanians 
have difficulties in handling protein and carbohydrate. However, the 
relationship between the observed hyperuricemia and hyperglycemia on Guam 
and the neurologic manifestations is not clear. 

Methods employed : A contract with the Amino Acid Laboratory of the 
Massachusetts General Hospital for the broad testing for inborn errors of 
metabolism was secured and blood and urine are being sent to the lab from 
Guam. 

Major findings : No major abnormalities in infants, retarded children 
and ALS or PD patients were encountered in the survey of blood and urine 
in 435 patients with various diseases and 574 normal infants and 20 normal 
adults in Guam and other islands of Micronesia. One girl apparently healthy 
but with a history of seizure disorder (apparently a febril convulsion) many 
years ago was found to excrete an unknown sulfur amino acid. We are 
reviewing her family clinically and collecting appropriate urine samples 
for confirmation of this finding. No specific changes were found in PD or 

33t 



Serial No, NDS (CF) - 67 E 1485 

ALS patients. B-amino isobutyric aciduria was detected in 56.8% of normal 
infants. Taurine excretion was prevalent in normal infants on the Caroline 
Islands; it was probably related to breast-feeding. Cystathioninuria was 
present in 9 normal infants . We have the impression that the infant 
population in Guam and other areas have relatively low rates of abnormal 
amino acids when compared with other populations . This cannot be claimed 
conclusively because sampling methods differed in our series. The Department 
of Public Health of the Government of Guam has routinely screened all new 
boms for PKU for the past 6 years and no case has been encountered. 

Significance to biomedical research and the program of the Institute ; 
This study contributed to knowledge of metabolic abnormalities of the 
Chamorro people of Guam. 

Proposed course : Collection of specimens is complete. We will follow 
up on the one unusual patient referred to above. 

Honors and Awards : None 

Publications: Shih, V.E., Brink, E.W., Peneva, P. and Brody, J. A.: Blood 

and urinary amino acid patterns in Guamanians and Micronesians . 
Amer. J. Pis. Child . In press . 



34 1 



Serial No. NDS (CF) - 67E 1486 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Torsion Dystonia - a clinical and genetic study 

Previous Serial Number: Same 

Principal Investigator: Roswell Eldridge, M.D. 

Other Investigators: Irving S. Cooper, M.D. 

St. Barnabas Hospital 
Morris B. Gross, M.D. 

Hunter College in the Bronx 
Wolfgang Zeman, M.D. 

University of Indiana 
Mary Bazelon Coleman, M.D. 

Children's Hospital 
Kathy O'Meara 



Cooperating Units: 



Department of Neurologic Surgery, St. Barnabas 

Hospital, New York 
Laboratory of Clinical Science, NIMH 



Man Years 




Total 


1 


Professional: 


2/3 


Other: 


1/3 



Project Description: 

Objectives : Torsion dystonia (TD) comprises a heterogeneous group of 
conditions characterized by disordered movement. TD may be due to either 
genetic or environmental factors. The present study has already defined 
further the nosology of these conditions. Further clinical family 
studies may suggest the basic defect in each. 

Methods employed : Initially, probands with a history of TD selected 
through 180 neurologic and neurosurgical centers provided the families 
for study. Recently, physicians and affected individuals themselves 
have contacted us requesting help. A detailed clinical family history 
is obtained. The latter stresses geographical origin of ancestral 
couples. The proband and all available relatives were given physical 
examinations. Patients from all areas of the U. S. treated by various 
methods are seen to avoid geographic, ethnic, and therapeutic bias. 



35t 



Serial No. NDS (CF) - 67 E 1486 

Maior findings: Clinical, genetic, psychometric and therapeutic 
aspects of dystonia have been evaluated in more than 200 patients in 130 
families. The results have appeared in publications indicated below. 
Among the conclusions are: at least two hereditary forms of dystonia 
exist; there is variation in clinical features and course of the 
hereditary types of dystonia; psychotherapy has a limited role as 
primary treatment; drugs reported to be helpful in the dystonias generally 
have been ineffective in most patients over a long period; and recent 
neurosurgical procedures offer hope. 

Significance to biomedical research and the program of the Institute : 
Elucidation of the fundamental defect in these forms of dystonia will be of 
practical importance. In addition to suggesting specific treatment, 
it should be possible to distinguish between the recessive and dominant 
forms chemically. The application to genetic counselling of such a test 
is obvious. As in other inborn errors of metabolism, such a study could 
provide basic, new information about central nervous system physiology. 

Parkinson's disease shares certain clinical features with dystonia and is 
relieved by the same operative procedure so that information gained from 
the dystonia study may bear on this important problem. 

Proposed course : Present efforts in this project are directed toward 
documentation of all abnormalities in the hereditary dystonias and search 
for appropriate therapy. In the latter connection we are working with 
other medical centers including the National Institute of Mental Health, 
Department of Neurology at Children's Hospital, Neurological Institute of 
New York City, St. Barnabas and Albert Einstein Hospitals, New York City, 
University Hospital, Cleveland and University of Montreal, Montreal. 

We are pleased with the ground work that has been laid in terms of under- 
standing the nosology and clinical course of the dystonias. 

Honors and Awards : None 

Publications: 



Eldridge, R, , Ryan, E. , Brody, J. A. and Cooper, 1,8.: Dystonia musculorum 
deformans: Evidence for two hereditary forms. Excerpta Medica International 
Congress Series No. 175, Progress in Neuro-Genetics , Vol, I of the Proceed- 
ings of the Second International Congress of Neuro-Genetics and Neuro- 
Ophthalmology, Montreal, September 1967, pp, 772-788, 1969, 



36t 



Serial No. NDS (CF) - 67 E 1486 

Eldridge, R. , Harlan, A., Cooper, I.S., and Riklan, M. : The Hereditary 
Torsion Dystonias (Dystonia Musculorum Deformans): Geographical 
distribution and I.Q. in dominant and recessive forms. In Transactions of 
the American Neurological Association, 94, 1969. 

Eldridge, R. , Harlan, A., Cooper, I.S. and Riklan, M. : Superior intelligence 
in recessively inherited torsion dystonia. The Lancet 1:7637, pp. 65-67, 
1970. 

Eldridge, R. , Edgar, A., and Cooper, I.S.: Genetics, Geography and 
Intelligence in the torsion dystonias. Proceedings of the Second 
Conference on the Clinical Delineation of Birth Defects, May 1969. The 
National Foundation-March of D imes (In press) 

Eldridge, R. : The Torsion Dystonias: Literature Review: Genetic and 
Clinical Studies. In The Torsion Dystonias (Dystonia Musculorum Deformans). 
Editor, Roswell Eldridge, Neurology suppl. 20:11, Part 2, November 1970. 

Eldridge, R. and Koerber, T. : The Torsion Dystonias: Some Genetic and 
Psychiatric Implications. The Psychiatric Forum . April, 1971. 



37t 



Serial No. NDS(CF) - 67 E 1487 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Genetic analysis of family data on Guam ALS cases 

Previous Serial Number: Same 



Principal Investigators: 



John M. Stanhope, M.D. 

NINDS Research Center 
Jacob A . Brody , M.D. 
Charles E. Morris, M.D. 

NINDS Research Center 
Roswell Eldridge, M.D. 



Other Investigator: 



Manuel T. Cruz 

NINDS Research Center 



Cooperation Unit: NINDS Research Center, Agana, Guam 

Man Years : 

Total: 1/4 
Professional: 1/6 
Other: 1/12 

Project Description: 

Objectives : To utilize the accumulation of 20 years of experience for 
an indepth genetic analysis of pedigree information of Guam ALS . 

Methods employed : Pedigrees were developed for the 370 definite cases 
of Guam ALS by Guamanian practitioners aware of actual biologic parents . Of 
these, 70 were suitable for segregation analysis. Also examination was made 
of sibs whose parents were both affected by ALS. 

Ma j or findings : The initial data indicate that Guam ALS may be inherited 
as a simple autosomal recessive trait. In 46 families suitable for test of 
the autosomal recessive hypothesis, 64 cases were observed while 74 cases 
would be expected from truncate analysis. The 95% confidence limits for 
such analysis cover the range from 64 cases to 84 cases so the observed 
number of cases is compatible at this level of significance although barely 
so. 

Two of 13 over 35 years of age whose parents both had ALS were found to 
have signs compatible with early ALS. If ALS is recessive all such offspring 
should eventually be affected. 



39t 



Serial No. NDS (CF) - 67 E 1487 

In testing for the autosomal dominant hypothesis in the 16 families 
suitable for this analysis, 9 cases were observed, 27 would be expected and 
19 to 35 cases would be the range at a 95% limit confidence. Therefore, 
autosomal dominant inheritance is possible only if one postulates the gene 
is not penetrant (i.e., there is no expressing of the disease) in 50% to 70% 
of those carrying it. 

Troublesome to any simple mode of inheritance postulated is the 2:1 male 
to female ration of ALS patients. A finding which may answer this discrepancy 
and possibly shed light on the basic process of the disease is that in these 
siblings there is an excess of female deaths under one year of age. In 
addition, 11 of our ALS or PD patients have only one Guamanian parent. We 
are also following the offspring of the first 100 ALS and PD patients and 
controls referred to above. 

Significance to biomedical research and the program of the Institute : 
Obviously establishing a genetic basis for Guam ALS and PD would have far- 
reaching consequences. The elusive question of the cause of Guam ALS and 
PD would be answered. A new genetic disease would be added to the expanding 
catalogue of inherited neurologic diseases. 

Proposed course : Complete information is being obtained on pedigrees 
of other Guam ALS and PD cases so as to increase the sample studied. 
Detailed study of offspring now at risk of specific mating types is underway. 

Honors and Awards : None 

Publications : None 



40 1 



Project Title: 



Serial No. NDS (CF) - 67 E 1488 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individiaal Project Report 

July 1, 1970 through June 30, 1971 

Serological studies of common viruses in cases of multiple 
sclerosis (MS) and controls 



Previous Serial Number: Same 



Principal Investigators; 



Other Investigators: 



Jacob A. Brody, M.D. 
John L. Sever, M.D. 

Perinatal Research Branch, NINDS 
Anne H. Edgar 
Jane McNew, R.N. 



Mark Dyken, M.D. 

Neurology Department, Indiana University Medical 

Center 
A. Donald Merritt, M.D. 

Department of Medical Genetics, Indiana University 

Medical Center 



Cooperating Units: Section of Infectious Diseases, PRB, NINDS 

Neurology Department, Indiana University Medical Center, 

Indianapolis 
Department of Medical Genetics, Indiana University 

Medical Center, Indianapolis 
The Wis tar Institute, Philadelphia, Pennsylvania 

Man Years : 

Total: 2 1/6 

Professional: 1 1/6 
Other: 1 

Project Description: 

Objectives : To test the hypothesis that MS may be caused by an unusual 
response to a common virus infection. To search for possible distortions of 
segregation and association among MS patients, siblings and controls. 

Methods employed : MS patients known to the Neurology Department, lUMC 
and to the Indiana Chapter of the National Multiple Sclerosis Society were 
contacted and asked to participate. In addition, for each case several 
controls with similar backgrounds and infectious disease experience were 
selected. Controls are classmate friends of the patient who grew up in the 
same community. Siblings of MS patients were also tested. A second sample 



4lt 



Serial No. NDS (CF) - 67 E 1488 

of MS patients and siblings was taken from the Washington, D.C. area. 
Patients and controls answered standard questions regarding infectioiis disease, 
environment, course of illness and family history, and blood specimens were 
taken. 

Serological analysis was conducted in the Section of Infectious 
Diseases, PRE, NINDS using a battery of common virus antigens by 
hemagglutination and complement fixation methods. Differences between the 
patient, his sibling and his controls were analyzed. 

A portion of frozen serum is being banked to test promising hypotheses 
in the future. 

Major findings : In the Indiana series we found that MS patients have 
higher titers than matched controls for measles, mumps, influenza C, 
parainfluenza type 3, varicilla and herpes virus hominus . In no case, 
however, were titers of MS patients higher than their siblings of the same 
sex. In the Washington series, female siblings had titers as high as 
female patients to measles while male siblings titer were lower than those 
of the patients . 

Significance to biomedical research and the program of the Institute : 
The observation that MS patients do have consistently higher titers against 
many viruses than controls supports an infectious or immune mechanism as 
being involved in the etiology of MS. The finding that higher titers also 
occur in siblings suggests that the phenomenon may be related to a common 
familial exposure or a familial immunologic defect. 

Proposed course : We are now testing gamma globulin levels of these 
sera in collaboration with Dr. Oldrich Kolar, Department of Neurology, 
Indiana University Medical Center, and testing the rabies titers in 
collaboration with Dr. Hilary Koprowski, Director, The Wistar Institute. 
We hope to extend our studies to the Shetland and Orkney Islands where MS 
occurs at a rate three times higher than elsewhere in the world. 

Honors and Awards : None 

Publications: Henson, I.E., Brody, J. A. and Sever, J.L.: Elevated measles 
antibodies in patients with multiple sclerosis and in their 
siblings. Presented at the 97th Annual Meeting of the 
American Public Health Association, Philadelphia, Pa., 
November 1969 . 

Henson, T.E., Brody, J. A., Sever, J.L., Dyken, M.L. and 
Cannon, J.M.: Measles antibodies in patients with multiple 

sclerosis and their siblings and controls. JAMA, 211:1985, 
1970. 

Brody, J. A.: Virus antibody titers in multiple sclerosis 
patients, siblings and controls. (An abstract.) (Presented 



42 1 



Serial No. NDS (CF) - 67 E 1488 

at the American Epidemiological Society Meeting in Seattle, 
Washington, April 1970.) 

Brody, J. A., Sever, J.L. and Henson, T.E.: Virus antibodies 
in the serum of multiple sclerosis patients and matched 
controls. Neurology , 20:389, 1970. (Presented at the 
American Academy of Neurology, May 1970. Proceedings to 
be published. ) 

Brody, J. A., Sever, J.L. and Henson, T.E.: Virus antibodies 
in MS patients, siblings and controls. JAMA. In Press. 



43t 



Serial No. NDS (CF) - 67 E 1489 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Neuropathological studies in veterans dying of ALS who 
served on Guam 

Previous Serial Number: Same 

Principal Investigators: Jacob A. Brody, M.D. 

R. Michael Scott, M.D. 

Other Investigators: Kenneth Earle, M.D. 

Armed Forces Institute of Pathology 
Asao Hirano, M.D. 

Montefiore Hospital 
Joseph Seggora, M.D. 

Veterans Administration Hospital 
F.A. Quadfasel, M.D. 

Veterans Administration Central Office 

Cooperating Units: Neuropathology Branch, Armed Forces Institute of 

Pathology, Washington, D.C. 
Department of Neuropathology, Montefiore Hospital 

New York 
Veterans Administration Hospital, Boston 
Neurology Section, Veterans Administration Central 

Office, Washington, D.C. 



Years : 




Total: 


1/3 


Professional: 


1/3 


Other: 






Project Description: 

Objectives : To determine if ALS in veterans who served on Guam is an 
acquired disease. 

Methods employed : Dr. Hirano has reported characteristic neurofibrillary 
changes in Guamanian ALS patients, but these changes are not seen in classical 
stateside ALS. Since a large number of U.S. servicemen were stationed on 
Guam during World War II, it is possible to examine the CNS of U.S. veterans 
who died of ALS who spent considerable time on Guam. The question to be 
answered is whether these men show the characteristic Guam-type neuro- 
pathological changes or the classical stateside ALS changes. 



45t 



Serial No. NDS (CF) - 67 E 1489 

Major findings : Brains from three veterans serving on Guam have been 
collected. In two, there were no neurofibrillary changes, while in one 
these changes were present, but not in the distribution observed in Guam 
ALS patients . 

Significance to biomedical research and the program of the Institute : 
These findings are evidence that Guamanian ALS does not result from short 
term exposure to an environmental agent. However, the brain of a Filipino 
dying of ALS after being on Guam for many years has shown typical 
neurofibrillary changes, suggesting that length of exposure to an environ- 
mental agent may be an important factor in subsequent development of ALS. 

Proposed course : This study is terminated and the data have been 
written up and accepted for publication. 

Honors and Awards : None 

Publications: Brody, J. A., Hirano, A. and Scott, R.M.: Recent neuropatho logic 
observations in amyotrophic lateral sclerosis and parkinsonism- 
dementia of Guam. Neurology . In Press . 



46t 



Serial No. NDS (CF) - 67 E 1490 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: The application of fluorescent antibody methods to the 
study of chronic neurological disorders 

Previous Serial Number: Same 

Principal Investigators: Jacob A. Brody, M.D. 

Minnie Toure, Biologist 
George Nemo, Ph.D. 

Other Investigator: None 

Cooperating Unit: None 

Man Years : 

Total: 1/3 
Professional: 1/4 
Other: 1/12 

Project Description: 

Objectives : To employ fluorescent antibody techniques using frozen 
sections of CNS tissue for detection of viral antibody. 

Methods employed : The two main staining techniques employed in 
fluorescent microscopy, the direct and indirect methods will be employed. 
Frozen CNS tissue sections 4-5u will be prepared using a microtome in a 
refrigerated cryostat. 

Major findings : None as yet. 

Significance to biomedical research and the program of the Institute : 
Fluorescent antibody methods are applicable to the study of chronic 
neurological disorders thought to be of autoantibodies as well as sites 
of delayed-type hypersensitivity reactions. A search for viral antigens 
and sites of viral replication in CNS tissue is also made possible using 
this technique. 

Proposed course : To be continued. 

Honors and Awards : None 

Publications: None 



47t 



Serial No. NDS (CF) - 67 E 1496 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Sequelae of CNS diseases in childhood 

Previous Serial Number : Same 

Principal Investigator: Jacob A. Brody, M.D. 

Other Investigators: Estelle Kornhauser, R.N. 
Otis D. Turner 

Cooperating Units: Office of Biometry, OD, NINDS 

Children's Hospital, Washington, D.C. 

Man Years : 

Total: 2/3 
Professional: 1/3 
Other: 1/3 

Project Description: 

Objectives : To determine if infection with viruses capable of 
penetrating the CNS cause permanent neurologic sequelae, particularly in 
those cases in which infection occurred under the age of two years. 

Methods employed : As the result of previoios studies we have decided 
to improve our methods and techniques by investigating populations in which 
known infections from encephalogenic viruses occurred. 

Ma j or findings : Although we have attempted to secure the necessary 
populations of children under age 1 in New York, Panama, St. Louis and 
Chicago, we have not yet encountered a situation suitable for testing our 
hypothesis . 

Significance to biomedical research and the program of the Institute : 
Although it is widely believed that infections of the CNS early in childhood 
produce brain damage, there are no definite patterns of brain damage 
or specific diseases which commonly are associated with brain damage. 
Documentation of specific viral-tropisms to learning and performance would 
be a major contribution to understanding and preventing minimal and major 
brain damage. 

Proposed course ; Well documented arbovirus outbreaks have been observed 
by MARU, NIAID and several hundred measles patients who were infected under 

49t 



Serial No. NDS (CF) - 67 E 1496 

age 1 are known in Chicago (Kenrad E. Nelson, M.D., Assistant Professor of 
Preventive Medicine, University of Illinois College of Medicine, Municipal 
Contagious Disease Hospital, Chicago). We plan to study these occurrences 
systematically . 

Honors and Awards : None 

Publications: Brody, J. A. and Wilner, E. : Measles, minor neurologic signs 
and intelligence. Developmental Med, and Child. Neurol . 11; 
449-454, 1969. 



50 1 



Serial No. NDS (CF) - 67 E 1497 
i 1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Guillain-Barre (GB) - Bell's palsy (BP) study 

Previous Serial Number: Same 

Principal Investigator: Paul M. Hoffman, M.D. 

Other Investigators: Lon R. White, M.D. 

Jacob A. Brody, M.D. 
George Nemo, Ph.D. 

Cooperating Unit: Perinatal Branch, NINDS 

Man Years : 



Total: 


1/4 


Professional: 


1/4 


Other: 






Project Description: 

Objectives : To determine if GB and BP are caused by abnormal 
immunological host responses to common viruses. Since during the immune 
response the virus may not be recoverable from the patient we are attempting 
to isolate the virus from materials collected from household contacts. 

Methods employed : Personal contact was made with the neurology 
residents or senior medical residents in a number of local hospitals and 
letters were sent to the practicing neurologists and neurosurgeons in the 
local area, so that prompt notification might be obtained whenever a case 
of GB or BP appeared. When notification of a case was obtained, we contacted 
the patient to learn of any contacts (preferably children) in the household. 
If there were contacts we proceeded to collect blood, throat swabs, and 
rectal swabs from the patient and his contacts. Blood specimens were 
centrifuged and serum stored at -20 °C. Throat swabs were placed in Hank's 
medium and PPLO medium; rectal swabs were placed in Hank's medium and vials 
were stored at -70 °C. 

When sufficient numbers of specimens had been collected it was planned 
to forward them for serological studies and attempts at virus isolation. 
Records were kept of patient's history and neurological status and of 
contact's exposure to infections. Lymphocyte transformation studies were 
also performed on the subject. The significance of serum factors depressing 
lymphocyte transformation and the interaction of the isolated virus with 
the patients lymphocytes as well as the reaction of the lymphocytes to 

51t 



Serial No. NDS (CF) - 67 E 1497 

peripheral nerve will be evaluated. In addition careful case-control 
studies of epidemiologic factors will be conducted in hope of encountering 
precipitating factors in these diseases. 

Ma.jor findings : None 

Significance to biomedical research and the program of the Institute : 
This disease has been associated with several abnormalities that suggest 
auto-immunity as an etiology. Our work with the hypothesis of a viral 
infection triggering an abnormal immune response is in keeping with our 
knowledge of 3SPE and possibly MS. 

Proposed course : This project will continue. 

Honors and Awards : None 

Publications: None 



52t 



Serial No. NDS (CF) - 68 E 1594 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Phenothiazine-induced neurological effects: A study among 
twins 

Previous Serial Number : Same 

Principal Investigators: James A. Schnur, M,D. 

Jacob A. Brody, M.D. 
Dean F. Young, M.D. 

Other Investigators: John D. Rainer, M.D. 

New York State Psychiatric Institute 

Cooperating Units: National Institute of Child Health and Human Development, 
Children's Diagnostic and Study Branch 
National Institute of Mental Health 

Section on Twin & Sibling Studies, Adult Psychiatry 
Branch 
National Academy of Science, National Research Council 
Columbia University, New York State Psychiatric Institute 
Spring Grove State Hospital, Catonsville, Maryland 

Man Years: 

Total: 1/2 
Professional: 1/3 
Other : 1/6 

Project Description: 

Objectives : The objective of this study is to assess whether the 
specific types of neurological side reaction induced by phenothiazine drugs 
are influenced by genetic factors. 

Methods employed : A preliminary study among 46 female geriatric patients 
on long-term phenothiazine treatment revealed 33% had dvsklnetic reactions 
and 13%, Parkinson-like reactions. The subjects for this study are twin 
pairs, concordant for the same psychiatric diagnosis, who have been on 
chronic phenothiazine therapy. Zygosity of the twin pairs is determined by 
history, appearance, and extensive blood typing. A single neurological 
examination was conducted on each pair to determine the patterns of 
neurological reactions . By comparing the patterns of reactions in monozygotic 
with those of fraternal twins, we can employ the usual methods of analysis to 



53t 



Serial No. NDS (CF) - 68 E 1594 

determine the relative importance of genetic factors in the manifestation of 
extrapyramidal signs resulting from phenothiazine induction. 

Mai or findings : Six pairs of twin patients, four monozygotic and two | 
dizygotic, on long time-high dosage phenothiazine treatment have been 
examined. The results indicated that the specific type of neurological side 
effect is not primarily determined by genetic factors, since identical twins 
may develop distinctly different patterns of reaction. 

Significance to biomedical research and the program of the Institute ; 
This work may help elucidate the presence or absence of a genetic contribution 
to the occurrence of the important neurological side reactions to phenothiazine 
drugs . 

Proposed course : We propose to add other twin pairs to the series, and 
analyze the data. 

Honors and Awards : None 

Publications: None 



54t 



Serial No. NDS (CF) - 68 E 1595 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individioal Project Report 

July 1, 1970 through June 30, 1971 

Project Title: An evaluation of the effect of successful thalamic surgery 
on the progress of unilateral Parkinson's disease 

Previous Serial Number: Same 

Principal Investigators: R. Michael Scott, M.D. 

Jacob A. Brody, M.D. 
Joyce M. Cannon, R.N. 

Other Investigators: Irving S. Cooper, M.D. 

St. Barnabas Hospital 
Robert S. Schwab, M.D. 

Massachusetts General Hospital 

Cooperating Units: Department of Neurosurgery, St. Barnabas Hospital, 
Bronx, New York 
Department of Neurology, Massachusetts General Hospital, 
Boston, Massachusetts 

Man Years : 

Total : 1 

Professional: 11/12 

Other : 1/12 

Project Description: 

Objectives : To evaluate Dr. Cooper's hypothesis that in patients with 
unilateral Parkinson's disease, thalamic surgery which succeeds in permanently 
abolishing the tremor and rigidity on the involved side will either stop or 
markedly delay the appearance of the symptoms on the other side of the body; 
and to determine the natural history of vinilateral Parkinson's disease. 

Methods employed : The charts of 1,700 consecutive thalamic surgical 
cases from January 1963 through September 1964 at St. Barnabas Hospital 
were reviewed to select 100 patients who came to surgery with symptoms of 
tremor and rigidity confined to one side of the body. Seventy-two patients 
were eventually contacted and examined. Two unoperated groups were studied. 
The first of these consisted of 15 patients who presented to Dr. Cooper with 
unilateral signs, who were accepted for surgery, but for various reasons were 
not operated upon. The second control group consisted of 20 patients seen 
by Dr. Schwab in Boston and subsequently treated medically. 



55 t 



Serial No. NDS (CF) - 68 E 1595 

Major findings : Successful thalamic surgery does not affect the progress 
of tremor and rigidity to the extremities of the opposite side of the body. 
The rate and frequency of spread in unoperated and operated patients were 1 
strikingly similar. Certain of these patients had a form of Parkinson's I 
disease characterized by unilateral tremor and rigidity of long duration. ' 
The average age of onset of these patients was earlier than that of "classical" 
Parkinson's disease patients, and tliey had a higher frequency of encephalitis 
or severe febrile illness prior to the onset of their illness. Thalamotomy 
was often extremely effective in these patients. 

Significance to biomedical research and the program of the Institute : 
This study further defines the role of surgery in Parkinson's disease. It 
emphasizes that thalamotomy does not alter the course of progressive Parkinson's 
disease, but can restore to normalcy certain patients with the benign 
unilateral syndrome. In addition, it suggests ways in which patients with 
the benign syndrome might be identified. 

Proposed course : Piablications have been prepared and the project 
terminated. 

Honors and Awards : None 

Publications: Cooper, I.S., and Scott, R.M. : The clinical and physiological 
implications of 10 year cure of unilateral movement disorders 
by thalamic surgery. In: Proceedings of the Ilird Parkinson's 
Symposium . Edinburgh, Livingstone, (in press). 

Scott, R.M., Brody, J. A., and Cooper, I.S.: The effect of 
thalamotomy on the progress of unilateral Parkinson's disease. 
J. Neurosurg . 32:286-288, 1970. 

Scott, R.M., Brody, J. A., Schwab, R.S., Cooper, I.S.: 
Progression of unilateral tremor and regidity in Parkinson's 
disease. Neurology , 20:710-714, 1970. 

Scott, R.M., and Brody, J.A. : Benign early-onset Parkinson's 
disease: A syndrome distinct from classic postencephalitic 
parkinsonism. (Presented at American Academy of Neurology, 
May 1970). Neurology , 21:366-368, 1971 



56* 



Serial No. NDS (CF) - 68 E 1597 

1. Collaborative & Field Research 

2 . Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Neurologic diseases in the Trust Territories and other 
Pacific areas 



Previous Serial Number: 



Same 



Principal Investigators 



Jacob A. Brody, M.D. 
James A. Schnur, M.D. 
NINDS Research Center 



Other Investigators; 



Manuel Cruz 

mms Research Center 
Jose Torres 

NINDS Research Center 
Francisco Leon Guerrero 

NINDS Research Center 



Cooperating Units: 



Ophthalmology Branch, NINDS 

Department of Public Health, Trijst Territory of the 

Pacific Islands, Saipan, Marianas Islands 
Population Genetics Laboratory, University of Hawaii, 

Honolulu, Hawaii 



Man Years : 



Total: 

Professional: 
Other : 



2/3 
1/3 
1/3 



Project Description: 

Objectives : To investigate neurologic illness occurring in the Trust 
Territories and the Pacific area. Last year this involved the study of 
leprosy patients in New Caledonia. This year it involved the study of 
congenital blindness among the people of Pingelap. 

Methods employed : A field trip was made to Ponape to examine the 
Pingelapese people in order to document the nature of their eye disease 
and their mode of inheritance and epidemiologic patterns which could 
suggest both genetic and environmental factors. 

Major findings : The congenital eye disease is apparently a form of 
achromatopsia characterized by nystagmus, blinking, photophobia, blindness, 
and impairment of color vision. The developiment of cataracts usually within 



57 1 



Serial No. NDS (CF) - 68 E 1597 

the first 5 to 10 years of life and frequent appearance of high myopia. The 
mode of inheritance appears to be recessive and no ns ex- linked. Our most 
significant finding was that the apparent trend noted in 1969 that children 
in age group 0-4 had a significantly lower rate of the congenital eye 
disease was not sustained. In a follow-up trip in 1971 we observed that two 
new patients were born with the disease since 1969 bringing the rate in the 
current population age - 4 up into the range of those rates for older 
populations. Six patients were brought to the Ophthalmology Branch, NINDS 
for study. The diagnosis appears to be achromatopsia with unexplained 
high myopia. 

Significance to biomedical research and the program of the Institute : 
The group of diseases which are included under tapetoretinal degenerations 
are poorly understood. By having a population isolate with a phenomenally 
high rate (10% of the population is blind) will give a unique opportunity 
for studying the full range of manifestations of this entity as the 
expression of a single abnormal gene. In addition, hopefully, a mechanism 
for prevention will develop. 

Proposed course : We will continue to follow this population and other 
populations as they become known which are of potential medical interest. 

Honors and Awards : None 

Publications: Brody , J. A., Hussels, I., Brink, E. and Torres, J.M.: A 
preliminary report on hereditary blindness among the 
Pingelapese people of the Eastern Caroline Islands. 
Lancet , 1:1253-1257, 1970. 



58t 



Serial No. NDS (CF) - 68 E 1598 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Epilepsy on Guam 

Previous Serial Number: Same 

Principal Investigator: John M. Stanhope, M.D. 

NINDS Research Center 

Other Investigators: Jacob A. Brody, M.D. 
Manuel Cruz 

NINDS Research Center 
Jose Torres 

NINDS Research Center 
Francisco Leon Guerrero 

NINDS Research Center 

Cooperating Unit: NINDS Research Center, Agana, Guam 

Man Years : 

Total: 1/2 
Professional: 1/4 
Other: 1/4 

Project Description: 

Objectives : To determine the incidence and prevalence of epilepsy on 
Guam. To investigate methods for field studies of epilepsy. To determine 
if previous reports of unusually high incidence of convulsive disorders on 
Guam are accurate. 

Methods employed : We are testing four basic approaches: (1) We are 
following-up a 1962 survey of convulsive disorders in Umatac and Merizo to 
determine the outcome of those children known to have had febrile convulsions 
6 to 8 years ago; (2) to determine the true incidence and prevalence in 
a sample population we are doing a house to house survey in the villages 
of Talofofo, Merizo, and Yona; (3) as referral neurologists on Guam we are 
updating all previous referrals of convulsive disorders to us and establishing 
a registry. To add to this registry we are contacting all medical and 
paramedical personnel on Guam to discover new cases. This registry will be 
permanent and permit us to conduct studies in the Guam population; (4) to 
further elaborate on methods for acquiring information we are following-up 
all births on Guam in 1958 and 1963 throughout the entire island to determine 
the rates of convulsive disorders in these preselected populations. 

59* 



Serial No. NDS (CF) - 68 E 1598 

Major findings : We have already published that the rates of "true" 
epilepsy and of febrile convulsions are higher on Guam than elsewhere. We 
are completing the first three phases of the Epilepsy Study outlined in 
previous reports . We have the information on the rate of "true" epilepsy 
(the fourth part of our program) and have established sf permanent Epilepsy 
Registry on Guam. There seems little doubt that the rates are high on Guam, 
but the reasons are not clear. Available data are now being analyzed by 
Dr. Stanhope. 

Significance to biomedical research and the program of the Institute : 
Further studies of epilepsy in a well-defined and accessible population 
will add to the understanding of this disease and contribute new information 
concerning epilepsy in a tropical environment. It will also yield important 
information on different survey techniques and their relative accuracy. 

Proposed course : The analysis will be completed as soon as possible and 
more detailed studies of epilepsy will develop from our basic information. 

Honors and Awards : None 

Publications : None 



60t 



Serial No. NDS (CF) - 68 E 1605 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Phenothiazine-induced parkinsonism in white and Negro 
patients with nonorganic psychoses 

Previous Serial Number : Same 

Principal Investigators: James A. Schnur, M.D. 

NINDS Research Center 
R. Michael Scott, M.D. 

Other Investigators: Jacob A. Brody, M.D. 

Cooperating Units: Spring Grove State Hospital, Catonsville, Maryland 
Crownsville State Hospital, Crownsville, Maryland 

Man Years : 

Total: 2/3 

Professional: 1/2 
Other: 1/6 

Project Description: 

Objectives : To determine whether increased skin pigmentation is 
associated with decreased prevalence of phenothiazine-induced Parkinson-like 
syndromes . 

Methods employed : This project was originally designed to investigate 
influence of skin pigmentation on the prevalence of naturally occurring 
Parkinson's disease by use of a questionnaire which was to be mailed to a 
sample of American physicians. A pilot study, however, showed this approach 
to be impractical, and therefore, it was abandoned. In view of the possible 
relationship between drug-induced and naturally occurring parkinsonism, we 
then decide to study comparable white and Negro populations who were on 
treatment with phenothiazines . Thus far, we have examined approximately 75 
patients in each group, samples sufficient in number for a comparative 
analysis . 

Major findings : Our initial results suggest that there is no difference 
in the prevalence of phenothiazine-induced parkinsonism between the two 
populations surveyed. 



6lt 



Serial No. NDS (CF) - 68 E 1605 

Significance to biomedical research and the program of the Institute : 
To further define the relationship between melanogenesis in the skin and in 
the pigmented nuclei of the basal ganglia. 

Proposed course : Analysis of data is now in progress. 

Honors and Awards : None 

Publications: Brody, J. A.: Genetic considerations in Parkinson's disease. 
Presented at the Laurentian L-Dopa Conference, Montreal, 
November 1969. In: L-Dopa and Parkinsonism , Edited by Andre 
Barbeau and Fletcher McDowell, F.A, Davis Co., Phila., 1970, 
pp. 27-30. 



62t 



Serial No. NDS (CF) - 69 E 1774 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Neurologic signs and symptoms associated with malabsorption 

Previous Serial Number : Same 

Principal Investigators: Paul M. Hoffman, M.D. 

Jacob A, Brody, M.D. 
Anne H. Edgar 

Other Investigator: None 

Cooperating Units: Central Veterans Administration Authority, Washington, 
D.C. 
Veterans Administration Hospital, Atlanta, Georgia 
Veterans Administration Hospital, Long Beach, California 
Veterans Administration Hospital, Durham, North Carolina 
Veterans Administration Hospital, Los Angeles, California 

Man Years : 

Total: 1 1/2 
Professional: 1 1/2 
Other : 

Project Description: 

Objectives : To determine if both clinical and subclinical malabsorption 
is associated with a high incidence of neurologic signs and symptoms. 

Methods employed : The Cooperative Veterans Administration Retrospective 
Study of surgery for peptic ulcer served as the source for patients in this 
study. A review of abstracts from this study showed that a given hospital 
was more likely to have done surgical procedure for the majority of its cases 
then another. For this reason, patients with 3 different surgical procedures 
performed at 4 hospitals were chosen. A fourth group of patients who had 
simple closures of perforated ulcers were selected from all 4 hospitals . 
Patients who had a subtotal gastrectomy with a Billroth II reanastomosis 
were selected from the Durham Veterans Hospital, patients who had a hemi- 
gastrectomy and vagotomy were selected from the Atlanta Veterans Hospital, 
and patients who had a vagotomy and pyloroplasty were selected from the 
Long Beach and Wadsworth Veterans Hospital. In order to insure that this 
population would be in the most susceptible age group, patients between the 
ages of 30 and 80 who had had surgery between 1952 and 1957 were selected 
from abstracts and hospital records where available. Patients were not 

63* 



J 

I 



Serial No. NDS (CF) - 69 E 1774 

with neurologic complications, any systemic disease with known neurologic 
complications, or a history of neurologic disease prior to their ulcer 
surgery. Patients were also excluded from the study if a revision of their 
original ulcer surgery or subsequent surgery for a recurrent ulcer had been 
performed. No attempt was made in this study to analyze the causes of death 
or those who died since surgery. A mortality study of the 2,800 original 
cases which includes this sample has also been undertaken. All patients 
included in the study had letters sent to their last known address asking 
them to report on one of several days to the outpatient clinic of the hospital 
where their surgery was performed. A complete history with special attention 
on the neurologic and gastrointestinal systems as well as a complete neurologic 
examination was performed on all patients by one of us (PMH) and a random 
sample of all patients as well as all patients with abnormal findings were 
examined by a staff neurologist. 

Major findings : No cases of motor neuron disease have been found in the 
examined group. A large number of cases of peripheral neuropathy were 
identified in the vagotomy and hemi-gastrectomy group. Only two cases were 
seen in the vagotomy and pyloroplasty group. Through evaluation of all 
patients with unexplained neurologic disorders in the Atlanta group showed 
that malabsorption, poor nutrition, chronic alcohol intake, and weight loss 
was all more prevalent in this group than in those without neurologic findings. 
In reviewing 500 death certificates of the original study group we encountered 
2 patients with multiple sclerosis, 2 with Parkinson's disease and 1 with 
amyotrophic lateral sclerosis. 

Significance to biomedical research and the program of the Institute ; 
There have been many clinical reports of cases of malabsorption who have 
shown signs and symptoms of nervous system disease. There have also been 
scattered reports in literature of patients who are known to have motor 
neuron disease, who had a history of having had a Billroth II type of gastric 
surgery performed many years prior to the onset of their disease. There has 
never been, however, a matched, controlled population study of the association 
of these two abnormalities . If this association is valid then further study 
into the mechanism of absorption of essential nutrients and their incorporation 
into nervous tissue may be a meaningful approach to the study of chronic 
neurologic disease. 

Proposed course : This project will continue with emphasis on causes of 
death within this population. 

Honors and Awards : None 

Publications : None 



64^ 



I 



Serial No. NDS (CF) - 69 E 1775 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Comparison of MS among Irish and Italian immigrants in 
Bos ton 

Previous Serial Number: Same 

Principal Investigators: Roger Detels, M.D. 

Jacob A. Brody, M.D. 

Other Investigator: None 

Cooperating Unit : None 

Man Years : 

Total: 11/12 
Professional: 1/6 
Other: 3/4 

Project Description: 

Objectives : To determine if people migrating from an area of low 
multiple sclerosis incidence to an area of high multiple sclerosis incidence 
retain the low rate of the country of origin or acquire the high rate of 
the area to which they migrated. 

Methods employed : Boston, a city of 2 1/2 million, has a large Irish 
and Italian population, the majority of which migrated about the turn of 
the century. Thus, individuals in these groups dying of multiple sclerosis 
would be expected to have died between 1920 and 1940. During these years 
an estimated 5,000 deaths occurred each year among the Irish and the Italians 
in Boston. Since these groups would use the same medical facilities it is 
possible to do a comparative study of the rates of multiple sclerosis between 
them. Italy is an area of low multiple sclerosis incidence while Ireland and 
Boston have high rates. Fatality rates for Irish and Italian immigrants and 
first generation Americans of Italian and Irish background can be determined 
by a review of death certificates during this period. As a further control 
the ratio of multiple sclerosis to amyotrophic lateral sclerosis deaths can 
be compared since amyotrophic lateral sclerosis rates are considered to be 
independent of latitude. 

Death certificates for the period 1920 to 1940 from the city of Boston 
will be reviewed for all deaths due to multiple sclerosis and amyotrophic 
lateral sclerosis and for all deaths occuinring among Italian and Irish 

est 



Serial No. NDS (CF) - 69 E 1775 

immigrants and first generation Americans of these descents. Information 
will also be obtained for years of residence in the United States and in the 
Boston area. 

Major findings : Preliminary analysis revealed that the Boston population 
is apparently not suitable for this study. 

Significance to biomedical research and the program of the Institute : 
It is known that populations migrating from high incidence areas of multiple 
sclerosis to low incidence areas, retain the high rate of the country of 
origin suggesting that events early in youth (possible infectious) cause 
multiple sclerosis. This study will provide the corollary data about 
migration from low risk areas to high risk areas and about multiple sclerosis 
rates among first generation migrants. 

Proposed course : This study will be terminated. 

Honors and Awards : None 

Publications: None 



66t 



Serial No. NDS (CF) - 69 E 1776 

1. Collaborative & Field Research 

2. Epidenilology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1969 through June 30, 1970 

Project Title: Cerebrospinal fluid amines in drug-induced extrapyramidal 
parkinsonism-like disorders . 

Previous Serial Number: Same 

Principal Investigator: James A, Schnur, M.D. 

Other Investigator: Thomas N. Chase, M.D. 
NIMH 

Cooperating Units: Unit on Neurology, NIMH, Spring Grove State Hospital, 
Catonsville, Maryland 
Crownsville State Hospital, Crownsville, Maryland 

Man Years : 

Total: 2/3 
Professional: 1/2 
Other: 1/6 

Project Description: 

Objectives : To study central nervous system amine metabolism in 
patients with drug-induced extrapyramidal disorders . 

Methods employed : Spinal fluids of clinically defined cases of drug- 
induced extrapyramidal disorders are analyzed by conventional biochemical 
methods for amine metabolites. 

Major findings : Levels of HVA and 5-HIAA in lumbar CSF were measured 
in 20 female psychiatric patients. Those receiving long-term treatment with 
antipsychotic drugs who remained free of extrapyramidal dysfunction had 
substantially higher concentrations of these monoamine catabolites than 
patients not taking these drugs. Individuals who developed parkinsonian 
or dyskinetic signs while receiving antipsychotic agents appeared to have 
significantly lower concentrations of both HVA and 5-HIAA than patients not 
manifesting these disorders despite similar drug exposure. 

Significance to biomedical research and the program of the Institute : 
The foregoing observations support the hypothesis that the compensatory 
acceleration of cerebral monoamine metabolism induced by antipsychotic drugs 
may be impaired in patients who develop extrapyramidal dysfunction and this 
is the mechanism of acquisition of this side affect. 



I 



67 t 



Serial No. NDS (CF) - 69 E 1776 

Propos ed cours e : This phase of the study is completed and project 
terminated . 

Honors and Awards: None 

Publications: Chase, T.N. , Schnur, J. A. and Gordon, E.K.: Cerebrospinal 
fluid monoamine catabolites in drug-induced extrapyramidal 
disorders. Neuropharmacology , 9:265-268, 1970. 



68* 



Serial No. NDS (CF) - 69 E 1777 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Beth es da, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: ALS among non-Chamorros after residence on Guam 

Previous Serial Number: Same 

Principal Investigator: Jacob A. Brody, M.D. 

Other Investigator: Estelle Kornhauser, R.N. 

Cooperating Unit: Bureau of Data Processing and Accounts, Social Security 
Administration, DHEW 

Man Years : 

Total: 1/2 
Professional: 5/12 
Others : 1/2 

Project Description: 

Objectives : This project was developed to determine if prolonged 
exposure (over one year) to the environment of Guam increases the likelihood 
of the development of ALS among statesiders. 

Methods employed : Through various workers in the Department of Defense 
we were put in contact with several construction companies which had 
maintained large staffs of statesiders on the island of Guam after World 
War II. These companies were asked to supply us with the names, birthdates, 
and social security numbers of all personnel employed on Guam and from these 
lists we selected only those workers who had spent more than one year on 
Guam.. This was a group of approximately 12,000 individuals. The Social 
Security Administration searched its records to determine which of these 
workers had died and where they had died. We are contacting the individual 
states to obtain the death certificates of the deceased workers and 
determine the cause of their death. 

Major findings : Of the approximately 12,000 cards submitted to the 
Social Security Administration, 450 were found to contain incorrect 
information making follow-up Impossible. 7,730 were considered as 
representing individuals still alive, and the remainder (4,000) were 
identified as to place of death. The names were submitted to the individual 
states and death certificates requested. About 80% of the requested 
certificates have been returned to date. Preliminary analysis indicates that 
there was no excess of ALS in this group. 



69t 



Serial No. NDS (CF) - 69 E 1777 

Significance to biomedical research and the program of the Institute ; 
Although results on this study are far from complete, the initial trend is 
that the rate of ALS among statesiders who have spent considerable time on 
Guam remains the same as that for statesiders who have never spent time in 
that environment. If this trend is borne out by subsequent findings, it 
would suggest that the environmental factors on Guam are less likely to be 
responsible for high rate of ALS among its native population. 

Proposed course : The study will be completed by submitting to Social 
Security approximately 600 punch cards in order to search for death 
certificates we were unable to locate through the individual states. These 
cards will then be coded and searched for possible ALS deaths. 

Honors and Awards : None 

Publications : None 



70t 



Serial No. NDS (CF) - 69 E 1778 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Familial cortical cerebellar degeneration 

Previous Serial Number: Same 

Principal Investigators: Paul M. Hoffman, M.D. 

William H. Stuart, M.D. 
Kenneth Earle, M.D. 

Other Investigators: Joyce M. Cannon, R.N. 
Anne Harlan Edgar 

Cooperating Unit: Armed Forces Institute of Pathology 

Man Years : 

Total: 1/2 

Professional: 1/3 , 

Other : 1/6 

Project Description: 

Objectives : To determine the genetic pattern and the incidence of 
cortical cerebellar degeneration in a family in the northern Georgia section 
of the Blue Ridge Mountains . 

Methods employed : A complete family history and a family tree were 
obtained from the proband and his family who are now living in Hiawasee, 
Georgia. Affected members were examined as well as those who were at risk 
but who have not shown the signs and symptoms of the disease. Hospital 
records and autopsy material, when available, was collected. An autopsy 
was performed on the index case. 

Major findings : The proband is an 80-year-old white male who demonstrated 
abnormal finger to nose and heel to shin tests as well as inability to maintain 
his balance, and persistent truncal titubation. He also exhibited scanning- 
type of cerebellar speech for the last two years. There was no evidence of 
long-track involvement or of abnormalities in position or vibratory sensation. 
The reflexes were described as normal. Further study of other affected 
members showed that the symptoms of ataxia, scanning spastic speech, and mild 
upper extremity involvement were constant throughout the family. Six affected 
members in two generations were examined. The neuropathology consisted of 
atrophy of the superior and anterior cerebellar cortex with secondary olivary 
degeneration. The disease was transmitted as an autosomal dominant. 



L 



7lt 



Serial No. NDS (CF) - 69 E 1778 

Significance to biomedical research and the program of the Institute : 
The hereditary cerebellar and spinocerebellar degeneration have been described 
as unique syndromes. Presently, however, it has been observed that symptoms 
within a family have been very variable. This family has similar symptoms in 
all affected members. Interest has now been shown in treating various types 
of spinocerebellar and cerebellar disorders with L-Dopa . The well-defined 
hereditary syndromes offer the greatest hope of finding an enzyme deficiency 
that might result in disordered biogenic amine metabolism. 

Proposed course : A report has been accepted for publication and this 
study was terminated. 

Honors and Awards : None 

Publications: Hoffman, P.M., Stuart, W.H., Earle, K.W.,and Brody, J.A. : 
Hereditary cerebello-olivary degeneration of late onset. 
Neurology , 20:400, April 1970. (Presented at the American 
Academy of Neurology, May 1, 1970.) 



72^ 



Serial No. NDS (CF) - 69 E 1779 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Ihe epidemiology of motor neuron disease in the United 
States 

Previous Serial Number: Same 

Principal Investigators: Paul M. Hoffman, M.D. 

Jacob A. Brody, M.D. 

Other Investigator: Joyce M. Cannon, R.N. 

Cooperating Units: Bureau of Disability Insurance, Social Security 
Adminis tration 
North Carolina State Department of Public Health 
University of North Carolina School of Medicine 
Duke University School of Medicine 
Charlotte Memorial Hospital 

Man Years : 

Total: 11/12 
Professional: 10/12 
Other: 1/12 

Project Description: 

Obj ec tives : To determine if the diagnoses on death certificates are 
an adequate reflection of the incidence of motor neuron disease in the 
United States. 

Methods employed : Hospital records were reviewed on those patients 
who had a diagnosis of motor neuron disease in years 1958 through 1962 at 
the three North Carolina Hospitals. Death certificates were then obtained 
on those patients who had died in the state of North Carolina. Those 
patients who are still living were followed-up through the County Public 
Health Departments within the state of North Carolina and information was 
obtained on these patients. 

Major findings : Of the death certificates obtained, the diagnosis of 
motor neuron disease appeared on 72 percent. Of these 70% were certified 
at death as having ALS . MS was certified in 10% of these patients, 
suggesting a possible systematic error of overdiagnosis of MS at death 
and underreporting of ALS. 



73t 



Serial No. NDS (CF) - 69 E 1779 

Significance to biomedical research and the program of the Institute : 
Most of the estimates of the prevalence of motor neuron disease are based 
on mortality reporting. Our study cast doubt as to the validity of this 
technique. 

Proposed course : A report has been prepared and accepted for 
publication. The project was terminated. 

Honors and Awards : None 

Publications: Hoffman, P.M. and Brody, J. A.: The accuracy of mortality 

statistics in clinically proven amyotrophic lateral sclerosis. 
Trans. Amer . Neurol. Ass . 95:261-263, 1970. 

Hoffman, P.M. and Brody, J. A.: The reliability of mortality 
statistics for amyotrophic lateral sclerosis. J. Chron. Pis . 
23, 1971. 



74t 



Serial No. ITOS (CF) 69 E I78O 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, I97I 

Project Title: Familial bilateral acoustic neuroma 

Previous Serial Number: Same 

Principal Investigator: Roswell Eldridge, M. D. 



Other Investigators: 



Cooperating Units: 



Dean F. Young, M.D. 

New York, New York 
Henry Hood, M.D. 

Danville , Pennsylvania 
W. J. Gardner, M.D. 

Cleveland, Ohio 
George T. Nager, M.D. 

Johns Hopkins Hospital 
Frank H. DeLand, M.D. 

Johns Hopkins Hospital 

Department of Otolaryngology, 
Department of Radiology, 
Department of Neurosurgery, 
Geisinger Medical Center 

Danville , Pennsylvania 
Johns Hopkins Hospital 
Baltimore , Maryland 



Man Years : 

Total : 2 

Professional: 1 
Other : 1 

Project Description: 

Objectives : To perform genetic, clinical and physiologic studies of a 
large family with hereditary bilateral acoustic neuroma. 

To clarify the relationship of this trait to other disorders with acoustic 
neuromas such as neurofibromatosis. 

Methods employed : Field studies were conducted in evaluating family 
members. On those seen personally, physical examinations were performed, 
stressing neurological and skin examinations. In addition, audiometric 
examinations including air and bone conduction and caloric examinations were 
conducted in the field. 



75t 



Serial No. NDG (CF) 69 E I78O 

Genealogic information was obtained from family members, family records, 
D.A.R. records, state and military records, census recordings. Medical 
history was obtained from family members, hospital and physician records, 
occasionally from school records or military records. 

On select patients extensive outpatient studies have included audiometric 
and vestibular testing, complete EIW and neurologic examinations, skull 
x-rays and brain scans using radioactive techniques. These were undertaken 
in cooperation with the Departments of Neurology, Radiology and Otolaryn- 
gology of Johns Hopkins Hospital. 

Major findings : The study of this, the largest kindred with hereditary 
neoplasm yet reported, has generated information which has been particularly 
useful to neurosurgeons and otolaryngologists since it casts considerably 
different light on prognosis and genetic risks to those individuals who 
develop such a disease at a yoiing age. 

Significance to biomedical research and the program of the Institute : 
The mode of inheritance to this trait has been confirmed and the place of 
this syndrome among those associated with neural sheath proliferation is 
clearer. Of primary importance is the establishment of appropriate diag- 
nostic techniques for early cases and treatment of these cases. 

Proposed course : Affected individuals are being followed to evaluate 
various forms of treatment. Relatives not known to be affected but at 
risk are being examined periodically to evaluate various diagnostic techniques, 

The initial phase of this project has been completed and resulted in the 
publications listed below. 

Honors and Awards : None 

Publications : 

Young, D.P., McNew, J. and Eldridge, R. : Hereditary Acoustic Neuroma - 
Clinical and Genetic Aspects. In Transactions of the American Neurological 
Association . 9i|, I969. 

Young, D.F., Eldridge, R., Na^er, G.T., DeLand, F.H., and McNew, J.: 
Hereditary bilateral acoustic neuroma (central neurofibromatosis). Pro- 
ceedings of the Second Conference on The Clinical Delineating of Birth 
Defects, May I969. The National Foundation -March of Dimes (in Press) 

Young, D., Eldridge, R., and Gardner, ¥. J. : Bilateral Acoustic Neuroma in 
a Large Kindred. JAMA. 21^1-, October 1970. 



76t 



Serial No. NDS (CF) - 69 E 1781 

1. Collaborative Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Twin Studies in Parkinson's Disease 

Previous Serial Number: Same 

Principal Investigators: Roswell Eldridge, M.D. 

Zdenek Hrubeck, M.D. 

National Academy of Sciences 

Other Investigator: Kathy O'Meara 

Cooperating Unit: The National Academy of Sciences - National Research 
Council Twins Registry, Washington, D. C. 

U. S. Veterans Administration 
Washington, D. C. 

Man Years: 

Total: 1/4 

Professional: 1/8 
Other: 1/8 

Project Description: 

Objectives : Parkinsonism is not a single entity but rather a symptom 
complex which may be idiopathic, or may be due to cerebral arteriosclerosis 
or may follow encephalitis. Genetic factors have been considered important 
by a number of authors but it is not yet established how important such 
factors are in any one form of Parkinsonism or if there is a form which has 
a simple genetic basis independent of acquired neurologic disease. The aim 
of the present proposal would be to use the twin method to evaluate genetic 
factors in various forms of Parkinsonism and/or to distinguish genetically 
determined Parkinsonism feom other forms. 

Methods employed : Survey of the VA twin registry in 1968 has revealed 
eight cases of Parkinsonism in one of a Veteran twin pair. Each of the 
pair were apparently discordant for Parkinsonism. Two twin pairs were 
monozygotic, five were dizygotic and in one the zygosity was unknown. 
We would contact each of these individuals and his co-twin, and others with 



77t 



Serial No. NDS (CF) - 69 E 1781 

che diagnosis who may be ascertained through updating of the registry, and 
arrange for an appointment with the individual in his home at a time when 
available relatives could also be present. During the family interview 
a pertinent medical history would be obtained and physician examination 
would be performed. Permission for review of hospital records would be 
secured and arrangements might be made for additional neurologic studies. 
To define zygosity, photographs would be taken of the twins, blood would be 
drawn for genotyping and dermatoglyphics might be recorded. (Personal 
examination of both twins and available relatives is important in order that 
mild cases not be missed). 

Major Findings : Interest continues in this project although there has 
been no additional ascertainment of cases since the last report. The 
National Science Foundation have not yet agree to our contacting patients on 
their registry but we expect twin pairs will be available to us in the 
future. 

Significance to biomedical research and the program of the Institute : 
In a chronic condition with late onset it is often difficult to determine 
the role of genetic factors in causation. The twin method provides a 
relatively simple method involving small numbers to answer this question. 

The nosology of Parkinson's disease is especially important now that the 
drug L-Dopa has been shown to help some with Parkinsonism. Is this drug 
most helpful in a specific form of the disease? Is it effective in 
hereditary Parkinsonism? 

Proposed course : See "Methods employed". 

Honors and Awards: None 

Publications: None 



78t 



I 



Serial No. KDS (CF) 69 E I782 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Twin Studies in Torticollis 

Previous Serial Wumher: Same 

Principal Investigators: Roswell Eldridge, M. D. 

Zdenek Hruhec, M. D. 

National Academy of Sciences 

Other Investigators: Kathy O'Meara 

Cooperating Unit: The National Academy of Sciences-National Research 
Council Twins' Registry, Washington, D. C. 

U. S. Veterans Administration 
Washington, D. C. 

Man Years : 

Total: l/U 
Professional: I/8 
Other : 1/8 

Project Description: 

Objectives : Torticollis may be broadly divided into infantile and post- 
infantile fonns. The former may be congenital or appear several weeks after 
birth but in either the cause appears due to events preceding birth. Post- 
infantile torticollis consists of a heterogeneous group of disorders which 
have been ascribed to a number of causes including trauma or inflammation of 
the cervical spine, myositis of nuchal musculature, functional illness, and 
disease of the peripheral or central nervous system. In addition torticollis 
on a hereditary basis, either as an isolated symptom or in association with 
other movement disorders such as torsion dystonia, has been the subject of 
numerous reports. The aim of the proposed study is to weigh the genetic 
factors in the post-infantile forms of torticollis and, if possible, to 
distinguish between discrete hereditary types. 

Methods employed : In I968 a review of the Veterans Administration twin 
registry disclosed 11 individuals with a diagnosis of torticollis. Each of 
the 11 pair were said to be discordant. Five were monozygotic, two were 
dizygotic and in the four the zygosity could not be established. We would 
contact each of these individuals and his co-twin, arrange for an appoint- 
ment with the individual in his home at a time when available relatives could 

79t 



Serial No, HDS (CF) 69 E I782 

also be present. (We would hope also to ascertain new cases by assisting 
in the up-dating of the twin registry). During the family interview the 
pertinent medical history would be obtained for all relatives and physical 
examination performed on those present. Pei-mission for review of hospital 
records would be secured and arrangements might be made for additional neurol- 
ogic studies. To establish zygosity^ photographs would be taken of the twins, 
blood drawn for genotyping, and dermatoglyphics might be recorded. 

Major findings : Interest continues in this project although there has 
been made no additional ascertainment of cases since the last report. The 
National Science Foundation has not yet agreed to our contacting patients 
on their registry but we expect twin pairs will be available to us in the 
future . 

Significance to biomedical research and the program of the Institute : 
Torticollis may be hereditary or acquired but under each of these headings 
there appear to be a number of discrete entities. The twin method presents 
a relatively simple means to distinguish genetically determined fonns. 
Concentration on torticollis which is simply inherited is worthwhile since 
such disorders should have a discrete biochemical basis which might be 
revealed by study with the neurochemical techniques now available. 

Proposed course : See "Methods Employed". 

Honors and Awards : None 

Publications: None 



80t 



Serial No. NDS (CF) - 70 E 1832 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Measurement of immune response after antigen stimulation 

Previous Serial Number: None 

Principal Investigators: George Nemo, Ph.D. 

Lon R. White, M.D. 

Other Investigators: Harrie Anne Sutton, Biologist 
Minnie Toure, Biologist 
Gary Cooper, Biologist 

Cooperating Unit: Unit on Congenital and Chronic Viral Infections, DB, NICHD 

Man Years : 



Total: 


5/6 


Professional: 


1/3 


Other: 


1/2 



Project Description: 

Ob j ec tives : To investigate the changes in mltogenic, cytotoxic and 
anti-viral capacities of lymphocytes in relation to the acquisition of 
specific immunity. 

Methods employed : Studies will involve both humans (immunized to 
vaccinia virus as part of routine health care practices) and experimental 
animals (mice, rats, guinea pigs). The antigens with which the animals 
will be inoculated include mycobacteria protein, vaccinia, sindbis, reovirus 
I, and the minute virus of mice. Lymphocyte transformation studies will 
be done with spleen, blood and lymph node cells. The effects of immune 
lymphocytes on the course of virus infection in vitro will be studies using 
monolayer tissue culture cells isogenic with the lymphocyte donor. Cyto- 
toxicity will be assayed visually and by Cr^l release. Virvises will be 
titered using standard techniques. Macrophage-migration inhibition will 
also be employed as an indicator of cellular immunity. 

Major findings : None as yet . 

Significance to biomedical research and the program of the Institute : 
It is widely thought that "autoaggressive" lymphocytes are involved in the 
etiology of chronic demyelinating diseases. It is further believed that 
lymphocytes may be important in recovery from viral infection. Experimental 
animal systems have suggested the possibility that certain chronic viral 

81t 



Serial No. NDS (CF) - 70 E 1832 

infections may owe their chronicity to impaired lymphocyte competence and 
that as this competence begins to be restored, host cells which bear viral 
antigen determinants on their surfaces may be injured, giving the appearance 
of an autoimmune disease. Such theories must be based and examined on the 
results of investigations such as those described. 

Proposed course : To be continued. 

Honors and Awards : None 

Publications : None 



82± 



Serial No. NDS (CF) - 70 E 1833 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Immune mechanisms in chronic and congenital viral infections 

Previous Serial Number: Same 

Principle Investigator: Lon R. White, M.D., DB, NICHD 

Other Investigators: George Nemo, Ph.D. 

Jacob A. Brody, M.D. 

Harrie Anne Sutton, biologist, DB, NICHD 

(HD DB - 5) 



Cooperating Units : 


None 


Man Years : 




Total: 


0.6 


Professional: 


0.3 


Other: 


0.3 



Project Description: 

Objectives : 1. To establish one or more models of chronic and/or 
congenital viral infection in experimental animals. 2. To define the 
development of cellular and humoral immunity to the same and to heterologous 
antigens . 

Methods employed : Neonatal and pregnant mice are injected with either 
a reovirus type 1 or the minute virus of mice (MVM) . The persistence of 
virus in animals is determined by isolation and flourescent and immuno- 
flourescent techniques. Serum levels of hemagglutination inhibition 
antibodies to both agents are followed. Lymphocyte transformation and 
cytotoxic activity is studied in spleen cell cultures in the presence of 
phytohemagglutinin on specific antigens . Any tumors which may develop are 
studied by light and electron microscopy. Extracts of MVM-containing tumor 
cells will be injected into newborn mice in addition to being studied in 
tissue culture. 

f Major findings ; Attempts to infect embryonic animals by maternal infection 

in the first week of pregnancy have failed, presumably because infected embryos 
are reabsorbed. Late gestation maternal infection of 8 animals has resulted 
in the live birth of 53 animals, 39 of which survived to adult life. All 
progeny have appeared to be normal, most having been observed for 1 year. Two 
such animals have died of solid tumors; one of these was lost to study and the 
other was found to have a tumor in the area of the right uterine horn with 



L 



^3t 



Serial No. NDS (CF) - 70 E 1833 

hepatic metastases. Extracts of both the tumor and liver yielded a hemagglu- 
tinating material presumed to be MVM antigen, suggesting that the animal was 
either reinfected late in life or had been chronically infected since fetal 
life. Cell-free materials from these tissues are now being studied both 
in vivo and in vitro in an attempt to define the possible role of MVM, either 
directly or indirectly, in the etiology of the tumor. 

Significance to biomedical research and the program of the Institute : 
Infection of the human embryo or fetus with rubella or cytomegalovirus is 
associated not only with developmental abnormalities and mental retardation, 
but also with infection persisting for months or years after, despite the 
presence of neutralizing antibody in the serum. There are several examples 
of related phenomena in experimental animals infected during prenatal or 
neonatal life. Previous studies have suggested that an impairment of the 
immune function of lymphocytes may be associated with such persistent 
infection. These phenomena provide insight into the question of how viral 
infection is normally terminated, and represent a challenge to older ideas 
on the role of specific cellular immunity in the natural history of virus 
infections. The investigation described represents a direct experimental 
approach to elucidating the cause and course of chronic infection following 
initial exposure to the agent during immunologic immaturity. The results of 
this investigation will be of immediate relevance to our understanding of 
other types of illness known or suspected to be associated with chronic viral 
infection. In addition, they may suggest new approaches to research in the 
role of congenital viral infection as a cause of diseases of unknown etiology 
such as prematurity, idiopathic growth failure, malignancy, mental retardation, 
developmental malformation, and autoimmune diseases. 

Proposed course : To be confined at present level of effort. 

Honors and Awards : None 

Publications : None 



at 



I 



Serial No, NDS (CF) - 70 E 183A 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Immune mechanisms in experimental allergic encephalomyelitis 
(EAE) 

Previous Serial Number: Same 

Principal Investigator: Lon R. White, M.D. 

Other Investigators: George Nemo, Ph.D. 

Marian Kies , Ph.D., Section on Myelin Chemistry, NIMH 

Cooperating Unit: Section on Myelin Chemistry, Laboratory of Cerebral 
Metabolism, DBBR, NIMH (HD DB - 6) 

Man Years : 

Total: 0.20 
Professional: 0.15 
Otiier: 0.05 

Project Description: 

Objectives : To demonstrate in vitro "activation" of lymphocytes from 
guinea pigs with EAE by a myelin derived basic protein. 

Methods employed ; Techniques of lymphocyte cytotoxicity and blastogenic 
transformation have been employed. 

Major findings : No cytotoxicity has been demonstrated. Transformation 
has been observed in some, but not all, experiments. These results seem to 
relate to the following variables: a) the "batch" of basic protein used, 

b) allotypic identity or dissimilarity between lymphocytes and target cells, 

c) strain of guinea pig utilized, d) interval between inoculation on animals 
and testing of lymph node lymphocytes. 

Significance to biomedical research and the program of the Institute : 
EAE is generally held to be pathogenically similar to certain human demyeli- 
nating diseases, particularly the post-infections encephalitides and multiple 
sclerosis, and may involve lymphocyte-mediated autoimmunity. The in vitro 
demonstration of a basic protein "activatable" lymphocyte would be of great 
value to a more complete londers tanding of the experimental disease and would 
suggest new approach to the human diseases. The knowledge gained relating 
to lymphocyte function would be of immediate relevance to problems of 



85t 



Serial No. NDS (CF) - 70 E 1834 

autoimmunity and chronic infection in certain diseases definitely or possibly 
due to viral infection during prenatal and neonatal life. 

Proposed course : No further experiments planned at this time. 

Honors and Awards : None 

Publications : None 



86 t 



Serial No. NDS (CF) - 70 E 1835 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



Project Title: 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Epidemiologic and immunologic study of families of subacute 
sclerosing panencephalitis patients and families of matched 
controls 



Previous Serial Number : Same 



Principal Investigators 



Roger Detels, M.D. 
John M. Stanhope, M.D, 
Jacob A. Brody, M.D. 
Jane McNew, R.N. 
Anne H. Edgar 



th er Inves t igato rs : 



John Sever, M.D. 

Head, Section on Infectious Diseases, Perinatal 

Research Branch, NINDS 
Richard L. Parker, D.V.M. 

Assistant Chief, Office of Veterinary Public Health, 

Center for Diseases Control, Atlanta 



Cooperating Units: Section on Infectious Diseases, Perinatal Research 
Branch, NINDS 
Veterinary Branch, National Communicable Disease Center, 
Atlanta 

Man Years : 



Total: 

Professional: 
Other : 



1 2/3 

1 1/2 

1/6 



Project Description: 

Objectives : To determine, through the use of a matched control study, 
factors related to the etiology of subacute sclerosing panencephalitis. 

Methods employed : Cases of SSPE were selected from the areas around 
St. Louis, Missouri, North Carolina, and California in order to evaluate 
the occurrence in distinctly different environments. 49 families with 
matched controls and an additional 8 families for whom a suitable control 
could not be obtained have been interviewed. Controls were selected on the 
basis of close friendship and similar background to the proband. A detailed 
history of events possibly related to the etiology of SSPE including episodes 
of measles, unusual illnesses, exposure to sick animals, etc. was asked of 



87 1 



Serial No, NDS (CF) - 70 E 1835 

probands, controls and their families. A 20cc sample of blood was drawn 
from each individual in the proband and control families . 

Major findings : Preliminary analysis of the 40 initial cases indicates 
that the median age of measles among probands was 18 1/2 months, 2 years 
younger than the median age of measles among controls . One-quarter of the 
probands, but none of the controls, had measles under one year of age and 
two of the probands were diagnosed by their physicians as having had measles 
twice. One-quarter of the probands had no history of measles. All of the 
patients came from middle or lower income families, and only one of the 
probands came from a truly urban environment. History of warts, herpes 
virus infection, allergies and neurologic disease were similar in proband 
and control families. All but 3 of the 40 probands had pets and 2 of these 
had had intimate exposure with animals. A history of exposure to sick 
animals was more than twice as frequent among probands (68%) as among 
controls . 

Significance to biomedical research and the program of the Institute : 
We have shown that the measles infection in cases was unusual and occurred 
frequently during a time when passive immunity was present. Further SSFE 
occurs in an unusual epidemiologic pattern with rates among males far 
exceeding those of females and rates in non-urban areas vastly exceeding 
those in urban areas much as would be expected with a zoonotic disease. 
Thus we have postulated that SSPE is caused by an unusual measles infection 
and a subsequent triggering event which is probably a zoonotic virus. 

Proposed course : Sera from 57 proband families and 49 control families 
will be analyzed for presence and titers of measles antibodies. Additional 
testing will be made as indicated by the results of the analysis of the 
questionnaires. Final analysis of the histories obtained from the proband 
and control families will be completed. Intensive search for the possible 
"zoonotic trigger" will be carried out. 

Honors and Awards : None 

Publications: Brody, J. A. and Detels, R.: Subacute sclerosing 
panencephalitis: A zoonosis following aberrant 
measles. Lancet, 11:500-501, 1970. 

Brody, J. A., Detels, R. and McNew, J.: Evidence that 
subacute sclerosing panencephalitis is caused by an 
aberrant measles infection followed by a zoonosis. 
Presented at Annual Meeting of the American Academy of 
Neurology, April 1971. To be published. 



Serial No. NDS (CF) - 70 E 1836 

1. Collaborative & Field Research 

2 . Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Epidemiologic factors in Jakob-Creutzfeldt 's disease 

Previous Serial Number: Same 

Principal Investigators: Jacob A. Brody, M.D, 

A. Roger Bobowick, M.D. 
Raymond P. Roos , M.D. 
C&FR, NINDS 

Other Investigator: Marjorie Matthews, R.N. 

Cooperating Unit: Laboratory of Slow, Latent and Temperate Virus 
Infections, C&FR, NINDS 

Man Years : 

Total: 1/4 
Professional: 1/6 
Other: 1/12 

Project Description: 

Objectives : Jakob-Creutzfeldt 's disease is one of the spongiform 
encephalopathies which has been transmitted to chimpanzees. Preliminary 
investigations suggest that two virus particles occur in the brains of 
patients. There are many forms of Jakob Creutzfeldt 's disease which are 
distinct clinically and to some degree neuropathologically . Nothing is 
known of the epidemiology of this syndrome. We wish to determine if there 
are common factors among patients who develop Jakob-Creutzfeldt 's disease 
and if there are specific factors related to distinct forms of this syndrome. 

Methods employed : Letters were sent to neuropathologists soliciting 
pathologically proven cases. With the cooperation of the referring physicians 
the families of the patients are contacted and asked to participate in the 
epidemiologic interview. At the time of the interview session a detailed 
questionnaire is completed on the relative who is the interviewee, on the 
patient, and on an age and sex matched friend of the patient who serves as a 
control. Blood samples are also taken for future seriologic studies. 

Ma j or findings : From the series of approximately 50 patients, we have 
interviewed, thus far, 12 families. The methods outlined above have proved 
workable. The families have been extraordinarily cooperative and in fact 
grateful. 



Serial No. NDS (CF) - 70 E 1836 

Significance to biomedical research and the program of the Institute : 
This neurologic disease is caused by a virus or a combination of viruses. 
Transmissible diseases must be studied epidemiologically if predisposing 
factors are to be elicited. The documentation of a predisposing factor to 
a virus-induced degenerative disease would be a major contribution to the 
understanding of the disease, its treatment and prevention. 

Proposed course : This series will be developed to include between 
30 and 50 cases and analysis of the data will begin. 

Honors and Awards : None 

Publications : None 



90 1 



Serial No. NDS (CF) - 70 E 1837 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Multiple sclerosis among native-born and migrants to 
California and Washington States 

Previous Serial Number: Same 

Principal Investigators: Roger Detels, M.D. 

Jacob A. Brody, M.D. 

Other Investigators: None 

Cooperating Units : None 

Man Years : 

Total: 2/3 
Professional: 1/3 
Other: 1/3 

Project Description: 

Objectives : To determine whether rates of multiple sclerosis among 
native-born in Washington and California are related to latitude, and to 
determine if migrants from high and low risk areas within and without the 
United States retain the rate from place of origin or acquire the rate of 
the place to which they migrated. 

Methods employed : Washington and California have a combined population 
of approximately 30 million of whom about 20 million are migrants. Decedents 
with multiple sclerosis and amyotrophic lateral sclerosis between 1954 and 
1964 can be analyzed by birthplace, race, sex, age and duration of life in 
California. ALS patients can be used as a partial control, since the ALS 
rate is thought to be stable regardless of latitude. Death rates can also 
be ascertained for both MS and ALS using 1960 census data as a baseline. 

Major findings : Death rates from MS were studied among 7 million 
native-born and 8 million migrant white, Japanese and Chinese in California 
and Washington. The death rate among native-born Washingtonians was higher 
than among native-born Calif ornians . Low death rates were found among 
American-born and foreign-born Japanese and Chinese in both states suggesting 
that racial factors may influence susceptibility to MS. Migrants to both 
states from areas reported to have low rates of MS had low rates suggesting 
that they had acquired protection before migration. Migrants to Washington 
from areas reported to have high rates had higher rates than similar groups 

91 1 



Serial No. NDS (CF) - 70 E 1837 

migrating to California. These findings suggest that causative factors may 
still be operative in the third and fourth decades of life in high risk 
areas and that a protective factor may also be involved in the etiology 
of MS. 

Significance to biomedical research and the program of the Institute : 
If these findings are substantiated they would lead to major insights in MS. 
This is the first large study of people moving from low risk to high risk 
areas and we have new evidence of a protective factor in MS. Data on possible 
changes in risk among those migrating from high risk areas suggests that 
causative factors persist after adolescence. The concept of separate 
protective and precipitating factors in MS may provide the clue in 
understanding the genesis of the disease. 

Proposed course : This study will be expanded through prevalence 
studies focusing on various migrant groups. 

Honors and Awards : None 

Publications: Detels , R., Brody, J. A. and Edgar, A.H.: Multiple 

sclerosis among American, Japanese and Chinese migrants 
to California and Washington. New Eng . J . Med . 
In press. 



92 1 



Serial No. NDS (CF) - 70 E 1838 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Study of the incidence of Parkinson's disease among offspring 
of Parkinsonians 

Previous Serial Number: Same 

Principal Investigators: Roger Detels, M.D. 

Jacob A. Brody, M.D. 

Other Investigators : None 

Cooperating Units: Church of Jesus Christ of Latter-day Saints, 

Salt Lake City, Utah 

The Genealogic Society, 

Salt Lake City, Utah 

Man Years : 

Total : 

Professional: 

Other: 

Project Description: 

Objectives : To determine if genetic factors play a part in the etiology 
of Parkinson's disease by examining the incidence of disease among offspring 
of probands . 

Methods employed : Fifty members of the Church of Jesus Christ of 
Latter-day Saints who died with the diagnosis of Parkinson's disease will 
be selected for review of death certificates in Utah between 1930 and 1940 
and an additional 50 members who had a hospital diagnosis of Parkinson's 
disease between 1930 and 1940 selected from a review of hospital records from 
the major hospitals in the Salt Lake City area. A matched index control will 
be selected for each proband by taking the next death certificate or hospital 
record of an individual of the same sex whose age was within 5 years of the 
proband at the time of death or illness. Offspring of probands and index 
controls will be identified, a review of hospital records, of obituary 
notices , and a review of the records of the Genealogic Society and of the 
Church files of the Church of Jesus Christ of Latter-day Saints. Addresses 
of offspring will be obtained from the Church and questionnaires will be sent 
out to the offspring. Cases of Parkinson's disease will be confirmed when 
possible by physical examination or in the case of death through available 
medical records . 



93t 



Serial No. NDS (CF) - 70 E 1838 
Major findings : None 

Significance to biomedical research and the program of the Institute ; 
In a chronic condition with late onset, such as Parkinson's disease, it is 
often difficult to determine the role of genetic factors. Members of the 
Church of Jesus Christ of Latter-day Saints maintain accurate, up-to-date 
genealogies . Thus , offspring of Parkinsonians can be located and the 
incidence of Parkinson's disease among them determined, providing a relatively- 
simple method for determining the genetic factor in the etiology of 
Parkinson's disease. 

Proposed course : See "Methods employed . " 

Honors and Awards : None 

Publications : None 



9h^ 



Serial No. NDS (CF) - 70 E 1839 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



Project Title: 



PHS-NIH 

Individiial Project Report 

July 1, 1970 through June 30, 1971 

Studies on the relationship of chromosomal abnormalities and 
certain viral infections in mice. (Alternate title: Effects 
of viruses on mitosis and meiosis in the mouse.) 



Previous Serial Number: Same 

Principal Investigators: Beverly J. White, M.D,, LEP, NIAMD 

Lon R. White, M.D., DB, NICHD 

Other Investigators: George Nemo, Ph.D. 

Jacob A. Brody, M.D. 

Cooperating Units: Laboratory of Experimental Pathology, NIAMD (LEP/NIAMD-17) 
Developmental Biology Branch (Unit on Chronic and 
Congenital Viral Infections), NICHD 

Man Years : 

Total: 1.5 

Professional: 0.9 
Other: 0.6 

Project Description: 

Objectives : a) . To localize within dividing mouse spleen cells in vitro , 
particularly in relation to chromosomes, the site of residence of the 
infecting viral genome (of the minute virus of mice - MVM) and the MVM 
template DNA. b) . To search for evidence of injury by virus to chromosomes 
of spermatogonia and somatic cells, c). To determine whether perinatal 
infection results in infection of the germ cells and, if so, to determine 
the duration of infection as well as the chromosomal and reproductive 
consequences of such infection. 

Methods employed : The viruses under study are two strains of MVM and 
two strains of reovirus type 1. The animal used is the Cumberland View 
Farms C57BL/6 mouse. The viruses are "grown" and quantitated in tissue 
culture. Intracellular MVM genome (infectious or template DNA) is localized 
by autoradiography using tritium "tagged" MVM DNA. Chromosome studies are 
carried out on short term spleen cell cultures as well as on cytologic 
preparations made directly from the spleen, testes, and bone marrow of 
infected and control mice. Viral antigen is localized using immuno fluorescent 
and immunoh is to chemical methods. 



95t 



Serial No. NDS (CF) - 70 E 1839 

Major findings : a). A "fate" study in mouse spleen cells using tritium- 
labeled virus preparations showed no difference in autoradiographic grain 
localization between control and virus infected cultures. These were carried 
out with "tagged" viruses of relatively low infectivity and specific activity 
(radioactivity), b). Meiotic preparations from the testes of adult mice were 
studied 4 days, 1 week, 3, 6, and 12 weeks following MVM infection. The 
only notable observation was of a relative decrease in number of certain cell 
types, suggesting an effect on the course of sp ermio genes is . 

Major improvements in methods involved with the propagation and 
quantitation of MVM have been accomplished. These have been used to 
produce viral "reagents" to be used in additional experiments. 

Significance to biomedical research and the program of the Institute : 
The role of viruses in the etiologies of chromosomal and developmental 
diseases is widely discussed, but negligibly investigated. The interaction 
of noncytolytic , non-oncogenic viruses with chromosomes and the spindle is 
also essentially undefined. The potential of these investigations is vast; 
basic information may be gained important to an understanding of human disease 
pathogenesis as well as to current concepts on the evolution of viruses and 
the basic nature of their interactions with dividing, differentiating cells. 

Proposed course : To be continued at same. level of effort. 

Honors and Awards : None 

Publications : None 



96t 



Serial No. NDS (CF) - 70 E 1840 

1. Collaborative &. Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Retinoblastoma, Clinical, Genetic and Psychometric Aspects 

Previous Serial Number: Same 

Principal Investigator: Roswell Eldridge, M.D. 

Other Investigators: Kathy O'Meara 

Jeffrey C. Allen, M.D. 
David Kitchen, M.D. 

Cooperating Units: Johns Hopkins Hospital, Baltimore, Maryland 

Children's Hospital of D. C, Washington, D. C. 
Department of Psychology, Downstate Medical Center, 
New York 

Man Years : 

Total: 4 
Professional: 3 
Other: 1 

Project Description: 

Objectives : Only a few traits are said to be associated with 
increased intelligence. Such a correlation has been documented for torsion 
dystonia, the recessive type. Retinoblastoma is another condition for 
which such an association has been reported. Because we have reservations 
about choice of controls and value of data obtained in a handicapped 
population, we wish to evaluate this association in retinoblastoma 
ourselves. 

Methods employed : Efforts will be concentrated on testing the 
psychometric performance of affected individuals with family history who 
are sighted. The unaffected sibs will form the control group. 

Major findings : To date over 20 affected individuals and 20 controls 
in 12 families have been evaluated by us. In addition, retrospective data 
from school groups testing is being obtained. 

Significance to biomedical research and the program of the Institute ; 
This work is of great potential significance. If there is an association 



97 1 



Serial No. NDS (CF) - 70 E 1840 

between the gene for retinoblastoma and higher intelligence, then 

finding the underlying chemical abnormality should add to our knowledge of 

intellectual development as well as suggest the cause of neoplasm. 

Honors and Awards: None 

Publications: None 



98t 



Serial No. NDS (CF) - 70 E 1842 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Mortality among Japanese- Americans in relocation camps 

Previous Serial Number: Same 

Principal Investigator: Roger Detels, M.D. 

Other Investigator: None 

Cooperating Units : None 

Man Years : 

Total : 1/2 

Professional: 1/6 
Other: 1/3 

Project Description: 

Objectives : To determine death rates among American and foreign-born 
Japanese in relocation camps in 1943 and 1944 from selected neurologic, 
cardio -vascular and malignant diseases. 

Methods employed : During the period 1942-45 there were over 120,000 
Japanese-Americans in the U.S., all of whom were under the authority of 
the War Relocation Board. The majority were in relocation camps. Copies 
of the death certificates of persons dying in these relocation camps are on 
file at the Bancroft Library of the University of California at Berkeley. 
Records, including medical charts for all internees, are on file at the 
National Records Center in Suitland, Maryland. Thus, it is possible not 
only to establish death rates from varioxis caxjses, but also to determine 
the accuracy of the diagnosis and the course of the disease by referring 
to the medical charts. Baseline statistics of Japanese-Americans in the 
U.S. in 1943-45 and of those in relocation camps were accurately maintained 
at 6 month intervals and are available at the National Archives. 

Major findings : Data were reviewed. No patients with MS or ALS were 
encountered. 

Significance to biomedical research and the program of the Institute : 
The availability of death certificates and medical charts allowed us to 
determine accurate death rates among American and foreign-bom Japanese- 
Americans and to correlate these rates to length of residence in Japan 
and the U.S. Thus, we should be able to comment on relative effects of 



L 



99 1 



Serial No. NDS (CF) - 70 E 1842 

environment and genetic make-up on such diseases as multiple sclerosis, 
amyotrophic lateral sclerosis, stroke and other cardio-vascular and 
malignant diseases. 

Proposed course : This study was terminated. 

Honors and Awards : None 

Publications : None 



loot 



I 



Serial No. NDS (CF) - 70 E 1843 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Causes of death among siblings of MS and ALS patients 

Previous Serial Number: Same 

Principal Investigator: Jacob A. Brody, M.D. 

Other Investigator: Estelle Kornhauser, R.N. 

Cooperating Unit: Department of Neuropathology, Montefiore Hospital, 
New York 

Man Years : 

Total: 1/3 
Professional: 1/4 
Other: 1/12 

Project Description: 

Objectives : Recently, we have developed serologic evidence that 
familial patterns of antibodies of MS patients differ significantly from 
controls. This suggests a familial process may be involved etiologically 
in MS. We wish to determine, therefore, if siblings of MS patients have 
unusual illnesses possibly related to immunologic defect. The diseases we 
are interested in include collagen diseases, thyroid diseases, and 
rheumatoid arthritis. 

Me th o ds empl oy ed : We will control the siblings of MS patients with 
the siblings of ALS patients . We have selected 100 MS patients and 100 
ALS patients from the autopsy file of Dr. Harry Zimmerman, Department of 
Neuropathology, Montefiore Hospital. We will contact surviving siblings 
to determine major illness in the sibships. 

Major findings : At present, we have information on 60 ALS patients 
and 70 MS patients in whom the diagnosis was confirmed at autopsy. Various 
attempts to locate survivors has been unsuccessful. 

Significance to biomedical research and the program of the Institute : 
There is no firm evidence that an autoimmune mechanism or a virus is the 
cause of MS. There is, however, accumulating suggestive evidence that this 
is the case. There is also evidence of altered serologic responses among 
siblings of MS patients. If we can document that the siblings of MS patients 



k 



lOlt 



Serial No. NDS (CF) - 70 E 1843 

have illnesses related to immunological mechanisms we would have valuable 
evidence that these mechanisms are involved in the etiology of MS. 

Propos ed co urs e : Attempts using other patient source will be made in 
hopes of getting better access to follow-up data on siblings. 

Honors and Awards : None 

Publications : None 



102 1 



Serial No. NDS (CF) - 70 E 1844 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Amyotrophic lateral sclerosis in veterans 

Previous Serial Number: Same 



Principal Investigators; 



Other Investigators 



Cooperating Units ; 



John Kurtzke, M.D. 

Chief, Neurology Service, Veterans Administration 

Hospital, Washington, D.C. 
Gilbert Beebe, M.D. 

Director, Follow-up Agency, National Research 

Council of the National Academy of Sciences 
Virginia C. Karl, M.A. 

Social Work Service, Veterans Administration 

Central Office, Washington, D.C. 



Jacob A. Brody, M.D. 
A. Roger Bobowick, M.D. 

Veterans Administration 

National Research Council of the National Academy of 
Sciences 



Man Years : 

Total: 1/3 
Professional: 1/4 
Other: 1/12 

Project Description: 

Objectives : We will determine if there are distinctive epidemiologic 
features related to amyotrophic lateral sclerosis using the veteran 
population. We will also determine if service in Guam and the Marianas 
Islands was a risk factor in developing ALS . Life-table estimates will be 
constructed to clarify prognosis as to survival with ALS. 

Methods employed : The methods paralleling those of the veterans study 
of multiple sclerosis will be used to obtain the "records sample." In 
addition, we will survey approximately 200 veterans with ALS and 200 controls 
with brain tumors using a detailed historical questionnaire. This population 
will be called the "interview sample." Finally, a prognostic study will be 
constructed from Veterans Administration Hospital admissions of the 1957 - 
1964 period for ALS and other motor neuron diseases. 



103 1 



Serial No. NDS (CF) - 70 E 1844 

Major findings : The records sample should number at least 425 World 
War II and Korean war veterans with ALS . With about 500 discharges annually 
for ALS, the large Veterans Administration Hospital system will supply 
sufficient case material to carry out the interview sample and construct 
the prognostic study. Dr. Franklin 0. Meister, Director, Neurology Branch, 
Veterans Administration Central Office, Washington, D.C. has endorsed the 
project and suggest that only VA Hospitals with neurologic units be used. 

The detailed historical questionnaire for the interview sample has been 
drafted and preliminary field tests indicate that it will be quite workable. 

Significance to biomedical research and the program of the Institute : 
Regional differences and socio-economic differences have been commented upon 
for amyotrophic lateral sclerosis in the United States. This study will 
determine for a large population if there are predisposing factors to this 
disease. Should we determine such factors, it would provide a valuable clue 
to the understanding, treatment and prevention of this disease. Prognosis 
for survival should also be clarified. 

Proposed course : With our collaborators, we will collect all known 
veterans with ALS and try to detect possible predisposing factors . 

Honors and Awards : None 

Publications: None 



IQl+t 



Serial No. NDS (CF) - 70 E 1845 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Natural history of Parkinson's disease and the effect 
of L-Dopa 

Previous Serial Number: Same 

Principal Investigator: Jacob A. Brody, M.D. 

Other Investigator: A. Roger Bobowick, M.D. 

Cooperating Unit: National Parkinson Foundation, Inc., Miami, Florida 

Man Years : 



Total: 


5/24 


Professional: 


1/8 


Other: 


1/12 



Project Description: 

Objectives : We wish to determine if L-Dopa affects the natural history 
of Parkinson's disease or is merely symptomatic treatment. 

Methods employed : We will study a cohort with diagnosed Parkinson's 
disease who had the disease between 1958 and 1964, prior to the advent of 
L-Dopa. We will then develop life tables to determine the life expectancy 
and specific cause of death of patients with Parkinson's disease before 
the advent of L-Dopa, We will then follow a matched series of patients who 
are receiving L-Dopa to determine life expectancy and cause of death. 

Major findings : Review of the records at the National Parkinson 
Foundation, Inc. in Miami revealed that the necessary information had been 
reliably recorded. A sample of 50 pre- and 50 post-L-Dopa era patients has 
been processed. Medical record data retrival forms and follow-up forms to 
physicians have been completed. The medical record data is remarkably 
complete. The follow-up information will require the usual multiple 
avenues of pursuit. The staff at the National Parkinson Foundation, Inc. is 
now working on the medical record data retrival forms for the remaining 450 
patients in the pre-L-Dopa cohort. 

Preliminary analysis of the first 100 patients suggest that in the 
population of Miami the mean age increased about 3 years from the early 
to the late 1960 's. This is consistent with the cohort theory of parkinsonism 
which supposes a common etiology of sub-clinical encephalitis lethargica 

105t 



Serial No. NDS (CF) - 70 E 1845 

around 1920 and predicts the disappearance of parkinsonism as a major 
clinical entity around 1980. Because of the profound implications, verificatior 
of this phenomenon is being sou^t in age data of Parkinson patients at 
other centers . 

Significance to biomedical research and the program of the Institute : 
It is not known whether L-Dopa provides only s3nnptomatic relief of Parkinson's 
disease or whether it actually affects the pathologic process v/hich produces 
this chronic illness. If indeed it does reverse the pathologic process, we 
would advance the understanding of the genesis of this disease. Since L-Dopa 
is a drug with known biohazards, we will be able to determine if these hazards 
shorten the life expectancy of treated patients or if they die of specific 
diseases such as cardiovascular disease or kidney disease which will permit 
us to focus medical attention on prevention of these specific complications 
among treated patients . 

Proposed course : The pre-L-Dopa cohort and the cohort receiving L-Dopa 
will be matched and appropriate follow-up information concerning major 
illness and death will be secured. 

Honors and Awards : None 

Publications : None 



I06t 



Serial No. NDS (CF) - 71 E 1917 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Twin study of multiple sclerosis: An epidemiologic inquiry 

Previous Serial Number: None 

Principal Investigators: A. Roger Bobowick, M.D. 

Jacob A. Brody, M.D. 

Other Investigators: John F. Kurtzke, M.D. 

Veterans Administration Hopsital, Washington, D.C. 
Zdenek Hrubec, Sc.D. 

Follow-up Agency, NAS-NRC 
Marjorie Matthews, R.N. 

Cooperating Units: Neurology Service, Veterans Administration Hospital, 
Washington , D.C. 
Follow-up Agency, National Academy of Sciences, National 
Research Council 

Man Years : 



Total: 1/4 
Professional: 1/6 
Other: 1/12 

Project Description: 

Objectives : The abundant epidemiological literature of multiple sclerosis 
has indicated beyond reasonable doubt that critical environmental factors 
influence the rate of disease. Prevelance and migration studies strongly 
suggest that the environmental factor(s) is operant about the time of puberty. 
In order to examine the nature of the etiologic exposure in multiple sclerosis 
then, it is most prudent to concentrate on events in patients and controls 
with comparable early life histories in whom host factors are equalized. 
Twins discordant for multiple sclerosis at an age beyond prime risk of 
acquiring MS offer this opportunity for study. 

Methods employed : The NAS-NRC twin registry has identified 23 pairs of 
twins one or both of whom has multiple sclerosis. This group has come from 
their population of 16,000 pairs of white male twins who are veterans of 
military service in World War II and born during the years 1917 to 1927. 
Initial contact with the twins will be a letter from the registry explaining 
the study. If the twins express an interest in the study, they will be 
contacted by phone by the NIH investigators to arrange an interview date. 
The information collected will be: 1) pertinent medical history and 

I07t 



Serial No. NDS (CF) - 71 E 1917 

neurological exam, 2) an indepth epidemiologic interview concentrating on 
events prior to age 20, and 3) blood samples. 

Ma.i or findings : Initial contact has been made with most of the twin 
pairs and thus far five pairs have agreed to participate. The first twin 
pair will be visited shortly. 

Significance to biomedical research and the program of the Institute : 
Although exhaustive studies have been conducted to invoke specific etlologic 
factors in MS, no definitive precipitative circumstances have been identified 
perhaps because of the subtlety of these factors or the difficulties of 
suitably controlling for numerous xmrelated circumstances. This novel 
application of the twin method of study offers an efficient technique for 
identifying these precipitating circumstances. Resolution of the cause and 
prevention of MS would be greatly enhanced by the identification of these 
precipitating factors . 

Proposed course : The twins will be visited over the course of the next 
year. 

Honors and Awards : None 

Publications : None 



108 1 



Serial No, NDS (CF) - 71 E 1918 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Follow-up of the Cooperative Measles Vaccine Field Trial 
in four communities 

Previous Serial Number: None 



Principal Investigators 



Jacob A. Brody, M.D, 
Anne H. Edgar 



Other Inves tigators ; 



Marion Dressier, M.D. 

DeKalb County Health Department, Decatur, Georgia 
Margaret I. Rathbun, M.D. 

Monroe County Health Department, Rochester, New York 
Edwin P. Isacson, M.D. 

State University of New York School of Medicine 
Russell E. Alexander, M.D. 

University of Washington School of Medicine, Seattle 
Philip S. Brachman, M.D.. 

National Communicable Disease Center, Atlanta 



Cooperating Units : 



DeKalb County Health Department, Decatur, Georgia 
Monroe County Health Department, Rochester, New York 
State University of New York School of Medicine 
University of Washington School of Medicine, Seattle 
National Communicable Disease Center, Atlanta 



Man Years ; 



Total: 

Professional; 
Other : 



1/4 
1/2' 
3/12 



Project Description: 

Objectives : To determine through contact with the participants in the 
killed measles vaccine field trial of the early 1960s, any possible sequelae 
to receipt of the vaccine. 

Methods employed : Review of the addresses of the study population in 
four communities (DeKalb County, Rochester, Buffalo and Seattle) provided 
definite addresses for 46% of the 2091 participants plus possible location 
of an additional 17%. In order to determine the likelihood of success of 
follow-up, questionnaires requesting history of childhood diseases, other 
infections, non-allergic diseases accompanied by a rash, and hospitalizations 
subsequent to receipt of the vaccine, were sent to the families of the study 

logt 



Serial No. NDS (CF) - 71 E 1918 

population in DeKalb County. To date, 52% of the definitely located families 
and 29% of the possibly located have responded. These dame methods are to be 
extended to the participants in the field trial in the other three cities. 

Major findings : To date the response to the study has been good but the 
small number of participants contacted allows no conclusions to be drawn. 

Significance to biomedical research and the program of the Institute : 
The isolation of a measles-like virus from brains of individuals with 
subacute sclerosing panencephalitis has suggested a relationship between 
these two diseases. Epidemiologic study has further indicated that these 
patients may experience measles at an unusually early age. Evidence of high 
measles antibody titers in patients with multiple sclerosis has raised the 
possibility that measles might play an etiologic role in this disorder. In 
addition reports have been published on an atypical measles illness occurring 
in children who have received killed measles virus vaccine several years 
earlier. In light of the increasing interest in measles as an etiologic 
factor in disease, and the specific sequela reported to occur in those who 
have received the killed measles vaccine, the medical history of this study 
population subsequent to vaccination is of particular interest. 

Proposed course : The participants for whom possible or definite 
addresses have been located in the other three communities will be contacted 
by mail and asked to supply the information requested on the questionnaire. 
Those not responding to the initial request will be contacted a second time 
in the hope of increasing the response rate. 

Honors and Awards : None 

Publications : None 



not 



Serial No. NDS (CF) - 71 E 1919 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Viability of white blood cells used for lymphocyte 
transformation 

Previous Serial Number: None 

Principal Investigators: Jeffrey C. Allen, M.D. 

George Nemo, Ph.D. 

Other Investigator: Jacob A. Brody, M.D. 

Cooperating Unit : None 

Man Years : 



Total : 


1/6 


Professional: 


1/12 


Other: 


1/12 



Project Description: 

Objectives : To develop a technique for studying lymphocyte transformation 
on blood specimens transported over extended periods of time so that the 
technique may be used in field studies. 

Methods employed : White blood cells are allowed to stand at room 
temperature in either minimal essential medium or autologous plasma over 
a 24-48 hour period. The ability of the white blood cells to undergo 
lymphocyte transformation when exposed to various antigens is studied by 
the standard technique of measuring lymphocyte transformation used in this 
laboratory. 

Major findings : There was considerable variability in measurements of 
lymphocyte transformation to vaccinia over a 24 hour period, however, there 
was no reduction magnitude of response of the cells in the interval studied. 
MEM was not as good as incubation mediums as autologous plasma. 

Significance to biomedical research and the program of the Institute : 
If this method can be perfected the technique of lymphocyte transformation 
may be used in field studies and large numbers of relevent individuals may 
be examined. It is also hoped that if some of the variability is removed the 
technique of lymphocyte transformation may become sensitive. In 
addition, if there is a preferential die-off of granulocytes over time, a 
relatively pure culture of lymphocytes may be obtained. 



lilt 



Serial No. NDS (CF) - 71 E 1919 

Proposed course : It remains to be determined how long white blood 
cells can remain in autologous plasma at room temperature and still retain 
their ability to undergo transformation. The variability in response is 
considerable. This variability should be studied. Such techniques as 
purifying the lymphocytes, drawing blood at a certain time of the day from 
the same individual, and more careful standardization of the procedure may 
control some of the variability. 

Honors and Awards : None 

Publications : None 



112 1 



I 



Serial No. NDS (CF) - 71 E 1920 

1. Collaborative & Field Research 

2 . Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: A study of pregnancies among nurses; high risk pregnancies 
due to transmissible viruses 

Previous Serial Number: None 

Principle Investigator: Estelle Kornhaioser, R.N. 

Other Investigator: Jacob A. Brody, M.D. 

Cooperating Unit : None 

Man Years : 

Total: 1/4 
Professional: 1/12 
Other: 3/12 

Project Description: 

Objectives : This project was developed to question the effects on the 
fetus of maternal virus infection. 

Methods employed : A questionnaire was designed using the usual backup 
history and pertinent questions relating to type of nursing service, year of 
pregnancy, normal or abnormal births, and all information relating to any 
type of exposure to a viral type disease. The questionnaire and letter of 
explanation was pre-tested in 1160 nurses registered in the Maryland and 
District of Columbia area. 

Major findings : Of the 1160 questionnaires sent we have received 
approximately 800 replies . To date there are 1353 pregnancies with 
approximately 250 miscarriages, stillbirths, and abortions resulting for 
many reasons. 

Significance to biomedical research and the program of the Institute : 
This study will determine if pregnant women working with people with infectious 
disease have an elevated risk of fetal damage. 

Proposed course : We plan to follow up with a second mailing and then 
continue our study to a larger group of respondents and continue to examine 
the risk factors involved in the care of patients who have infectious diseases. 

Honors and Awards : None 
Publications : None 

113t 



Serial No. NDS (CF) - 71 E 1921 

1. Collaborative & Field Research 

2. Epidemiology Branch 
/ 3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Measles infection of mice: Effect of maternal immunity 

Previous Serial Number : None 

Principle Investigators: Jacob A. Brody, M.D. 

George Nemo , Ph . D . 

Other Investigators: Lon R. White, M.D., DB, NICHD 
Gary Cooper, Biologist 

Cooperating Unit: None 

Man Years : 

Total: 1/6 
Professional: 1/12 
Other: 1/12 

Project Description: 

Objectives : To develop an experimental animal model to study the 
pathogenesis of subacute sclerosing panencephalitis (SSPE) . 

Methods employed : Adult female mice previoiosly immunized with measles 
virus will be bred and their offspring challanged with measles at various 
times after birth. Measles antibody levels will be determined periodically 
and a thorough analysis of mouse tissues for measles virus and measles- 
related antigens will be performed. 

Major findings : None 

Significance to biomedical research and the program of the Institute ; 
An agent which by most virological criteria resembles measles virus has been 
isolated from brains of SSPE patients . A papova-like virus has been visualized 
in electron micrographs of specially treated cell lines inoculated with SSPE 
brain material suggesting the possibility of a dual infection. The exact 
role these agents play in the pathogenicity of the disease is unclear. An 
experimental animal model may aid considerably in elucidating the mechanism. 

Proposed course : See "Methods employed . " 
Honors and Awards : None 
Publications : None 

I15t 



Serial No. NDS (CF) - 71 E 1922 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Search for a negative DNA strand in cells infected with the 
minute virus of mice (MVM) 

Previous Serial Number: None 

Principle Investigator: Lon R. White, M.D., DB, NICHD 

Other Investigators: Harrie Anne Sutton, biologist, DB, NICHD 
George Nemo , Ph . D . 
Gary Cooper, biologist 
Jacob A. Brody, M.D. 

Cooperating Unit : None (HD DB - 15) 

Man Years : 

Total: 0.7 
Professional: 0.7 
Other: 0.0 

Project Description: 

Objectives : a). To demonstrate a "template" DNA (negative strand) 
complementary to the single stranded DNA (positive strand) of the MVM 
virion, b) . To determine if the negative strand is associated with the 
cell's chromosomal DNA. c). To investigate the kinetics of negative strand 
synthesis relative to cellular DNA replication, cellular division, virus 
infection, and virus production. 

Methods employed : These studies will utilize H or C "tagged" viral 
DNA with annealing demonstrated by: a) liquid scintillation detection of 
retained isotope on a filter, b) autoradiography with virus infected cells 
in metaphase. Some experiments will use purified chromosomes and chromosomal 
DNA from mitotically phased cells in culture. 

Major findings : None 

Significance to biomedical research and the program of the Institute : 
Characterization of virus-cell interactions using a single-stranded DNA virus 
(parvovirus group, of which MVM is representative) is expected to advance our 
understanding of the mechanisms by which a virus infection might alter the 
genetic potential of the host cell (either a somatic or germ cell) as well 
as of mechanisms involved with the establishment of latent and chronic viral 

iiyt 



Serial No. NDS (CF) - 71 E 1922 

infections. These may be directly relevant to the etiologies of certain 
reproductive, developmental, and chronic diseases whose causes are now 
unknown . 

Proposed course : To be continued at an increased level of effort. 

Honors and Awards : None 

Publications : None 



Il8t 



Serial No. NDS (CF) - 71 E 1923 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Effect of measles virus on specific and non-specific 
stimulation of lymphocyte transformation 

Previous Serial Number: None 

Principal Investigator: Jeffrey C. Allen, M.D. 

Other Investigators: George Nemo, Ph.D. 

Jacob A. Brody, M.D. 

Cooperating Unit : None 

Man Years : 



Total: 


1/6 


Professional: 


1/12 


Other: 


1/12 



Project Description: 

Objectives : To define the effects live and UV- inactivated measles 
virias have on the ability of human peripheral lymphocytes to respond to 
apecific and non-specific stimulation in vitro . 

Methods employed : Measles virus from both Ostereich and Edmonston 
strains will be grown to high titer in tissue culture. Lymphocytes will be 
obtained from healthy volunteers and prepared in the usual manner. A 
battery of specific antigens will be used including vaccinia, PPD , Candida, 
diptheria and mumps. PHA will be the only non-specific antigen used. 

Lymphocytes will be cultured in the usual way and incubated for the 
number of days appropriate to the particular antigen. Measles virus of 
varying titers, either live or killed, will be added simultaneously, before 
or after the particular antigen is added. Tritiated thymidine will be 
added one day before the harvest and uptake will be measured by liquid 
scintillation spectrotometry . Virus cultures and antibody measurements 
will be made from lymphocyte cultures at the time of harvest. In certain 
experiments fetal calf serum will be used to control for the effects of 
varying titers of measles antibodies of volunteers. 

Major findings : Preliminary experiments suggest that measles virus 
(Ostereich) will inhibit non-specific lymphocyte PHA stimulation at titers 
greater than or equal to 10 PFU/cc even though there is less than 1 virus 
per lymphocyte in the original culture. 

119 1 



Serial No. NDS (CF) - 71 E 1923 

Significance to biomedical research and the program of the Institute : 
Measles virus has been known for some time to suppress specific cellular 
immune responses in vivo . Controversy exists with regard to its effect on 
lymphocytes in vitro and current reports suggest that the virus inhibits 
specific but not non-specific stimulation. The mechanism of this inhibitory 
effect is unknown. It is not known what effects humoral or cellular immunity 
have on this inhibitory effect. Children with thymic aplasia and absent 
cellular immunity are subject to aberrant and severe measles infections. In 
vitro studies may suggest the role cellular immunity plays in lost defense 
against viruses. 

Proposed course : After the basic work is completed, measles antigen 
will be broken down to its basic protein and RNA components, and the effects 
of these substances will be evaluated. Other viruses will also be investigated 
such as rubella and mumps. Various disease states will be investigated such 
as SSPE and MS. A more complete study of the effects of measles antibody 
and antigen- antibody complexes is anticipated. Investigation at the cellular 
level for the site of measles operation will be studied with the use of 
autoradiography or fluorescent staining. 

Finally, the supernatort of the measles-lymphocyte cultures will be 
investigated for the presence of inhibitory factors. 

Honors and Awards : None 

Publications: None 



120t 



Serial No. M)S (CF) 71 E I92U 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Biochemical Studies in Torsion Dystonia 

Previous Serial Number: None 

Principal Investigators: Roswell Eldridge, M.D. 

Thomas Chase, M. D. 

Other Investigator: Kathy A. O'Meara 

Cooperating Unit: Laboratory of Clinical Science, NIMH 

Man Years : 

Total: 2 
Professional: 1 l/2 
Other : 1/2 

Project Description: 

Objectives : Torsion dystonia has been shown recently to consist of at 
least two hereditary conditions as well as an acquired form. Through 
appropriate biochemical investigation it should be possible to determine 
the precise biochemical defect (or abnormal gene product) in the recessive 
form of torsion dystonia. 

Methods employed : Patients are selected from the nationwide registry 
of torsion dystonia cases we have compiled and admitted to Dr. Chase's 
service for one or more weeks of the intensive evaluation. In addition 
to routine neurologic and chemical study, metabolism of biogenic amines is 
evaluated as completely as possible. Following this, there is often a 
period lasting several months in which drug trial is carried out. 

Major findings : Initial results on the small series of patients suggest 
that there may be an increased turnover of catecholamines in those with 
the recessive form of dystonia. This in in marked contrast to the levels 
seen in Parkinson's disease and Parkinson's dementia on Guam. 

Significance to biomedical research and the program of the Institute : 
Demonstration of the molecular lesion in one of the torsion dystonias would 
be a noteworthy achievement since another disease would be added to the 
small series of inborn errors of metabolism for which the precise lesion 
is recognized. In addition, undoubtedly important information with the 
forthcoming regarding biochemistry and physiology of movement control. 

1^1 1 



Serial No. NDS (CF) 71 E I92I1 

In addition, because of the recognized association of increased intelligence 
and the recessive form of dystonia, it is possible information regarding 
the development of intelligence would also result. 

Proposed course: Continued investigation of patients from the genetics 
registry. Addition of more sophisticated techniques as they become 
available. 

Honors and Awards : None 

Publications: Chase, T.N. : Biochemical and pharmacologic studies of 

dystonia. In The Torsion Dystonias (Dystonia Musculorum 
Deformans). Editor, R. Eldridge. Neurology suppl. 20:11, 
Part 2, November 1970. 



122 1 



Serial Wo. WS (CF) 71 E 1925 

1. CollalDoratlve & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-WIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Intelligence in Israeli Patients with Torsion Dystonia 

Previous Serial Number: None 

Principal Investigators: Roswell Eldridge, M.D. 

Other Investigators: Morris Gross, PhD. 

Richard Goodman, M. D. 

Cooperating Unit : Department of Education 

Herbert H. Lehman College 
Bronx, New York 

Man Years : 

Total : 1/2 

Professional: 3/8 
Other : 1/8 

Project Description: 

Objectives ; The recessive form of torsion dystonia has recently been 
shown to be associated with increased intelligence. However, the series 
which this information is based was small and individuals were scattered 
over a large area of the northeastern part of the United States. In Israel, 
where we suspect there to be over 20 patients with the recessive form of 
torsion dystonia, a more homogeneous testing situation prevails so that 
results should be more meaningful. If this association is true the implications 
are dramatic so that confirmation to this Israeli study is clearly in order. 

Methods employed ; Bona fide cases of torsion dystonia in Israel would 
be selected by the principal investigator on the basis of personal examination. 
Psychometric data would be obtained in a retrospective manner through the 
Israeli Department of Education and similar data would be obtained for 
unaffected sibs. A comparison would then be made of the performance in 
these two groups. 

Major findings ; Although we have known for several years of the 
presence of over 20 cases of torsion dystonia in Israel we have not yet been 
able to arrange for visitation. Neurologists, psychologists, and educators 
in Israel have been alerted and indicated their interest in participating 
in this study. 



I23fc 



Serial Wo. EDS (CP) 71 E I925 

Significance to biomedical research and the program of the Institute : 
The positive association between torsion dystonia and intelligence would 
suggest that the chemical abnormality producing dystonia may also enhance 
intellectual development. Elucidation of this basic chemical abnormality 
would suggest a method for enhancing intellectual development;, particularly 
those from the retarded population. 

Proposed course : Several weeks in Israel would be necessary to make 
the necessary home visits and arrange for the collection of appropriate 
psychometric data. 

Honors and Awards : None 

Publications : None 



12 



i^t 



Serial No„ KDS (CF) 71 E 1926 

1. Collaborative & Field Research 

2. Epidemiology Branch 
3- Bethesda, Maryland 

FES-KLE 

Individual Project Report 

July 1, 1970 through June 30, 197I 

Project Title: The Difficulty of Diagnosing Acoustic Neuroma in Young 
Patients 

Previous Serial Number: None 

Principal Investigators: Jeffrey C. Allen, M.D. 

Roswell Eldridge;, M.D. 

Other Investigator: Kathy O'Meara 

Cooperating Units: Department of Neurosurgery 

Columbia Presbeyterian Hospital 
New York 

Man Years : 

Total : 1 
Professional: 1/2 
Other: l/2 

Project Description: 

Objectives : To document and publicize the difficulty of diagnosis 
when acoustic neuroma occurs in young patients. 

During a course of clinical and genetic study of acoustic neuroma in young 
individuals we have been impressed with the unusual length of time and 
unusual number of specialists consulted before diagnosis of acoustic 
neuroma became a consideration. This period was often one of unusual 
lonnecessary stress and expense to the patient and his family and often 
resulted in loss of valuable time in many instances. 

Methods employed : Patients who have early onset of acoustic neuroma 
were questioned in detail regarding physicians consulted and diagnostic 
procedures employed. 

Major findings : In general, the younger the individual when symptoms 
of acoustic neuroma begin, the longer the period before diagnosis and the 
greater the number of physicians consulted and the more expense to the 
family. 



12 5t 



Serial No. WDS (CF) 71 E I926 

Significance to "biomedical research and the program of the Institute : 
Such documentation should impress the medical community ahout the resistance 
of this condition in young individuals thereby reducing time, expense and 
tuCTiioil to patient, family, and physician when a new case develops. Through 
such publicity it is possible that our acoustic neuroma registry will be 
expanded . 

Proposed course : Documentation of similar experience in the remainder 
of our acoustic neuroma patients. Publication of results. 

Honors and Awards : None 

Publications : None 



126t 



Serial No. KDS (CF) ?! E I927 

1. CollalDorative & Field Research 

2. Epidemiology Branch 

3. Bathes da, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, I97I 

Project Title : Family Studies in Young Patients with Acoustic Neuroma 

Previous Serial Number : None 

Principal Investigators: Jeffrey C. Allen , M. D. 

Roswell Eldridge, M. D. 

Other Investigators: Kathy A. O'Meara 

George T. Nager, M. D. 
Johns Hopkins Hospital 

Cooperating Units : Department of Otolaryngology, 
Johns Hopkins Hospital 
Baltimore, Maryland 

Man Years : 

Total: 1 1/2 

Professional: 1 
Other: l/2 

Project Description: 

Objectives : To determine the nosology of acoustic neuroma by studying 
clinical features, the age at onset of symptoms and presence of a family 
history of the disorder in young patients with unilateral and bilateral 
acoustic neuroma. 

Methods employed : Field studies were conducted in evaluating family 
members. On those seen personally, physical examinations were performed, 
stressing neurological and skin examinations. In addition, audiometric 
examinations including air and bone conduction and caloric examinations 
were conducted in the field. 

Patients were selected through chart review of neurosurgeons' files. All 
patients selected were alive at the outset of this study and experienced 
onset of symptoms before age U5. 

Genealogic information and medical history was obtained from family members 
and family and hospital records. (Medical history was obtained from family 
members, hospital and physician records.) 



127t 



Serial Wo. M)S (CF) 71 E I927 

On select patients extensive outpatient studies have included audiometric 
and vestibular testing, complete ENT and neurologic examinations and skull 
x-rays. These were undertaken in cooperation with the Departments of 
Neurology, Radiology and Otolaryngology of Johns Hopkins Hospital. 

Major findings : Patients with bilateral acoustic neuroma are more likely 
to have earlier onset of symptoms and a positive family history of either 
central, peripheral or mixed neurofibromatosis. Nine of the 5I propositi 
seen so far had bilateral acoustic neuroma. Three of these had a definite 
family history of the trait. Of the Ul propositi with unilateral acoustic 
neuroma, a definite family history was noted in only one. 

Most patients in the study commented on the length of time and number of 
physicians consulted before a definite diagnosis was made. Ttie average 
number of years between onset of symptoms and surgical intervention was 
7.0 for the bilateral cases and ^.h for the unilateral cases. The larger 
niimber of years for bilateral cases may reflect the failure of physicians 
to consider this diagnosis in younger individuals. 

Significance to biomedical research and the program of the Institute : 
The syndrome of bilateral acoustic neuroma seems to be distinct for unilateral 
acoustic neuroma using parameters such as age of onset of symptoms, likelihood 
of having a positive family history, presence of stigmata of neurofibromatosis 
and mode of inheritance. Unilateral acoustic neuroma on the other hand 
probably is not a distinct syndrome and may represent the manifestations of 
a sporadic dominant mutation or the expression of a recessive trait. Nearly 
^0 percent of our patients with unilateral acoustic neuroma were of Ashkenazic 
Jewish extraction. '^ 

Proposed course : The project will be completed after 70 kindreds have 
been examined. 

A large body of descriptive information has been collected on these diverse 
kindreds. These kindreds could provide a fertile source for laboratory 
investigation. Pathologists have not been able to differentiate between 
an acoustic neuroma from a patient with unilateral or a patient with bilateral 
disease. Perhaps there are differences at a biochemical level. 

Patients with bilateral disease may have a generalized neoplastic diathesis 
and laboratory investigation may reveal this in the patient and certain 
members of their family. 

Honors and Awards: Presentation at Middle Atlantic Neurosurgery Society 

Publications : None 



128 1 



Serial No. KDS (CF) 71 E I928 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda^ Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, I97I 

Project Title : Chromosome Studies in Retinoblastoma 

Previous Serial TJumber : None 

Principal Investigators: David Kitchen, M.D. 

Roswell Eldridge, M.D. 

Other Investigators : Kathy 'Meara 

Cooperating Unit: Cytology Laboratory, Eye Institute, 
Columbia Presbyterian Hospital 
New York, New York 

Man Years : 

Total: 2 
Professional: 1/2 
Other: 1 I/2 

Project Description: 

Objectives : Retinoblastoma is a malignant tumor of infancy and 
childhood which fortunately is often amenable to surgery. There appear to be 
several causes for retinoblastoma. Cases are generally sporadic but in 
approximately 10 percent there is a family history suggesting genetic basis. 
In addition, a number of reports have now appeared of retinoblastoma 
associated with gross chromosomal abnormality, generally of the D group. In 
these cases there is generally an associated physical abnormality in contrast 
to the other forms of retinoblastoma. Chromosome studies will be performed 
on select individuals and families to add further documentation to the role 
of gross chromosomal change and retinoblastoma. 

Methods employed : Blood is drawn on selected patients and their 
relatives during a home visit for our psychometric evaluation of retinoblastoma. 
Material is sent to Dr. David Kitchen's laboratory for karyotype analysis. 

Major findings : In none of the familial cases of retinoblastoma have 
karyotype analysis has there been evidence of chromosomal abnormality of a 
gross nature. In contrast several sporadic cases of retinoblastoma associated 
with other physical defects have been found with abnormal feryotype by Dr. 
Kitchen. 



ia9t 



Serial No. NDS (CF) 71 E I928 

Significance to biomedical research and the program of the Institute : 
Confirmation of the role gross chromosomal change makes to retinoblastoma 
is vital in pinpointing the precise etiologic event of this form of cancer. 

Proposed course : See "Methods employed". 

Honors and Awards : None 

Publications : None 



130^ 



Serial Ro. WS (CF) Jl E I929 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Genetic Study of an Irish Isolate in South Carolina 

Previous Serial Rixmher; None 

Principal Investigator: Roswell Eldridge, M. D. 

Other Investigators: Jeffrey C. Allen, M.D. 
Kathy 'Meara 
Charles Still, M.D. 

Cooperating Unit : Department of Neurology, 

University of South Carolina 
Columbia, South Carolina 

Man Years : 

Total: 2 
Professional: 1 
Other: 1 

Project Description: 

Objectives : The Irish Travellers of Murphy Village, South Carolina are 
a unique Catholic isolate whose exact origins are unknown. The present 
population is approximately 1,100 of which U20 are under 20 years of age. 
The men receive income from spraying barns and laying linoleum. They secure 
their business by travelling in small trucks throughout the eastern United 
States. 

The Village is of particular interest to us because of the apparent high 
rate of inbreeding which predisposes autosomal recessive traits. 

Methods employed : At present the need is for precise demographic data 
such as population break down by sex and age. Patterns and family size 
would be of interest . 

Genetic information may be best secured through conversation with responsible 
elders in the Village. Undoubtedly the group traces from a small founding 
population and precise information about this as well as information about 
members added to the group in recent years would be important. 

The genetic relationship of this population to the present day Irish might 
be established by noting the gene frequencies with ABO, Rh and other 
accessible footmarkers. 

131t 



Serial No. WDS (CF) 71 E I929 

Medical genetic studies most certainly focus on existing recognized familial 
problems such as mental retardation and skin disease. In addition, a 
comprehensive medical evaluation of the community and historical account of 
disorders in earlier generations would be worthwhile. Screening for newborns 
looking for recognized metabolic and chromosomal abnormalities might be 
productive. 

Major findings : Preliminary information has already been secured 
regarding the background, demography, economy, social organization, 
education, medical problems and genetics. Several familial traits are 
present among the Villagers suggesting that a comprehensive survey would 
reveal other genetic traits. 

Significance to biomedical research and the program of the Institute : 
Studies of such isolates have been unusually productive in the delineation 
and understanding of genetic entities. Examples include Victor McKusick's 
study of the Amish and Carl Witkop's study of the Wiesorts. 

Proposed course : Considerable ground work has been laid for this 
project and now results await investment of time and personnel. 

Honors and Awards : None 

Publications : None 



132^ 



Serial No. M)S (CF) 71 E 1930 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-WTH 

Individual Project Report 

July 1, 1970 through June 30, I97I 

Project Title: Von Hippel-Lindau's Syndrome - Clinical^ Genetic and 
Biochemical Aspects 

Previous Serial Number: None 

Principal Investigator: Roswell Eldridge^ M.D. 

Other Investigators: Jeffrey C. Allen, M. D. 
Kathy O'Meara 

Cooperating Units: Department of Neurology 
Johns Hopkins Hospital 
Columbia Presbeyterian Medical Center 

New York 
Man Years : 

Total : 2 
Professional: 1 I/2 
Other : 1/2 

Project Description: 

Objectives: In I9II Eugen von Hippel described a number of patients 
with retinal angiomatosis. In I926 Arvid Lindau described a number of 
patients, who in addition to the retinal angiomatosis, had such intracranial 
lesions as cerebellar cysts and cerebellar medullary angioblastic tumors 
as well as pancreatic cysts, renal cysts and adrenal tumors. The familial 
nature of this syndrome has since been noted by numerous authors. 

What has impressed us is the infrequency with which these tumors are 
bilateral. In contrast to other hereditary traits associated with neoplasm 
of paired structures the involvement is generally bilateral. Is the 
tendency to unilateral involvement in the Von Hippel-Lindau's syndrome real 
and, if so, does it reflect a different etiologic mechanism than seen in the 
other hereditary neoplasms? Or is the unilateral involvement only apparent, 
reflecting failure to scrutinize the presumably healthy member of the 
paired organs? 

Methods employed : The propositi with Von Hippel-Lindau's syndrome will 
be ascertained through neurologic departments of selected medical centers. 
Contact with individuals and their families will be made through their 
personal physician, and home visits will be arranged for physical examination 
and detailed history. 

133^ 



Serial No. KTOS (CF) 71 E I93O 

Major findings : None 

Significance to biomedical research and program of the Institute : 
The first step will be to document whether or not this syndrome tends to be 
unilateral in its involvement. Later more sophisticated laboratory studies 
can be undertaken to determine more closely the mechanism of abnormal gene 
action which produces the neoplasias. 

Proposed course : See "Methods employed". 

Honors and Awards : None 

Publications: None 



I3^t 



Serial Wo. WDS (CF) 71 E I93I 

1. CollalDorative & Field Research 

2. Epidemiology Branch 
3- Bethesda, Maryland 

FES-Wm 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Genetic study of population isolates in Maine 

Previous Serial Number : Rone 

Principal Investigators: Jeffrey C. Allen, M. D. 

Roswell Eldridge, M. D. 

Other Investigators: Morris Lambdin, M. D. 

Ellsworth, Maine 
Thomas Roderick, M. D. 
Jackson Laboratory 
Bar Harbor, Maine 

Cooperating Units: Division of Public Health Nursing, 

State of Maine Dept. of Health & Welfare 

Maine Genetics Counselling Center 
Ellsworth, Nb,ine 

Maine Medical Association; Lincoln, Sagadahoc, 
Washington, Waldo and Hancock Counties 

lyfen Years : 

Total : 1/2 

Professional: l/k 
Other : l/k 

Project Description: 

Objectives : To investigate numerous genetic subisolates along the 
coast of Maine for evidence of hereditary neurological disorders. 

Methods employed : A preliminary trip was made to establish contacts 
and examine the feasibility of the study. Thereafter an introductory 
letter and brief questionnaire was sent to every registered physician and 
public health nurse in the five major coastal counties. The questionnaire 
was designed to provide information on the awareness of any unusual 
familial disorders; diagnosed and undiagnosed, which are prevalent in 
remote areas of the state, especially on some of the offshore islands. 

If feasible, demographic and historical data will be collected from State, 
County and local sources to corroborate the prevalence of genetic isolation. 



135' 



Serial No. M)S (CF) 71 E I93I 

Patients and their families will "be ■visited and an appropriate referral 
will be made to local diagnostic facilities. 

Major findings : So far^ (^S of II9 (58 percent) questionnaires have 
been returned. Several interesting leads exist. 

Significance to biomedical research and the program of the Institute : 
Maine's rugged eastern coast contains innumerable islands of varying sizes. 
Several of these have been inhabited for several generations and 
populations vary from 50 "to 200 persons. Inbreeding is commonly practiced. 
On several islands, most of the inhabitants have only two or three 
different surnames » Because of the remoteness of the islands inhabitants 
have not availed themselves of the usual medical facilities. They are 
visited rather infrequently by a public health nurse or physician. The 
likelihood of finding recessive hereditary disorders is high in these 
populations . 

Proposed course : Personal visits will be made to selected families. 
Genetic and demographic surveys will be conducted wherever appropriate. 

Honors and Awards : None 

Publications : None 



I 

I 




136^ 



ANNUAL REPORT 

July 1, 1970 through June 30, 1971 

Special Projects Branch 
Collaborative and Field Research 
National Institute of Neurological Diseases and Stroke 
National Institutes of Health 

Following the previous fiscal year's reorganization of the Special Projects 
Branch, four professional personnel were added to the staff: a fully trained 
neurologist and a staff associate to the Branch, a staff associate to the 
Head Injury Section, and a staff associate to the Epilepsy Section. Broad 
responsibilities of the staff included anticonvulsant drug program, support 
of activities of the Secretary's Advisory Committee on the Epilepsies, its 
sub-committees and task forces, the NINDS Ad Hoc Committee on Cerebral Death, 
the NINDS Ad Hoc Committee on Anticonvulsant Drugs, and the Head Injury and 
Epilepsy Information Services. 

Section on Epilepsy 

Meetings of the NINDS Ad Hoc Committee on Anticonvulsant Drugs were held in 
October 1970 and February 1971. The Fall meeting was held in Seattle, 
Washington, to enable the committee members to observe progress on the NINDS 
sulthiame research contract, and the potential for other related neuropharma- 
cological research there. The Committee has continued to function in a highly 
efficient and useful manner providing review of research in progress, of 
proposals for new research contracts, and of the pharmacology of investigational 
anticonvulsant drugs. They suggested the priorities for proposed studies and 
other guidance to the section. One of the most beneficial effects of the 
Committee is the origination of suggestions from individual members--as the 
background of each member differs, a variety of views and ideas are presented 
for Institute follow-up. 

During fiscal year, the pharmaceutical industry continued to cooperate with 
the section. Data on drugs from seven companies was provided and discussed 
by the NINDS Ad Hoc Committee on Anticonvulsant Drugs. Priorities were 
established for support of clinical trials for two investigational drugs. 
One will be conducted in a state hospital for patients with generalized 
epilepsy; the other in university clinics for patients with absence (petit 
mal) epilepsy. Preliminary studies will be employed to perfect anticonvulsant 
blood level methodology and determine metabolic information prior to the 
outset of full scale trials. 

Research contracts for the study of ethosuximide in absence epilepsy at the 
University of Virginia and The Medical College of Wisconsin were completed 
in fiscal year 71. The study of ethosuximide in absence epilepsy has 
examined several hypotheses. The hypothesis that ethosuximide is not 
effective in control of approximately one-half of those patients who suffer 
from absence (petit mal) epilepsy is apparently correct when one defines 

lu 



control as complete (100%) control. Ethosuximide has, however, improved 
virtually every patient in whom the drug has been tolerated. The hypothesis 
that failure of response to ethosuximide occurs in the presence of thera- 
peutic blood levels has been demonstrated when response is defined as 
complete control. The hypothesis that patients who respond to ethosuximide 
will show no evidence of focal brain lesions is apparently true although some 
patients with brain lesions may also respond. Another hypothesis is that 
ethosuximide does not impair psychometric performance; evidence so far has 
shown that indeed ethosuximide does not impair such performance when thera- 
peutic blood levels are not exceeded. There is no evidence to the present 
time that ethosuximide alters clinical manifestations of absence seizures; 
there is also evidence that many conventional measurements of seizure 
frequency do not provide a reliable assessment of seizure control. 

An ancillary technique, that of EEG telemetry, has been developed during this 
study, and found to be the best criterion for measurement of control of 
paroxysmal abnormal discharge. The reliability of this technique is related 
to the duration of the individual seizure, the longer ones being more easily 
recognized. Currently, the telemetry study consists of continuous twelve- 
hour EEG recordings; the information is taken from FM tapes and evaluated at 
NIH. 

The telemetered EEG has correlated quite well with the best clinical method 
used in the ethosuximide study to date, that is, observation by a trained 
observer. In almost eyery case, however, the number of seizures estimated 
by the observer is less than that ascertained by telemetered recording, 
strongly suggesting that the observer, regardless of experience, does not see 
every seizure. This telemetry technique will be utilized in subsequent 
studies. Detailed analysis of the data from the ethosuximide research contracts 
will be made at the conclusion of the study. An investigational anticonvulsant 
will be evaluated in this model upon the completion of the present phase. 

Pursuant to a recommendation of the NINDS Ad Hoc Committee on Anticonvulsant 
Drugs, a contract was awarded for the clinical evaluation of the investigational 
anticonvulsant, sulthiame, in June 1970. The pilot study has been completed 
and a double-blind comparison of sulthiame and di phenyl hydantoin is now 
underway. Sixty patients with a diagnosis of partial epilepsy will take part 
in the study. Each patient will be evaluated over a fourteen-month period. 
Important features of this research are accurate reports of seizure incidence, 
other drug effects, anticonvulsant blood level determinations, and measures 
of psychological performance and social functioning. 

The Institute will establish a pharmacology laboratory within the Section on 
Epilepsy early in the coming fiscal year. Much planning has taken place to 
prepare for this activity, following its endorsement by the Ad Hoc Committee 
on Anticonvulsant Drugs. Harvey J. Kupferberg, Ph.D., has been recruited to 
head the laboratory. The metabolism and interactions of many anticonvulsant 
drugs will be studied initially. 

Staff members collaborated with officials of the National Institute of Mental 
Health to plan an epidemiology survey for the prevalence and incidence of 



2u 



convulsive disorders in seventh grade students in Washington County, Maryland. 
The children are to be examined in the summer of 1971; data will be evaluated 
in the coming fiscal year. 

Two meetings of the Secretary's Committee on the Epilepsies were held. It 
was decided to sponsor two small closed workshops to examine in depth two 
neglected areas of interest to workers in epilepsy. A workshop on laboratory 
methods for preparing animal models of epilepsy will be held in November 1971. 
Another workshop to be held in November 1971 will examine many aspects of the 
epidemiology of epilepsy. A third event jointly encouraged by the Secretary's 
Committee and the Ad Hoc Committee will be a detailed examination of the 
pharmacology of anti epileptic drugs. World leaders in the field will present 
papers to an audience of clinicians so their treatment of patients will be 
enhanced. Unfortunately, much of the researchers' efforts have not yet been 
applied to the treatment of epileptic patients. A monograph will be published 
following the symposium. 

The Section issued two publications for workers in the epilepsy field; a 
review of the literature on the effectiveness of marketed anticonvulsants, 
by James J. Coatsworth, M.D., a consultant to NINDS, and a review of the 
literature on anticonvulsant blood level determination methods and clinical 
importance by members of Branch personnel. Editorial support was provided in 
both instances by Branch personnel. Dr. Coatsworth 's report is of especial 
interest, for it highlights the meager information available on anticonvulsants 
in use today, and the paucity of well -documented clinical studies that have 
been conducted in the past. NINDS-sponsored studies on anticonvulsants have 
been designed to overcome these shortcomings and will provide useful information 
on the drugs' efficacy and safety. 

Epilepsy Abstracts is now in its fourth year of publication. The Excerpta 
Medica Foundation has assumed all publication and distribution responsibilities 
with modest support from the Institute. Epilepsy Abstracts Retrieval System 
(EARS), a free text search and retrieval system based upon Epilepsy Abstracts 
was proven a powerful research tool. Every word in the abstract is a key 
word permitting rapid thorough searches of the literature from any of a 
variety of terminals. EARS was demonstrated at the meetings of the American 
Epilepsy Society, American Academy of Neurology, and American Neurological 
Association. In each case, reception was most favorable. It is planned to 
have this retrieval system available to research workers throughout the 
country in the coming year. 

A computer-based bibliography on epilepsy research from 1900 to 1959, with 
some 8,000 index citations, has been developed. It will be issued in the 
coming fiscal year and will compliment the published Epilepsy Abstracts 1947 
to the present time. 

The investigational anticonvulsant albutoin was compared with two marketed 
anticonvulsants, primidone and diphenylhydantoin, in a double-blind study at 
New Castle State Hospital, New Castle, Indiana, September-December 1970, 
under a NINDS research contract. The design of the trial was improved, 
based upon experiences of the 1969 trial, when albutoin was compared with 
phenobarbital . Accurate seizure records, blood level determination, and 



3u 



other laboratory and ward data were recorded through the NIH WYLBUR 
computer system for ease in analysis. Plans are underway for a trial of 
another investigational anticonvulsant, carbamazepine, using the patient 
population at New Castle State Hospital, and methodology developed during 
the albutoin study. 

Section on Head Injury 

In response to a recommendation of the National Advisory Council, Neurological 
Diseases and Stroke, the NINDS Collaborative Study of Cerebral Death, a 
directed study under contract has been established. With guidance and support 
from the Office of the Associate Director for Collaborative and Field Research, 
the staff prepared comprehensive bibliographies and reprint collections with 
translations of the literature on cerebral death. These documents were 
provided to the NINDS Committee on Cerebral Death, and to prospective 
principal investigators who would collaborate under the contract mechanisms. 

The overall objectives of the collaborative study of cerebral death are to 
verify or modify the current clinical and electroencephalographic (EEG) 
criteria for cerebral death in relation to patient age and cause of coma: to 
determine the minimal time that clinical and EEG criteria must be operative 
to indicate cerebral death in relation to age and cause of coma; and to 
assess the neurologic deficit of patients who recover after having fulfilled 
all criteria for some length of time. In addition to the clinical and EEG 
evaluations of patients, several contractors will conduct ancillary studies 
of cerebral death; these will include computer analysis of electroencephalo- 
grams, bedside EEG monitor, averaged evoked potential responses, electro- 
retinograms, analysis of cerebrospinal fluid for chemical indicators of 
brain death, and detailed neuro-ophthalmological examinations. 

Experience dictates that prior to embarking on a large-scale collaborative 
project of this type a pilot or feasibility study is needed to pretest 
procedural and technical elements of the overall study, to develop the 
statistical model, and to establish executive capability; specifically, its 
aims include the following: to determine whether the contemplated neuro- 
logical, EEG, and other appropriate examinations can be adequately handled 
in a collaborative study, and to establish a basis for achieving standardiz- 
ation; to determine whether, in a collaborating hospital, any cases of 
suspected cerebral death would be missed, and if so, why? to develop means of 
determining when a clinical and EEG picture consistent with cerebral death is 
the result of sedative drug intoxication; to develop procedures for collecting, 
reviewing, editing and processing data for the later or definitive study; to 
evaluate means for guaranteeing uniformity in handling patients, making the 
necessary observations, and recording data for analysis; to determine the 
minimal significant set of clinical, EEG, laboratory and demographic observa- 
tions that will be needed for each patient in the definitive study; to determine 
the best statistical model for describing patients suspected of cerebral 
death, so that the risks associated with an operating rule for determining 
brain death can be estimated most efficiently from data gathered in the 
study; to determine the sample size needed in the later study in order to 
obtain desired precision of estimates, with due regard for variability in the 



4u 



patient population with respect to age and etiology of coma; and to create 
an executive capability that could be extended to the definitive study. 

To date, the project has been in a planning phase and will not become 
operational until about September 1, 1971. Preliminary findings are expected 
to accrue in the coming fiscal year and major findings in subsequent years. 

A thorough search of available literature in the area of cerebral death has 
clearly indicated that there is currently much disagreement as to which 
criteria are necessary and sufficient for the diagnosis of cerebral deatti 
and that so far no systematic attempt has been made to refine the existing 
criteria with respect to patient age, cause of coma, or length of time that 
the criteria must be satisfied. There are cogent medical, legal, and social 
reasons for establishing valid and refined criteria for cerebral death, and 
it is anticipated that these criteria will become widely accepted as a result 
of the collaborative study. Hence, families could be spared the emotional 
distress and financial burden of an unnecessary delay in the declaration of 
death in appropriate circumstances and at the same time be protected against 
undue haste in cessation of efforts at resuscitation. It is further possible 
that potential organ donors could be more readily identified and at an 
earlier time, resulting in more viable organs for transplantation. 

Contract negotiations for the pilot phase of the study are scheduled for 
completion in June 1971. The project will then become operational on or 
about September 1, 1971, and run for approximately one year. At the end of 
the pilot phase it should be possible to make a decision concerning the 
advisability of continuing with the collaborative study. If a decision to 
continue is made, the investigators will meet and agree to a protocol for the 
remainder of the study. This later or definitive phase will proceed until 
a sufficient number of patients are studied to give valid statistical results, 
all data is collected and analyzed, and a final report is drafted. 

Contractor's Project Director : 

Dr. A. Earl Walker (University of New Mexico) 

Dr. Reginald G. Bickford (University of California-San Diego) 

Dr. David C. Poskanzer (Massachusetts General Hospital) 

Dr. Francis E. McGee, Jr. (Medical College of Virginia) 

Dr. Julius Korein (New York University Medical Center) 

Dr. Benjamin Boshes (Chicago Wesley Memorial Hospital) 

Dr. Wigbert C. Wiederholt (Ohio State University) 

Dr. Lorenzo G. Runk, III (University of Pennsylvania Graduate Hospital) 

Dr. Donald R. Bennett (University of Utah) 

The head injury model construction program has been phased out. The data 
collected during the past three years is being reviewed and will provide the 
basis for a monograph on the mechanical properties of the head and neck. 

The final report on the survey of head injured veterans of the Korean campaign 
has been submitted to the Director of the Institute. 



I 



5u 



The section is continuing its collection and classification of head injury 
literature. Its recurring classified bibliographies are being shared with 
more than one thousand interested investigators in the field. 



6^ 



CONTRACT NARRATIVE 

Special Projects Branch--Section on Epilepsy 

July 1, 1970— June 30, 1971 

NEW CASTLE STATE HOSPITAL (PH-43-68-1310) 

Title : Study of the Anticonvulsant Properties of Albutoin 

Contractor's Project Director ; Joseph T. Brock, M.D. 

Current Annual Level : $90,000 

Objectives : To study the relative anticonvulsant properties of albutoin, 
diphenylhydantoin, and primidone administered to patients with seizures 
refractory to treatment, and to evaluate the possible side effects of the 
drugs and drug blood levels. 

Course of Contract : Patient trials began September 14, 1970. Forty-nine 
patients began the thirteen-week Latin cube experiment. The investigational 
drug albutoin was compared with the two marketed drugs, each for three week 
periods with two week intervals of regular medication. Clinical data was 
collected and sent to the Section on Epilepsy, NINDS , Bethesda, for review 
and preparation for computer-aided analysis. 

Major Findings : Detailed analysis is now underway. A preliminary review of 
the clinical data shows when compared to patients' ordinary medications, 
single drugs are less effective in preventing seizures. There was a greater 
incidence of seizures and side effects with the investigational drug than 
with either diphenylhydantoin or primidone. 

Significance to NINDS program and Biomedical Research : The pharmaceutical 
industry has demonstrated a little interest in developing new anticonvulsant 
agents. Aside from the economic factors involved, one of the industry's major 
problems is to obtain satisfactory clinical studies of anticonvulsant drugs. 
Through this contract and others, NINDS has supported clinical studies of 
anticonvulsant drugs. Well controlled studies will be significant indicators 
of therapeutic merit of new anticonvulsant drugs. It may encourage industry 
to develop its promising new agents for clinical trial. It is anticipated 
that through NINDS sponsored studies anticonvulsant drugs will reach the 
market more readily. 

Proposed Course of Contract ; The contract will be amended without additional 
funds to allow performance in FY72 . Additional anticonvulsant trials are 
planned . 



7u 



CONTRACT NARRATIVE 

Special Projects Branch — Section on Epilepsy 

July 1, 1970--June 30, 1971 



MEDICAL COLLEGE OF WISCONSIN (NIH-69-2169) 

UNIVERSITY OF VIRGINIA SCHOOL OF MEDICINE (NIH-69-2196) 

Title : Study of Anticonvulsant Properties of Ethosuximide 

Contract Project Directors ; Philip T. White, M.D., (MCW) 

Fritz E. Dreifuss, M.D., (UV) 

Current Annual Level : $ 65,462 (Medical College of Wisconsin) 

$ 53,474 (University of Virginia School of Medicine) 

Objectives : To study the effect of ethosuximide on the frequency and inten- 
sity of petit mal epileptic seizures in patients previously untreated for 
this disease; to evaluate the effect of ethosuximide therapy on physiologic, 
psychometric, and other functions in human subjects. 

Course of the Contract : Due to the difficulty in acquiring previously un- 
treated petit mal patients, the contracts were extended to June 30, 1971. 
Each center will be successful, or nearly so, in reaching its goal of 20 
patients. Data is collected for each patient during the initial hospita- 
lization period, during an outpatient and during the final hospitalization 
period, and sent to the Section on Epilepsy, NINDS , Bethesda, for review 
and entry for computer-aided analysis. 

Major Findings : The procedures and protocol established during the colla- 
borative study of epilepsy were proven valuable in conducting an evaluation 
of an anticonvulsant agent. Detailed analysis of the clinical results will 
be made at the end of the study. EEG telemetry has been developed as an 
ancillary technique. 

Proposed Course : The study will be extended to permit comparison of etho- 
suximide with an investigational anticonvulsant. 



9u 



CONTRACT NARRATIVE 

Special Projects Branch--Section on Epilepsy 

July 1, 1970— June 30, 1971 



UNIVERSITY OF MINNESOTA (NIH 70-2269) 

Title: Development of a Gas-Liquid Chromatographic Analysis of Blood 

Contract Project Director ; Harvey J. Kupferberg, Ph.D. 

Current Annual Level : $7,819 

Objectives : To develop a specific GLC method for analysis of blood and 
CSF for levels of sulthiame, and for levels of carbamazepine. 

Course of the Contract : The contractor has completed the project, pro- 
viding methodology for blood level determinations of the anticonvulsants 
which are or will be studied in research contracts for clinical evaluation 
of sulthiame and carbamazepine. 

Major Findings ; The GLC methodology was readily adapted from, the contrac- 
tor's animal studies to preliminary patient studies. Information on 
absorption characteristics and biological half-lives of the drugs was also 
obtained. 

Proposed Course : The contract has been completed. 



llu 



w^ 



CONTRACT NARRATIVE 
Special Projects Branch — Section on Epilepsy 
July 1, 1970— June 30, 1971 



UNIVERSITY OF WASHINGTON (NIH 70-2281) 

Title : Study of Sulthiame in the Treatment of Partial Epilepsy 

Contractor's Project Director : John R. Green, M.D. 

Current Annual Level : $106,597 

Objectives : To study the relative anticonvulsant properties in partial 
epilepsy of sulthiame and diphenylhydantoin; to measure these drugs in 
patients' blood, and to assess patients' psychological competence and so- 
cial function. 

Course of Contract : A preliminary study of hospitalized patients was com- 
pleted and enabled the contractor to undertake the long term study on 
October 20, 1970. More than half of the goal of 60 patients have been ac- 
quisitioned; each will be evaluated over a 14-month period. 

Major Findings : The contractor will review and analyze the data at the con- 
clusion of this two-year double blind study. The bioavailability of the 
specially prepared diphenylhydantoin was found greater than the commercial 
preparation the patients had been receiving. This necessitated a revision 
in dosage schedules to prevent toxicity. 

Proposed Course ; The contract will be amended with additional funds to 
allow completion of the second year planned when the contract was awarded. 



I 



13' 



I 



CONTRACT NARRATIVE 

Special Projects Branch — Section on Epilepsy 

July 1, 1970— June 30, 1971 



EXCERPTA MEDICA FOUNDATION (NIH-71-2018) 

Title : Epilepsy Abstracts , Volume 4 

Current Annual Level : $39,220 

Objectives : To scan serial publications and periodicals from approximately 
3000 of the world's biomedical journals, select appropriate articles to be 
included in Epilepsy Abstracts in accordance with the guidance of the Project 
Officer and his editorial advisors; prepare abstracts with appropriate trans- 
lations into English from foreign languages, classify, index, and store the 
abstracts in a computer retrievable form; and produce a 9-track computer tape 
in the NIH-PRS format, to be delivered when Volume 4 is completed. The 
Excerpta Medica Foundation will produce camera-ready copy for each monthly 
issue of Epilepsy Abstracts , which includes an Index of subjects and authors, 
and will print and distribute the journal monthly, including a cumulative 
index at the end of the volume. In order to pay for the production of the 
camera-ready copy, the printing, and distribution, the Excerpta Medica 
Foundation will sell subscriptions to recover the cost of production of 
camera-ready copy, printing, and distribution. 

Course of Contract : Following award of the contract, the Excerpta Medica 
Foundation promoted the subscription of Epilepsy Abstracts , and acquired 
900 subscriptions at $15 each. At the close of this report period, subscrip- 
tions are still being acquired. The Excerpta Medica Foundation produced 
monthly issues as directed and have increased slightly the number of original 
articles abstracted. 

Proposed Course of Contract : It is anticipated that all the remaining issues 
will be distributed as scheduled and that the computer tape will be delivered 
in accordance with the contract. 



15 u 



Serial No. NDS (CF)-71 SP 1932 

1. Collaborative and Field Research 

2. Epilepsy Section 

3. Bethesda, Maryland 

PHS--NIH 

Individual Project Report 

July 1, 1970--June 30, 1971 

Project Title: Long-term electroencephalographic telemetry of patients 
with absence (petit mal) seizures. 

Principal Investigator: J. Kiffin Penry, M.D. 

Other Investigators: Roger J. Porter, M.D. 

Fritz E. Dreifuss, M.D. 

Cooperating Units: Department of Neurology, University of Virginia School 
of Medicine, Charlottesville, Virginia. 

Man Years : 

Total: 1.33 
Professional: 0.33 
Other: 1.0 

Facilities and Equipment: 

The telemetry equipment and FM tape recorder were provided by the 
Section on Epilepsy. An oscilloscope and the clinical research unit were 
provided by the University of Virginia Hospital at Charlottesville. 

Project Description: 

Objectives : The purpose of this study is to evaluate patterns of 
paroxysmal abnormal discharge in patients with absence (petit mal) seizures. 
Another primary objective is to determine if this method can be established 
as a primary mode of evaluating any absence drug. 

Methods employed : Each patient has a 12 hour telemetered electroencepha- 
lographic recording on the clinical research unit of University of Virginia 
at Charlottesville. The telemeter equipment is placed on the patient's head 
by an EEC technician who also applies the electrodes. The patient is free to 
move about the ward and and the signal from the transmitters is taken to a 
dipole antenna and then to two receivers where two channels of EEC are fed 
to an FM tape recorder. Quality control is maintained by observation of the 
incoming data on the oscilloscope. The 12-hour recordings are taken back to 
the laboratory of the Epilepsy Section in Bethesda where they are played back 
at four times recording speed: Each 12-hour record is then read by a physi- 
cian. Patterns of discharge and effects of anti-absence drugs are evident 
when the data is processed by a 360 IBM computer and the Cal Comp plotter. 

17^ 



I 



Serial No. NDS (CF)-71 SP 1932 

Major Findings : The initial 12 patients with absence epilepsy have all 
shown some improvement on ethosuximide . Many patients have shown complete 
remission of paroxysmal activity. It was felt that this criterion of seizure 
control is probably the most objective available, and it is the intent of the 
Section on Epilepsy to incorporate this method in evaluation of investigational 
anti-absence drugs. 

On further evaluation 16 patients with absence seizures have shown a 
decline in seizure duration with increasing age. While this is a confirma- 
tion of previously suspected phenomena, this is the first time that such 
information has been collected in other than an anecdotal way. 

Significance to Biomedical Research and to the Program of the Institute : 
This study has applied telemetry techniques to improve the evaluation of 
clinical research in anti-absence pharmacology. The study has further sug- 
gested patterns of paroxysmal discharge in patients with absence seizures and 
points a way for further investigation of the mechanisms of absence epilepsy. 

Honors and Awards: None 

Publications : 

Penry JK, Porter RJ, Dreifuss FE: Quantitation of paroxysmal abnormal 
discharge in the EEGs of patients with absence (petit mal) seizures, 
for evaluation of antiepileptic drugs. Epilepsia (Amst) 12: 1971. 

Penry JK, Porter RJ, Dreifuss FE: Patterns of paroxysmal abnormal 
discharges in twelve-hour telemetered EEGs of untreated children with 
absence (petit mal) seizures. Neurology (Minneap) 21: 392, 1971. 



Serial No. NDS (CF)-71 SP 1933 

1. Collaborative & Field Research 

2. Epilepsy Section 

3. Bethesda, Maryland 



PHS— NIH 

Individual Project Report 

July 1, 1970— June 30, 1971 



Project Title: Quantitation of clinical manifestations of spike-wave 
activity by a reaction time method. 

Principal Investigator: Roger J. Porter, M.D. 

Other Investigators: J. Kiffin Penry, M.D. 

Fritz E. Dreifuss, M.D. 

Cooperating Units: Department of Neurology, University of Virginia School 
of Medicine, Charlottesville, Virginia. 

Man Years : 

Total: 0.5 
Professional: 0.25 
Other: 0.25 

Facilities and Equipment: 

Electroencephalographic equipment and space for testing were provided 
by the University of Virginia Hospital at Charlottesville. The reaction time 
equipment including seizure detection device and digital timer were designed 
and built by the Section on Epilepsy. The video-recording apparatus was 
provided by the Section on Epilepsy. 

Project Description: 

Objectives : The purpose of this study is to determine whether reaction 
time in absence patients is or is not impaired in a gradual fashion from the 
point of spike-wave initiation as has been suggested by some authors but dis- 
puted by others. There is some evidence for a "trough-like" pattern decrease 
of consciousness. The onset of decrease clinical functions during spike-wave 
paroxysms is evaluated by the reaction time method. 

Methods employed : A device is employed which gives instantaneous recog- 
nition by voltage criteria that a spike-wave burst has started. This burst 
is of much higher than normal background, and this factor alone is used to 
electronically trigger the reaction timer. On instantaneous recognition the 
reaction timer is triggered and a tone is delivered to the subject. The sub- 
ject responds by turning off the high pitch tone with a telegraph key. Be- 
tween paroxysms the patient is maintained in a state of alertness by a program 
of approximately 10 random stimuli per minute. All the data is collected 

19U 



Serial No. NDS (CF)-71 SP 1933 

by television, including a portion of the screen reserved for the reaction 
time from the digital clock. There was no age limits in selecting patients, 
but they must all have spike-wave paroxysmal discharge. 

Major Findings : The study is not yet complete but preliminary findings 
suggest that patients maintain some ability to respond early during the 
paroxysmal burst; this responsiveness is frequently not seen 1-2 seconds 
after onset. Analysis of responsiveness during short bursts suggests that 
patients may retain a normal reaction time during euch paroxysms. 

Significance to Biomedical Research and to the Program of the Institute ; 
This study has applied video recording techniques and sophisticated electronic 
methods to improve the quality of clinical research. Specifically, this study 
is an analysis of the relation of the patient's behavior to his EEG during 
paroxysmal electroencephalographic events. An understanding of this rela- 
tionship is import ant --not only as a guidepost for further research in the 
mechanism of epilepsy, but also in determining the day-to-day therapeutics 
of the epileptic patient. 

Proposed Course ; The study will be continued with additional patients 
in the coming fiscal year. 

Honors and 'Awards: None 

Publications: None 



20u 



CONTRACT NARRATIVE 
Special Projects Branch — Section on Head Injury 
July 1, 1970— June 30, 1971 



NATIONAL ACADEMY OF SCIENCES (PH 43-64-44. Task Order 11) 

Title: A 15-Year Follow-Up of Head-Injured Veterans of the Korean Campaign 

Contractor's Project Director : Seymour Jab Ion, M.S. 

Current Annual Level : No additional funds 

Objectives : 1. To provide NINDS with the current addresses of the head- 
injured veterans participating in the study. 2. To select and locate 
matched controls. 3. To code the Red Cross interview schedules. 4. To 
key punch the Red Cross coded forms. 5. To key punch the original coded 
acute data for the so-called Meirowsky cases. 6. To obtain General 
Classification Test scores from retired records in St. Louis. 7. To prepare 
a data tape embodying original (acute) and Red Cross Interview information. 
8. To prepare tabulations and analyses from the total material, relating to 
characteristics of the original wound and its treatment. 

Major Findings : All 8 objectives were accomplished. A report has been 
submitted to the director, NINDS. 

Proposed Course of Contract : This project will be concluded in fiscal 
year 1971. 



21 u 



i\ 



I 



CONTRACT NARRATIVE 
Special Projects Branch — Section on Head Injury 
July 1, 1970— June 30, 1971 



WEST VIRGINIA UNIVERSITY (PH 43-67-1137) 
CASE WESTERN RESERVE UNIVERSITY (NIH-69-2201) 
UNIVERSITY OF WASHINGTON (NIH-69-2232) 

Title : Determination of the Physical Properties of Tissues 

Contractor's Project Director : Dr. Russell R. Haynes (West Virginia Univ.) 

Dr. Albert H. Burstein (Case Western 

Reserve Univ.) 
Dr. Colin H. Daly (Univ. of Washington) 

Current Annual Level : $41,000 (West Virginia University) 

$27,000 (Case Western Reserve University) 
$30,000 (University of Washington) 

Objectives : To determine certain physical properties of the tissues of the 
head (scalp, skull, dura, brain, fluids); and to develop mathematical 
solutions for various simple mechanical force problems relating to effect of 
force upon the head. These objectives represent Phase I and preliminary 
work on Phase IV of a program to construct and test accurate physical models 
of the head. Various physical properties such as bulk modulus, shear 
strength, tensile strength, compressive strength, etc., are being determined 
using specimens from human cadavers, autopsies, biopsies, and similar 
specimens from the Macaca mulatta. 

Major Findings : This project period represents the fourth year of this 
collaborative study. The first year was generally devoted to developing 
test equipment, determining test parameters, establishing mechanisms for 
securing test specimens, and designing standardized test procedures. The 
second year saw the beginning of data collection. During the third year, 
both the University of Michigan and West Virginia University completed their 
determination of the relevant properties of the head and made considerable 
progress in the identification of substitute materials. In order to fill 
certain vital gaps in the data, two smaller contracts were let last year: 
to the University of Washington to determine the dynamic material properties 
of the cerebral vasculature and to Case Western Reserve University to 
evaluate the mechanical properties of the head-neck junction. During the 
current year preliminary data on the latter two contracts have been collected, 
and a final report is due in June 1971. A final contract was awarded to the 
University of West Virginia for a coordinated analysis of all the data 
obtained in this program. This data analysis and review is currently being 
studied by the project officer and will form the basis of a monograph on the 
mechanical properties of the head and neck. 

Significance to NINDS Program and Biomedical Research : The data collected 
provide an essential basis for the study of cell and tissue reaction to 
mechanical forces, which is essential for understanding head injury. 

23U 



Serial No, NDS (CF)-69 SP 1785 

1. Collaborative and Field Research 

2. Special Projects Branch 

3. Bethesda, Maryland 



PHS— NIH 

Individual Project Report 

July 1, 1970— June 30, 1971 



Project Title: A 15-Year Follow-Up of Head Injured Veterans of the Korean 
Campaign 

Previous Serial Number: None 

Principal Investigators: A. R. Taylor, M.B., F.R.C.S. 

S, Jablon, M.S. 

Other Investigators: W. F. Caveness, M.D. 
A. M. Meirowsky, M.D. 
A. C. Dresser, M.S.W. 
R. W. Hurt, M.D. 
C. Kretschmann 

Cooperating Units: National Research Council Follow-Up Agency, Washington, B.C. 
American National Red Cross Service to Military Families, 
Washington, D.C. 

Man Years: 

Total: 0.4 
Professional: 0.3 
Other: 0.1 

Projection Description: 

Objectives : 

1. To obtain information on employment patterns and the incidence of 
posttraumatic sjrmptoms and epilepsy over the past 15 years in a group of 
head-injured veterans of the Korean Campaign. 

2. To relate these findings to the initial data on the severity, therapy, 
and sequelae of the injuries; and to compare the status of the injured men to 
that of a group of non-injured control subjects. 

3. By determining the incidence of seizures in the parents, siblings 
and children of the veterans, to see if a genetic factor might be involved 
in predetermining epilepsy after head trauma. 

4. To identify those veterans who would be willing to participate in a 
comprehensive 15-year follow-up hospital evaluation. 

25 ^ 



Serial No. NDS (CF)-69 SP 1785 

Methods Employed : Acute data were supplied by Dr. Caveness and 
Dr. Meirowsky. The National Research Council Follow-Up Agency took a 30 
percent sample of the injured group and selected non- injured controls who 
were in Korea in the same units as the injured men at the time the head 
injuries were sustained. Current addresses, induction AGCT scores and pre- 
induction employment of the injured and non-injured men were obtained by the 
National Research Council Follow-Up Agency. The staff of the Section on Head 
Injury obtained permission of the veterans for interviews. American National 
Red Cross Service to Military Families workers interviewed the veterans, 
members of their families, and their employers. The medical data were edited 
by the staff of the Section on Head Injury. The National Research Council 
Follow-Up Agency edited the employment information, coded the interview 
schedules, transferred all acute and follow-up data to a seven-track tape 
file, and prepared tabulations on which the final report was based. 

Major Findings : The final report has been submitted to the director, 
NINDS . The following deductions were made about the future treatment of 
combat head injuries: 

1. Investigation should be made into the possible ways of treating 
memory and concentration defects in line with present methods employed in 
speech therapy. 

2. Positional vertigo, the common posttraumatic variety, should be 
intensively studied to uncover possible lines of treatment. 

3. Head injured men should be educated, from the time of their first 
reception, not to fear the outcome and not to regard the brain as the "master 
organ". 

4. They should not be segregated from limb and trunk injuries. 

5. There should be a national organization to continue education in 
civilian life and be ever present to sustain the veteran when his sjmiptoms 
occur or recur in response to stress. 

Significance to Biomedical Research and the Program of the Inst itute: 
This study has provided data on employment and the posttraumatic state 
fifteen years following head injury. 

Proposed Course : With submission of the final report, this project has 
been concluded. 

Honors and Awards : None 

Publications: None 



26^ 



ANNUAL REPORT 

For Period July 1, 1970 through Jvine 30, 1971 

Perinatal Research Branch 

National Institute of Neurological 

Diseases and Stroke 

National Institutes of Health 

TABLE OF CONTENTS 



I. General Sunnnary 
II. Specific Summaries 



Pajge No . 



A. Section on Obstetrics 9 

B. Section on Pediatric Neurology 11 

C. Section on Behavioral Sciences 13 

D. Section on Infectious Diseases 15 

E. Section on Pathology 21 

F. Section on Epidemiology and Genetics 29 

G. Section on Data Management and Retrieval 35 
H. Section on Project Services 

Medical Literature Services 37 

I. Section on Speech, Language and Hearing 41 

III. Contract Narratives 

The Wayne State University (PH-43-68-669) 

Maternal ajnino acid level as related to fetal 

birth weight of the infant. 45 

The Johns Hopkins University School of Medicine 
(PH-43-68-710) Long-range effects on the fetus 

of certain maternal infections during pregnancy. 47 

The University of California (NIH 69-4) 

Cytomegalovirus infections in pregnancy. 49 

IV. Individual Project Reports 

Section on Infectious Diseases 51 

Office of the Chief 93 

Section on Obstetrics 101 

Section on Pediatric Neurology 117 

Section on Behavioral Sciences 139 

Section on Epidemiology and Genetics 157 

Section on Pathology 193 

Section on Speech, Language and Hearing 225 



.^•.oiD\-riq ti ANNUAL REPORT 

For Period Jiily 1, 1970 through June 30, 1971 

Perinatal Research Branch 

National Institute of Ne\irological 

Diseases and Stroke 

National Institutes of Health 

SUMMARY OF PROFESSIONAL OR SCIENTIFIC ACC0t4FLISHMENTS 

A. FINDINGS 

A number of scientific findings that came to light from COLR data during 
this fiscal year can be highlighted in reference to the development of 
mental and motor performance in the growing child: 

The proportion of variability in four-year IQ attributed to the 
genetic component (heritability) is lower among Negroes than among 
-•'■ whites. 

Any measure of social class is an important predictor of Stanford- 
-Binet IQ at age four and is a significant determinant of mental and 
motor performance. Within both white and Negro children, those with 
the higher socio-economic index do better on the Binet at age four 
than those with the lower index. Higher IQ (Binet at age four) in 
either race is associated with higher educational level of the mother. 
Females have a higher IQ (Binet) at age four than males in either 
- race. 

The IQ in singletons seems to decline in the interval between ages 
four and seven years vJaereas the IQ of twins seems to show an 
improvement, suggesting that twins who usually perform more poorly 
•' than singletons during early life are able to "catch up" diiring 
childhood. 

The offspring from interracial matings do not differ significantly 
in birth weight or birth length or in mental or motor performance 
•■' at eight months from the offspring of intraracial matings of either 
race. However, by age four months and at one year the interracial 
offspring were significantly smaller than the controls, and at age 
four years, the IQ's of interracials were lower than either control. 
' This suggests an adverse environmental rather than an adverse genetic 
influence on the interracial offspring which is not experienced by 
intraracial progeny. 

Birth weight is better than gestational age in predicting mental and 
motor performance at eight months of age and of IQ at four years, 
independent of race, social class, or sex. 

Head size at one year of age may be a good predictor of IQ at age 
four. There is a 50^ chance of an IQ being less than 80 at age four 
for the one year male with a head size less than ^3 cm, and for the 
one year female with a head size less than k2 cm. 



A number of additional studies on twins and sibs speak to the physical 
development of the growing child: 

Half sibs compared to full siblings show more RH inconipatibility, 
seizures, congenital heart defects, mental retardation, club foot, 
and Polydactyly. Evidence suggests that genetic factors play an 
important role in seizures and mental retardation and environmental 
(prenatal) factors in congenital heart defects and club foot. 

Billington's hypothesis that sensitization of the mother during 
pregnancy against paternal antigens leads to non-pathological 
placental hypertrophy and increased birth weight in succeeding 
pregnancies has been supported and confirmed. 

Respiratory Distress Syndrome occurs more frequently in twins than 
in singletons. 

The relationship between type of twin placentation and zygosity was 
studied in 569 sets of twins. The perinatal death rate is lk% 
in either race. The frequency of congenital malformations was not 
higher in twin than in singleton deaths. Malformations in mono- 
chorionic deaths were multiple and lethal. Heavy infants and twins 
with large intra-pair birth weight differences were common in 
separate dl amniotic -dichor ionic groups; the fused di-di group had 
light infants and the lowest death rates. 

In comparing somatic vs. visceral growth rates, histological study 
of the kidneys of 51^ neonatal deaths and stillbirths revealed that 
the growth in birth weight and body length is faster in the Negro 
than in the white during the early phases of pregnancy, but visceral 
growth, as of the kidneys, proceeds at similar rates in both races, 
independent of whether the growing fetus is small or large for dates. 

A few studies focused on the blood and vas.c\ilar conditions in the growing 
child: 

High RH antibody titers are not as highly associated with serious 
morbidity in the Negro as in the white progeny. 

Neonatal polycythemia is associated with longer gestation, lower 
birth and placental weights, less placental pathology, and lower 
socio-economic status than controls. Polycythemic neonates do not 
differ from controls in psychological scores at eight months or in 
neurological findings at age one, but showed, especially among the 
Negro females, a lower four year Binet IQ. 

Chorangioma cases show an increased association with neonatal throm- 
bocytopenic purpura, toxemia, fetal hemangioma, erythroblastosis and 
single umbilical artery. The condition is commonest in whites and 
females. The prematurity rate of survivors is not different from 
that of single live births. 



2v 



I 



Neonatal Retinal Hemorrhage occurs in approximately 20^ of vertex 
births. The possible relationship of such hemorrhage to birth 
weight, mental and motor performance at eight months, Binet IQ, at 
four years and birth position is being investigated using matched 
non-retinal hemorrhage cases as controls. 

Generalized skin hemorrhages at birth seem to be associated with 
subsequent marked hearing loss, speech and behavior problems and 
possible mental retardation. This apparent association is being 
investigated to assess whether it is spurious or valid. 

Among 306 cases of single umbilical artery, in utero and neonatal 
deaths occurred in 13.8^ of cases. Many of these deaths are due to 
congenital malformations. Compared to controls matched for race, 
sex, institution, birth weight, gestational age, and socio-economic 
index, the single umbilical artery group had six-fold more occurrences 
of velamentous and marginal insertions of the cord, slightly more 
neurologic ally abnormal cases but similar mean mental and motor 
scores at eight months and four year IQ scores. 

The cumulative fertility rate, the perinatal death rate, birth 
weight of offspring, and gestational age of mothers with sicklemia 
were the same as in non-sickling mothers. The infant and child 
death rates, however, were higher in the sickling group. 

An important finding with reference to cerebral palsy revealed that 
spastic and non-spastic prematures of equal immaturity do not differ 
in their Apgar scores, bilirubin levels, or history of birth trauma. 
Cerebral hemorrhage, however, is one suspected cause of spastic 
diplegia. In the effort to elucidate the etiology of cerebral palsy 
in the prematures, this carefully designed study lends no support 
either to the genetic or the anoxic or toxic (hyperbilirubenia) or 
traumatic or nutritional (intrauterine blighting) hypotheses. The 
only statistically significant difference between spastic and non- 
spastic prematures was a low post-natal hematocrit level in the 
spastic s, and reason for this difference should be explored. 

A few studies on virology and immunology pointed out that patients who 
had carcinoma of the uterine cervix were found to have increased amounts 
of antibody against Type II Herpes Virus two years before the clinical 
emergency of cancer. The Australian Antigen was detected in the blood 
of 600 cases of infectious hepatitis studied during an epidemic in the 
United States. 

B. STUDIES m PROGEESS 

In addition, a considerable number of studies are currently in progress 
and are specifically described in the Individual Project Reports. It 
must be pointed out that all this research activity is separate from and 
has anteceded the research activity of the Ad Hoc Task Forces that came 
into being during the last fiscal year. Under the aegis of these Task 
Forces, the main data analysis activity may be highlighted as follows: 

3v 



I. TASK FORCES 

1) Labor and Delivery — This group is analyzing the characteristics 
of labor and delivery and its complications in relationship to 
perinatal loss, child development and neurological and intellec- 
tual deficits. 

2) Infectious Diseases — The main objective here is the investigation 
of the role of infectious agents and particularly viruses which 
occur in pregnancy and diiring the perinatal period in the fetal 
deaths of subsequent child development. 

3) Genetics and Congenital Malformations — The focus is the pursxdt 
of genetic factors related to human development and especially 
neurological and intellectual deficits. Further, this group is 
assimilating information regarding congenital malformations and 
its epidemiology. The unique population of twins born to mothers 
in the Study will be followed to at least fifteen years of age 

so that observations under mental and physical development can 
be made to at least through puberty. The development of special 
protocols for the yearly examination of these twins and for the 
special neiirological and psychological examinations at ages 
twelve and fifteen are currently under way. Furthermore, a 
chromosomal study of children from five Collaborating Institutions 
(Boston, Philadelphia, Buffalo, Tennessee, and Oregon) to relate 
major and minor chromosomal aberrations in these children to the 
outcome of pregnancy has been launched with the University of 
Colorado acting as the coordinator. In addition, a comprehensive 
study of sickle cell anemia to relate the occurrence of sickling 
in the mother with the outcome of pregnancy and to study the 
growth and development of children who are themselves sicklers 
is also iinder way. A proposal by Dr. J. V. Neel of the University 
of Michigan has been endorsed to establish genetic markers used 
in a variety of polymorphic blood loci for correlations with 
pregnancy outcomes; the necessary blood samples will be drawn 
and stored to be typed later. A request by Dr. Osborne is 
currently being considered by more than one NIH Institute to 1 
collect dental cast impressions on the seven year olds in the 
Collaborative Study with a view to assessing the timing of the 
occurrence of an insult in utero and its correlation with the 
emergence of specific morbidity in the child; this is part of a 
larger study of morphological and developmental asymmetries of 
interest to geneticists and teratologists. I 

^) Drugs in Pregnancy — This group studies the large number of drugs 
that are taken by women during pregnancy to explore whether these 
relate to any observed differences in fetal outcome including 
congenital malformations or subsequent child development. 

5) One Year Task Force — The concern is the analysis of pre- and 

perinatal factors related to neurological abnormalities identified 
in one year old children. The one year outcome was classified in 

4v 



ovt o;)Ti.specific categories in a mutually exclusive hierarchical order 

of disease conditions; these categories along with neurologically 
normal cases will he compared for frequencies of a large array 
of pre- and perinatal variables to detect associations. Instances 
where strong associations exist will he subjected to more 

■ • definitive analysis. 

6) Four and Seven Year Task Force — The objective is the analysis of 
pre- and perinatal factors related to neurological and intellectual 
deficits identified from four to seven years of age. The intent 

is to discover predictors of IQ at four years of age by means of 
multiple regression analysis. The number of variables will be 
reduced by obtaining a simple correlation of study variables 
with four year IQ. Variables lacking specific correlation will 
be eliminated from the regression analysis or analyzed separately. 
In addition a few composite indices made up of IQ and other four 
year performance tests will be subjected to regression analysis 
against the whole array of antecedent factors. 

7) Three and Eight Year Speech, Language and Hearing Task Force 
This group is concerned with analysis of pre- and perinatal 
factors related to speech, language and hearing performance as 
judged in three year olds, and then again in ei^t year old 
children. 

8) Physical Growth — This task force is concerned with the development 
of the physical growth profiles based on the physical measurements 
obtained on Collaborative Study children from birth on through 
age eight. Further, the group is looking at physical growth 
characteristics in children who differ by socioeconomic back- 
ground and other developmental characteristics. 

"^'"■'f^) Toxemia — This task force is concerned with the evaluation of the 
utility of the clinical signs and symptoms which are part of the 
toxemia complex and its significance vis-a-vis perinatal mortality 
and prematurity. The First International Workshop on Clinical 
Diagnostic Criteria of Toxemia of Pregnancy scheduled in 
December 1971 is sponsored by the Perinatal Research Branch sind 
shall present data from the Study as well as a proposed set of 
guidelines for the development of the theory of toxemia and its 
implications in clinical practice. 

10) Pathology Task Force — This task force is concerned with the 
analysis of the brain specimens from children dying during the 
perinatal period to get insight into the kind and frequency of 
apparent brain injury identified from postmortem examination. 

11) Basic Document — Frequency and cross tabulations of continuous, 
discrete and dichotomous variables are used to reveal the 
prospective relationship, if any, between selected obstetrical 
variables and mortality, birth weight and one year neurological 
outcome by race and institution. 



5v 



12) Anesthe sia-Analges ia — Gravida have been categorized into two 
grotrps, "normaJ." aind those that are "not normal" as defined. 
The effect of anesthesia-analgesia in various combinations, as 
defined, will be tabulated in relation to selected outcomes from 
these two populations, to assess the effect, if any, of selected 
anesthesia-analgesia agents as administered. The protective 
effect of barbiturate against lowering Apgar by anesthesia will 
be investigated. 

13) Epidemiological and Statistical Advisory Committee — This was 
established by the Perinatal Research Committee to offer merit 
review of study proposals as submitted by the various task 
forces prior to final approval. 

II. OTHER STUDIES 

1) The relationship between weight gain during pregnancy and the weight 
of the baby is to be investigated by the Primate Nutrition Study 
currently being funded jointly by the National Institute of Child 
Health and Human Development, the National Institute of Neurological 
Diseases and Stroke, and Johns Hopkins University. The pilot phase 
is to explore through experimental design the lead that came out 
from COLR data, confirming previous findings, that increase in 
maternal weight gain reduces the frequency of low birth weight 
children. 

2) The Collaborating Institutions at Minnesota and at Oregon have been 
funded by the Office of Education to investigate the relationship, 
if any, between elementary school performance and antecedent pre-, 
peri-, and postnatal information available in the Project. 

3) Dr. Rosenblith, in Providence, has established a neonatal behavioral 
scale working on the Providence sample of the Project population 
and is now validating it against the parameters of data collected 
at various endpoints in the Study. This behavioral scale, if validated 
on the population of the Project as a whole, will serve as an 
additional source of identification of high risk infants, especially 

in areas where more efficient or more accurate methods are needed 
than the medical examinations can provide. 

h) In cooperation with the Collaborating Institution at Boston and 

through funds made available through the National Institute of Child 
Health and Human Development, under the scope of the COLR contract for 
data collection, a study of dermatoglyphics is iinder way to find out 
if normal progeny can be thus distinguished from abnormal progeny. 

5) In cooperation with a member of NICHD, the Branch will carry out a 
multi-factor analysis of the contribution of various preciirsors to 
the occurrence of minimal brain dysfunction. 



OTHER FUNCTIONS 

The Branch, through the Office of the Chief, continues to monitor and overview 
the flow of activities in the following eight areas: 

Sample maintenance, overdue forms, early monitoring of data collected, 
quality control (inter- and intrainstitutional), form processing 
(editing, coding, punching, storage and magnetic tape), study proposals 
from the task forces as well as from individual investigators, merit 
review of manuscripts prior to publication, the Project Officer's role in 
contract negotiations and supervision of data collection in the 
Collaborating Institutions. 

The Branch is initiating and exploring various modsilities and patterns of 
enlisting and involving other Institutes within NIH in design and funding of 
a nimber of studies in cooperation with one or more Institutions in the 
field. The Institutes thus far involved are: National Cancer Institute, 
National Heart Institute, National Institute of Infectious Diseases and 
Allergy, National Institute of Dental Health, National Institute of Mental 
Health. 

The Branch is investigating and exploring mechanisms of merit review and 
funding of studies based on local data ansilysis by the various Collaborating 
Institutions. This is needed after the sharp fiscal stringency that has 
come about during Fiscal Year 1972. 

The Branch is making contact with several organizations to carry forward 
cooperative efforts. A Joint Conference on Minimal Brain Dysfunction has 
been agreed upon among the CIBA Corporation, NICHD, and NINDS and has 
received approval for presentation under the auspices of the New York 
Academy of Science in March 1972. At this conference, the present status of 
theory, research, etiology, management, and treatment will be explored and 
will receive broad dissemination throiigh the Academy's annals publication. 

The Branch has also been negotiating with the Office of Education to carry 
out a study relating pre- and perinatal variables to learning disability in 
early grades. It has been agreed by the two units involved that the actual 
study be carried out during FY 1972. It is now being planned. 

The Branch has held discussions with the Bureau of Indian Health, HSMHA, in 
regard to research on communication research — including audiology, treatment 
of the deaf, and prediction of stroke by analysis of dysarthria. Interaction 
and agreement on our prospective areas of interest and capability will 
continue. 

The Branch has received a number of requests for reviews of grant applications 
by Branch members who are specialists in the area of perinatal research. 

The Branch has carried out an analysis of its own and NINDS -wide research on 
the child aged 0-5, following a request by the Division of Research Grants, 
which is pursuing the question for the Director of NIH in response to the 
President's proposed initiative on children's health in that age group. 

7 V 



The Branch has received, and has either satisfied or is in process of 
accommodating, a niunber of requests for different kinds of data emerging 
from the Collaborative Study. Among these are The Bureau of Economdc 
Research; Division of Epidemiology, Columbia University Medical School; 
Faculty of Law, Tel Aviv University; Vocational Counseling Associates; 
School of Public Health, North Carolina University; School of Business 
Administration, University of Washington; and Downstate Medical Center, 
State University of New York. 

The Branch has carried out an advisory function for certain organizations. 
The Branch Chief was appointed to the Advisory Liaison Committee, American 
College of Obstetricians and Gynecologists, and to the World Health 
Organization's Expert Committee on the Prevention of Perinatal Mortality 
and Morbidity. 

PROBLEMS 

The imposed arbitrary cut of $2.1 million within the Perinatal Project dxiring 
the coming fiscal year was at long last negotiated and absorbed. With the 
help of the Perinatal Research Committee, the Branch was able to adjust to 
this unwarranted cut both within its own internal operation and throuighout 
the Collaborating Institutions. The collaborators showed an acute sense of 
responsiveness and loyalty despite the growing sense of anxiety and low morale 
amongst them. To adjust to this cut was an experience which involved 
considerable effort and time, and was most pa3.nful, to say the least. 

PROGRAM DEVELOPMENT 

During the fiscal year, the Branch has undertaken an analysis of research 
needs and capabilities for the purpose of developing a research program to 
follow the termination of data collection in the Collaborative Study. A 
five year plan is being prepared in this context to explore leads from the 
current phase of the Perinatal Program and to pursue new and other leads in 
the whole area of developmental neiirology. This plan is to be reviewed for 
design and budget prior to Fiscal Year 1972. In its targeted research, the 
plan utilizes laboratory as well as field studies to explore in depth the 
effect of genetic, biologic, physical, and psycho-social factors on the 
developing nervous system. Implementation will not be limited to any one 
group of institutions or to many institutions at a time, nor will it be 
necessarily limited to any one funding mechanism. 



8v 



A. SECTION OW OBSTETRICS 
Report for the Period July 1, 1970 through June 30, 1971 

I. SUMMARY OF SdEEfTIFIC OR PROEESSIOHAL ACCOMPLISHMEMTS 

With the active cooperation of 9 experts in clinical research on toxemia 
of pregnancy and selected members from the Collaborating Institutions, 
data from the Perinatal Research Study were analyzed for the association 
of toxemia of pregnancy as reported in the Study protocol with fetal and 
neonatal mortality and on the reporting of edema, proteinuria and blood 
pressures during pregnancy. This information was then used to design a 
new study, utilising the original data on the clinical diagnostic criteria 
of toxemia. A chronological data basis for specified periods of pregnancy 
was established for blood pressures, proteinuria, edema, and their 
combinations, in association with the Immediate and long-term outcomes of 
pregnancy. These data, in turn, are being used to define "iso-risk zones" 
of blood pressures, edema, proteinuria, and combinations of the same in 
the gravida for specified outcomes of pregnancy: live birth, birthweight, 
fetal death, neonatal death. From these findings, critical limits will 
be set for identification of cases with toxemia in the Study pregnancies. 
Our preliminary findings indicate that the epidemiologic approach to the 
study of toxemia may contribute to the resolution of some of the complex 
riddles of toxemia. The methodological problems involved in this study 
were presented and discussed at the annual meeting of the Swiss Obstetric 
Society. We are cooperating with Dr. E. Hughes, Chairman of the Committee 
on Nomenclature of the American College of Obstetricians and Gynecologists, 
on the definition and nomenclature of the toxemias. 

Together with the members of the Task Force on Labor and Delivery, a 
study has been designed to determine the influence of specific labor and 
delivery factors on the fetus in terms of immediate outcome and later 
neurologic and psychologic outcome. In this study, a methodology -" 
developed by Dr. E. Friedman for the quantification of uterine activity 
and its deviations in relation to the phases of labor will be used to 
measure possible damage to the fetus, dependent on the dyscoordination or 
changes in the duration of any one or combinations of the phases of labor. 
The identification of gravidae with specified types of labor will produce 
several standard cohorts that may be used by other Study sections and 
task forces as variables for their respective special studies. The design 
of this study was completed in three workshops of the Task Force on Labor 
and Delivery. 

Through the cooperation of Dr. K. Benirschke, the karyotypes of the 
surviving children with Down's syndrome in the Perinatal Research Study 
have been identified through skin and leucocyte cultures. 

II. PROBLEMS 

No problems except for the shortage of professional and clerical- 
I statistical man power. 

9v 



Ill, PROPOSED FUTUEE OBJECTIVES 

The studies on toxemia and labor and delivery will more than fully 
occupy the staff of the Section during the next fiscal year. 

The experience gained in the collection of obstetric data will be 
employed in the planning of a comprehensive prenatal care organization. 



10, 



B. SECTION ON PEDIATRIC KEUROLOGY 
Report for Period July 1, I97O through June 30, I97I 



I. SUMMARY OF SCIENTIFIC OR PROFESSIONAL ACCOMPLISHMENTS : 

A. Logistics : 

Editing of data gathered from examinations of Perinatal Project 
children continues to occupy most of the effort of the Section on 
Pediatric Neurology. This data is provided from pediatric and neuro- 
logic examinations performed at seven years of age and is recorded 
on forms (PED-7^, PED-75, PED-76). In addition, diagnostic summary 
forms are prepared (lDC-77). To date, 21,711 IDC-77 forms (the 
final step in the data recording process) have been completed. 
There is a small "backlog of incomplete forms; 19I forms are in prog- 
ress and kkl are ready for final, hrief review. 

Staff members serve resource and recorder functions on the Four- 
and Seven-Year Outcome Task Force of the Perinatal Research Committee 
(PRC). This Task Force is engaged in comprehensive analysis of 
perinatal and postnatal factors as they may affect the child's 
intelligence at four years of age. 

Tvfo staff members are serving resource and secretarial functions on 
the Physical Growth and Development Task Force of the PRC. 

Members of the Section are participating in the One-Year Outcome 
Task Force. This task force, originating in the Office of the 
Chief, Perinatal Research Branch, will study associations between 
prenatal and perinatal events and the neurologic status of the 
infant at one year of age. 

One staff member has been serving on the Basic Document Committee 
and for other task forces of the PRC. 

Attending Quality Control examinations, conducted at the Collabora- 
tive Study insitutions, occupies 25^ of the time of one staff 
physician. 

The Section continues the task of maintaining a postcard inventory 
of examinations given in order to prevent cases from being lost to 
the Study, 

B. Research Progress : 

Accomplishments in research have been meager in the past year be- 
cause priority for resources to analyze data has been assigned to 
task forces of the Perinatal Research Committee, and to the Basic 
Document effort. 

11 V 



A procedure for definitive analysis of Perinatal Research Branch 
data, designed in 1969? is still being programmed in the Office of 
Biometry. 

The study of electroencephalograms of Project children was termi- 
nated by Perinatal Research Committee action. 

II. PROBLEMS : 

None can be mentioned. 

III. PROPOSED FUTURE OBJECTIVES : 

The axialyses of the One-Year Outcome Task Force and the Four- and Seven- 
Year Outcome Task Force will occupy much attention in the coming year. 

A research design to study spastic diplegia of premature infants is 
being developed. This study might be launched after completion of the 
present commitment to analyze project data which has been accumulated. 



12 V 



C. SECTION ON BEHAVIORAL SCIENCES 
Report for the period July 1, 1970 through June 30, 197I 

S UMVIARY OF SCIENTIFIC OR PROFESSIONAL AGGOMPLISHMENTS 

A? Research 

I. Task Force . The major research effort has been devoted to the 
planning and execution of a study relating findings dirring pregnancy, 
delivery, infancy and early cliildhood to intellectual and motor per- 
formance at four years of age. This research is being carried out in 
collaboration with the Four and Seven Year Neurological and Psychological 
Task Force, Two hundred and seventy one precxarsor variables are being 
related to: (l) Stanford-Binet IQ; (2) the condition of low IQ (mental 
retardation); (3) a factor score representing overall perfonnance on the 
four year battery which includes tests of concept formation and fine and 
gross motor abilities in addition to IQ; (^) a second score representing 
overall test performance which is characteristic of "brain damaged" 
children. 

Completion of all analyses related to the first two outcomes is expected 
by the end of this fiscal year. 

Individual research efforts are continuing in the areas of (l) item 
analyses of the four year battery, (2) heritability of intelligence as 
estimated from the study of twins and sibs, (3) outcomes in children 
from interracial and from consanguineous matings, (h) follow-up of 
children rated as dysfluent at three years of age. A study of the 
relationships of birthweight and gestational age to Bayley scores at 
eight months and IQ at four" years has been con^ileted and is being 
prepared for publication. 

In the area of methodological studies, test-retest reliabilities have 
been established for all sub-tests and overall ratings in the four and 
seven year psychology batteries on two small random samples of children. 
(Ns = lij-0 and 228 respectively) The four-year results are now coxi5)lete. 
This data is collected through the procedure of Inter-institutional 
Quality Control. All children are retested after an interval of 
approximately three months by an examiner from another institution. All 
trials (15 per year) are supervised by a psychologist from this Section. 
He observes the re-testing, records the data and discusses the results 
with the psychologists involved. In the four-year battery, the test- 
retest reliability for the Stanford-Binet is high and the retest-observer 
reliability, which reflects scoring agreement among examiners from 
different institutions, is very high. For the Graham- Ernhart Block 
Sort Test these reliability coefficients are moderate and very high 
respectively. For the three gross motor tests many of the relatively 
small n^umber of children who fail on the first test, pass on the second 
test. This is also true for the fo-ur fine motor tests. For the Overail 
Behavior Rating, the "suspect" category appears to be quite unstable, 
(it is also small), with most of these children being rated as normal on 
the second test. This same tendency is evident in the "suspect" category 

13V 



for the Overall Test Impression Rating. 

In the seven-year battery test -ret est reliabilities for the three WISC 
IQs, the Bender-Gestalt Test, the Aaditory-Vocal Association Test, the 
Goodenough-Harris Drav;-A-Person Test and the three sub-tests of the WHAT 
are high and the retest-observer reliabilities are very high. For the 
Tactile Finger Recognition Test, the test-retest reliability is moderate 
and the retest-observer reliability is very high. The same tendency 
observed in the four year sair^jle for children who were rated initially 
as "suspect" in Overall Behavior and in Overall Test Impression to be 
rated the second time as normal appears in the seven year sample. 

B. Data Collection 

From the period 7-1-70 to 3-l-71j 5^7 four-year psychology examinations 
have been received in the Section. Data collection with this instrument 
has been completed. As of 3-1-71^ S5^ four -year examinations have been 
processed (edited). The current backlog of these examinations is three. 

From the period 7-1-70 to 3-1-71^ 3872 seven-year psychology examinations 
have been received. Projected to 6-30-71^ 5^72 such examinations will 
have been received. As of 3-1-71^ ^015 seven-year examinations have been 
processed (edited). The current backlog of these examinations is 2U2. 

II. Proposed Future Objectives 

These are completion of the four-year study described above and planning 
in collaboration with the Task Force for a second comprehensive study 
using the results of the seven-year psychological test battery as end 
points. 

III. IVtLscellaneous 

Organizational Changes 

The position of research psychologist vacated by Dr. Lee Willerman has 
been filled by Dr. Paul Nichols. 



Ik 



D. SECTION ON INFECTIOUS DISEASES 

Report for pwlod July 1, 1970 through June 30, 1971 

I. SOMKART OF SCIENTIFIC OR PROFESSIONAL ACHIEVaiENTS 

Thft Section la organized into eight Independent but Integrated Units. 
The research activities are divided Into four broad areas: 

A. Large serological surveys of perinatal infections in support of the 
Periaat&l Research Study. 

B. Extended perinstal iiiivestigatlons, including both clinical studies 
and laboratory investigations based on leads froa the Perinatal 
Research Study. 

C. Brain tissua culture studies of perinatal infections and chronic 
neurological diseases. 

D. Hutritioa and infection in aan and prlaates. 

The S^tion published approxisately 38 nanuscripts and presented 42 
papers during the present fiscal year. 

A. Serological Survei^ of Perinatal Infections 

1. Serelogical Investigations Using Co«ple«ent Fixation and 
HsaagglntinatioB Methods 

tephasis has been centered on selected studies of pregnancies with 
abnonal outcones and Matched controls. Tests have been coapleted 
on patients with abortions, stillbirths, cataracts and ■icro- 
cephaly, along with aatched controls, to detemiae the possible 
laflnaace of virus infections in relation to these eutcoaes. 
Several of these studies have now been published; others 
are in press. In each ease the Multiple senn speclnens froa 
apprexlaately 100 patients and 200 natched controls* are used and 
■ere than 20 virus antigens are generally es^loyed. 

Sladlar studies now in progress include cancer of the cervix, 
repeat abortions and stillbirths. 

Studies have Included the use of Australia antigen to provide 
Now inforaation on hepatitis related infections. Recent tests 
have been initiated for Au antibody deteralnations. 

2. Serological Tests Bnploying Tissue Cultures 

Specific tests for cytomegalovirus, rubella, EB virus and other 
antigens are conducted in the laboratory for tissue culture investi- 
gations. This laboratory, under the direction of Dr. Fuccillo, 



15 V 



pcrfonu tlic •pacific neutralization taats as well as flsvraacant 
aatibady determiaatioiui for SB Tlrtts and has daralopad specific 
hcaagflatlnation tasts foz harpcsvims, typas 1 aad 2» as wall as 
cjtf Bgel iMnrir Tha laboratory has alao bean actlvaly lovolTad 
in tha Tims isolation stndias to conf Im the sarologlcal 
obsarrations. This iavolvad tha attcaptad isolation of 
▼irasaa froa ovar 2500 placental spacisans and an oi|«al 
auBbar of throat swab spociaans as wall as tha laboratory 
support for tha isolation of cytonagalovims froa the pregnant woaen 
under study at the Kaiaer Hoapital is Los Angeles and the children 
being atudied in laltlnore and Pr«lerick, Maryland. Ve are alao 
actively involved in isolating rubella virus as well as other 
viruses fron the tissue speclaens obtained fron experiaentally 
infected anlnala and wenen Inannixed for rubella. Keperts of 
these investigations are in press. 

3. laManoglebttlia Peterninations 

To help identify cengenitally Infected children in the Perinatal 
Research Study, we are testing the 30,000 cord sera which are avail- 
able for li^ levela. More than 2/3 of these have been tested and 
the reaaining 1/3 should be conpleted in the next fiscal year. It 
would appear that approzinately 1000 of the 30,000 will have elevated 
1^ levels. With these identified we will proceed with nore specific 
testing for viral antibody in the Iffl conponeut of the serum. This 
will require the use of fluorescent antibody techniques and ultra- 
centrifuge tion . 

B. Extended Perinatal Inveatigations 

Clinical studies have been in progress in four areas. These represent 
an extension of perinatal inveatigations based on leads obtained in the 
Collaborative Perinatal Research Study. Two of these investigations 
are funded by contracta. 

1. Cytaaegalovirus Infections in Pregnant Wonen 

This is a three year study lAich is now In its third year of active 
work. Serial urine apeclneaa are being collected at the Kaiser 
Hospital la Loa Angeles. Pregnant weaen are studied for antibody 
as well aa virus excretion throughout pregnancy and the children 
are ezaained and atudied for the presence of infection at birth. 
Specimens are aent to the Section where the virua isolation and 
antibody detemlnatlons are actually conducted. Virus isolationa 
are being aade and the study is proceeding aa expected. The 
investigation la supported under contract and is in collaboration 
with Dr. Margaret Jones, Professor of Pediatrics, UCLA. 



16 V 



2. Mycoplasiiui Infections In Pregnant Women 

This investigation is now In its third year and is conducted in 
collaboration «d.th the Naval Medical Center, Dr. Melvin Moseles. 
Serial vaginal samples are obtained daring pregnancy and tested in 
our laboratory for the presence of Mycoplasma and T-Strain infection. 
In addition, throat swabs and blood samples are obtained from the 
^ children at birth. Approximately 15Z of babies are found to have 
Mycoplasm infection as are their mothers. The study also involves 
the attempted recovery of Mycoplasma from the ovaries of women 
undergoing hysterectomy. The first phase of work is being 
prepared for publication. 

3. Serial IgM Determinations in High Risk Infants 

High risk and low birtfaweight infants at the University of Tennessee 
are being studied under a contract arrangement with Dr. Sheldon 
Korones. Serial determinations of IgM are being evaluated as a 
means for identifying children with perinatal Infections. The 
study is now in its third year. Reports from this study were 
published this year. 

A. Longitudinal Study of Rubella-Damaged Children 

This is an ongoing study of children at the Johns Hopkins Medical 
Center who are identified as having congenital rubella « The 
longitudinal observations have permitted the association of early 
second trimester rubella with deafness and peripheral pulmonic 
stenosis in the children. Follow-up is being continued to permit 
further identification of more subtle defects, such as mental 
retardation in association with perinatal infection. The study la 
funded under contract and the principal investigator is Dr. Janet 
Bardy. Several publications from this study appeared this year. 

5. Experimental Animals 

Investigations using pregnant rabbits with vertically transmitted 
rubella Infection have demonstrated not only the transmission of the 
infection but the particular predilection of the vims for repli- 
cation in the cartilaginous growing tissue of the fetus. These 
studies are being extended and because of the observations particu- 
lar emphasis is being placed on human clinical material to deter- 
mine if there is an unusual involvement of the cartilaginous tissue 
In fetal specimens of man as well as In rabbits. Recent work on 
the experimental production of vaginal herpesvirus homlnis I 
infection in the Cebus monkey has been published. This animal 
•yatem appears to be an excellent model for the study of vaginal 
and congenital herpes Infections. 



6. Volunf^r Studies 

Huaan Toluntcer studies are being conducted with the use of low 
teapersture adapted vaccine material. The antigenic response and 
shedding characteristics of this vaccine preparation are being 
studiad. 

The effect of aannitol on the excretion of hunan cytomegalovirus was 
tested in volunteers. The drug was found to produce no increase in 
viurea or virus in the nasopharynx. 

7. Drug Evaluation Studies 

Because of the severe damaging effects of cytomegalovirus and Herpes 
simplex virus, studies have been conducted in conjunction with the 
Children's Hospital of the District of Columbia - Dr. Gordon Avery on 
the possible value of drugs for these diseases. These Investigations 
are being reported. In addition investigations on the use of 5-IDU 
for congenital herpes Infection continue in conjunction with Dr. A. J. 
Nahmias. We arc planning a joint collaborative study with Dr. Nahmlas 
and others. 

8. Clinical and Experimental Transmission of Toxoplasmosis 

Because of recent evidence on possible importance of the cat in the 
transmission of toxoplasmosis, an lovestlgatlon was carried out in 
Puerto Rico on the possible shedding of toxoplasmosis by children in 
this incidence population. There was no shedding of T. gondii among 
these children, however serological studies demonstrated the high rate 
of infection which occurred primarily between one and three years of 
age. This report is now in press. 

9. Hepatitis 

Antigen studies utilizing the Australian antigen with complement 
fixation and gel diffusion tests demonstrated the high frequency of 
antigen among children with mongolism at the Pacific State Hospital. 
Comparative studies in other populations are now in progress as are 
investigations of the experimental transmission of the virus In monkeys 
and the development of improved techniques for antigen and antibody 
determinations. Congenital transmission of Australia antigen has 
been found to occur. An epidemic of Infectious hepatitis involving 
over 300 patients has been studied. Au antibody has been tested for 
in mothers of mongoloid children. Reports of these studies are in press. 

10. Immune Mechanisms in Perinatal Infections 

It has become increasingly apparent that specific studies on the Inmune 
mechanisms of the developing fetus and newborn are needed so that we may 
better understand the deficiencies which permit congenital infection and 



18 V 



chroalc Infection to occur in the fetus and persist In the child. Experl- 
■ental studies utilizing the haaster, rat and monkey are In progress In 
vfalch coRblned antibody, delayed hypersensitivity and interferon deteral- 
natloQS are nade serially In Infected and non-Infected anlnals as well as 
In association vlth antigenic stlaalatlon. 

11. Cytoacgalovlrns - Serotypes Associated with Clinical Disease 

At least three strains of cytoaegalovlrus appeared to be Identifiable. 
The iaportance of the various strains In clinical disease is unknown. 
Cosparatlve serological tests are now in progress in our laboratory to 
try to distinguish these strains and correlate this with the clinical 
findings In the patients. In addition because of the high frequency of 
infection and disease with c3rtoaegalovirus in patients receiving iasrano- 
suppresslve drugs and/or patients with leukemia, collaborative studies 
with investigators In the National Cancer Institute are in progress in 
which strains of cytomegalovirus are being isolated and compared to each 
other and the clinical findings in the patients. Reports are being 
prepared for publication. 

C. Brain Tissue Culture Studies 

With the isolation of measles virus from the brains of patients with 
subacute sclerosing panencephalitis, particular eaq>hasls has been placed 
on the comparison of various strains of virus isolated from these patients 
and characterization of these strains in relation to wild measles virus 
and vaccine virus. In addition Imswnologlcal studies have proceeded and 
doMnstrate no specific lanme deficiency in the patients with the disease. 
Additional reports on these topics have been published by the laboratory 
this year. Epidemiologic investigations conducted in conjunction with 
Dr. Jabbonr at the University of Tennessee demonstrated that approximately 
one-half of the patients identified are in the southeastern part of the 
United States. Additional investigations are emphasizing Creutzfeldt- 
Jakob Disease, progressive multifocal lenkoencephalopathy, multiple scler- 
osis and Parkinson's Disease r utilizing both biopsy and autopsy specimens. 
Work being reported this April desonstrates the presence of chronic 
suppressed measles infection in the Ijrmph nodes of children with SSPE. 
This finding significantly changes the concept of the infection from 
one localized in the central nervous system to a generalized Involvement. 
T— ime suppression may be related to this disseminated infection. 

II. PROBLEMS 

The past year has seen a considerable turnover of personnel. Difficul- 
ties arose because of the loss of technicians «uid statisticians in the 
face of an increasing demand for testing. 

Further, losses of technical personnel, either through resignation, 
maternity leave, or leave without pay, has slowed down the effort in the 
exploration of serum samples from the Collaborative Perinatal Program. 



1<) 7 



E. SECTION ON PATHOLOGY 
Report for the Period July 1, 1970 through June 30, 1971 
I. SUMMARY OF SCIENTIFIC OR PROFESSIONAL ACCOMPLISHMENTS 
A, Project Activities and Material 

1. Pathology Task Force 

On October 7, 1970 the first meeting of the Pathology Task 
Force was held in Bethesda under the co- chairmanship of 
Drs. Stanley Aronson (Providence, Rhode Island) and 
Lewis E. Lipkin. The following pathologists, in addition 
to the co-chairmen, agreed to serve as members: 

Dr. Shirley G. Driscoll (Boston, Massachusetts) 

Dr. Floyd Gilles (Boston, Massachusetts) 

Dr. Bernard Klionsky (Pittsburgh, Pennsylvania) 

Dr. Haruo Okazaki (Rochester, Minnesota) 

Dr. Edward P. Richardson, Jr. (Boston, Massachusetts) 

The project neuropathologic material was considered in 
considerable detail by the Task Force. Its characteristics 
and the implications for data development from such specimens 
were explored. It was agreed in general that, in addition to 
modifications in the then current analysis and reporting 
procedure, the material warranted several in-depth studies. 
The latter are under exploration at this writing and will be 
noted under "III. Proposed Future Objectives." 

2. Case Integration and Analysis 

Following the last Task Force meeting, considerable effort was 
devoted to completing the procedural modifications begun last 
year (c.f. I,A,1. FY 69-70). In designing these changes, 
particular attention was paid to meeting the needs of the other 
project sections and the contributing institutions, while at the 
same time accelerating the initial review process. The danger 
inherent in such partial workup of an individual case is always 
the possible necessity for repeating the work at a later date. 
In reformulating procedures, relaxation of previously established 
standards was necessary in several areas. In particular: 

a. The deliberate maintenance of ignorance of clinical aspects 
by the examiner was sacrificed. 

b. The usual multiple stain examination of the same structure 
was deferred. 



21 V 



At this time only a limited number of tissue blocks are checked 
and only the H&E sections examined, rather than as previously 
all blocks and all four stained slides for each. A procedure 
for block selection based on gross photographs has been developed 
as has a new histologic protocol. The latter not only 
facilitates the examination, but serves to document the scope 
of the current examination, thus reducing future duplication. 

3. Tissue Processing and Preservation 

Almost the entire technician effort in this laboratory is 
devoted for the most part to: 

a. The preparation of histologic slides from the paraffin 
embedded blocks. 

b. The preservation of the remaining wet tissues which are 
regarded as "public documents" in the same sense as project 
forms or sera in other sections. 

For long-term efficiency, we have been preparing a full 
complement of three special stains and one unstained blank for 
each block of every case. In order to meet the requirements 
of the accelerated review, it has been necessary to dispense 
with all blanks and special stains except in urgent instances, 
and to prepare only a single H&E slide for each block. During 
the first phase of this effort more than 100 cases were so 
processed making nearly 3,200 H&E sections available for 
examination later in the calendar year. 

The remaining technician effort is largely devoted to regular 
checking and filling of storage containers (specimens are 
preserved in 807o alcohol when total embedding in paraffin is 
not feasible). Despite diligent efforts, no fully satisfactory 
bagging method is available. We therefore are in the process 
of transferring all specimens to specially selected and sealable 
polyethylene containers which will reduce the need for 
continual checking and filling. 

B. Special Studies 

1. Biologic Pattern Data Processing 

This project was subject to two commendatory reviews during the 
past calendar year. The planned-for quantitative analysis of the 
parameters of chromatolysis has been begun and data regarding 
cell size and shape, quantity and dispersion characteristics 
of Nissl substance, nuclear and nucleolar eccentricity, 
nuclear size and shape, etc., are being determined on hundreds 
of chromatolytic and contralateral control neurons which have 
been automatically scanned. The simultaneous determination of 



22 V 



DNA content and synthesis rate in Feulgen stained radio- 
autographs is proceeding in tissues of a transplanted lymphoma, 
and about to be begun in embryonic neural tube. The marrow 
and blood cell identification component of the grain counting 
system being specified for NCI is in progress. 

Extensive equipment additions and modifications have been 
accomplished without preventing the scanning microscope from 
being used for data acquisition on the above mentioned projects. 
These are detailed in the relevant individual project report, as 
are the numerous computer programs created, debugged and made 
available for general image processing use. (Serial No. NDS 
(CF)-65 PR/P 1278) 

2. Twin Placentation in Relation to Zygosity 

The relationship between type of twin placentation and zygosity 
was studied in 569 sets of twins in the Collaborative Study. 
Perinatal death rates, pathologic findings on deaths, birthweight, 
gestational age and birthweight differences were evaluated in 
relation to twin placentation. This paper has been published 
and the study is completed. (Serial No. NDS (CF)-68 PR/P 1650) 

3. Kidney Malformations in Fetuses of AxC Line 9935 Rats 

The rate of spontaneous genito-urinary malformations in AxC 
line 9935 rats is being determined. Randomly selected 
pregnant rats are killed at or near term and the mothers and 
fetuses were examined in detail for malformations involving 
the genito-urinary system. This paper has been published and 
the study is completed. (Serial No. NDS (CF)-69 PR/P 1763) 

^- Placental Study of Abortion Material (Obtained by an Induced 
Abortion ) 

A detailed histological review of more than 100 specimens of 
induced abortion material was undertaken by Dr. Fujikura in 
cooperation with the Department of Anatomy at Kyoto University 
in Japan. The material was compared with the products of 
spontaneous abortions. The paper has been accepted for 
publication. (Serial No. NDS (CF)-69 PR/P 1764) 

5. The Interrelationship Between Selected Congenital Malformations 
and Major Pathologic Findings 

The relationship between selected malformations in autopsied 
deaths and major pathologic factors in the baby as well as in 
the mother and placenta are being analyzed. (Serial No. NDS 
(CF)-69 PR/P 1765) 



23 V 



6. Reproductive Ability of the American Negro with Sickling 
and its Public Health Implications 

The reproductive performance of 654 sicklers and 1,890 
nonsicklers in the Collaborative Study was compared. The 
cumulative fertility rate of mothers with sicklemia was the 
same as that of nonsickling mothers. Because of evidence 
that mothers with sickling have normal reproductive abilities, 
the gene will continue to propagate and a plea is being made 
that sickling tests be done on all Negroes. (Serial No, NDS 
(CF)-69 PR/P 1766) 

7 . The Significance of Chorioangiomas 

A review of 81 histologically verified cases of chorioangioma 
showed an increased association with neonatal thrombocytopenic 
purpura, toxemia, fetal hemangioma, erythroblastosis, and 
single umbilical artery. There is a distinct female preponderance 
and the condition is more common in whites. The prematurity rate 
in the surviving group is not different from that of single live 
births in the Collaborative Study. The paper has been published 
and the study is completed. (Serial No. NDS (CF)-69 PR/P 1769) 

8. Pathologic Effects of Ligation of the Anterior Spinal Artery 
and/or the Great Radicular Artery in Monkeys 

Spinal cords of monkeys which had been subject to ischemia by 
means of surgical ligation of the anterior spinal artery and/or 
great radicular artery. The ligation of the anterior spinal 
artery just below its anastomosis with the arteria radicularis 
magna of Adamkiewicz produced clinical paraplegia and severe 
necrotic changes in the spinal cord caudal to this level. The 
ligature of either the arteria radicularis magna or the anterior 
spinal artery above the anastomosis produced anoxic changes which 
might be only temporary and were not reflected by serious clinical 
deficits. These findings are significant and of practical 
application in case one of these arteries must be sacrificed or 
is involved in a pathologic process. The publication is in 
preparation. (Serial No. NDS (CF)-69 PR/P 1772) 

j 

9. Thrombocytopenic Purpura and Placental Hemangioma 

This combination heretofore unreported and found in two cases in 
the Collaborative Study is discussed. The paper has been 
published and the study is completed. (Serial No. NDS (CF)-70 
PR/P 1858) 



2k 



I 



10. Mental and Motor Development in Monozygotic Co-Twins with 
Dissimilar Birthweights 

Eight-month mental and motor series as well as four-year I.Q.'s 
of 125 sets of identical twins in the Collaborative Study who 
had unequal intrapair birthweights showed no difference in these 
parameters between co-twins. Since the lighter of monozygotic 
twins presumably suffers drastic nutritional setbacks in utero 
compared to its co-twin, these findings suggest that the human 
brain is fairly resistant to the effects of intrauterine 
malnutrition. Submission of this paper for publication is 
being suspended pending data analysis of the seven-year I.Q. 
(Serial No. NDS (CF)-70 PR/P 1859) 

11. A Follow-up of Children with Single Umbilical Artery 

A follow-up study of 355 cases of single umbilical artery is 
being undertaken with particular emphasis on the somatic, 
mental and motor development of those who have reached four 
years of age. (Serial No. NDS (CF)-71 PR/P 1911) 

12. Birthweight in Relation to Renal Glomerular Development and 
Gestational Age in Whites and Negroes 

A histologic evaluation of the kidneys of 514 neonatal deaths 
and stillbirths in the Collaborative Study was made and the 
findings assessed in relation to birthweight and gestational 
age. Racial differences are discussed particularly in regard 
to birthweight and in infants whose birthweights do not match 
gestational age. (Serial No. NDS (CF)-71 PR/P 1912) 

13. The Clinical Signifiance of Generalized Petechiae at Birth 

In a pilot study about a quarter of the infants diagnosed as 
having had generalized skin hemorrhages at birth, had significant 
changes in the form of marked hearing loss, speech and behavior 
problems and, one instance of mental retardation accompanied by 
distinct visual impairment. On the hypothesis that significant 
skin petechiae could be accompanied by hemorrhages in internal 
vital structures such as the brain, inner ear, retina, etc., a 
careful evaluation of the balance of these children is being 
made and findings compared with those of matched controls. 
(Serial No. NDS (CF)-71 PR/P 1913) 

Other Activities 

There have been numerous presentations of work done within the 
section. Presentations at the first Biological Congress, the 
SPIE, and the New York Microscopical Society have covered various 
aspects of the work in image processing. The last meeting in 
May 1971 listed three individual talks by involved personnel. 



25V 



The Head of the Section has served as contract officer for the 
collaborative program with the Artifical Intelligence Group of 
the National Bureau of Standards as described in the individual 
project report. The Section Head continued his working relation- 
ship with the Image Processing Group, DCRT and the National Cancer 
Institute. 



II. PROBLEMS 



A. Personnel 

The basic problem of a contracting, financial and personnel base 
and the concurrent desire to evaluate the overall nature of the 
pathological material has made for a serious problem. The need 
for additional professional personnel is undoubted, and can only 
partially be met by developing work-type task forces. Additional 
professional personnel will require additional filing and clerical 
help since slides alone are examinable at times at faster rates 
than they can be refiled even with our present relatively minisucle 
professional staff. 

PROPOSED FUTURE OBJECTIVES 

A. The accelerated overall case review has and will continue to receive 
first professional and technical priority within the section. At 
this writing, the results of the first 35 microscopic examinations 
completed under this revised procedure are under review, in order to 
assure ourselves that no information essential for other sections or 
the Collaborating Institutions is likely to be slighted. 

B. Several in-depth studies are in the planning stage. These include: 

1. A population study of selected obstetric variables within the 
autopsy population in order to elicit suggestive structures, 
lesions or patterns to be specifically searched for within the 
material. Dr. Rudolf Vollman, Head, Section on Obstetrics, has 
agreed to collaborate with us in this study. 

2. We are hoping to interest members of the Pathology Task Force 
in participating in studies of such characteristic lesions of 
the newborn period as the Virchow or Banker lesions of the 
periventricular white matter. These lesions are of particular 
interest, and our material represents a unique opportunity for 
a definitive study of such changes. 

3. Collaboration with the Task Force on Genetics in an extensive 
study of sickling. This will largely be under the direction of 
Dr. Froehlich who has for several years pointed out the importance 
of such work in the context of our placental material. 



26 V 



We continue our plans to apply the technics developed by the 
biologic image processing project to the project neuropathologic 
material. Current work on neurocellular degenerations such 
as chromatolysis will provide the basis for such future 
applications. 



I 



27 V 



F. SECTION ON EPIDMIOLOGY AND GEHETICS 
Report for the period Jiily 1, 1970 through June 30, 19T1 

I. SUMMARY OF SCIEIiynFIC MP PROFESSIONAL ACCOMPLISHMENTS 

During the past fiscal year the major effort of this Section was 
directed towards organizing the task force on Genetics and Congenital 
Malfoimations; implementing the comprehensive plan for analysis of 
genetic and socioeconomic data which was developed hy a panel of genetic 
experts last year and was adopted and extended by the task force on 
Genetics and Congenital Malf oimations ; reorganizing and reorienting the 
previous data analysis efforts; improving the quality of the data and 
completing oux data file. 

The Section has continued to receive, edit and code infoimation on the 
Family Health History Review form (FHH-9). Daring this period, a-pproxi- 
mately 6,000 FHH-9 forms were received and approximately 5^000 have been 
processed and sent to punch. The FEIH-9 is being used to derive a new 
socioeconomic index for comparison with that derived for each family 
seven years earlier. For this, the family income item is being rescored 
with a more realistic ceiling of $15,000 rather than $10,000 used on the 
Socioeconomic Interview foim (SE-l). This will require recalling of all 
schedules already processed but it is anticipated that they can be re- 
scored without undue delay. 

The most significant event in the operations of this Section diiring the 
last fiscal year has undoubtedly been the creation of task forces to plan 
euid direct all data analysis in particular areas, . The task force respon- 
sible for the analysis of genetic and socioeconomic data is that of 
Genetics and Congenital Malformations, which at present is composed of 
Drs. Richard Osborne, Kuxt Hirschhom, Walter Nance, William J, Schull, _, ■ 
and Ntinos C. Myrianthopoulos, assisted by professionals from the Office 
of Biometry, and the Perinatal Research Branch. The task force idea is 
not new to this Section. As reported in last year's Annual Report, this 
Section had already convened a pan.el of genetic experts to discuss problems 
of genetic analysis and help develop a comprehensive and workable analysis 
plan. That group had been most successful in developing such a plan and 
the newly-created task force adopted it into, and has proceeded to im- 
plement it and extend it in special areas outside the protocol. 

Thus the efforts of this Section toward data analysis are vigorous and 
the prospects seem good. Some projects have been completed, some are 
now in progress, some have been approved for immediate implementation, 
and some are now being planned. These are described in detail in the 
indiATidual project reports. One part of the genetic study of intellectual 
and motor perfoimance which is being done in collaboration with investi- 
gators at the University of Minnesota has been completed. Among some of 
the interesting preliminary results of this study are that the within 
family environmental variance is larger among Negroes than among whites, 
which means that the heritability of mental perfoimance is lower among 

i 29v 



among Negroes than among whites; and that the coiirparison of the if-year 
to the T-year IQ scores shows aji improvement in twins hut a decline in 
singletons which may mean that the twins who usijally perfoim poorer than 
singletons during their early period of life, may he ahle to catch xjp 
during childhood and adolescence. Dr. Kaylor's study with Dr. Warbiirton 
on the effect of parity, matenaal age and change of mate on placental 
weight and birthvreight has also heen completed and appeared in the 
January issue of the American Journal of Humsui Genetics, 

The task force, operating always on the premise that the major contri- 
bution of genetic ajialysis \-rlll be in the area of special studies, has 
approved a nijmber of new projects. Among these are, a comprehensive 
study of congenital malformations, a genetic study of obstetric variables, 
a study of ABO and Rh incompatibility in pregnancy wastage and infant 
survival and a study of heritability of twinning in man. 

The task force during their meetings and discussions took the position 
that analysis of Collaborative Study data should not constitute an end 
in itself but should be used to locate areas in which specific problems 
lie. These problems shouJ.d then be pursued as logical extensions of the 
objectives ajid scope of the Collaborative Study, In accordance with these 
guidelines the task force recommended to the Perinatal Research Committee 
that the unique population of twins bom to mothers in the Collaborative 
Study be followed to at least 15 years of age so that observations on 
their mental and physical development can be made at least through puberty. 
The Perinatal Research Committee approved the proposal and the task force 
is now in the process of developing special protocols for yearly examina- 
tion of these twins and special neurological and psychological examinations 
at ages 12 and 15 years. 

The task force has approved the participation of this Section in a 
chromosomal study of children from five collaborating institutions to 
relate major and minor chromosomal aberrations to abnormalities found in 
these children. The participating institutions are in Boston, Mass., 
Philadelphia, Pa., Birffalo, N.'Y,, Memphis, Tenn,, and Portland, Ore, Dr, 
H, Lubs of the University of Colorado is acting as a coordinator for the 
study. The task force has approved the participation of this Section in 
a comprehensive study of sickle-cell anemia to relate the occurrence of 
sickling in the mother with the outcome of pregnancy and to study the 
growth and development of children who are themselves sicklers. The task 
force has also endorsed a proposal by Dr, J.V,' Weel of the University of 
Michigan to establish genetic markers using a variety of polymorphic blood I 
loci for correlations with pregnancy outcomes. Because of difficulties 
with funding of such an extensive project it was decided to draw and 
store the necessary blood samples now and type them at a more convenient 
time later. Since there is a reasonable probability that the effects of 
these genes as well as tha-t for sickle-cell anemia may be influenced by 
the degree of black ancestry the task force recommended that a simple 
estimate be made of skin color as an index of this ancestiy for control 
purposes. 

The task force on Genetics and Congenital Malformations has been in 

30 v 



constant contact and collalDoration with other task forces with common 
inte2re,its in data analysis. Dr. Myrianthopovilos and Dr. K. French of 
the University of Oregon are collaborating with the task force on four 
and seven year examinations to provide the demographic and socioeconomic 
description of the population; a joint study is planned with the task 
force on physical growth and development of the physical growth and de- 
velopment of twins; advice and help has "been provided with the analysis 
of some of the speech and hearing data to the appropriate task force; 
and suggestions have been made to the task force on infectious disease 
concerning new approaches for analysis of the relationship between vital 
antibodies and congenital malformations. 

Dr. Waylor has been instrumental in completing three files of special 
genetic interest. One is the consajiguinity file whose codes have been 
exhaustively hand-reviewed. All efforts are now being made to resolve 
code discrepancies, especially in second and subsequent pregnancies. 
Another is the record linkage file which involves over 6,000 women who 
reported having close relatives participating in the Study. This has 
now been completed and all the infoimation on the women and their repor- 
ted relatives has been put on tape. A hand review of another file, that 
of interracial matings has also been completed and this file now contains 
199 cases with known outcomes. 



II. PROBLEMS 



The major problem which has confronted the Section during the past year 
has been that of the shortage of trained professionals to direct and 
participate in data analysis. As in past years, the Section has tried 
to resolve this problem by inviting qualified professionals from within 
and without the Collaborative Study to participate in data analysis. 
This plan has for the most part been successful but is no substitute for 
the constant presence of the investigator at the Section headquarters 
and his immediate access to all data. 



III.- FUTUKE OBJECTIVES 

The future objectives of the Section are to implement the analysis of 
the genetic and socioeconomic data of the Collaborative Study according 
to the plan of genetic analysis which was developed with the help of 
genetic experts and the task force on Genetics and Congenital Malformations, 
Although the plan calls for collaboration with investigators and insti- 
tutions outside the Collaborative Study, there is no doubt that the plan- 
ning and direction of research must come from this Section. A second 
objective, therefore, is to strengthen our professional staff in the 
Section, 

At the same time the Section, with the advice and guidance of its task 
force is continuously trying to identify specific areas of fruitful 
■ research from the first round of analysis, which, although not within 
the protocol, should be pursued as logical extensions of the objectives 

3IV 



of the Collaborative Study. Considerable progress in this area has 
already been made. 

MISCELLANEOUS 

A« Personnel 

Tlie personnel of the Section on Epidemiology and Genetics consists at 
present of the following: Professional, Dr. N.C. Myrianthopoulos, Head, 
Dr. A.F.I Waylor, geneticist. Miss T.L.' Martin, statistician; one secretary, 
five statistical assistants. Daring the last year this Section lost two 
positions, that of Miss A. Baszynski, fieldworker, and that of one sec- 
retary. The Section is now recruiting for a fieldworker whose duties 
vrould be to assist in the follow-up of the t-(-7ins to age 15 years. 

B. Activities of the Section Head and the Professional Personnel 

The Head of the Section, Dr. Myrianthopoulos, in addition to his foimal 
duties has continued his independent investigations in the genetics of 
neurological disorders, especially the lipidoses, and has contributed 
chapters in several textbooks on these topics. Dr. Myrianthopoulos has 
maintained his affiliation with George Washington University School of 
Medicine as a Clinical Associate Professor of Neurology and Director of 
the Genetic Counseling and Research Center. He is a member of the Medical 
Advisory Board of the Huntington's Chorea Foundation, a private orgajii- 
zation which supports research in Huntington's chorea, and of the Commit- 
tee upon Huntington's Chorea of the V/orld Federation of Neurology. Dr. 
Myrianthopoulos has been elected a member of the Academy of Medicine of 
Washington, D.C 

In September, 1970, Dr. Myrianthopoulos attended the Fourth Annual Work- 
shop of the World Federation of Neurology Eesearch Group on Huntington's 
Chorea in Munich, Geimany, where he chaired the session on Genetics and 
Epidemiology and discussed the current status and future prospects of 
research in Huntington's chorea. In January, 19T1, Dr. Myrianthopoulos 
also attended a meeting of a special committee of this research group in 
Louvain, Belgium, during which plans were made for the Huntington Centen- 
nial Symposium in Columbus, Ohio for April, 1972 and for the preparation 
of a complete bibliography on Huntington's chorea. In addition. Dr. 
Myrianthopoulos helped to set up a special study of Huntington's chorea 
in the low countries and the north of Fi-ance. Dr. Myrianthopoulos also 
attended the annual meeting of the American Society of Human Genetics in 
Indianapolis in October, 1970, where he presented a paper on respiratory 
distress syndrome in twins. 

Dr. Naylor participated in all of the meetings and discussions of the 
task force on Genetics and Congenital Malformations and has been instru- 
mental in designing many of the ongoing studies, often doing the program- 
ming himself. As mentioned earlier, he also designed and completed three 
data files of special genetic interest. Dr. Naylor attended the annual 
meeting of the American Society of Human Genetics in Indianapolis in 
October, 1970, where he presented a paper based on Collaborative Study 

32 V 



data adducing evidence for a powerful parity effect in spontaneous 
a.bortion. 



Bibliogi-aphy 

Myrianthopoulos, N.C,', Naylor, A.tF.I and Aronson, SoMJ: Tay-Sachs disease 
is probatly not increasing. Nature 22^:60^, 19T0» 

Myrianthopoulos, W.'tiJ: A survey of twins in the population of the 
Collaborative Study. Acta Genet. Med. Gemellol. 19:15-23, 19T0o 

Mjrrianthopo-ulos, W.C,'': Review of "Genetic Counseling. A Guide for the 
Practicing Physician" "by W, Fuhrmann and F. Vogel. Social Biology 
17:244-246, 1970. 

Myrianthopoulos, W.C.': Genetic aspects of multiple sclerosis. Handbook 
of Clinical Neurology Vol, 9, Ch, 5^ Amsterdam, Worth -Holland 
Publishing Co,, 1970, 

Aronson, S,M,* and Myrianthopoulos, N.C: Epidemiology and Genetics of 
the Sphingolipidoses, Handbook of Clinical Neurology Vol. 10, Ch. 24, 
Amsterdam, North -Holland Publishing Co., 1970. 

Willeiman, L,, Naylor, A.F,' and Myrianthopoulos, N.C: Intellectual 
development of children from interracial matings. Science 170:1329- 
1331, 1970. 

Myrianthopo-ulos, N.CJ: An epidemiologic survey of twins in a large, 
prospectively studied population. Am. J. Hum. Genet .. 22:611-629, 
1970. 

Warburton, D. and Naylor, A.F.^: Effect of parity on placental weight 
and birthweight: An immunological phenomenon? A report of the 
Collaborative Study of Cerebral Palsy. Am. J. Hum. Genet . 23:4l-54, 
1971. 

Myrianthopoulos, N.C,^: Respiratoary distress syndrome in twins. Acta 
Genet. Med. Gemellol . in press. 



33 V 



G. SECTION ON DATA MANAGEMEIW AND RETRIEVAL 
Report for Period July 1, I97O through June 30, I97I 



I. SUMMARY OF ACCOMPLISHMEMS 

This Section is responsible for recei-\n.ng, coding, filing and storing 
forms in accordance with a system designed to facilitate form and/or 
data retrieval. The Acting Head of this Section has continued in this 
assignment from the Office of Biometry umtil June 1, 19?!^ at which 
time Dr. B. H. Fox Asst. to the Chief, Perinatal Research Branch 
became the Acting Head. He also supervises the systems analysts and 
programmers providing services to PRE from the Office of Biometry to 
aid in data analysis. The current data file consists of 58,806 
registrants -- of this number 5^,177 were core study cases. Of these 
core study cases 53^837 have delivered. 

Approximately 3'^ million forms will have been submitted by June 30, 
1971' The available data from approximately 3-1 million of these forms 
has been processed into over 5*9 million piinch cards in I5I card 
formats and converted into active computer tape files, h-9,631 births 
have been reviewed at 1 year of age, ^3^888 at k years of age and 
32,194 at 7 years of age. 

Personnel staffing, with the expected decrease in volume of study forms 
received, decreased from 33 "to 27 . Personnel have been encouraged to 
take advantage of training being offered through career counselling and 
self -improvement requests in an effort to permit a continued smooth 
transition. 

During the year, work was continued in extending the development and 
use of complete program packages to provide an extended Variable File 
System with update and editing in conjunction with PRB operated master 
files and data banks, thus reducing future cost of operations. In 
addition several standard programming packages have been utilized 
during this fiscal year period for both studies and statistical analysis 
of data greatly facilitating this type of work previously done by other 
methods. This included the use of Express III System, Data Check System, 
Table Mak.er 2, Inquiry Response System, and Remote Terminal programs. 
Personnel have been trained in remote terminal operation for providing 
appropriate services. 

The research activities of PRB were supported with special infonnation 
retrieval for case selection, tabulations, and creation of subfiles of 
the Variable File System. Also a major effort has been accomplished 
with the production of quantities of tabulations for large-scale studies 
involving considerable amounts of data, utilization of dedicated 
equipment, extensive systems analysis and complex data processing. A 
major project was the production of Monograph II in addition to major 
assignments received from the Research Task Forces organized last year. 
•Supplementary production of tables for the Basic Doc\jment was also 
produced during the year. 

35^ 



The Forms Accountability Program for review of study forms was 
continued with the objective of maximizing of all recoverable data for 
inclusion in the PRE master files and data banks. A quality review 
system of file folders has continued with review of coding, organizing 
and binding of forms in the file folders which facilitate their use 
while preventing loss or misfiling of individual study forms. This 
has been extended by the initiation of file clerk review of folders 
for organization of forms in a standard order and identification and 
refiling of misfiled forms. An inventory control card system is 
completed summarizing the status of each file folder and included in 
the file. 

II. PROBLEMS 

The Section, which contains a large number of non-professional staff 
has continued shifts of assignment of personnel including retraining 
where required to meet program needs. Continued efforts for 
retraining and placement of personnel will help in problems that can 
be expected to arise with a reduced workload in data collection. 

III. PROPOSED FUnmE OBJECTIVES 

Continuation in the development of sophisticated information retrieval- 
systems to supply research needs more rapidly is planned with the 
integrated use of dedicated computer equipment and maximation of remote 
terminal operation. The conversion and retraining of staff will 
continue so that present personnel can be reassigned to functions as 
required. New systems development are planned to be continued which 
will fijTther effect reduced turnaround time on study requests as well 
as economy of operation. For example, improved file structure systems 
tailored to groups of studies with increased flexibility of use are 
planned. 



36 



H. SECTION ON PROJECT SH^VICES 
Report for the period July 1, 1970 through June 30, 1971 

SUMMARY OF ACCOMPLISHMENTS 

The Section on Project Services, Medical Literature Unit, is responsible 
for the acquisition of selected medical and statistical textbooks, 
abstracts, scientific directories, atlases, annual publications of books, 
and periodicals, and the organizing and indexing of published literature 
on all medical- scientific subjects, which in any way contribute to the 
various studies undertaken by the Perinatal Research Branch. The present 
book collection in the PRB Reading Room numbers 1082. Subscriptions to 
leading journals and abstracting periodicals pertinent to the program now 
total 102. In the interest of economy, a number of journals not being 
utilized to the optimum were discontinued during the past year. Up-to-date 
listings of journals and new book acquisitions are now in the process of 
compilation and will soon be circulated to Branch personnel. In addition, 
our locator file, containing a listing of books by author and title, now 
numbers 1109, and controls the books housed in the various sections of the 
Branch, readily retrievable upon request. 

Approximately 1200 visits were made to our Reading Room this past year. 
Its facilities were utilized by PRB personnel, consultants, task forces, 
and others. 

The activities of the Unit are wide and varied and encompass the following 
functions: 

Provides a Reading Room and maintains a lending library. 

Processed books and/or journals borrowed from PRB Reading Room collection; 
processed book, journal and photocopy requests, through NIH-NIM Library,^ 
procuring for Branch personnel, items not available in PRB Reading Room; 
xeroxed from journals and/or books, articles of interest pertinent to the 
program, and circulated these to the staff; provided a quick service for 
acquisition of literature, urgently needed and not available in-house, 
from NIH Library. In most instances material was procured the same day. 

Xeroxing on the premises of approximately 25,100 pages of material, 
requested by PRB Staff. 

Dissemination of material and information pertinent to the Program and in 
answer to a variety of inquiries and requests from the scientific and lay 
community. This was accomplished by correspondence, by telephone and by 
mail. 

Periodic mailing of NINDS publications and reprints relevant to the Study 
and of possible interest to PRB Staff, Project Directors, Consultants, 
Perinatal Research Committee and Task Forces. 



37v 



Annual distribution of prepared lists regarding new acquisitions of 
books, journals and abstracts, etc. to those utilizing our facilities. 

Literature searches for specific articles, subjects or authors as 
requested by investigator. 

Compilation and maintenance of mailing list for annual distribution of 
Bibliography of Collaborative Perinatal Project Publications. 

Coordination and compilation of Annual Report in proper format for 
submission to NINDS Director. 

Book ordering for PRE Reading Room. 

Compilation of Bibliography of Collaborative Perinatal Project Publica- 
tions. This is updated annually at the end of each fiscal year and 
includes publications based on pooled core data, local core data, and 
non-project studies of special interest to authors at PRB and those 
from collaborating institutions who are paid in part or in toto from 
Project funds. During the past year a Quarterly Bibliography of all 
COLR publications, with summaries, was initiated to enhance the visibility 
of our research effort on a more concurrent basis than is possible with 
the Annual Bibliography. 

Collection of publications by authors from the collaborating institutions i 
prior to 1965 which fall into the following categories: publications 
using Project population, publications of personnel supported in part 
or in toto by Project funds, and ancillary studies. 

Indexing reprint collection. These are reprints in the perinatal area 

of critical interest to our scientists. They are retrievable on demand 

by author, subject and number. Subject headings parallel those used in 
Index Medicus. 

Semi-annual preparation of bibliography of Branch personnel publications 
for inclusion in Scientific Directory and Annual Bibliography, NIH. 

Clearinghouse for scheduling of meetings and conferences in the PRB 
Reading Room. During the past year 84 meetings were hosted and on 
occasion information was gathered and material xeroxed and collated for 
use at a specific meeting. 

Reorganized our Collaborative Project reprint file for instant accessi- 
bility and retrieval of material. 

Arranged tours of National Library of Medicine and Clinical Center for 
those foreign visitors to the Branch who were interested and also 
collected literature of interest for them; arranged for the loan of 
equipment through NIH Audio Visual Unit for use by PRB Staff members, etc. 



38 



II . PROBLEMS 



Backlog is the big issue and continues in the following areas: 

Subject indexing of reprints for Reprint Retrieval File 

Cutting, annotating and filing of reprints 

Collection of COLR reprints published prior to 1965 from 

collaborating institutions in our continuing effort 

to update the file. 



III. PROPOSED FUTURE OBJECTIVE S 

Will continue to concentrate on giving service and meeting the needs of 
the Branch in literature retrieval, special reference searches, 
compilation of bibliographies, etc., as requested. 



I 



39 V 



I. SECTION ON SPEECH, LMGUAGE AMD HEARING 
Report for the Period July 1, 1970 through June 30, 1971 

I. SUMMARY OF ACTIVITIES AND DEVELOPI/JENTS 

A. Routine 

1. Processing of 3-year Speech, Language and Hearing (SLH) forms. Status: 
essentially completed (15I forms received last year). 

2. Collection of 8-year SLH data. Status: continuing at six 
collaborating instititions. 

3. Processing of 8-year SLH forms. Status: continiiing (l3,711 forms 
edited to 2-28-71). 

k. Supervision of inter-institutional quality control. Status: 
continuing on trimester basis. 

5. Project site visits by Section Head as Project Officer. Status: 
continuing. 

6. Collection of data for Language Organization Scales. Status: 
continuing at q.uality control visits. 

B. New 

1. Development of forms and method for machine tabiilation and analysis 
of quality control results. Status: essentially completed. 

2. Organization of SLH Task Force for data analysis. Status: completed 
and in operation. 

a. Section Head f\mctions as coordinator. Speech Pathologist as 
secretary for SLH Task Force. 

3. Development of data analysis plan. Status: first steps taken. 

h. Analysis of concordance in case identification by exams at 8-years, 
7-years and 3-years. Status: corripleted. 

5. Tabulation of first 5000 8-year exams by test, institution, race, sex, 
birthweight, I.Q. and education of gravida. Status: completed. 

6. Tabulation of cases with dysfluency, preliminary to devising study 
proposal. Status: completed. 

7. Preliminary look at cases with significant petechiae and. abnormal 
speech, language or hearing findings. Status: in operation. 



I 



i4-lv 



C. Discontinued 

1. Eight-year SLH exams at 6 institutions (New York Medical College, 
Columbia-Presbyterian Medical Canter, Medical College of Virginia, 
Philadelphia Children's Hospital, Brown University, Charity Hospital 
of New Orleans). Reason: recommendation by the Perinatal Research 
Committee. 

2. Updata of 8-year SLH manual and forms. Reason: completed. 

D. Non-Project 

1. Consultant and training activities by Section Head serving the National 
Center for Health Statistics regarding hearing tests in the National 
Health Survey. 

2. Active participation by the Speech Pathologist as Chairman and member 
of the NIWDS EEO Advisoiy Committee, and member of the EEC Task Force 
whose task is to organize and implement an Affirmative Action Plan 
for NINDS. 

3. Participation by the Speech Pathologist as panel member at the annual 
convention of the American Speech and Hearing Association in New York, 
November l8, 1970. 

k. Appointment of Section Head as NINDS member of the Committee on Hearing, 
Bioacoustics, and Biomechanics (CHAEA) of the National Research 
Council-National Academy of Sciences and attendance at its annual 
meeting at Cape Canaveral, March-April, 1971* 

5. Project Head serves as resource person in Speech, Language and Hearing 
areas for NINDS. 

II. PROBLEMS 

A, Personnel needs . 

1. Language Specialist (Speech Pathology or Psychology) 

2. Secretary 

3. Statistical assistant (one statistical assistant was transferred to 
another branch to achieve promotion; this section now is comprised 

of one statistical assistant, one Speech Pathologist, one Audiologist- 
Section Head). 

III. OBJECTIVES 

A. Development of an effective plan for data analysis, leading to the 
disclosure of relationships between: 

1. Perinatal conditions sind communicative disorders: 

42 V 



2. Test results at 3 -years and at 8-years; 

3. Inter-instutional disparities and causative factors; 
h. Communicative disorders and learning disabilities. 



43 V 



CONTRACT KAEEATIVE 
Perinatal Research Branch. — Section on Pediatric Neurology 
July 1; 1970 through June 30, 1971 

WA^IE STATE UNIVERSITY (PH U3-68-669) 

Title: Maternal Amino Acid Level as Related to Fetal Birthweight of the Infant 

Contractor's Project Director ; Kamram S. Moghissi, M.D. 

Current Annual Level ; $18,000 

Objectives ; 

To investigate relationships in pregnancy between dietary intake of protein, 
blood amino acid, protein and globulin fraction and weight gain of gravida 
and dimensions of newborns and Bay ley test. 

Course of Contract : 

The last contract year of the study has ended as of January 25, 1971» I^e 
workscope of the project was more than accomplished. There remains only to 
complete developmental testing of babies as they reach 8 months of age; and analy- 
sis of all individual amino acids on those cases tested at 8 months as they are 
completed. 

A new finding of particular interest is an association between branch chain 
amino acids and the 8-month Bay ley test scores. 

Major Findings : 

The following publications have resulted from the study: 

1. Churchill, J.A., Moghissi, K.S., Evans, T.W. and Frohman, C. : 
Relationships of Maternal Amino Acid Blood Levels to Fetal 
Development. Obstetrics and Gynecology . 33; kS2-k9^, I969 

2. Moghissi, K.S., Churchill, J.A., and Frohman, C. ; Relationships of 
Maternal Amino Acid Blood Levels to Fetal Development. In Perinatal 
Factors Affecting Human Development . Proceedings of the Special 
Session held during the Eighth Meeting of the PABO Advisory Committee 
on Medical Research, Washington, D.C., June 10, I969, Washington, D.C., 
Pan American Health Organization, Sci. Publ. No. I85, I969, pp. 16-I9. 

Significance ; 

The study may furnish a means by which to identify gra-vlda's providing sub- 
optimum amounts of nutrients to the developing infant. Nutrient supply may 
improve the outcome of these pregnancies. 

Proposed Course of Contract ; 
Study is in last contract year. 

i+5v 



i 



CONTRACT NABRATIVE 

Perinatal Resaarch Branch - Section on Infectious Diseases 

July 1, 1970 — June 30, 1971 

JOHNS HOPKIMS PMIVERSITY SCHOOL OF MEDICINE (PH-4 3-68- 710) 

Title ; Long-range effects on the fetus of certain aaternal Infections 
during pregnancy 

Contractor's Project Director ; Janet B. Hardy, M. D. 

Current Annual Level ; $22,207 

Objectives ; The project Is designed to study the long-range effects of 
certain maternal Infections during pregnancy on fetal outcome with observa- 
tion of the identified children for a period of at least seven years. 
Serologic speciaens and pertinent clinical information relating to perinatal 
infections and pediatric clinical findings are being obtained longitudinally 
from approximately 2100 children. Handprints tor dermatogl3rphlcs are 
available from approximately 800 children with particular eiphasis on 
children with congenital infections. Serial serum specimens are available 
from approximately 300 children with definite or suspect congenital infections 
and complete data and specimens are available from the Johns Hopkins 
Comprehensive Care and the Frederick County Study of cytomegalovirus, 
rubella and toxoplasmosis. Clinical data and longitudinal follow-up Is 
available for approximately 200 non-Collaborative Study children with 
congenital rubella. 

Major Findings ; A list of the publications assisted at least in part from 
this contract Includes a number of papers. Of particular importance is the 
data published on cord Immunoglobulin levels from 2750 cases. Additional 
important data has been generated concerning age specific distribution of 
certain infections in the Inter-dty population as compared to the Frederick 
Cevaty population. This study Included one phase of analysis of buccal 
smears for sex chromatin. The rubella studies have been extremely pro- 
ductive. 

Significance te HIHBS Program and Biomedical Research ; The present study 
of the long-range effects of certain maternal Infections on the fetus 
provides the longitudinal observations necessary to define the complete 
spectrum of fetal damage caused by these agents. Combined clinical and 
laboratory observations permit the analysis of the long-term follow-up 
data. 

Proposed Course of Contract ; The total contract period is designed to 
complete the collection of clinical and laboratory data pertaining to 
cord immnnoglobullns ; age specific distribution of certain infections 
(cytoaegalovirus, rubella and toxoplasmosis); effect of perinatal toxo- 
plasmoals; dermatoglyphlcs and Barr Body studies; and longitudinal studies 
of children with congenital rubella. 



GONTSACT NASKATIVE 

P«rlAatal R«stt«rch Branch - Section on lafoctloos Dlsoasos 

July 1, 1970 — JttBO 30, 1971 

OWIVKRSITT OP CALIPOBMIA (MIH 69-4) 

Tltlo: CytOBOgnloTlrao lnfoc£ions In prognnncy 

Contractor *■ Projoct Director ; Dr. Hargaret Jonaa 

Currant Annnal Laval ; $ 33 , 300 

ObjactlTae ; Study of watamal and contaaltal Infactlona with eytooMgalorlrua 
and harpearlrua honinla utilizing aerologlc tacfanlquaa aa well aa Tlrua 
laolatlon frea urine aad tlaaua apeclaena. 

Major Plndlmta ; During the aecond year of the contract approzlaately 1287 
petlenta have been enrolled. To date 3140 urine apeclMena aad 4308 blood 
apeclaana hare been collected. In addition, 7 abortion tlaane apeclaeaa 
have been obtained. A total of 685 throat airab apeclaena have been collected 
along with 410 cerrlcal awaba aad 159 cervical aaeara. The aera are being 
teated for antibody to cytonega 1 ovlrua and herpesvlrua honlnla. 

Serology apeclaena taken through Deceaber 1970 have been teated for antibody 
to cytoaagalovlrua. The rate of aeroconveralona for CKV approaehea IZ In 
the flrat 1500 woaen. Theae reaults are being recenf Iraed ; checked for 
Herpea I and II; teated for cord Ig^; analyzed la relation to vlrua aheddlag, 
tlae of pregnancy aad clinical flndlnga In the child. 

Slgnlflcaace to NIMDS Prograa and Bleaedlcal Reaearch ; Congenital cytoaegalo- 
vlrua lafectlona can cauae aevere fetal daaage aad death. The preaeat atudy 
la dealgaed to deteralne the tlae durlag pregnancy In which aatemal lafectlona 
aay reault la congenital dlaeaae. The data will alao def lae for the flrat 
tlae the frequency aad peralateace of aatemal aad fetal lafectlona. 

Propoaed Courae of Contract ; The coatract will provide the coatlaued atudy 
of pregaaat woaea. We aatlclpate belag able to atudy 400 patleata durlag 
the third coatract year, brlaglag the total aoaber of petlenta studied to 
2000. Speclaeas of urlae, blood aad tlaaua will be forwarded, aa la the paat, 
for laboratory atudy of the tlae of lafectloa during pregnancy and the tread 
of coagealtal lafectloa. 



49 V 



Project Title: 



Serial No. KDS (CF)-57 TR/TD U02 " 

1. Perinatal Research Branch 

2. Section on Infectious Diseases 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

Jull^ 1, 1970 through June 30, I97I 

Serological and Virus Isolation Studies of Infectious 
Diseases in the Collaborative Study on Cerebral Palsy, 
Mental Retardation, and Other Neurological and Sensory 
Disorders of Infancy and Childhood. 



PreAd-Ous Serial Number: Same 



Principal Investigators; 



Other Investigators: 



Dr. 
Dr. 
Dr. 



Dr. John L. Sever, PRE, NINDS 
Dr. David A. Puccillo, PRE, NINDS 
Mrs, Anita Ley, PRE, NINDS 
Mrs. Renee Traub, PRE, NINDS 
Mrs. Maiy Ruth Gilkeson, PRE, NINDS 

Gabriel Castellano, MBA 

Janet Hardy, Baltimore, Maryland 

Sheldon Korones, Memphis, Term. 



Cooperating Units: 



Collaborative Institutions in the Perinatal 

Research Study 
Laboratory of Infectious Diseases, NIAED 
NICHD 
Cooperating Institutions in California and Havaii 

("With the Section on Infectious Diseases) 
Microbiological Associates 



Man Years ; 



Total: 

Professional: 
Other : 



3.5 
5-5 



Project Description: 

Objectives : The purpose of the infectious disease investigations in the 
Perinatal Research Study is to determine insofar as possible the role of 
infections in the production of abnormal pregnancy outcomes. To accomplish 
this, serial specimens are taken throughout pregnancy, at delivery, and at 
six weeks postpartum. These sera are being tested with antigens to determine 
the antibody responses of the patients during pregnancy and postpartum, and 
then to relate this serological infonnation to the clinical data for the 
pregnancy and the child. In addition, serum specimens from the children at 
one year of age were obtained from the last 10,000 study pregnancies and 



51' 



Serial No. NDS (CF)-57 PR/ID U02 

these are now being studied. In special cases when congenital infection was 
suspected on the basis of clinical or laboratory findings, throat swabs and 
blood specimens were obtained from the children. 

Methods Employed: To accomplish this program, blood specimens were 
obtained from pregnant women at set intervals throughout pregnancy and post- 
partum. Initially, during 19^9^ "the collection of blood specimens was made 
once every trimester. Late in I96O, the requirements for the collection were 
increased and strengthened so that blood specimens were taken at the time of 
registration, at set intervals of approximately every two months throughout 
pregnancy, at delivery, and at six weeks postpartum. At the same time, a 
uniform method for the collection and processing was adapted as a required 
study procedure. The use of special vacutainers, sterile technique, special 
sterile vials, and new shipping containers were all required. Intensive 
laboratory training sessions were held at NIH by the Section on Infectious 
Diseases for all technicians in the collaborating institutions. These sessions 
were repeated each year and a training film was prepared and sent to collab- 
orating groups on request. Careful, complete control was mainatined on all 
collection and processing of serum. At the Serum Center of the Section, 
personnel checked every vial of serum as received for quality and quantity. 
Regular reports of this information were sent every three months to each 
study institution. Telephone calls were made immediately to Project Directors 
whenever there was a decrease in the quality and quantity of sera received. 
Completeness of sets of specimens from each patient was also reviewed by 
Serum Center personnel. To improve the collection of complete sets of sera, 
a special reporting form was devised for the collaborating institutions and 
monitored by the Section. 

By the spring of I96I all institutions attained the minimum requirement 
of 90^ satisfactory specimens for quality and quantity. Satisfactory quality 
was defined as straw colored sera with no hemolysis. Satisfactory quantity 
was a minimum of h vials, each with 3 iilL of serum and proper labels. At each 
meeting of the Project Directors a report was given concerning the quality 
and quantity of specimens submitted by each institution. This detailed in- 
tensive review was conducted throughout the entire time of sampling of sera 
from the mothers and continues at present with specimens being received from 
children at one year of age, special study specimens, and sera from cooperating 
groups . 

Completeness of sets of serial specimens was determined from the sera 
submitted to the Serum Center of the Section and the special reporting form 
sent to the Section. Data for the first 28,386 patients showed that there 
was at least one specimen submitted for 9^.2^ of the patients. The majority 
of the omitted specimens occurred during the first months of 1959. A computer 
analysis of specimen set completeness was prepared. 

With the development of a firm base for obtaining the required specimens 
and tissues, the program embarked on a large commitment for the development 
of necessary antigens, new techniques, and the training of a competent 
laboratory staff for the study of the specimens. Antigen development was 



52v 



Serial Wo. NDS (CF)-5T PR/ID il02 

conducted by the personnel of the Section and experienced investigators under 
contract. Professional laboratory personnel were selected for the Section's 
Units on Statistics and Design^ Serology^ Virology, Experimental Animal 
Research, Immunological Studies, Epidemiology, and Experimental Pathology and 
Neurology. Three years ago the Unit on Cytogenetics was added. Ihis program 
is jointly sponsored by the National Institute of Child Health and Human 
Development and the National Institute of Neurological Diseases and Stroke. 

To document the occurrence of an infection, two approaches are available: 
l) Isolation of the microorganism from the patient, or 2) Detection of an anti- 
body rise in serum specimens. Both approaches are being used in the present 
studies. Appropriate isolation procedures for virus, bacteria, and protozoa 
are being used with throat swab specimens from the children. This approach 
was not used for detecting infections in the mothers since it would have 
required obtaining throat swab and anal specimens at the time of each infec- 
tion. When this has been tried in the past, it has been unsuccessful because 
the women are usually unable to come to a laboratory for the collection of 
specimens when they have minor illnesses. Furthermore, a great many of the 
infections under consideration frequently do not result in significant illness 
of the women. These subclinical infections go unnoticed and \inrecognized. 
For these reasons, the serological approach was selected. By collecting 
serial serum specimens, antibody levels for various viruses and protozoa can 
be determined. The development of antibody to a microorganism in a patient 
who was previously antibody negative provides indirect evidence for infection. 
The presence of specific antibody indicates prior exposure to that antigen or 
mi cr oorgani sm . 

The serological test most frequently employed in these studies is the 
complement fixation method. This basic method has been used for many years 
as the Wasserman test for the diagnosis of syphilis. With the use of viral 
antigens the test is very versatile, performed rapidly, and provides broad 
coverage of a great many of the more than 125 viruses which are known to be 

of importance to man. Antigens were prepared for most of these viruses. Tests 
of specificity were conducted with animal sera. For man, considerable data is 
available from our studies and those of many other laboratories to indicate 
that both group and specific reactions occur with these antigens. The adeno- 
virus CF (Type 2) antigen, for example, is group reactive and provides evidence 
for adenovirus infections in general. To date, there are 31 adenoviruses 
recognized for man. Rubella CF antigen on the other hand is very specific 
and detects infection with rubella only. The sensitivity, specificity, and 
persistence of the test is also known. With this type of information, it is 
pissible to design serological studies for the -viruses and protozoa. Bacterial 
antigens are usually not available for specific serological tests. Only direct 
evidence for these infections can be used. The history of infection as 
reported by the patient has proven to be quite unreliable in most cases and 
is used only in general information. 

The majority of initial serological studies are conducted with the use 
of the complement fixation method. All tests are reproduced completely and 
a minimum of 90^ agreement within twofold variation is required. All sera 



i 



53^ 



Serial No. NDS (CF)-5T PR/ID U02 

showing significant change in antibody, together with any sera which did not 
reproduce, are tested a third time. For more specific testing or confirmation 
of these resialts the hemagglutination inhibition and neutralization methods 
cs,u be used. These latter serological procedures are very specific and are 
also employed for follow-up testing whenever the initial studies with the 
complement fixation method suggest the need, for further investigation. 

Major Findings : A total of lUo complement fixing antigens have been 
developed. Approximately three-fourths of the antigens have been thoroughly 
evaluated and are now being applied in routine testing of study sera. The 
development and maintenance of large quantities (l,000 ml) of satisfactory 
antigens for 50 viruses is an integral part of the investigation being carried 
on by the study. The other antigens are receiving intensive developmental 
work and 20 of these antigens are under test for specificity. Specific con- 
trol antisera have been prepared for 90 microorganisms. In addition, to 
provide improved safety, extensive work has been conducted on the inactivation 
of the live virus antigens . 

The serological studies are being conducted in accordance with three 
major study designs: First is the epidemiological studies to determine the 
frequency of virus experience among study populations. Specimens from 
representative patients at study hospitals are being tested for evidence of 
antibody. By studying these specimens, it is possible to establish the 
frequency of antibody and change in antibody titer to each virus . The data 
for each hospital is then analyzed in relation to other information from the 
Collaborative Perinatal Study, and in relation to other infonnation from 
epidemiological data concerning the seasonal occurrence of abnormal preg- 
nancies and children. 

The second and most active category of study involves the selection of 
particular microorganisms for intensive testing. Studies of this type have 
involved, for example, the testing of sera from a large number of patients for 
antibody to toxoplasmosis or rubella. Intensive studies are now being con- 
ducted utilizing antigens for Australia antigen influenza A, miimps, cytomegalo- 
virus, herpes simplex, and rubella. Patients identified as having serological 
evidence for an infection are then grouped and clinical data for these groups 
and the remaining patients and their children are compared and analyzed. 

Third, studies are designed to obtain maximum data concerning the virus 
experience of patients with abnormal pregnancy outcomes, and "matched controls.' 
The results of this type of study are then analyzed in terms of differences in 
frequency of antibody and antibody change among the abnormals and matched 
controls. The matching of the patients include factors which are known to 
influence virus experience, such as time of the year during which the specimens 
were obtained, race, age, niaiber of living children in the family, and 
geographic location of the patients . These studies are conducted when a 
sufficient number of abnormals of a particular type have been identified so 
that statistical analysis might establish valid information. The initial 
studies were directed at abnormalities which are relatively frequent, such as 
abortions, stillbirths, and neonatal deaths. The less frequent abnormalities 

5I1V 



Serial No. NDS (CF)-57 PR/ID U02 

or those which cannot be recognized in infancy or early childhood are being 
studied as greater numbers of these patients are identified in the Collabora- 
tive Study population. These studies have included spastic diplegia, CWS 
malformation, cranio-facial abnormalities, and mongoloid infants. 

Collaborating Studies : The primary deficiency of data in the Study has 
long been recognized as the late registration of Study patients. Since only 
20^ of the patients register during the first trimester of pregnancy, it is 
impossible to document adequately the infectious diseases experience of 
patients during the first trimester. To provide data on the first trimester 
of pregnancy, one additional Collaborative Study was joined with the program 
of the Section on Infectious Diseases. 

Study of Viral Infections in Pregnancy 

Dr. Margaret Jones, UCLA and Kaiser Hospital in Los Angeles, Calif. 

In addition to this study, the collaboration with the Perinatal Study in 
the Kaiser Hospital in Oakland with Dr. Yerushalmy has been an integral part 
of the program since its initiation in 1959- 

Significance to the Program of the Institute : The use of micro-serological 
techniques for a large group of new viruses provides an opportunity to inves- 
tigate the course of human disease caused by viruses which are either diffi- 
cult to isolate or are resistant to evaluation because the clinical effects 
are delayed until a long time after infection has subsided. This is particu- 
larly true in the case of birth defects . The application of this tool of 
analysis is providing valuable information on the epidemiological aspects of 
viirus infections. 

Proposed Course of the Project : The serological program will continue to 
be expanded in terms of antigenic materials and the performance of tests . 

As additional abnormal pregnancy outcomes are reported, these will be 
added to existing studies on abortions, stillbirths, neonatal deaths, con- 
genital malformations, and mongols . New studies will include congenital mal- 
formations of various types and low I.Q. at four years of age. 

A specific two-year commitment has been made to complete tests on U,000 
abnormals and 4,000 controls using 10 antigens. The PES will Identify the 
specifically abnormal children and the laboratory will match controls and 
perform the necessary tests. The cord sera from 30,000 children are being 
tested for IgM levels. This work will be completed next year (if materials 
are provided by the PRC) . The 1,000 children with high IgM are being tested 
and studied in detail. 



I 



55' 



Serial No. NBS (CF)-57 PR/lD 402 

Publications: 

Matsen, J. M., Jones, M. H., Sever, J. L. , Goldenberg, E. D., Gilkeson, 
M. R., and Justus, K. M. : Family rubella study in Los Angeles. 
Calif. Med . 112: 14-19, 1970. 

Henson, T. E., Brody, J. A., Sever, J. L. , Dyken, M. L., and Cannon, J.: 
Measles antibody titers in multiple sclerosis patients, siblings and 
controls. JAMA 211: 1985-1988, 1970. 

Korones, S. B., Todaro, J., Roane, J. A., and Sever, J. L. : Maternal 
virus infection after the first trimester of pregnancy and status of 
offspring to 4 years of age in a predominantly Negro population. 
J. Pediat . 77: 245-251, 1970. 

Asbed, R. A., Masland, M. W., Weinberger, M. M., and Sever, J. L.: 
Early case finding of children with communication problems. Part I. 
Report of a community screening program. Volta Review . 72: 23-49, 1970. 

Mendez-Cashion, D., Sever, J. L. , Nazario, R., and Gilkeson, M. R. : 
Frequency of rubella antibody in Puerto Rican adiilts. Bol. Asoc. Med . 
P. Rico . 61: 406-408, 1969. 

Sever, J. L., and Terasaki, P. I.: Maternal -fetal incompatibility. 
III. Central nervous system and cardiac anomalies. In Terasaki, P. I. 
(Ed.): Histocompatibility Testing . Copenhagen, Denmark, Munksgaard, 
1970, pp. 495-500. 

Weinberger, M. M., Masland, M. W., Asbed, R. A., and Sever, J. L.: 
Congenital rubella presenting as retarded language development. Amer . J . 
Pis. Child . 120: 125-128, 1970. 

Sever, J. L., Kurtzke, J. F., Alter, M. , Schumacher, G. A., Gilkeson, M. F 
and Ellenberg, J. H. : Virus antibodies and multiple sclerosis. 
Arch. Neurol . , In press. 

Newman, S. J., McCallin, P. F., and Sever, J. L. : Attempts to isolate 
H-1 virus from spontaneous human abortions. A negative report. 
Teratology . 3: 279-281, 1970. 

Sever, J. L. , Gilkeson, M. R., Chen, T. C, Ley, A. C, and Edmonds, D.: 
Epidemiology of mongolism in the Collaborative Project. Ann. N.Y. Acad . 
Sci. 171: 328-341, 1970. 

Sever, J. L. : Persistent central nervous system infection by a virus: 
Rubella. Res. Publ. Ass. Res. Nerv. Ment. Pis ., In press. 



56v 



Serial No. NBS (CF)-57 PR/ID 402 

Sever, J. L. : Virus infections and malformation. Fed. Proc . 30: 114- 
117, 1971. 

Sever, J. L. : Infectious agents and fetal disease. In Waisman, H. A., 
and Kerr, G. (Eds.): Fetal Growth and Development . New York, N.Y., 
McGraw-Hill Book Company, 1970, pp. 221-233. 

Sever, J.L. : Vlroses e o feto, Revlsta de Atualizacao en Ginecologia 
et Obstetricia . Vol. IV, Wo. 10, 1970, pp. 36-54. 



I 



57v 



4 



Serial No. NDS (CF)-61 PR/ID 835 

1. Perinatal Research Branch 

2. Section on Infectious Diseases 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Clinical Investigations in Human Volunteers and Other 

Populations of Virus Effects and Production of Prototype 
Human Antisera, Vaccines, and Other Agents. 

Previous Serial Number: Same 

Principal Investigators: Dr. John L. Sever, PRB, NINDS 

Dr. Earl Matthew, PRB, NINDS 
Dr. Donald Henson, PRB, NINDS 
Dr. Dale Dietzman, PRB, NINDS 

Other Investigators: Dr. David A. Fuccillo, PRB, NINDS 

Cooperating Units: Bureau of Prisons, Department of Justice 
(Dr. Myrl Alexander, Director) 
Petersburg Federal Reformatory 
(Dr. Joel Rascoff , Chief Medical Officer) 



I 



Man Years: 



Total: 1.00 
Professional: .50 
Other: .50 



Project Description: 

Objectives : To study the efficacy of prophylactic and therapeutic 
materials for the prevention and control of infectious diseases. To study 
the safety, antigenicity, communicability and immunogenic ity of candidate 
rubella vaccines. To determine whether intravenous mannitol causes 
cytomegalovirus . 

Methods Employed : Human volunteer studies are conducted in collabora- 
tion with the Federal Bureau of Prisons. These studies are reviewed and approvec 
by the Clinical Research Committee and the Medical Board of the National 
Institutes of Health, and the Vaccine Development Board of the National 
Institute of Allergy and Infectious Diseases. Intravenous mannitol was given 
to human volunteers who were studied for CMV virus shedding excretion and 
antibody. Vaccine studies with low temperature adapted rubella vaccine are 
scheduled . 



59' 

i 



Serial No. NDS (CF)-61 PR/ID 835 

Major Findings ; Volunteer studies were performed to evaluate the 
effect of mannitol on excretion of CMV and antibody response. The mannitol 
proved to be safe in that it did not increase excretion or antibody. 

Significance to the Program of the Institute : Volunteer studies 
provide the basic data necessary to evaluate potential rubella vaccines. 
These studies should provide important and necessary information on the 
effectiveness and safety of such preparations. It will also elucidate the 
epidemiology of CMV in man. 

Proposed Course of the Project : Additional studies will be 
performed when necessary or appropriate. 

Honors and Awards : None 

Publications: None 



60 V 



i 



S«rl«l H«. n>S (CF)-62 PR/ID 972 

1. P«rlaatal lascarch Braoch 

2. Section on Infectious Dlseaaes 

3. Bethesdn, Haryland 

FHS-4IIH 

ladlvldvBl Project leport 

July 1, 1970 through June 30, 1971 

Project Title: Experlaentel Anisel Tlsaue Culture, Hlstopethologlcel and 

Serological Inreatlgatlona of the Role of Ylruaea and Other 
Microorgaalsna in the Perinatal Period. 

Previous Serial Knaber: Saaa 

Principal Investigators: Dr. VUllaa T. London, PKK, HINDS 

Dr. David ▲. Fueclllo, PRB, HINDS 

Dr. Johm L. Sever, P&B, HINDS 

Mr. Villiaa Helas, OB, HINDS 

Other Investigators: Mrs. Anita C. Ley, PRB, HITOS 

Mrs. Blanche Curfnan, PRB, HIHDS 

Cooperating Units: Dr. Andre J. Hahnlas, Departnent of Pediatrics, 
Bnory University S^iool of Medicine, Atlanta, Ga. 

Dr. J. M. Rice, Bioassay Section, EPB, NCI 

Dr. Donald B. Cheek, Departnent of Pediatrics 

Johns Hopkins University Medical School, Baltiaore, Md. 

Dr. Louis W. Catalano, Jr., Departnent of Neurology, 
College of Physiciana and Surgeons, Celunbia University, 
Hev York, Hev York 

Man Tears: 

Total: 8 
Professional: 2 
Other: 6 

Project Description: 

Objectives : To study the role of viruses and other nicroorganlsna 
in the perinatal period, the infection of gravid and nengravld aninals of 
several different species by parenteral routes with various viruses and 
ether nicroorgaaisns to detemlne the effects of these agents on the aninals 
and their fetal tissues. 

Attssqpt to recover inoculated agents froa the various aninals and fetal 
tlasues and the correlation of these relsolationa with tine (In gestatloc) of 
inoculation, and dosage given. 

6iv 



,i» 



Serial Mo. NDS (CF)-62 PR/ID 972 

Correlate these findings with gross and hlstopsthologlcal findings. 
Correlate all of this inforaatlon with serological findings. 

Initiate a nutritional study of non-hoaan prlnates using pregnant 
rhesus nenkeys to test the hypothesis that there Is no causal relation 
between aatemal nutrition during pregnancy and certain sensory, patho- 
logical, Imunologlcal and biochemical characteristics of the Infant. It 
Is considered that a nonkey trial In which the concept of randoaness is 
peraitted is a necessary preliminary to a human trial. 

Methods Employed t An investigation of the role of vimses and other 
microorganisms in the perinatal period by the continual use of experimental 
animals; tissue culture techniques; histopathological studies; and sere- 
logical testing. 

Pregnant mice, rabbits and monkeys are being Inoculated by various 
routes with viruses and other microorganisms. These animals are being 
observed and checked for evidence of disease and/or effects on fetal tissmes. 

Virus isolation investigations utilizing tissue cultnre to recover 
viruses from tissues and fluorescent antibody technique to study the location 
of virus infection produced in experimental animals. 

Histopathological and gross anatomical studies are conducted en ^ 
specimens obtained from the experimental animal studies. 

Extensive serological studies are conducted with the many viral antigens 
developed for the Collaborative Study and new antigens with materials being 
studied previously mentioned. 

In the non-human primate nutritional study, pregnant rhesus menke3rs 
will be maintained throughout pregnancy en one of four diets restricted la 
either calories or protein. One diet will be deficient in both. The 
pregnant animal will be delivered at 158 days of gestation by cesarean 
section and the Infant's tissues will be processed for biochemical analysis 
by the Hopkins Unit. The natritionally deprived female monkeys will be 
continued on their respective diets and studies for Immunological responses 
to various antigens such as tuberculin, rubella and mumps virus will be 
performed. 

Major Findings ; The Inoculation of rabbits with low passage wild 
rubella virus strains and vaccine strains gave variations In antibody 
response and virus shedding. These variations can be useful as markers of 
strain differences. The lew passage wild virus strains produce high anti- 
body levels and variable amounts of virus shedding. The vaccine strains 
resulted in little or no antibody and no virus shedding. 



62 V 



Serial No. RDS (CF)-62 PR/ID 972 

Genital Infection vna readily eatabllehed In 10 feaale Cebua Bonkeye 
when Inoculated Intrayaglnally with type 2 Herpearlrus hoslnla. Infection 
«aa protren by vlrtts laolatlone and aerologlcal atvdlea and the denonatratlon 
of herpetic Tealdea and/or ulcers on the Tul^a and ^raglna vith noderate 
cerrldtls. Relnoculatlon with HVH type 2 was perfomed In all 10 anlaals 
with resulting vaginal reinfection being established In 3 aonkeys despite 
the presence of serua aeetrallzlng antibodies. The lesions produced In the 
Cebus aonkey appear to parallel closely the disease observed In huaan genital 
Infections. 

Polylnoslnlc - polycytldyllc ribonucleic acid (Poly I:C) coaplexed 
with poly-D-lyslne was studied as a prophylactic and therapeutic naterlal 
In nice Inoculated with herpes simplex virus. With a treatnent reglaen 
over a period of several days alee treated with Poly I:C > poly-D-lyslne 
had significantly less Mortality than anlaals treated with Poly I:C alone 
or saline only. This enhanced protection was still detectable If treat- 
aent was begun as late as 4 days following Inoculation of HSV, at which 
tlae an occasional aouse was showing signs of early CHS illness. 

A pilot nutrition study using 38 rhesus aonkeys has been started. 
To date 8 anlaals have becoae pregnant and at least one anlaal is on each 
of the four diets. It appears that the anlaals will eat the diets as 
prepared and aalntain pregnancy. 

Significance to the Prograa of the Institute ; A prograa using ezperi- 
aental anlaals, tissue culture techniques, and hlstopathologlcal investiga- 
tions coapleaents the strict serological approach being used on huaan sera 
obtained froa the Collaborative Study and thus balances the Investigations 
of the role of viruses and other nicroorganisas in the perinatal period. It 
presents the direct neans of Investigation of these agents «rhich may contribute 
to perinatal pathology. 

Proposed Course of the Project ; Further studies using tissue froa 
patients with subacute sclerosing panencephalitis, Creutzfeldt-Jakob disease, 
aayotrophlc lateral sclerosis and progressive aultifocal leucoencephalitis , 
Inoculated intracerebrally into the fetus of pregnant rhesus aonkeys are 
now In progress. 

The Cebus herpes aonkey aodel provides an experlaental approach for the 
study of congenital infection and possible prevention or control of congenital 
disease. We propose a pilot study, first, to deteraine if congenital infec- 
tion can be produced and, second, to explore aethods of prevention or treat- 
aent if this pilot is successful. 

If the above studies were successful In producing congenital herpes 
type 2 Infection in the Cebus, we would propose to proceed with a study of 
chemotherapy using cytoslne arablnoslde, 5-IDU, Poly I:C, and Poly Lysine 
for clearing the aatemal vaginal infection prior to delivery of the baby. 



L 



63. 



Serial Ho. MDS (CF)-62 PR/ID 972 

Secoad, ve would Inroatigaeo Innmlzatloa with herpes type 2 antigen for 
Its efficacy la ellailnatlat vaginal Infections and, third, we would study 
cesarean section aa a «eans of hy-passlng the Infected vaginal area and 
preventing congenital Infection. 

At the ceapletlon of the pilot priaate nutrition study, the data will 
he analyzed and any correction In aethods and diets will be aade. The aaln 
phase will then be started ualng 136 rhesus nonkeys. 

Honors and Awards: None 

Publications: 

Nabaalas, A. J., London, W. T., Catalano, L. W., Fucclllo, D. A., and Sever, 
J. L.: Genital Herpesvirus hoalnis type 2 Infection: An cxperiaental nodal 
in Cebus aonkeys. Science 171: 297-298, 1971. 

London, W. T., Catalano, L. W. , Hahaias, A. J., Fucclllo, D. A., and Sever, 
J. L.: Genital Herpesvirus homlals type 2 infection of aonkeys. Obstct. & 
Gynec . 37: 1971. (In press). 

London, W. T., Fucclllo, D. A., Andersen, B. , and Sever, J. L.: Concentration 
of rubella virus antigen in chondrocytes of congenltally Infected rabbits. 
Nature 226: 172-173, 1970. 

London, V. T., Fucclllo, D. A., Ley, A., and Sever, J. L.: Antibody response 
of yarious strains of rubella rirus when inoculated into rabbits. Proc. Soc . 
Exper. Biol. Med . (In press). 

Catalano, L. W. , London, W. T., Rice, J. M. and Sever, J. L.: Treataent of 
herpesvirus encephalitis with Poly I:C - Poly-D-Lyslne complexes. Obstet. t 
Gynec . (In press). 



6k 



Serial No. WDS (CF)-65 PR/ID 1270 

1. Perinatal Research Branch 

2. Section on Infectious Diseases 

3. Bethesda^ Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Toxoplasmosis: Serological and Clinical Studies. 

Previous Serial Number: Same 

Principal Investigator: Dr. John L. Sever^ PRE;, NIKDS 

Other Investigators: Dr. Joseph S. Drage, PRE;, NINDS 

Cooperating Units: Section on Pediatric Neurology^ PRE, NIKDS 

Man Years: 

Total: .2 
Professional: .2 
Other: .2 

Project Description: 

Objectives: This study relates rises in antibody titer to abnormal 
pregnancy outcomes. 

Methods Employed : Computer analysis of multiple variables relating to 
injection with toxoplasma and pregnancy outcome. 

Major Findings : Within the group of 47 patients with titer elevations 
of greater than 409 b^ or significant increases in antibody titer, five were 
found to have definite toxoplasmosis and ten were suspected of having 
toxoplasmosis. The ten included six with motor retardation, two pregnancies 
resulting in stillbirths, and two in neonatal deaths. The sera from ten of 
the remaining 32 apparently normal children were tested for antibody to 
toxoplasmosis and one was found to have a high titer. 

Proposed Course of the Project : Publication of findings. 

Honors and Awards : None 

Publications : None 



65x 



Serial Wo. NDS.(CF)-6t PR/ID I503 

1. Perinatal Research Branch 

2. Section on Infectious Diseases 

3. Bethesda^ Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 19 T^ 

Project Htle: Epidemiologic Studies of Perinatal Infections 

Previous Serial Number: Same 

Principal Investigators: Dr. John L. Sever, PRE, NINDS 

Mrs. Dorothy M. Edmonds, R.N., PRE, NllffiS 

Other Investigators: None 



Cooperating 


Units : 


None 


Man Years: 






Total: 




1.50 


Profess 


sional: 


1.50 


Other : 








Project Description: 

Objectives: To utilize information from the Collaborative Perinatal 
Research Study and other cooperative studies to identify pregnancies compli- 
cated by maternal infections or infections in childhood; to further delineate 
these cases by serologic testing of the stored sera; to utilize Collaborative 
Study and related data to determine outcome of these pregnancies in relation 
to the outcomes of matched controls and the general study population in order 
to gain information on the frequency of maternal infections during pregnancy 
and their effects on the developing fetus. Studies include: Microcephaly, 
Hydrocephaly, Abortions, Cataxacts, and Bacterial and Protozoal Infections. 

Methods Employed: Primary material utilized for these studies comes 
from the Perinatal Research Study and other cooperative studies . For this 
reason the serologic data developed by the Section on Infectious Diseases is 
correlated with the clinical information available either from print-outs or 
direct hand review of the charts stored in the Perinatal Research Branch. 

Major Findings: The frequency of some virus antibodies has been found to 
be increased in several of the abnormal patient groups . These findings are 
being followed with additional testing of similar groups of patients. 

Significance to the Program of the Institute : The development of the 
serologic data requires the further analysis in relation to the epidemiology 
of infections in the Perinatal Research Study as well as the epidemiology of 
perinatal infections with other collaborating groups. This data then provides 

67 V 



Serial No. NDS(CF)-67 PR/ID 1503 

the basis for correlating serologic and clinical information. Special studies 
are initiated in populations where high frequencies of infection or abnormal 
pregnancy outcomes have been noted. 

Proposed Course of the Project : Special emphasis will be placed on the 
possible association of neoplastic diseases and other tumors with herpes 
simplex infections. We will analyze the clinical data for the 5500 reported 
infections in the Perinatal Research Study. Detailed epidemiological studies 
will be conducted on stillbirths, congenital malformations of various types, 
and children with low IQ's at 4 years of age. 

Honors and Awards: None 

Publications : 

Sever, J.L, : Viruses and embryos. In Fraser, F.C. and McKusick, V.A. 
(Eds.): Congenital Malformations , Proceedings of the Third International 
Conference, The Hague, The Netherlands, September 7-13, 1969, Amsterdam, 
Excerpta Medica Foundation, International Congress Series No. 204, 1970, 
pp. 180-186. 

Sever, J.L. : Viral Teratogens: A Status Report. Hospital Practice . 
5: 75-83, 1970. 

Sever, J.L. : Viruses and The Fetus. Int. J. Gynaec. Obstet . 8: 763-769, 
1970. 



6a V 



Serial He. MDS (CF)-67 PR/ID 1506 

1. Perinatal Reaearch Branch 

2. Section on Infectloes Dlaeaaea 

3. Betheada, Maryland 

PHS-NIH 

Individual Project Report 

Jvlj U 1970 through June 30, 1971 

Project Title: Maternal Infection and Pregnancy Outcoae 

Prerloua Serial Noaber: Saae 

Principal Investlgatora : Dr. David A. Fucclllo, PRB, HIHDS 

Dr. John L. Sever, PRB, HINDS 

Other Inveatlgatora: Dr. Wllllaa T. Leaden, PRB, HIHDS 
Mra. Ranee Trauh, PRB, HIHDS 
Kra. Mary Rath Cilkeaen, PRB, HIHDS 
Kra. Plera Hader, PRB, HIHDS 

Cooperating Units: DnlTersity of California (Dr. Margaret H. Jonea) 

Hagaaii Peraaaente Medical Croup (Dr. Paul F. McCallla) 
Univeraity of Puerto Rico, School of Tropical Medicine 

(Dr. Dolerea Mendez-Cashlon) 
Peanaylvania Hospital (Dr. Coraon and Dr. Belogneae) 

Man Tears: 

Total: 4.5 
Prof eaaional : 1.0 
Other: 3.5 

Project Deacrlptioa; 

Objectivea : To utilise varioua virological technlquea in an iatenslTe 
study of Tiruses to deteralne their role in the production of birth defects 
and related abnoraalitiea. To develop Tirologlcal technlquea neceasary for 
the iJiTeatigatioa of the natural courae of the diaeaae aa cauaed by the 
infectloua ageata. 

Methods Eaployed : Hea ▼Irus Isolation techniques and aerua neutralisa- 
tion teata are uaed for large acale teating in the study of pregnant uoaen and 
their children. The derelopaent of new technlquea, such as the HAI teat for 
Herpea hoainia Typea I and II and a new test for cytoaegalorlruses will now 
peralt the deteralnation of the frequency of Tlrua experience aaong atudy 
populations along with the presence of antibody and change in antibody titer 
to the Tirus. These tests are being uaed to eatablish the reliability of 
other teata and to deteralne their aenaitivity and apeciflclty. A aerologic 
atudy utilizing theae tests on sera collected on the Collaborative Study 



69 V 



Serial H«. HDS (CF)-67 PR/ID 1506 

population was conductad to dataralne the fraquancy of aatlbody chansaa daring 
pregnancy and tha effact of these Infections on the developing aad>ryo. Wa 
have coapletad the testing of 2500 placentas for rubella vims for the 
purpose of Identifying children with congenital rubella by the presence of 
rubella In the placental material. These reports are being prepared for 
publication. The use of a fluoreacent, specific antl-lgM teat la proving 
to be a valuable method for ldentlf3rlng congenital Infections. 

Vaccina for rubella la being given to pregnant women (at tha Pennsyl- 
vania Hospital) who are subsequently being aborted. This will help determine 
the effect of vaccine on the fatua. 

Major Plndlnga ; Three new tests were developed for specific virus 
serology. These were hemadaorptlon tests for Harpea I» Herpes II and 
cytomegalovlrua. Tha tests now provide highly specific, reliable, rapid, 
mlcromathods for virus serology with these aganta. This Is of particular 
value becauae prevloua matboda were particularly laborloua and expensive or 
not specific. Also, since these are hemagglutination tests, the anti- 
complementary effects found In many Perinatal Kasearch Study sera can be 
avoided and tha sera are now usable. 

Vlrua Isolation atndlaa of woman for cytomegalovirus show transient 
as well as persistent Infections. Studies of the children of these pregnancies 
are now In progress. 

Tha fetal abortion studies, after rubella vaccinations, have been 
completed for about 75 caaaa and results will be reported at a later date. 

Rubella vaccina atudlaa are continuing with the temperature adapted 
vaccine material. 

Significance to the Program of tha Institute ; The results from these 
studies help determine what effect virus infection has on abnormal pregnancy 
outcomes and alao provide valuable Information on the epidemiological aspects 
of virus Infections. 

Proposed Course of the Project : Studies are now in progress on 
cytomegalovlrua infactiona during pregnancy and In acveral population 
groupa in Maryland. Additional atudlaa are being conducted on Herpea Typea 
I and II infections in women. Drug evaluation of Cytoslne Arabinoside for 
CMV and 5-IDU for Harpea heminia la being conducted in monkeys. 

Honors and Awarda; Nona 

Pttbllcationa : 

Puccillo, D. A., Modar, P. L., Traub, R. G., Hanaen, S. and Sever, J. L.: 
Micro Indirect Hamagglutlnatlon Teat for Cytomegalovlrua. Appl . Microbiol. 
21: 104-107, 1971. 



70 V 



8«rl«l ■•. »8 (eF)*67 PK/ID 1506 

CmtMlMa», L. W., FkcUIq, 9. ▲., Tr««b, K. C, ami t«r«r, J. L.: 
laolatiM ef l«b«ll« Tlrw fraa Plaewtas ami Thrwt CsltWM •£ lafaats. 
i—r. J. Ob«f t. ftr— c . la prMs. 



71 V 



Serial No. NDS (CF)-69 PR/ID 1729 

1. Perinatal Research Branch 

2. Section on Infections Diseases 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 throttgh June 30, 1971 

Project Title: Stndy of the Possible Transalsslon of Toxoplasaosls In 
Huaans and Ma— als 

Previous Serial Noaber: Saae 

Principal Investigators: Dr. John L. Sever, PRE, NINDS 

Dr. Stephen J. NcvBan, PRB, HINDS 
Dr. David A. Fucclllo, PRB, HINDS 
Dr. Dolores Mendez-Cashlon, Puerto Rico 
Dr. Wllllaa T. London, PRB, HIHDS 

Other Investigators: Hone 

Cooperating Units: Departneat of Pediatrics 
CeBtro-4fedlco 
San Joan, Puerto Rico 

Study has been coapleted. 

Honors and Awards: Hone 

Publications: 

Hevaan, S. J., FuccUlo, D. A., Sever, J. L., London, W. T. and Mendez- 
Cashlon, D.: Tozeplasnosls In Puerto Rico. II. A serological and 
epidemiological study In Puerto Rlcan children. Boletln de la Asoclaclon 
Medlca de Puerto Rico, In press. 



73v 



Serial Ho. HDS (CF)-69 PR/ID 1731 

1. Pcrliuital Research Branch 

2. Section on Infections Diseases 

3. Bethesda, Maryland 

PHS-HIH 

Individual Project Report 

Jnly 1, 1970 throngh Jane 30, 1971 

Project Title: Experimental in vitro and In vivo techniques for Isolation 
of Infectious agents fron chronic diseases. Use of zonal 
centrlfugatlon and Isopycnlc ultracentrlfugatlon for 
purification of the suppressed form of neasles virus fron 
SSPE hralns and FML papovavlrus fron progressive nultlfocal 
lenkoencephalopathy brain speclaens. Study of the role of 
Interferon In chronic Infection. Csaparatlve study of the 
role of delayed hypersensitivity, hunoral lanunlty and 
Interferon In neurotropic neasles Infections In rats. 
Study of the fetal lanmologlcal cenpetence In rhesus 
■onkeys. Sevelopnent of Inaunologlc methodology In 
evaluating Intra-uterlne Infections and CHS Infections. 

Previous Serial Huniber: Same 

Principal Investigators: Dr. Lulz Herta-Barhosa, PRB, HIHDS 

Dr. Dale Dletsman, PRB, HIHDS 
Dr. David A. FuccUlo, PRB, HIHDS 
Dr. William T. London, PRB, HIHDS 
Dr. John L. Sever, PRB, HIHDS 

Other Investigators: Mrs. Rebecca Hamilton, PRB, HIHDS 

Mrs. Barbara Wit tig, PRB, HIHDS 

Miss Helen Krebs, PRB, HIHDS 

Mrs. Anita Ley, PRB, HIHDS 

Cooperating Units: University of Tennessee (Dr. J. T. Jabbour and Dr. 

Charles Cape) 
University of Vermont (Dr. George Schumacher) 
Indiana University Medical Center (Dr. Wolfgang Zeaan) 
University of Calif emia L.A. (Dr. Gary Gltnlck) 
Wilmington General Hoapital (Dr. George Bolnes) 

Man Teara: 

Total: 5 
Professional: 2 
Other: 3 

Project Description: 

Objectives ; To establish ufaether persistent or tolerant viral 
infections are associated with chronic diseases such as polymlositis , 

75 V 



Serial He. NDS (CF)-69 PR/ID 1731 

ulcerative enterocolltla, aajro trophic lateral sclereela, Creutzf eld t~ Jakob 
dlaeaee, pregreaslTe anltlfocal leekoeacephalepathy, and Multiple sclerosis. 

To purify and concentrate PML and SSPE ▼Iruses and examine these 
agents electron alcroscoplcally, blochenlcally, and antlgenlcally In 
parallel with the conventional formm of these viruses. 

To determine Interferon levels In patients with ulcerative entero- 
colitis, hepatitis, and SSPE, as well as to establish whether their 
peripheral white cells are capable of producing Interferon to normal levels 
when compared with cells from normal subjects. 

To evaluate the role of cellular laannlty, humoral lamunlty, and 
Interferon In protecting experimental animals against neurotropic measles 
virus. 

To elucidate the maturation of the lomune system In sobryonlc life 
and correlate this process with Intra-uterlne Infections. 

To separate IgM ab^ IgG from cord sera from children with congenital 
diseases and determine the serological specificity of the 19S Immunoglobulin 
using fluorescent antibody assays In an attempt to develop a diagnostic test 
for fetal Infections. 

Methods Employed ; Brain specimens from well-documented cases of MS, 
FML, ALS, SSPE, etc, were homogenized to a lOZ suspension and Inoculated 
Intracranlally In groups of 5 rhesus monkey fetuses during the first third 
of gestation when the animals are expected to be inmnologlcally immature 
and hence more susceptible to Infectious agents. These fetuses were carried 
to term and, immediately after birth, one animal was killed and examined 
hlstopathologlcally for CNS lesions. The remaining animals were carefully 
observed for clinical symptoms, abnormal behavior, and antibody pattern. A 
2-year follow-up has been scheduled. 

Brain tissue provenlent from patients with encephalopathies, intes- 
tinal mucosa from patients with ulcerative enterocolitis, muscle from 
patients with polymiositls , and lymph nodes from Individuals with SSPE were 
examined by electron microscopy, fluorescent microscopy and, whenever 
possible, cultured In vitro . These tissue cultures were submitted to 
vlrologic and serologic assays, inoculated into experimental animals, and 
co-cultivated with human diploid and heteroploid cell lines in efforts to 
isolate and identify intracellular agents. 

Brain autopsies from patients with FML and known to contain intra- 
nuclear virus were frozen and thawed three times, the nuclei extracted and 
disrupted by physical means. Nuclear extracts were isopycnically banded in 
CsCl and fractions examined under the electron microscope. Fractions 
containing the papovavlrus were used to IflBsunize guinea pigs in order to 



jGv 



Serial No. NDS (CF)-69 PR/ ID 1731 

produce aonospeclflc antisera Intended to Identify the virus. Slallar 
approach, but using zonal centrlfugatlon to purify the virus, was planned 
for SSPE and Is awaiting approval of the necessary contract. 

Interferon levels In serua of hepatitis and enterocolitis patients 
were obtained by Vk Inhibition test using Slndbls virus as the challenge 
virus. SSPE patients' white cells were grown in vitro , infected with 
different viruses (including aeasles), and Interferon on supematants 
measured by the HA Inhibition method. Adequate controls were utilized. 

Levis* rats were Inomnized with eeasles vaccine conblned with 
Freund's adjuvant and after antibodies developed and delayed hypersensitivity 
becaae ■aaifest as measured by skin tests with PPD, these animals served 
as serum and lymphocyte donors to groups of histocompatlble, non-Immunized 
animals. Interferon was produced by ^ vitro grown macrophages and 40,000 
units were inoculated Into other groups of rats. Passively Immunized animals, 
i.e., those rats receiving pre-determlned amounts of antibody, competent 
lymphocytes and interferon were challenged with 10 U>so neurotropic measles 
virus and protection conferred measured by comparing death rates with those 
of control groups. 

Mumps and chyknagunia viruses were inoculated into groups of rhesus 
monkey fetuses at first, second, and third periods of gestation. At 
appropriate Intervals the fetuses were removed and blood collected for 
Immoaological evaluation. Antibody curve was determined by CF test; 
lymphocyte transformation was measured by uptake of tritlated thymidine in 
the presence of mumps antigoi; Interferon production was established by HA 
inhibition assays. Concomitantly, the pathogenesis of «imps virus was 
studied in some fetuses to determine the fetal susceptibility in terms of 
stage of gestation and to correlate these observations with the immuno- 
logical data. 

lamBnoglobttlins from cord sera provenlent from cases of congenital 
infections were precipitated by the ammonium sulfate method and concen- 
trated 10 times in saline. These preparations were ultraccntrlfuged in 
linear gradients of sucrose and 7S gamma globulins separated from 19S 
molecules. The latter fractions were serologically evaluated against 
different antigens (Toxoplasma, rubella, QfV) to determine antibody 
specificity and thus the identity of the causative agent. 

Sera and spinal fluid from patients with SSPE were compared with 
specimens from patients with other neurological diseases using the immuno- 
diffusion technique in gel plates. Highly concentrated measles virus 
antigens were utilized in the test. 

Major Findings ; Virologlcal studies with SSPE lymph node biopsies 
resulted in the isolation of measles virus and in the demonstration of an 
unidentified bullet-shaped virion in the cytoplasm of several stroma cells. 
The measles virus was isolated in mixed cultures with HeLa cells. This 



77V 



SarlAl Ho. RD8 (eP)-69 PK/ID 1731 

finding attggaata that SSPE la a ayataoalc Infection with poaalble apaclf Ic 
defoctlyenaaa of the cellolar Ijanmlty. 

A alBple dlagnoatlc teat for SSPE «aa developed by the Ovchterlony 
technique uaing the patient 'a aplnal flnld and concentrated meaalea antigen. 
The aaaay proved highly apeclflc and hence of conalderable dlagnoatlc value. 

A herpea-llke vlrua waa found aaaoclated with caaea of ulcerative 
enterocolltla. The vlrua partldea were vlauallzed by electron mlcroacopy 
and tlasue culturea of nucoaa and aub-mucoaa apeclaena ahoved acattered 
Intracellular vlrua when aaaayed by lamunofluoreacence. 

Circulating Interferon waa not found In patlenta with chronic or 
acute hepatltla nor In patlenta with enterocolltla. Serua and aplnal fluid 
froa patlenta with SSPE alao failed to ahow detectable levela of Interferon. 
However, the peripheral white cella from theae patlenta reaponded to In 
vitro Infection by producing noraal aaounta of Interferon, Indicating tiiat 
the IF ayaten la not Impaired In theae condltlona. 

Plconavlma-llke cryatala were aeen by electron alcroacopy In aeveral 
■nade blopalea froa patlenta with polyaloaltla . tUxed culturea of auade 
cella and alalan cella were prepared In atteapta to laolate vlruaea. A 
papovavlrua waa purified froa a FML brain autopay and aubaequently uaed to 
Inoculate laboratory anlaala. The purification waa achieved by banding the 
vlrua In CaCl (1.6 gra/al). Spectrophotoaetry and electron alcroacopy 
monitoring resulted In a rather pure vlrua preparation of approximately 10' 
Intact partlclea/5 graas of tlaaue. 

Significance to the Prograa of the Inatltute ; The developaent of 
alzed cell culturea to unnaak latent Infectlona provldea an excellent 
methodology for the atudy of chronic dlaeaaea of poaalble viral etlologlea. 

The auccesaful development of a almple dlagnoatlc teat for SSPE 
warrants further Immunological atudlea with cerebrospinal fluid from patlenta 
with other neurological dlaeaaea In an effort to define antibody pattema 
of dlagnoatlc value. 

The preaence of a latent. Intracellular measles virus In the lymph 
node of patlenta with SSPE auggeata a tolerant Infection with deficiency of 
the delayed hypersensitivity. This finding should lead to detailed examina- 
tion of the Immune competence of SSPE patlenta aa well as that of Indlvldnala 
with other CNS Illnesses. 

The understanding of alow vlrua infectlona of the CRS will depend upon 
purification of the auppr eased form of these agenta followed by careful 
biochemical and blophyalcal analyala. To accompllah thla goal, the uae of 
zonal centrlfttgatlon for cellular fractionation ahould be oiphaalzed. 



78^ 



Serial H*. HDS (CF)-69 PR/ID 1731 

Ffpo— J C»Mf of th« Proj«ct ; Special ai^hasls will be placed epen 
a brala caltvre reaearch prograa. lanreatlgatlea of the aeehaalaa ef patlM>- 
genesla aad poaalble Ibmom def Iclenclea In patlenta vlth nenrologlcal diseases 
will be conducted. Our selection of patients with aoltlple scleroels, 
Parkinson's Disease, progresslre wtltlfecal leukoencephalopatby, and aayo- 
trophlc lateral sclerosis Is supported by the existing data which suggest 
possible Tlral etiologies for each of these diseases. Tissue speclnens and 
blood from patients will be provided through collaborative-contract arrange- 
■ents with Investigators throughout the country. 

Utilising the nlzed culture technique «e hope to detenlne If It Is 
possible to release suppressed virus from these chronic neurologic diseases 
and to gain a further understanding of the pathogenesis of latent Infections 
of the CMS. Antibody levels and conpctence of lynphocytes fron patients will 
be exsnlned using the standard technlquea. 

The SSPE strain ef aoasles virus will be sti|dled In laboratory anlnals 
and efforts will be directed at the developnent of an anlaal aodel systen for 
this disease. 

Honors and Awards: Hone 

Publications: 

Horta-Barbosa , L., Fuccllle, D. A., Haailton, R., Traub, R. , Ley, A., and 
Sever, J. L.: Sene characteristics of SSPE ncasles virus. Proc. Sec. Exper . 
Biol, h Med . 134: 17-21, 1970. 

Vernon, M. L., Horta-Barbosa, L., Fucclllo, D. A., Sever, J. L., Barlnger, 
J. R. , and Blmbaun, G.: Virus-like particles and nucleoproteln-type 
fllsnents In brain tissue fron two patients with Creutzfeldt- Jakob Disease. 
Lancet 1: 964-967, 1970. 

Horta-Barbosa, L., Krebs, H., L^, A., Chen, T. C, Gllkeson, M. R., and 
Sever, J. L.: Progressive Increase In cerebrospinal fluid measles antibody 
levels in subacute sclerosing panencephalitis. Pediatrics in press. 

Horta-Barbosa, L., Fucclllo, D. A., and Sever, J. L.: Viral and protozoan 
infections of the newborn. In Abramson, H. (Ed.): Resuscitation of the 
Newborn Infant . St. Louis, Mo., C. V. Mosby Co., In press. 



79V 



i 



Serial He. MDS (CF)-69 PR/ID 1732 

1. Perlnetel Research Branch 

2. Section on Infections Diseases 

3. Bethesda, Maryland 

PHS-NIH 

Indlvldoal Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Investigation of the Role of Mycoplasma spp , and Other 
Microerganisas in the Perinatal Period. 

Previous Sorial Huaber: Same 

Principal Inrestigators ; Dr. David L. Madden, PRB, KINDS 

Dr. Williaa T. London, PRB, MINDS 
Dr. John L. Sever, PRB, NINDS 
Dr. Earl B. Itatthew, PRB, NINDS 
Dr. Dale Dietnuin, PRB, HINDS 

Other Investigators: Dr. Melvin Museles, NNMH 

Mr. Kenneth Meats, PRB, NINDS 

Cooperating Units: National Naval Medical Hospital, Bethesda, Md. 

Man Tears: 



Total: 


1.60 


Professional: 


.60 


Other: 


1 



Project Description: 

Objectives : To study the role of Mycoplasna spp . in perinatal diseases 
of aan. To develop aniaal aodels to detemine the pathogenesis of tiiese 
diseases. To develop nev test systems for the rapid diagnosis of perinatal 
diseases. 

Methods Employed : Yaginal and oral swabs are obtained from pregnant 
vomen and monkeys at the time of labor or C-section. Swabs are obtained from 
infants and mothers. These specimens are cultured in standard media. Isolated 
cultures will be identified by the metabolic-inhibition (M.I.) test. Serum 
samples obtained prior to delivery, at time of delivery and post-delivery are 
studied for presence of antibodies by the M.I. test, indirect iDmunofluorescence 
and agglutination tests. Pregnant monkeys found to be free of Mycoplawna 
infections will be Inoculated with various strains of Mycoplasma to detemdne 
the effect of these strains upon mother and infant. The use of newer techniques 
for determining antibodies in body fluids will be used in an effort to develop 
quicker and more accurate tests for detection of viral antigens and antibodies. 
He will utilize new virus strains and tissues in an attempt to grow higher 
tltered virus antigens. He will use new techniques such as counter-lnmnmo- 
electrophoresis and radioinsmnoprecipitation tests. 

8iv 



Serial Ho. HDS (CF)-69 PR/ID 1732 

Major Fiad<«B* i Santa froa 100 aothar-baby pairs, whose Mycoplaaaa 
flora and antibody tltara for Mycopla— a had been deterained, were atvdied 
for abnoraal aaoants of IgG and Ii^ antibody in an effort to deteralne if 
in utero infection occurred. None aaa found. Post-natal follow-up on 
children known to have been infected vith Myeoplasaa at birth has indicated 
that they vere not adversely affected as jndged by birth veight, weight 
gain or increased susceptibility to disease. 

Inoculation of M. hoainis into the vagina of pregnant aonkeys did 
not cause fetal infections. The pregnant aonkeys soon eliainated this 
agent froa their bodies. In non-pregnant aonkeys, a severe vaginitis 
occurred and the organisas have persisted in the vaginal tract for a long 
period of tiae. It was also observed that soae of the serological character- 
istics of this organisa were altered. Concurrent with this study, a natural 
spread of M. orale II in the aonkey colony was observed. 

An SSPE virus antigen was grown to high titers in spinner cultures 
and then concentrated 100 tiaes in a Spinco centrifuge. A aethod to 
concentrate spinal fluid was developed by utilizing evaporation of fluid 
through cellulose dialysis tubing. This concentrated antigen and spinal 
fluid in a gel diffusion or counter-iaaunoelectrophoresis test was of 
coaparable sensitivity to the C? or HI test on uaconcent rated spinal fluid 
and was auch less coaplicated to perfora. 

Significance to the Prograa of the Institute ; A prograa devoted to 
studying the effect of Mycoplawm in perinatal infections coapleaents the 
virological studies currently being dene. This study and the support given 
to other investigators aay aore accurately define the role of Myeoplasaa in 
disease. The developaent of additional techniques for the serological 
diagnosis of perinatal diseases which are as sensitive or aore sensitive 
than current tests being used, and which by-pass the probleas of anti- 
coapleaentary sera, will increase the usefulness of the stored perinatal 



Proposed Course of the Project ; Further studies are being done on 
the association of Myeoplasaa in noraal deliveries. Study of the pa biogene- 
sis of Myeoplasaa in aonkeys will be continued to see if an aniaal aodel can 
be developed for septic and non-septic abortions and other perinatal infec- 
tions. Atteapts to define the association of Myeoplasaa in neurological 
diseases will be aade. Continued effort will be aade to apply these tech- 
niques to other antigen-antibody coabinations in order to develop additional 
serological methods to detect perinatal and neurological diseases. 

Honors and Awards: None 



82a 



S«rlAl Ho. HDS (CF)-69 PK/ID 1732 

PvblleatloBs: 

Itaddea, D. L., Barton, R. E. and MeCullough, N. B.: SpontanooiM infection 
in Ex-gomf roft guinea plga due to Cloatrldlan porf rlngena . Lab Aninal Care 
20: 45^455, 1970. 

Kaddan, D. L., Hlldarbxaadt, R. J., Monlf, 6. R. 6., London, V. T., Sorer, 
J. L. and McCnllongh, H. B.: The laelatlon and Identification of Mycoplaana 
fron Macaca aulatta. Lab Anlnal Care 20: 467-470, 1970. 

Madden, D. L., HUderbrandt, R. J., Monlf, 6. R. 6., London, W. T., 
McCullottgh, N. B. and Sever, J. L.: The laolatlon and Identification of 
Mycoplaaaa fron Cercoplthecoa aethlopa. Lab Anlnal Care 20: 471-473, 1970. 

MBtthev, B. B., Dletnan, D. E., Madden, D. L., Sever, J. L., Mattl, A. and 
Dodaen, W. E.: The poaalble abaence of Atiatralla antigen In 6/6 trana- 
locatlon type of Down 'a a3mdrone. Lancet. (In preaa) 



i 



83V 



Serial Wo. NDS (CF)-69 PR/id 1733 

1. Perinatal Research Branch 

2. -Section on Infectious Diseases 

3. Bethesda, Maryland 

PHS-WIH 

Individual Project Report 

July 1, 1970 through June 30, I97I 

Project Title: Viral Diseases of the Nervous System 

Previous Serial Number: Same 

Principal Investigator: Dr. David A. Fuccillo 

Other Investigators: Dr. J. L. Sever, PRB, NINDS 
Dr. S. Baron, LVD, NXAID 
Dr. W. T. London, PRB, NINDS 

Cooperating Units: Laboratory of Viral Disease, NXAID 

Georgetown University School of Medicine, 

Washington, D.C., Dr. J. A. Bellanti 
Harvard Medical School, Boston, Massachusetts, 
Dr. L. Johnson 

Man Years: 

Total: 2.0 
Professional: 1.0 
Other: 1.0 

Project Description: 

Objectives : The main objective of this project is to establish the 
clinical and biological significance of the two different strains of hearpes 
simplex viioas (Type I, "oral" and Type 2, "genital") in the causation of 
human disease including carcinoma of the cervix. 

Methods Employed : The principal methods employed are: (l) the quanta! 
cross -mi croneutralization test, by which type specific herpesvirus antibody is 
identified; (2) mass serological surveys of materials from selected patients 
with specific disease entities; (3) virus isolation, titration, and charac- 
terization procedures; (h) interferon titration via plaque reduction assays. 

Major Finding : Patients with carcinoma in situ of the uterine cervix 
were demonstrated to have increased amounts of antibody to type 2 herpes-virus . 
Antibody was also elevated in these patients two years before they developed 
cancer (collaboration -with Dr. Johnson). 

Significance to Biomedical Research and the Program of the Institute : 
These investigations attempt to elucidate the pathogenesis of AnLral infections 
of the adult and fetus using immunological and v:.rological techniques. Herpes 

85 V 



Serial No. NDS (CF)-69 PR/ID 1733 

simplex virus has been one agent which has received particllar attention in 
these studies, since it has significant neurotropic capabilities in tenns of 
newborn and adiolt encephalitides. There is considerable spe ciiilation that 
this virus may have latent, "slow" virus potential in relationship to chronic 
diseases of humans, including carcinoma and central nervous system infection. 
Investigation of the clinical and biological properties of the two strains of 
herpes simplex virus have permitted more definitive establishment of such 
capabilities. Furthermore, the therapeutic potential of artificial interferon 
inducers as anti-viral agents was investigated with herpesviarus in animals and 
one human, particularly in relationship to central nervous system Infections. 

Proposed Course of Project : Expanded study of antibody and isolation of virus 
from women with carcinoma in situ and carcinoma of cervix in the Collaborative 
Study . 

Honors and Awards : None 

Publications: 

Catalano, L. W. , Jr., Fuccillo, D. A. and Sever, J. L.: Piggy-Back Micro- 
transfer Technique. Appl. Microbiol . l8: 109ij-1095, I969. 

Catalano, L. W. Jr. and Sever, J. L.: Role of Viruses as Causes of Con- 
genital Defects. Ann. Rev. Microbiol ., In Press. 

Catalano, L. W. Jr., Fuccillo, D. A., Traub, R., and Sever, J. L.: 

Isolation of Rubella Virus from Placentas and Throat Cultures of Infants 

in a Prospective Study Following the 196^-65 Epidemic, J. Pedlat. . In Press, 

Fuccillo, D. A., Moder, F., Traub, R., Hensen, S. and Sever, J. L.: 

Micro Indirect Hemagglutination Test for Cytomegalovirus. Appl. Microbiol . 

21: 104-107, 1971. 



86v 



Serial No. NDS (CF)-70 PR/ID 1848 

1. Parlaatal Rsaearch Branch 

2. Section on Infection* Diseases 

3. Bethesda, Maryland 

PHS-RIH 

Individual Project Report 

July 1, 1970 through June 30 » 1971 

Project Title: Delayed Hypersensitivity In Chronic Viral Diseases 

Previous Serial Nunber: Sane 

Principal Investigator: Dr. Earl B. Matthew 

Other lavestlgators: Mrs. Mary Krasay 
Dr. Donald Hanson 
Dr. David A. FuecUlo 

Cooperating Units: Hone 

Man Tears: 

Total: 1 
Professional: 1/2 
Other: 1/2 

Project Description: 

Objectives : To detenlne the role of delayed fajrpersensltlvlty 
(cellular Inmnlty) In chronic viral Infections. 

Methods Biployed : The nacrophage migration Inhibition test (MI) , 
lynphocyte transfonatlon and lynphotozln assay vere used In tiie study. 

Major Findings : The nacrophage migration Inhibition test» lynphocytlc 
transfomatlon and lynphotoxln assay vere all attenpted In studying the 
nottse cytomegalovirus Infection. Hone vere found to give reliable results 
and no correlation could be made. This vns due In most part to the type of 
animal used, the mouse not developing good delayed hypersensitivity unless 
an abnormal stimulus Is utilized such as Freund's adjuvant. Unfortunately, 
this distorts the natural disease picture. The mouse CMV system Is vorth- 
vhlle for more refined study vhen other techniques become available. 

Development of the lymphocyte transformation test as a marker of 
cellular Immunity in human virus infections has been a major effort In the 
past year using various viral antigens. The studies of these In subacute 
sclerosing panencephalitis and multiple sclerosis are In the last stages of 



L 



87^ 



S«rl«l Ho. IDS (CF)-7e PR/ID 1848 

caaplatloA. Th* 9tmij of theso In patients with acnto lyaphocytlc lottkeala 
is coBpleto. 

Slgniflcnncc to tho Progr«« of the Inotltute ; Prior to the develop- 
■ent of the above testa, no good tests for aeasurlng cellular looimlty In 
himans to rlral antigens existed. Antibody teats to viral antlgena have 
been developed and used In this and other laboratories for nany years. 
This represents a test for only part of the body's 1— iim response. The 
developaeat of lyaphocyte transfonatlon Into a clinically uaable test for 
virus Infection now allows both fons of the body's protective laensne 
■echanlsns, fauaoral and cellular, to be teated alnultaneously . 

Proposed Course of the Project ; To complete the above mentioned 
specific projects prior to July 1971 lAen the principal Investigator Is 
tendnatlng his stay In NIHDS. 

Honors and Awards: Hone 

Publications: Heme. 



S«rlal No. NDS (Cr)-70 PR/ID 1849 

1. Perinatal Kcsearch Branch 

2. Section en Infectious Diseases 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Chronic Infection with Cytoaegalovlruses In Man and Anlnals 

Prerleus Serial Nuaber: Same 

Principal latest Iga tor : Dr. Donald Benson, PRE, MINDS 

Other Investigators: Mr. Leonard Moore, PRB, NINDS 

Dr. John L. Sever, PRB, NINDS 

Dr. David A. Fucclllo, PRB, NINDS 

Dr. Earl B. Matthew, PRB, NINDS 

Cooperating Units: NCI, Dr. Edward Henderson, Dr. Ronald Yankee, 
Dr. Stuart Slegel, and Dr. Arthur Levine 
Araed Forces Institute of Pathology, Dr. A. J. Strano 

Man Tears: 

Total: 2 
Professional : 1 
Other: 1 

Project Description: 

Objectives : Study chronic cytonegalovlrus infection, antibody levels, 
virus excretion, lymphocyte response, disease in nan and alee. Relate 
specific pathologic processes to antibody levels and virus excretion patterns. 

Methods Enployed : Children with congenital CMV and patients with 
leukemia with CMV infections are being followed for virus excretion, antibody 
response and lynphocyte response. Techniques for CMV isolation have been 
perfected. Antibody response is being followed with CF, HAI and neutralization 
■ethods. Lymphocyte transformation is determined with trltiated thymidine. 

Major Findings ; Chronic infection of the tissues of the mouse have 
been demonstrated. Results have shown that the response of the host to 
infection depends on the anatomy of the organs Infected. 

Eighty-seven children with acute lymphocytic or acute myelocytic 
leukemia have been longitudinally studied for 3 to 18 months. Results 
Indicate a correlation between clinical symptoms and rises In antibody titers 
to CMV. There is no association between antibody titers and frequency of 



Serial Ho. NDS (Cr)-70 PR/ ID 18A9 

virus Isolation froa urine or throat or langth of virus excretion In these 
children. Basic problea now Is to develop tests that discriminate between 
CH7 rilssealnatlon and focal chronic Infection. 

Significance to the Program of the Institute ; Cytomegalovirus causes 
congenital disease and death as well as significant Infection In patients 
with nallgnandes. An understanding of this chronic Infection and the 
laatune responses of the host should be of great value In the prevention and 
treatment of these diseases. 

Proposed Course of the Project : This Is a new study. We will 
continue along the lines outlined above. 

Honors and Awards: Hone 

Publications: 

Henson, D. and Sever, J. L.: Effects of Viruses on the Fetus. In Marcus, S. 
L. and Marcus, C. C. (Ed.): Advances In Obstetrics and Gynecology . 
Baltimore, Md., Williams and WUklns Co., Vol. 2, 1971. In press. 



Stflal Ho. HDS (CF)-71 PR/ID 1903 

1. Parlnatal lessarch Branch 

2. Section on Infoctlooo Dlaeascs 

3. Bothesda, Maryland 

PHS-NIH 

Individual Project Report 

Jttly 1, 1970 throttgh Jane 30, 1971 

Project Title: Inveatlgatlon of the Etiology and Effect of Serun and 
Infectious Hei»atltls In the Perinatal Period. 

Previous Serial Nunber: None 

Principal Investigators: Dr. David L. Madden, PRB, HINDS 

Dr. John L. Sever, PRB, HINDS 
Dr. Dale E. Dletznan, PRB, NIHDS 
Dr. Earl B. Matthev, PRB, HINDS 

Other Investigators: Dr. Benedict Hagler 

Cooperating Units: Lynchburg Training School and Hospital 

Man Tears: 

Total: 1.75 
Professional: 1.75 
Other: 

Project Description: 

Objectives : To detemlne the etiology of Australia antigen associated 
(serua) and infectious hepatitis. To detemlne the relationship of hepatitis/ 
congenital jaundice and postnatal jaundice. To develop aninal Bodels and new 
diagnostic tests for these diseases. 

Methods E>n>loyed : A large epidenic of infectious hepatitis (600 cases) 
occurred in the Lynchburg Training School and Hospital during the suaner of 
1970. Over 5000 blood and 400 fecal sanples were collected during a 4-aonth 
period. These sanples represent serial speclnens obtained prior to the 
developnent of disease, at tine of acute disease and post-infection. A 6- 
nonth follow-up has now been carried out. Australia antigen deteminations 
were perfomed on 1200 patients. A controlled study of the effect of ganaa 
globulin on this strain of infectious hepatitis was initiated. Serun and 
fecal sanples will be cultured in a variety of tissue culture systens and 
inoculated into experinental anlnals. Material fron patients with hepatitis 
hospitalized at Stdmrban Hospital has been collected and is being utilized 
in coaparison studies. 



I 



91V 



S«rlAl Ho. HDS (CF)-71 PR/ID 1903 

MajT Fladl«K« ; TIm rasults of the opldoalologleal atudy uadortokoa 
at tho LyncUbsrg TralBlag Scliool aad Hoopltal ladlcoto that the dlaoaao 
waa not aaaoclatad with Aaatralla antlgan. It waa foand that the dlaeaac 
vaa tranaBlttod fnm vard to ward by infected patient workera or through 
contact vlth the patlenta. It appeara that thia outbreak la the largeat 
nen-ceaawn •ovree of Infoctiewi preaently known. In thla outbreak, the 
uae of ganaM globulin waa not effective unleaa given 14-21 daya before the 
firat dlnlcnl cane. Thla waa partly related to the occurrence of aub- 
cllnlcnl cnaea which una followed by waaa contaalaatlon of the ward. The 
occurrence of clrcnlntlng Au antigen In chronic carrlcra did not alter the 
courae of the Infectloua hepatltla. Prellnlnary reaulta auggeat that the 
incidence of Au antigen in trlaoiqr 21 karyotype patlenta waa wnch higher 
than in patlantn with Down 'a ayndroae dne to other karyotypea or in patlenta 
with noraal karywtypea. 



Significance to the Progran of the Inetltnte ; The finding of an 
etlologlcnl agent or an infectloua hepatltla aaaoclated antigen would be 
an iaportant atep in the pranrentlon of thla dlaoaae which affecta aatveral 
thowaand ialiwiduala each year. The naterlal collected fron the Lynchburg 
outbreak la probably the largest accunulntion of pre-jaundlce» acute, and 
convnlaeeent natenals erer collected fron a aingle outbreak. It will be 
aactraoMly valuable for future reference. 

Propoaed Courae of the Project ; Attcnpta to define the role and 
cause of aeruw and infections hepatitis will be continued. The tlaaue 
culture atndy has Just been initiated. Attcnpta to detect specific antigen 
and antibody in these feces and seruas will be nade. 

Honora and Awarda: None 

Publications : None 



92 V 



Project Title: 



Serial No. NBS (CF)-63 PR/OC 1144 

1. Perinatal Research Branch 

2. Office of the Chief 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

An Instrument For The Conduct Of A Retrospective Study Of 
Seizures, Cerebral Palsy, Mental Retardation and Other 
Neurological and Sensory Disorders of Infancy and Childhood. 



Previous Serial Number: Same 



Principal Investigators: Z.A. Shakhashiri, M.D., PRE, NINES 

Leonard V. Phelps, Clearwater, Florida 
Glen S. Bartlett, M.D., Case Western Reserve Univ., 

Cleveland, Ohio 

Other Investigators: Lenore Bajda, M.D., PRB, NINDS 

John R. Day, M.D., Chevy Chase, Maryland 

Blanche L. Vincent, S.N.O., Greensboro, North Carolina 

Zula C. Meekham, B.S.N. , PRB, NINDS 

Rose R. Tortorella, PRB, NINDS 



Cooperating Units ; 



Georgetown University Hospital, Retarded Children's Clinic, 
Selected Maternity Hospitals and Physicians in Metropolitan 
Washington. 



Man Years: 



Total: 

Professional: 

Other: 



.35 
.30 
.05 



Project Description : 

Objectives: Design an instrument for the conduct of a retrospective study of 
seizures, cerebral palsy, mental retardation and other neurological and sen- 
sory disorders of infancy and childhood in order to test certain basic and 
important hypotheses concerning the occurrence of neurological damage. 

Methods employed : Recognized damaged outcomes of pregnancy, such as seizures, 
diplegias, hemiplegias and choreoathetoids are to be studied and related to 
defined perinatal or postnatal events. These outcomes were selected because 
they were construed to be related to or manifestations of or involved in the 
biological or psycho-sociological mechanism underlying the following hypoth- 
eses: (1) anoxia, (2) toxic influences on the brain, (3) metabolic influences. 
(4) trauma to the head, (5) infection of the brain, (6) dehydration of the 
child, (7) genetic or familial patterns, and (8) socioeconomic status. 



I 



93V 



Serial No. NDS (CF)-63 PR/OC U44 

Proposed Course of the Pro.ject ; The abstraction of the maternity and nursery 
records and interviewing of the mothers have been completed. The physicians 
of sib controls have been contacted cind information attesting to the health 
status of these sibs has been obtained. All forms have been edited and coded, 
and IBM cards have been keypimched. Partial analysis of the data has been 
carried out. Comparison of findings in this project with findings in the 
Collaborative Study will be carried out at a later time. 

Preparation of the manuscript has been delayed until the end of FY '72. 

Honors and Awards: None 

Publications: None 



94v 



I 



Serial No. NDS (CF)-63 PR/OC 1146 

1. Perinatal Research Branch 

2. Office of the Chief 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Public Health Implications Study of Perinatal Mortality in 

the Collaborative Study and in the Collaborative Study Cities, 

Previous Title: Revision and Expansion of Previous Project Entitled A 

Commentary On The Appropriateness Of The Use Of Certain 
Tabular Data, For Formulating Generalizations Concerning 
Popvilations In The Same Cities As Those In Which The 
Collaborative Study On Cerebral Palsy, Mental Retardation 
And Other Neurological And Sensory Disorders Of Infancy 
And Childhood Is Being Conducted. 

Previous Serial Number: Same 

Principal Investigators: Z.A. Shakhashiri, M.D., FRB, NINDS 

Leonard V. Phelps, Clearwater, Florida 
Glen S. Bartlett, M.D., Case Western Reserve Univ., 

Cleveland, Ohio 

Other Investigators: None 

Cooperating Units: The Census Bureau and the National Center for Health 
Statistics cooperated in the furnishing of necessary 
statistical infoimation for the United States and cities. 
The respective state or city health departments are 
providing natality and mortality data for the Project 
cities. 

Man Years: 

Total: 0.50 

Professional: 0.30 

Other: 0.20 

Project Description : 

Obj ectives : To evaluate fetal and infant mortality of the Collaborative 
Study population and of the cities from which that population is drawn, with 
the aim of comparing the two popiolations, city by city, and institution by 
institution, on mortality characteristics. 

Methods employed : In addition to the data previously available from the 
National Center for Health Statistics for perinatal events, detailed data on 
natality and perinatal mortality are being sought for the study cities from 



95v 



Serial No. NBS (CF)-63 PR/OC 1146 

either the state or city health departments, whichever has jurisdiction for 
these records. The data include figures for livebirths, stillbirths, and 
deaths under 24 hours, 1 day to 7 days, 8 days to 28 days and 1 month to 
12 months, evaluated by birthweight, length of gestation, race and sex, and 
plurality for the years 1959 through 1966. Corresponding data will be com- 
piled by institution for the PRB study population. The state or city health 
departments have been asked to furnish either completed tabulations or raw 
data to be tabiilated (arrangements finalized) by the PRB Section on Data 
Management and Retrieval. 

Considerable effort has been and is being expended, in connection with the 
Section on Data Management and Retrieval, PRB, and the Office of Biometry, 
NIKDS, to create a usable data file of the external data being obtained in 
connection with this study. The aim is to provide a file with more general 
utility than the limited scope of this study. When such a file is created, 
the information necessary to make use of the file will be made available to 
interested persons in PRB. 

Ma.jor Findings : Evaluation of birthweight and length of gestation data for 
all core live births (first and subsequent pregnancies) reveals differences 
between races and between sexes that are generally persistent among all the 
institutions. Birthweights are lighter among non-whites than among whites, 
and among females than among males. With vdiite males the heaviest, the order 
of decline is next white female, then, at about the same weight, non-white 
males, then non-white females. There is a particular excess of non-whites at 
low birth weights (2500 grams or less). Length of gestation is shorter among 
non-white than among whites by about 1 week, with an excess of both short- 
gestation and long-gestation deliveries among non-whites. Length of gestation 
is slightly shorter among males than among females in both races, but less 
consistently so among whites. 

Perinatal mortality has declined in the study population since its first year, 
with a transient elevation in 1962. Group I mortality (fetal deaths 1001 
grams and over plus deaths under 7 days of age per 1000 total births) declined 
from 28.5 in 1959 to 17.6 in 1966 (mean 21.7) among whites and from 33.2 to 
21.8 (mean 28.4) among non-whites. Group II mortality (fetal deaths 501 grams 
and over plus deaths under 28 days per 100 total births) declined from 34.9 
in 1959 to 19.1 in 1966 (mean 26.0) among whites and from 39.8 to 25.5 (mean 
33.3) among non-whites. These trends tended to persist from institution to 
institution, though to varying degrees. 

Previous deadline could not be met because of delays in receipt of data from 
the cities and in preparing these data from their various sources into a single 
program for the computer. 

It is anticipated that the file will be completed and the present phase of the 
study reported by the end of June 1972. 

Honors and Awards : None 
Publications: None 



Serial No. NDS (CF)-66 PR/OC 1378 

1. Perinatal Research Branch 

2. Office of the Chief 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Maternal Factors Affecting Birth Weight. 

Previous Title: The Prediction of Birth Weight-Multivariate Analysis. 

Previous Serial Number: Same 

Principal Investigator: Heinz W. Berendes, M.B., PRB, NINDS 

Other Investigators! W. Weiss, Office of Biometry, NINES 

J. Deutschberger, Office of Biometry, NINDS* 
E. Jackson, Office of Biometry, NINDS 

Cooperating Units: Perinatal Research Branch, NINDS ■ 

Man Years: 

Total: 

Professional: 

Other: 

This study has been completed. 

Honors and Awards: None 

Publications: Weiss, W., and Jackson, E.G.: Maternal factors affecting 

birth weight. In Perinatal Factors Affecting Himian Development . 
Proceedings of the Special Session held during the Eight Meeting 
of the PAHO Advisory Committee on Medical Research, Washington, 
D.C., June 10, 1969. Washington, D.C., Pan American Health 
Organization, Sci. Publ. No. 185, 1969, pp. 54-59. 

* Deceased 



97 V 



Serial No. NBS (CF)-68 PR/OC 1618 

1. Perinatal Research Branch 

2. Office of the Chief 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

Jioly 1, 1970 through June 30, 1971 

Project Title: The Effect of Labor on the Outcome of the Child. 

Previous Serial Number: Same 

Principal Investigators: Z.A. Shakhashiri, M.D., PRE, NINDS 

W. Lawrence Holley, M.D., Children's Hosp., Akron, 

Ohio 
A. A. Lilien, M.D., Somerville, New Jersey 

Other Investigators: None 

Cooperating Units: Perinatal Research Branch, NINDS 

Man Years: 

Total: .08 

Professional: .06 

Other: .02 

Project Description ; 

In order to single out the effect of labor, if any, on fetal growth and 
development, a study is made of outcomes of normal pregnancies terminated 
spontaneously, compared to outcomes of similar pregnancies terminated by 
elective induction of labor and to outcomes of similar pregnancies terminated 
by elective cesarean section. 

Methods Employed : Project infants of normal pregnancies terminated as 
described above will be compared for selected outcome variables such as 
mortality, Apgar, bilirubin, mental and motor scores at eight months and I.Q. 
at four years. All three groups will be controlled for race, birth weight 
and gestation length. 

Ma j or Findings : A search is under way to ascertain whether cases are available 
in adequate numbers to make the design feasible. Additional non-Project cases 
might be needed. 

The scope of this Project has been referred to and is being undertaken by the 
Ad Hoc Task Force on Labor and Delivery. See Serial No. NDS (CF)-71 PR/OB I90U. 

Honors and Awards : None 

Publications: None 



99^ 



( 

I 



Serial No. NDS (CF) -66 PE/oB 1331 

1. Perinatal Research Branch 

2. Section on Obstetrics 
3- Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 19T0 through June 30, 1971 



Project Title: Obstetric Factors in Twin Pregnancies. 

Previous Serial Number: Same 

Principal Investigator: Rudolf F. Vollman, M. D. , PRB, WINDS 

Other Investigators: Jose G. Marmol, M.D. , PRE, NIWDS 
Irene B. Ross, PRE, HINDS 

Cooperating Units: All institutions participating in the Collaborative Study 

Man Years 

Total: 0.2 
Professional: 0.1 
Others : 0.1 

Project Description: 

Objectives : The early diagnosis of a twin pregnancy remains still an 
important problem on which depend the prenatal care and the management of 
the labor and delivery. The study has two objectives: 

1. To study the outcome of twin pregnancies in relation to the time 
the diagnosis was first established. 

2. To make a comparison of the obstetric problems presented by the first 
versus the second twin and their effect upon the fetal outcome. 

Methods- : With the help of a computer printout, an established case and card 
file, and an additional revie-w of a current file on abortions and fetal deaths, 
the twins delivered in the Collaborative Project through December 31, 1965, 
have been identified. All case records were reviewed and additional informa- 
tion or clarification was solicited from the collaborating hospitals as needed. 
The mothers' medical, family and reproductive history, together with informa- 
tion on the course of the study pregnancy, intercurrent diseases, drugs, 
obstetric complications, labor and delivery, and outcome of pregnancy were 
abstracted. These data have been used to prepare a set of tabulations for the 
study. of the variables specified above. This study has been completed and 
the variables tabulated. The preparation of the manuscript has been delayed 



I 



101 V 



Serial No. WDS (CF)- 66 PR/OB 1331 



due to a shortage of manpower. 

Honors and Awards: None 
Publications: None 



102 V 



Serial No. WDS (CF) -66 PR/oB 1333 

1. Perinatal Research Branch 

2. Section on Obstetrics 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 



Project Title: Distribution of Abortions by Chronologic and Gynecologic Age 
of the Gravida. 

Previous Serial Number: Same 

Principal Investigator: Rudolf F. Vollman, M.D. , PRB, NINDS 

Other Investigators: Jose G. Marmol, M.D., PRE, NINDS 
Irene B. Ross, PRB, NINDS 

Cooperating Units: All institutions participating in the Collaborative Study 

Man Years 



Total: 


0.3 


Professional: 


0.1 


Others: 


0.2 



Project Description: 

Objectives : Information is accumulating which demonstrates that endocrine 
and morphologic conditions for optimal reproductive performance are reached 
only several years after menarche. The conventional association of pregnancy 
outcome by chronologic maternal age will be compared with the mother's 
gynecologic age, based on the age at menarche. 

Lfethods : For study pregnancies terminating in abortion, a number of important 
maternal variables have been abstracted from the study record: race, 
chronologic and gynecologic age, marital status, gravidity and parity, length 
of the menstrual cycle, medical and obstetric complications of the study 
pregnancy, duration and outcome of the study pregnancy. These variables 
serve as controls in the analysis of chronologic versus gynecologic maternal 
age. 

Based on the preliminary findings, this study has been extended to include all 
fetal deaths in the Perinatal Research program. Work is progressing very 
slowly due to a shortage of manpower. 



103 V 



Serial Wo. NDS (CF) -66 PR/oB 1333 



Honors and Awards: None 
Publications: None 



104^ 



Serial Wo. NDS (CF)-68 PR/oB 1623 

1. Perinatal Research Branch 

2. Section on Obstetrics 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 



Project Title: Prenatal Drugs. 

Previous Serial Number: Same 

Principal Investigator: Alan L. Scriggins, M. D., University of Vermont 

Hospitals 

Other Investigators: Rudolf F. Vollman, M.D., PRE, WINDS 

Seymour Katsh, Ph.D., University of Colorado Medical 

Center 
S. Barbara Katz, Office of Biometry, WINDS 

Cooperating Units: All institutions participating in the Collaborative Study 



The study has been completed. 
Honors and Awards: None 
Publications : None 



105 



Serial No. NDS (CF)-68 PR/oB 1625 

1. Perinatal Research Branch 

2. Section on Obstetrics 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 



Project Title: Menstruation and Ovulation in the Monkey. 

Previous Serial Number: Same 

Principal Investigator: Rudolf F. Vollman, M.D. , PRE, NINDS 

Other Investigators: Irene B. Ross, PRE, NINDS 

Cooperating Units: Department of Embryology, Carnegie Institution of 
Washington 

The study has been discontinued. 



^ 



107V 



Serial No. NDS (CF)-TO PR/oB I85O 

1. Perinatal Research Branch 

2. Section on Obstetrics 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 



Project Title: Toxemia in Pregnancy and Its Relationship to the Outcome of 
Pregnancy. 

Previous Serial Number: Same 

Principal Investigator: Rudolf F. Vollman, M.D. , PRB, NINDS 

Other Investigators: Leon C. Chesley, M.D., Downstate Medical Center, 

Brooklyn, New York 
Russell R. de Alvarez, M.D., Temple University School 

of Medicine, Philadelphia, Pa. 
R. Gordon Douglas, M.D. , Providence Lying- In Hospital, 

Providence, R.I. 
Emanuel A. Friedman, M.D. , Harvard University Medical 

School, Boston, Mass. 
Adolph H. Sellmann, M.D., Charity Hospital, New Orleans, 

La. 
Gilbert J. Vosburgh, M.D., Columbia University, College 

of Physicians and Surgeons, New York, N.Y. 

Cooperating Units: All institutions participating in the Collaborative 
Project 

Man Years 

Total: 1.6 
Professional: 0.8 
Others: 0.8 

Project Description: 

The purpose of this study on toxemia of pregnancy is threefold: 

1. To establish quantitative gradients for defined areas of the individual 
signs of toxemia versus outcome of pregnancy. 

2. To measure the relative weights of the clinical signs of toxemia -- 
hypertension, proteinuria, edema -- singly and combined, in their 
contribution to perinatal mortality and birthweight. 



109 V 



Serial No. NTS (CF)-TO PR/oB 185O 

3. To define a cohort of cases based on 1 and 2 as "pregnancies 

complicated "by toxemia" in the Perinatal Research Study for further 
research in association with the long-term development of the children. 

Methods : It was demonstrated in a preliminary study of the clinical reports 
of toxemia in the Study that the rates of toxemia vary widely between the 
participating institutions and the 2 different Study forms used, even when the 
populations of the gravidae were controlled for race, age, and parity. It was 
concluded that this large variability in the incidence of toxemia must be 
related to the use of different diagnostic criteria by the individual 
collaborating hospitals. It was therefore decided to use the original (raw) 
data of the clinical symptoms of toxemia and to analyze them individually and 
in combination to establish a uniform cohort of cases with toxemia for the 
study. The study will be carried out in cooperation with a special ad hoc 
Task Force on Toxemia and will absorb all of the professional and none 
professional time available. 

Some preliminary findings were presented at the International Symposium on 
Gestosis in Basel, Switzerland, April 25-26, 1969. 

Honors and Awards: None 

Publications: Vollman, R.F. : Rates of toxemia by age and parity. In 
Rippmann, E.T. (Ed.): Die Spaetgestose (EPH-Gestose) . 
Basel, Switzerland, Schwabe & Co. Verlag, 1970, pp. 338-3ij-2. 



110 V 



Serial No. WDS CCF)-TO PR/OB 185I 

1. Perinatal Research Branch 

2. Section on Ohstetrics 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1910 through June 30, I97I 

Project Title: Low Eclampsia Rate in the Collaborative Project. 

Previous Serial Number: Same 

Principal Investigator: Jose G. Marmol, M.D. , PRE, NINDS 

Other Investigators: None 

Cooperating Units: None 

The study has been completed. 

Honors and Awards: None 

Publications: Marmol, J.G. : Cases of eclampsia in the Collaborative Project. 
Schweiz. Z. Gynaek. Geburtsh. 1: 169-I80, 1970 



f 



1 

I 



lliv 



Serial No. WDS (CF)-71 PR/OB 190U 

1. Perinatal Research Branch 

2. Section on Obstetrics 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 3O;, 197I 

Project Title: The Study of Labor and Delivery 

Previous Serial Number: None 

Principal Investigator: Rudolf F. Vollman, M.D., PRE, NINDS 

Other Investigators: Emanuel A. Friedman, M.D. , Harvard Iftiiversity Medical 

School, Boston, M^ss. 
Luke Gillespie, M.D., Boston Lying-in Hospital, Boston, 

Mass. 
lyfe-rvin Green, M.D., Nev York Medical College, New York, 

N.Y. 
Esther Jackson, Office of Biometry, NINDS 
Schuyler G. Kohl, M.D., State University of New York, 

Downstate Medical Center, Brooklyn, N.Y. 
Kenneth R. Niswander, M.D. , University of California at 

Davis, Calif. 
David Rubinstein, Office of Biometry, NINDS 
Vincent Tricomi, M.D., Brooklyn- Cumberland Medical Center, 

Brooklyn, N.Y. 

Cooperating Units: All institutions participating in the Collaborative Project 

Man Years 

Total: 0.3 
Professional: 0.2 
Others: 0.1 

Project Description: 

This study has two objectives: 

A. To deteiroine the influence of specific labor and delivery factors on 
the fetus in terms of immediate outcome and later neurologic and 
psychologic development . 

B. To develop a reference standard of actuarial, obstetric, and labor and 
delivery features that will result in optimal outcome. 



113 V 



Serial No. NDS (OF) -71 PR/OB 190U 



Labor is usually reported in quantitative units of hours and minutes and is 
conventionally divided into short, normal, and long labor by arbitrarily 
selected time intervals. These intervals, in turn, have been associated with 
certain complications of labor. FKEEDMAU has demonstrated in a long series 
of publications that these associations do not necessarily reflect the under- 
lying physiology or pathology of uterine activity. He devised a methodology 
to quantify uterine activity and its deviations in relation to the phases of 
labor, which are based upon the observed progress of labor in time. Possible 
damage to the fetus is not necessarily dependent upon the total duration of 
labor, but on the dyscoordination or changes in the du3ra.tion of any one or 
combinations of the phases of labor. 

The identification of gravidae with specified types of labor will produce 
several standard cohorts that may be used by other Study sections and task 
forces (Pediatric Neurology, Pathology, Physical Growth and Development etc.) 
as variables for their respective special studies. 

Msthodology: 

A reference cohort of gravidae with "normal labors" is defined, for which the 
specified outcome will be tabulated. This reference cohort will be compared 
with cohorts of specified types of dysfunctional labor and their respective 

outcomes. 

Those gravidae with incomplete data on labor and delivery in the Study record 
will be identified and their general characteristics (race, age, parity, sex 
of fetus) and outcomes will be tabulated separately. 



Honors and Awards: None 
Publications: None 



114 V 



Serial No. NDS (CF)-Tl PR/OB 1905 

1. Perinatal Research Branch 

2. Section on Obstetrics 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

Jxily 1, 1970 through June 30, 1971 

Project Title: Reference Bibliography on Human Teratology. 

Previous Serial Number: None 

Principal Investigator: Rudolf F. Vollman, M.D. , PRB, NINDS 

Other Investigators: None 

Cooperating IBiits: Reference and Interlibrary Loan Sections of the NIH 
Library 
Reference Section of the National Library of Medicine 

ysan Years 

Total: 0.3 
Professional: 0.2 
Others: 0.1 

Project Description: 

The scientific publications on human teratology during the past 170 years 
have been reviewed and approximately 4000 items have been selected, analyzed 
gind coded by content. The criteria for selection were: 

1. originality of observation reported 

2. morphological classification of malformations 

3. morphogenesis of malformations in association with embryological stages 
k. hypothesis on etiology of malformations in time and teratogenic factors 

5. experimental teratology 

6. review articles. 

Findings : The study of teratology was initially stimulated by the curiosity 
to explore the range of the lusus naturae from which very early concepts on the 
embryological potential were deducted. It thus contributed to the development 
of systematic research in normal human embryology. From experimental embryology, 
critical times, effective factors, their quantity and quality of interaction 
with the formative process were Identified and have been extrapolated to explain 
possible mechanisms in human teratology. 

The manuscript is ready for publication. 

1115 V 



Serial No. WDS (CF)-Tl PE/oB I905 



Honors and Awards: None 
Publications: None 



116 V 



Serial No. NBS (CF)-63 PR/PN 1163 

1. Perinatal Research Branch, NINES 

2. Section on Pediatric Neurology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
Jvily 1, 1970 through June 30, 1971 

Project Title: An Investigation into the Relationship Between Congenital 
Heart and Great Vessel Anomalies and Selected Factors as 
Recorded in the Collaborative Perinatal Research Project. 

Previous Serial Number: Same 

Principal Investigator: Lenore Bajda, M.D., PRB, NINDS 

Other Investigators: Heinz W. Berendes, M.D,, PRB, NINDS 
John L. Sever, M.D., PRB, NINDS 

Cooperating Units: Office of the Director, NHI 

Man Years: 

Total: 

Professional: 

Other: 

Project Description ; The primary objective of this study is to assess 
relationships between certain maternal variables and congenital heart-great 
vessel anomalies. 

Additional objectives include investigating relationships between early 
signs of abnormality and the existence of definitive congenital heart lesions 
and determining the existence of congenital heart-vessel anomaly in conjunction 
with mental retardation as recorded in the eight-^nonth psychological examina- 
tion, the one-year summary records, four-year psychological examination, and 
seven-year neurological and psychological examinations. 

To date, maternal parameters analyzed included age of gravida, parity, prior 
pregnancy outcome, prior and current health status, ABO blood group, current 
smoking pattern, and viral antibody status. 

Study data was obtained from Collaborative Study records received by PRB 
from the onset of the Study (January 1959) through December 1964. These 
records provided 112 live and stillbirth cardiac cases for study out of a 
population pool of approximately 38,000. Analysis of an expanded Study 
cohort through 1965, with a population pool of approximately 55,000 providing 
additional cases, is underway in anticipation that the additional cases will 
support earlier findings and perhaps provide further clues for identifying 
etiological relationships. 



I 



117 V 



Serial. No. NDS (Cr)-63 PR/PN 1163 

There was a definite preponderance of mothers over 30 in the C-V Study group. 
Controling for race, and removing cases with chromosomal aberrations, there 
were more white mothers in the 30 and over age group than expected at the 
.05 level. This trend is also noted among Negroes. There was a greater than 
expected number of gravida with systemic disease complications and prior 
pregnancy loss among the mothers of the cardiacs. A breakdown of these 
factors for greater specificity is pending. 

Because the number of patients with each specific cardiac abnormality was 
small, specific associations between serological findings and clinical 
observations were not possible, although several interesting trends were 
noted. Analysis on the larger cohort is nearing completion. 

A preliminary report on the 1964 cohort study was presented at the 1966 
Annual Meeting of the Teratology Society. 

The urgent priority of processing the entire Collaborative Project study data 
has temporarily delayed further work on this study. 

Honors and Awards: None 

Publications: None 



118 V 



Serial No. NDS (CF)-63 PR/PN ll6k 

1. Perinatal Research Branch, HINDS 

2. Section on Pediatric Neurology 

3. Bethesda, tferyland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Early Signs as Predictors of Death and Neurological Abnor- 
mality Among Premature Infants Weighing 1000-2000 Grams 

Previous Serial Number: Same and incorporating Serial No. NDS (CF)-69 

PR/PN YJUO 

Principal Investigator: Joseph S. Drage, M.D., PRE, NINDS 

Other Investigators: Karin B. Nelson, M.D., PRE, NINDS 
Heinz Berendes, M.D., PRE, NINDS 

Cooperating Units: None 

Wfen Years: 

Total: .00 

Professional: .00 

Other: .00 

Project Description : A group of I365 single livehom premature infants with 
"birth-weights of 1000-2000 grams has been prospectively studied relating the 
presence of specific neurological signs during the nursery period to neuro- 
logical status at one year of age. Of the I365 infants, 917 were examined 
at one year, 27^ died during the first year, and 173 were lost to follow-up. 
Ihere were 201 neurologlcally abnormal Infants among the 917 examined at 
one year. The first fovir PED-2's (Neonatal Pediatric Examinations) were 
reviewed for the presence or absence of early signs, A positive early sign 
was defined as the occurrence, on at least one examination of the specific 
early sign being studied. Specific signs studied included cry, suck, palmar 
grasp, traction response, Moro reflex, eye movement, muscle tone, local 
convulsions, general convulsions, highest serum bilirubin. Coombs* test, 
procedures of resuscitation, etc. Each of these signs was considered 
separately, and those showing significant association with death and abnormal 
outcome were then combined. When three signs were considered in combination, 
the. more that were positive the greater the association of neurological 
abnormality and death, and of neurological abnormality alone. 

A preliminary manuscript has been prepared. Because of Task Force study 
assignments, this project is in abeyance. 

Honors and Awards: None 

Publications : None 



119 V 



Serial No. EDS (CF)-66 PR/PN 1335 

1. Perinatal Research Branch, NIHDS 

2. Section on Pediatric Neurology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, I971 

Project Title: Mortality and Morbidity Among Infants Weighing 1000-2000 Grams 

Previous Serial Number: Same 

Principal Investigator: Joseph S. Drage, M.D., PRE, NINDS 

Other Investigators: Karin B. Nelson, M.D., PRE, NINDS 
B.H. Williams, M.D., PRE, NINDS 



Cooperating Units: None 




Man Years: 




Total: 


.00 


Professional: 


.00 


Other: 


.00 


Project Description: A group 


of 1 



A group of I36U liveborn infants, with birthweights 
ranging from 1000-2000 grams, has been studied regarding outcome within the 
first year of life. This group of I36U infants represents approximately 
2^ of the group of 55^000 single live births from which they were drawn. 

Within the group of I36U infants, 9IT were examined at one year of age, ajid 
of these 201 were considered to have definite neurological abnormality. There 
were 27^ deaths during the first year of life. Thus, U75 of the infants either 
died during the first year of life or were considered neurologically abnormal 
by examination at one year. There were 173 infants lost to follow-up and 
this represents 12^ of the original I36U cases. Over 50^ of the deaths 
occurred during the first 2k hours and over 90^ occurred during the first 
28 days. 

The 201 infants neurologically abnormal at one year were classified in the 
following way: isolated motor retardation, diplegia, hemiplegia, quadri- 
plegia, monoplegia, athetosis, motor retardation with neurological signs 
insufficient for other diagnosis, and other (infants with neurological signs, 
but not fitting a specific diagnostic category). Congenital malformations 
were also monitored for this group. 

Outcome at one year (death, abnormal, normal, and lost) was then tabulated 
within each 100-gram birthweight interval. In general for I36U infants, 
as the birthweight increased, the percent of deaths decreased. For infants 



121V 



Serial Noi NDS (CF)-66 PR/PN 1335 

in jeopardy (dead or abnormal at one year), the same relationship held. The 
percent of lost cases within each 100-gram birthwelght interval varied 
slightly. Data has been obtained and is undergoing analysis 

Because of Task Force study assignments, this project is in abeyance. 

Honors and Awards: None 

Publications: None 



:it2a 



Serial No. KDS (CF)-66 Pr/pN 1338 

1. Perinatal Research Branch, KIKDS 

2. Section on Pediatric Neurology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, I97I 

Project Title: The Association of Mental Sub normality with Head Circumfer- 
ence, Congenital IVfelformations, and Other Conditions of the 
Newborn Term Infant 

Previous Serial Number: Same 

Principal Investigator: Lenore Bajda, M.D., PRB, NINDS 

Other Investigators: Karin B. Nelson, M.D., PRE, NINDS 

Cooperating Units: Office of Biometry 

Man Years: 

Total: 
Professional: 
Other : 

Project Description : The objective of this study was to determine the 
relationship between head size and certain other physical features of the 
Collaborative Study child noted shortly after birth, at the one-year examina- 
tion and at k year upon completion of the psychological examination. The 
project was in abeyance pending updating of the data file. Meanwhile 
Dr. Nelson and Mr. Deutschberger have completed their study on "Head Size at 
One Year as a Predictor of Four- Year IQ" (PR/PN I5O9) using a sample of the 
Collaborative Study population including a partially selected pediatric group 
of 9} 379 children. They concluded that there is approximately a ^O'fo chance 
of an IQ of less than 80 at k years of age for the one-year male with a head 
size less than k3 cm. and a one-year female with a head size of less than h2 cm. 
Plans are under way to examine in detail the low and high head measure sample 
of the Nelson-Deutschberger study for other factors which might account for the 
correlation between head size and the four-year IQ values. Depending on the 
results of this analysis, the original study proposal will then move either 
forward with a larger sample, or terminate. 

The -urgent priority of processing the entire Collaborative Project study data 
has temporarily delayed further work on this study. 

Honors and Awards: None 

Publications : None 



I 



123 



Serial No. EDS (CF)-68 PR/pN I628 

1. Perinatal Research Branch, NIMDS 

2. Section on Pediatric Neurology 

3. Bethesda, IVfe-ryland 

PHS - NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Effects of Prenatal Protein Deprivation on Behavior and 
Brain Structure of- Mice 

Previous Serial Number: Same 

Principal Investigators: John Ao Churchill, M.D., PRB, NINDS 

J. H. Carleton, M.D., Miami, Florida 
W. lawrence Holley, M.D., Children's Hosp., Akron, 

Ohio 
Other Investigators: None 

Cooperating Units: None 

Man Years: 

Total: 
Professional: 
Other 

Ten^jorarily discontinued pending a time -when brain specimens can he processed 
and examined. The brains are now in celloidin blocks. Dr. Jack Carleton, 
Co- investigator, has left the Branch. 

Honors and Awards: None 

Publications : None 



i 



125 



Serial Wo. HDS (CF)-68 PR/pn 1633 

1. Perinatal Research Branch, KINDS 

2. Section on Pediatric Neurology 

3. Bethesda, tferyland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Neuropsychologic Outcome of Children with Retinal 
Hemorrhages at Birth 

Previous Serial Number: Same 

Principal Investigators: Arthur L. Rosenhaum, M.D. , U.C.L.A., Los Angeles, Cal. 

John A. Caiurchill, M.D., PRB, NINDS 

Other Investigators: None 

Cooperating Units: None 

Man Years: 

Total: .05 

Professional: .05 

Other: .00 

Project Description : Retinal hemorrhages in the newborn occur in approximately 
20^0 vertex births. Much work has been done to elucidate the etiology of 
these hemorrhages, but little is known about their significance in terms of 
long term follow-up. This study was designed to study the neuropsychologic 
outcome of children with retinal hemorrhages, and also the relationship 
between laterality of the hemorrhage and birth position. 

Approximately 200 cases of retinal hemorrhage in the newborn were studied, 
and matched with "non-hemorrhage" controls. These matched pairs were then 
compared in rel^itionship to the following outcome variables: a) Birthweight, 
b) Bay ley mental and motor scores given at 8 months of age, and c) The Binet 
IQ given at k years of age. 

The retinal hemorrhage cases were also reviewed in an attempt to analyze the 
possibility of a relationship between birth position and the eye in which 
the hemorrhage occurred. 

This material is still in the process of being prepared for submission for 
publication. 

Honors and Awards: None 

Publications: None 



I 



127 V 



Serial Wo. UDS (crF)-69 Er/PW 17^+8 

1. Perinatal Research Branch, MIKDS 

2. Section on Pediatric Wetirology 

3. Bethesda, lyfe-ryland 

PHS-MH 
Individiial Project Report 
JvCLy 1, 1970 through Jime 30, I97I 

Project Title: Neonatal Polycythemia: I. A Manifestation of Chronic 
Injury During Distress 

Previous Serial Number: Same 

Principal Investigator: Miles M. Weinberger, M.D., Nat*l, Jewish Hosp. 

Denver Colorado 

Other Investigators: Arthur Oleinick, M.D., EPID, NCI 

John A. Churchill, M.D., PRE, EIITOS 

Cooperating Units: None 

Ifen Years: 

Total: .05 

Professional: .05 

Other: .00 

Project Description : The objectives of the study were to study demographic 
and maternal factors associated with neonatal polycythemia. 

As part of the protocol dtiring the Collaborative Study on Cerebral Palsy, 
capillary hematocrits were obtained as near to k8 hours as possible (generally 
between 36 and 60 hours) on hhj6&3 newborns, or SGfo of all 77 and over were 
identified, and controls, matched for institution and year of birth, race 
sex, socioeconomic index, and presence or absence of U-year follow-up examina- 
tion were selected randomly from all infants with i+6-hour hematocrits 50 
through 65. Various demographic and maternal factors were identified on both 
subjects and controls, and differences were statistically evaluated. 

The subject cases (those with polycythemia manifested by hematocrits 77 and 
over) were found to have been the product of longer gestation, but were 
smaller in weight than the control population. There was an increase in 
incidence of placental pathology and the placenta of the subject cases was 
significantly lighter than the weights of the controls. One-minute and five- 
minute Apgar scores were both lower in the subject cases and there was a 
greater incidence of dysmaturity diagnosed in the subject cases. When compared 
with the whole Collaborative Study population, infajits with polycythemia were 
noted to come from lower socioeconomic groups. Data suggest that neonatal 
polycythemia may be manifestation of chronic intrauterine distress. 



L 



129^ 



Serial No. KDS (CF)-69 PR/pn rjkS 

This material was presented at a Perinatal Research Branch (nIKDS) seminar 
m April 1970. Considerable progress has been made toward completing this 
study for submission for publication. However, no additional work will be 
done on it until after June 1971. 

Honors and Awards: None 

Publications : None 



130 V 



Serial No. EDS (CF)-69 PR/PN 17^9 

1. Perinatal Research Branch, NIKDS 

2. Section on Pediatric Neurology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Neonatal Polycythemia: II. Outcome 

Previous Serial Number: Same 

Principal Investigators: Miles M. Weinberger, M.D., Nat'l. Jewish Hosp., 

Denver, Colorado, John A. Churchill, M.D., PRE, NINDS 

Other Investigators: Arthur Oleinick, M.D., EPID, NCI 

Cooperating Units: None 

Man Years: 

Total: .05 ■ 
Professional: .05 
Other: .00 

Project Description : The objectives of the study were to determine polycythemia 
in the neonatal period as an adverse effect on neuropsychological outcome. 

See project report on Neonatal Polycythemia: I. A Manifestation of Chronic 
Injury During Distress covering demographic and maternal factors. In addition, 
the neuropsychological status of these infants was examined at various 
intervals through four years of age. 

Examining only the outcome of those infants without recognized congenital 
malformation and gestational ages of 36 weeks and greater, it was found that 
while these infants tended to be small for gestation age and have lower Apgar 
scores, there was no significant difference in other findings in the newborn 
period. There was also no difference in psychological scores at eight months. 
At one year, there was no difference in abnormal neiorological findings between 
subjects and control cases. At four years, however, the subjects did manifest 
a statistically- significant lower score on the U-year Stanford-Binet examina- 
tions than the controls. This difference was greatest among Negroes, especially 
among the Negro females. White males did not manifest any difference in k-yeax 
IQ. 

Significance is not so much that some differences in U-year IQ were found among 
some of the subgroups of the polycythemic Infants, but that contrary to the 
expectations from the literatiore the differences between polycythemic infants 
and non-polycythemic controls were so small. 



131 V 



Serial No. NDS (CF)-69 PR/PN 17^9 

This material is still in the process of being prepared for submission for 
publication. However, no additional work will be done on it until after 
June, 1971. 

Honors and Awards : None 

Publications : None 



132 V 



Serial Wo. NDS (CF)-69 PR/PN 1750 

1. Perinatal Re search Branch, KLWS 

2. Section on Pediatric Neurology 

3. Bethesda, Iferyland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, I97I 

Project Title: Congenital IVferrow Dysfunction in Down's Syndrome 

Previous Project Title: Abnormal Hematopoiesis in Newborns with Down's 

Syndrome 

Previous Serial Number: Same 

Principal Investigators: Miles M. Weinberger, M.D., Nat'l Jewish Hosp. 

Denver, Colorado 
Arthur Oleinick, M.D., EPID, NCI 

Other Investigators: None 

Cooperating Units: None 

Man Years: 

Total: .05 

Professional: .05 

Other: .00 

Study has been completed. 

Honors and Awards : None 

Publications : Weinberger, M.M. and Oleinick, A. : Congenital marrow 

dysfunction in Down's syndrome. J. Pediat . 77: 273-279, 1970 



133 V 



Serial No. NDS (CF)-70 PR/PN I853 

1. Perinatal Research Branchy KIKDS 

2. Section on Pediatric Neiirology 

3. Bethesda, tfe-ryland 

PHS-KEH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: The Effects of Protein Malnutrition on Ontogeny of the Brain 

Previous Serial Number: Same 

Principal Investigator: John A. ChTirchill, M.D., PRE, NINDS 

Other Investigators: None 

Cooperating Units : None 

Man Years: 

Total: 
Professional: 
Other : 

Project Description ; A review was made of the literature pertaining to the 
effect of protein-calorie malnutrition of the mother during pregnancy on 
the nervous system structure and function of the offspring. From this 
review, a critique was prepared and presented at the meeting of the American 
Association for the Advancement of Science, held in Boston, Massachusetts 
on December 26-31, 1969* Study has been completed. 

Honors and Awards: None 

Publications : None 



I 



135 V 



Serial No. NDS (CF)-70 PR/PN 1854 

1. Perinatal Research Branch, NINES 

2. Section on Pediatric Neurology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: The Etiology of Cerebral Palsy in Prematures. 

Previous Serial Number: Same 

Principal Investigator: John A. Churchill, M.D., PRE, NINES 

Other Investigators: None 

Cooperating Units: None 

Man Years: 

Total: .30 

Professional: .25 

Other: .05 

Project Description : Theories of the etiology of cerebral palsy were studied 
in a group of 1364 singleton liveborn infants weighing 2.0 Kg. or less at 
birth, the group comprising all such small babies in the Collaborative Project 
totalling 54,370 births. Subjects diagnosed as having cerebral palsy at 
1 year of age numbered 45. 

No support for the genetic theory was obtained by a study of siblings of the 
spastics. 

The anoxia theory gained no support in that Apgar and other indirect measures 
of asphyxia differed insignificantly between the spastic and equally premature 
non-spastic groups. 

Hyperbilirubinemia did not appear to cause spastic diplegia since bilirubin 
levels differed very little between spastics and non-spastics. Of 18 cases 
with erythroblastosis, none had cerebral palsy. 

Intrauterine blighting of the fetus was considered unlikely. Intrauterine 
disturbances often produce growth retardation. None of the small-for-dates 
infants had spastic diplegia. Furthennore, 5 spastics were born by "elective" 
caesarean section, one being remarkably free from abnormal prenatal events. 

Evidences of cranial injury or of factors conducive to birth trauma were no 
more frequent in spastic than in other prematures. The spastic cases born 
by caesarean section, where travmia should be minimized, weighed against the 
traimia theory. 

Cerebral hemorrhage as the cause of premature spastic diplegia was found to 

137 V 



Serial No. NDS (CF)-70 PR/PN 1854 

desei*ve attention. Hematocrits obtained in the first few postpartim days 
were significantly lower in spastic than in non-spastic prematures. 
HemodJlution did not explain the observation. Others have shown that low 
hematocrits occur in prematures who have cerebral hemorrhage. 

Compelling and direct evidence that cerebral hemorrhage causes spastic 
diplegia of prematurity cannot be derived from this study. However, the 
results make clear a need to investigate hemorrhagic mechanisms in prematures. 

This has been submitted for publication. 

Honors and Awards: None 

Publications: None 



138 V 



Serial No. NDS (GF)-68 PR/bS l6kO 

1. Perinatal Research Branch 

2. Section on Behavioral Sciences 

3. Bethesda, Iferyland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: The relationship of demographic, perinatal and other 

developmental characteristics to intellectual and motor 
performance, of pre- school children. 

Previous Serial Number: Same 

Principal Investigator: Sarah H. Broman, Ph.D., PRB, NINDS 

Other Investigators: Jaswant Khanna, Ph.D., University of Tennessee 

Joseph Weber, formerly Office of Biometry, NINDS 

Cooperating Units: University of Tennessee 

Office of Biometry, NINDS 

Man Years: 

Total: .25 
Professional: .20 
Other: .05 

Project Description : 

30 characteristics of some 13,000 study children have been related to their 
pass-fail performance on individual items in the psychology battery admin- 
istered at four years of age. Preliminary analyses relating four demographic 
variables to performance on the Stanford-Binet show that: (l) differences 
among white, Negro and Puerto Rican children in IQ, socioeconomic index, 
educational level of mother and proportion of males to females are highly 
significant. Whites exceed the other two groups in mean IQ and socioeconomic 
index. Differences among the groups in mean number of years of mother's 
education are more evenly spaced with whites ^ Negroes p' Puerto Ricans. 
Sex ratios in these groups follow a different order with Puerto Ricans :?' 
whites 7 Negroes; (ll) Highly significant differences in percent pass by 
ethnic group occur on all 28 of the Stanford Binet items examined. These 
items cover the test levels from 2 years 6 months through 6 years. Among 
whites percent pass is significantly higher than among the total group on 
27 items. Among Negroes percent pass is significantly lower than the total 
group on 26 items. Among Puerto-Ricans, percent pass is significantly lower 
than the total group on I6 items, does not differ from the total group on 
nine items and is higher on three items: (ill) Within both the white and 
Negro groups, children who pass have a significantly higher socioeconomic 
index than those who fail on all of the 28 Binet items. Among Puerto- 
Ricans however these differences occur on only 10 items; (iV) Similarly, 
among both whites and Negroes, educational level of mother is significantly 

139 V 



Serial No. KDS (CF)-68 PR/bS l6kO 

higher for passers than for those who fail on all 28 items. However for 
Puerto Ricans this occiirs on only five items; (v) Among whites, se:x; 
differences in performance occur on 13 of the 28 Binet items with 
proportionally more males than females failing these items. Among Negroes 
sex differences in favor of femaJ.es occur on 15 items and in favor of males 
on one item. Among Puerto Ricans sex differences are significant on only 
five items, again with more males than females failing these items. 

The content of the items on which all of the above differences occur is of 
course highly relevant and is now being categorized. The majority of test 
items administered at this age are verbal. However the following trends in 
ethnic group differences have been noted. Using McNemar's classification 
of items, verbal item differences are largest between white and Puerto 
Ricans; memory item differences are largest between whites and Negroes, non- 
verbal item differences are relatively small among all groups. Using five 
factors identified by Valett, whites are consistently higher than both 
Negroes and Puerto Ricans on groups of items representing the three factors 
of general comprehension, vocabulary and verbal fluency, and judgement and 
reasoning. For the factor of visual motor ability, whites consistently 
exceed Negroes but only slightly exceed Puerto Ricans. For the last factor 
considered, memory and concentration, whites are higher than Negroes and 
slightly higher than Puerto Ricans. When Negro-Puerto Rican differences 
are examined, the two groups differ very little on the factors of general 
con^jrehension and memory and concentration. Puerto Ricans are higher on 
the visual motor items and Negroes are higher on vocabulary and verbal 
fluency and to a lesser extent on judgement and reasoning items. 

Subseq^uent analyses will explore the relationship of perinatal, neur-ologi cal 
and other psychological variables to performance at four years of age. 

Honors and Awards: None 

Publications: None 



1^40 V 



Serial No. M)S (GF)-69 PR/bS 1752 

1. Perinatal Research Branch 

2. Section on Behavioral Sciences 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, I97I 

Project Title: Duration of Membrane Rupture and Psychological Outcome 

Previous Serial Number: Same 

Principal Investigator: Dr. Lee Willerman, PRB^NINDS 

Other Investigator: Dr. John A. Churchill, PRB^NINDS 

Cooperating Unit: Section on Pediatric Neirrology, PRB,NINDS 

This project has been discontinued. 



l4iv 



Serial No. EDS (GF)-69 PR/BS 1753 

1. Perinatal Research. Branch 

2. Section on Behavioral Sciences 

3 . Bethe sda, Maryland 

PHS-NIH 
Individual Project Report 
July 1^ 1970 through June 30, 1971 

Project Title: Growth and Intellectual Development of Children from 
Consanguineous Matings 

Previous Serial Number: Same 

Principal Investigator: Dr. Lee Willerman, PRB_, NINDS 

Other Investigators: Dr. Ntinos C, ]\fyrianthopoulos, PRE, NINDS 

Dr. Alfred F. Naylor, PRB, NINDS 

Cooperating Unit: Section on Epidemiology and Genetics, PRE, NINDS 

Man Years: 

Total: .10 
Professional: ,05 
Other: .05 

Project Description : 

Objectives ; Consanguineous matings have been reported to be associated 
with a higher Incidence of fetal and neonatal mortality as well as mental 
retardation in the offspring. The present study will examine outcomes in 
the offspring of l48 sets of parents in the Collaborative Study who are 
second cousins or closer. Cases coded as consanguineous are now being 
reviewed for validity of such codes in the Section on Epidemiology and 
Genetics. Following this review, analysis of the outcome variables is 
expected to proceed quickly. 

Honors and Awards: None 

Publications: None 



143 V 



Serial No. KDS (CF) -69 PR/BS YJ^k 

1, Perinatal Research Branch. 

2, Section on Behavioral Sciences 

3, Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Growth and Intellectual Development of Children From 
Interracial IVlatings 

Previous Serial Number: Same 

Principal Investigator: Dr. Lee Willerman, PRB, NINDS 

Other Investigators: Dr. Alfred R, Naylor, PRB, NINDS 

Dr. Ntinos G. jy^rrianthopoulos, PRE, NINDS 
Dr. John A. GhurchiU., PRB, NINDS 

Cooperating Units: Section on Epidemiology and Genetics, PRE, NINDS 

Section on Pediatric Neurology, PRE, NINDS 

Man Years: 

Total: .40 
Professional: .35 
Other: .05 

Project Description : 

Objectives : Offspring from Negro-white matings (n=17l) were individually 
matched to children from white-white and Negro-Negro matings on hospitaJ. 
of birth, socioeconomic index, and marital status. Though not significantly 
different from the controls in either length or weight at birth, by four 
months of age the interracials were significantly smaller than the controls. 
At one year the interracials were still smaller, but the magnitude of the 
differences had diminished considerably. On psychological test performance 
at eight months, no differences were observed. At four years the IQs of the 
interracial children were significantly lower than the white controls, but 
not significantly lower than the Negro controls. Birthweights and lengths 
of the interracial children were intermediate to the larger white and 
smaller Negro children regardless of the race of the interracial mother. 
These results suggest a genetic basis to the findings that Negro children 
weigh less and are shorter than white children at birth. 

Honors and Awards: None 

Publications: 

Willerman, L, , Naylor, A. F., ly^nrianthopoulos, N. C. : Intellectual 
Development of Children from Interracial Matings. Science , I97O, 
Vol. 170, pp. 1329-1331. 

145 V 



Serial No. NDS (CF)-69 PR/BS I756 

1. Perinatal Research. Branch. 

2. Section on Behavioral Sciences 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Preschool Stuttering and Early Maternal Attitudes 

Previous Serial Number: Same 

Principal Investigator: Raymond H. Holden, Ed.D. , Brown University 

Other Investigator: Paul J. LaBenz, Sc.D., PRB, NINDS . 

Cooperating Unit: Child Development Study, Brown University 

Man Years: 

Total: .35 
Professional: ,30 
Other: .05 

Project Description : 

This study evaluates the relationship of dysfluency in speech noted in 
children at age three years, and attitudes of their mothers as reported on 
the Parental Attitude Research Instrument (PARl) when the children were 
8 months old, A sample of 1100 children in the Providence Child Development 
Study was screened for speech, language and hearing problems at three years 
of age. Twenty six children were identified as dysfluent. Two random 
sanrples consisting of 26 cases showing no dysfluency and 2.6 cases rated as 
unknown were selected from their respective groups within the sample. 

No significant differences were found between the dysfluent and normal 
groups with regard to total PARI scores, socioeconomic status or mean IQ 
(Revised Stanford-Binet Form L-M, I960) at age \ years. The group rated 
unknown was I9 points lower than both of the other groups in mean IQ. 
Review of their records indicated that they could not be rated for dysfluency 
because they were uncooperative, ixntestable and/or mentally retarded. 
Follow-up on partial samples of the dysfluent group revealed a marked drop 
in mean verbal IQ at age 7 years, and a high proportion of abnormal 
articulation ratings at age 8 years. This study will be completed when 
additional follow-up data is available. 

Honors and Awards : None 

Publications: None 



1^7a 



Serial No. NDS (CF)-69 PR/BS 1757 

1. Perinatal Research Branch. 

2. Section on Behavioral Sciences 

3. Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Social Class and Outcome in the Neurologically Abnormal 
Infant 

Previous Serial Mumber: Same 

Principal Investigator: Raymond H, Holden, Ed.D., Brown University 

Providence, Rhode Island 

Other Investigator: Lee Willerman, Ph.D., PRE, NINDS 

Cooperating Unit: Brown University, Providence, Rhode Island 

This study has been conjjleted. 

Honors and Awards : None 

Publications: 

Holden, R. H. and Willerman, L. : Social class and outcome in the 
neiirologically abnormal child. In Trapp, E. P. (ed.) Readings on the 
Exceptional Child. 2nd Ed. App let on-Century- Crofts, I97I. In Press. 



1^4-9 ■ 



Serial Wo. NDS (CF)-69 PR/BS 175 8 

1. Perinatal Research Branch 

2. Section on Behavioral Sciences 

3. Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1, 1970 through June 30, I97I 

Project Title: Neonatal, Ethnic and Social Class Factors in Infant and 
Pre- school Test Performance 

Previous Project Title: Matiirity at Birth, Mental and Motor Performance at 
Eight MDnths and Intelligence Quotient at Four Years 

Previous Serial Number: Same 

Principal Investigator: Sarah H. Etroman, Ph.D., PRB, WINDS 

Other Investigator: Heinz W. Berendes, M.D., PRB, WINDS 

Cooperating Unit: Office of the Chief, PRB, WINDS 

Man Years : 



Total: 


.25 


Professional: 


.20 


Other: 


.05 



Objectives : This study examined the effects of two measures of maturity at 
birth on mental and motor development at eight months and on a subsequent 
measure of intelligence at four years in two large samples of children who 
are being followed longitudinally in the Collaborative Study in the National 
Institute of Neurological Diseases and Stroke. Birthweight and gestational 
age were related to Bayley Mental and Motor scores at eight months and to 
Stanford- Binet IQs at four years within the context of ethnic, social class 
and sex classifications. The relationship of Bayley scores to Binet IQ was 
also examined. A multiple linear regression model was used. 

The major findings were as follows: 

1. Among infants tested at eight months (W=31,678) there is a highly 
significant association between the group of predictor variables of ethnicity 
(white and Wegro) sex, social class, (four levels of maternal education), 
birthweight and gestational age and the criterion of Bayley mental score. 
However only seven percent of the variance was explained in the criterion 
measiire. The most "useful" predictor was birthweight, followed by gestational 
age. 

2. A similar pattern resulted from the analysis of Bayley motor scores. Nine 
percent of the variance was explained and again birthweight, followed by 
gestational age, was the best predictor. 

3. Among children tested at four years (N=16,658) there is a highly signi- 

151 V 



Serial No. NDS ((IF)-69 PR/BS 175 8 

ficaxLt association between the group of predictor variables of ethnicity, 
sex, social class, birthweight, gestational age, eight month mental scores 
and eight month motor scores and the criterion of Stanford- Bine t IQ. Twenty- 
eight percent of the variance was explained in the criterion measure. The 
most useful predictor was ethnicity followed by social class (maternal 
education) . 

The inplications of these findings and other derived from analyses of the 
relationship of the predictors to the criterion measures within the four 
separate ethnic-sex groups are discussed in the context of the characteristics 
of the samples studied. This stxidy has been completed. 

Honors and Awards: None 

Publications : None 



152V 



Serial Wo. WDS (CF)-70 PR/BS 1856 

1. Perinatal Research Branch. 

2. Section on Behavioral Sciences 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1970 through June 30, 1971 

Project Title: Biosocial Influences on Euxnan Development 

Previous Serial Number: Same 

Principal Investigator: Dr. Lee Willerman, PRB, NIHDS 

Other Investigator: None 

Cooperating Unit: None 

This paper has heen completed and was presented at the k-Jth Annual Meeting 
of the American Orthopsychiatric Association, San Francisco, California, 
March 23-26, 1970. The abstract has been published in the American 
Journal of Orthopsychiatry, kO: 324, I97I. 

Honors and Awards: None 

Publications : None 



153v 



Serial No. NDS (CF)-70 PR/BS I857 

1. Perinatal Research Branch 

2. Section on Behavioral Sciences 

3. Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1^ 1970 through June 30, 1971 

Project Title: The G-enetics of Intellectual and Motor Performance 

Previous Serial Number: Same 

Principal Investigator: Sarah H, Broman, Ph.D., PRB, NINDS 

Other Investigators: N. C. l\fyrianthopoulos, Ph.D., PRE, NINDS 

Lee Willerman, Ph.D., PRE, NINDS 
V. L. Anderson, Ph.D., University of Minnesota 
Paul Nichols, Ph.D., University of Minnesota 

Cooperating Units: Section on Epidemiology and Genetics 

The Dight Institute for Human Genetics, University 
of Minnesota 

Man Years: 

Total: .25 
Professional: .20 
Other: .05 

Project Description : 

Objectives : The objective of the study is to assess the contribution of 
genetics to the variance of behavioral measures, particularly intellectual 
and motor performance, by the calculation of heritability in twins. The 
results of the psychological tests given at ages eight months, four years 
and seven years have been analyzed for all twins of known zygosity and for 
all sibs from first and second study pregnancies. It is also planned to 
analyze these data for an appropriate sample of unrelated child pairs. As 
a physical parallel, the heritabilities of height, weight and head cir- 
cumference at several age levels have been computed. 

Some selected findings are as follows: (l) heritability of IQ at age four 
does not appear to be lower than in older populations; (2) the between family 
variance componant of the four-year IQ test was nearly twice as large in the 
white population as in the Negro population suggesting that heritability 
is lower in Negroes than in whites; (3) an analysis of varisjice of weight 
and height measured at four years revealed that, unlike IQ, the variance 
Goniponants were not different in whites and Negroes; (k) at seven years, 
a high correlation (••86) was found between 12 subtests social class loading 
and the Negro-white differences on the test, while no relationship was 
found between Negro-white differences and heritabilities of the subtests 

155 V 



Serial No.' NDS (CF)-70 PR/BS I857 

after controlling for the subtests' social class loading; (5) the 
discrepancy in IQ between twins and singletons decreased in both races 
from four to seven years. 

Analyses of these data are continuing. 

Honors and Awards: None 

Publications: None 



156 V 



Serial No.NDS (CF)-63 PR/EG 11T4 
lo Perinatal Research Branch 
2o Section on Epidemiology 

and Genetics 
3. Bethesda^ Maryland 



PHS 
Individual Project Report 
July 1, 1970 through June 30, 19T1 

Project Title: Birthweight in Relation to Selected Socioeconomic Variables 

Previous Serial Number: Same 

Principal Investigator: Dr. N.C. Myrianthopoulos, PRB, imroS 

Other Investigators : Dr. Joshua Lederberg, Stanford University 

Cooperating Units : None 

Man Years : 

Total: .10 
Professional: .05 
Other: .05 

Project Description : 

Objectives : This is a continuation of a study to relate birthweight to 
socioeconomic and medical variables. The results of the first part of the 
study have already been published (Ann. Htm. Genet. 31:71-83, 196t)<> 

Proposed course : In the second paii: the plan is to compare children in the 
100th percentile of birthweight with children in the 50th percentile of 
birthweight and children of diabetic mothers, in teims of several socioeconomic, 
biological and medical variables. A magnetic tape with all the necessary data 
has been prepared. Analysis is done at Stanford University under Dr. 
Lederberg's direction. No further progress has been made in this study. 

Honors and Awards: None 

Publications : None 



157 V 



Serial No. EDS (CF)-63 PR/EG 1175 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Bethesda^ Maryland 

PHS-WIH 

Individual Project Report 

July 1, 1970 through Jvme 30, 1971 

Project Title: Detennination of the Zygosity of Twins Bom to Mothers 
in the Collahorative Study 

Previous Serial Number: Same 

Principal Investigator: Dr. N.C. Myrianthopoulos, PKB, NINDS 

Other Investigators : None 

Cooperating Units : All Institutions participating in the Collaborative Study 

Man Years: 

Total: .15 
Professional: .15 
Other: ,00 

Project Description : 

Objectives ; This is a continuing project to deteimine the zygosity and 
epidemiologic characteristics of twins bom to Study mothers. 

Methods employed : Twin zygosity is determined by comparison of sex, 
placentation, blood groups, and finger and palm prints. This information is 
forvarded by all Institutions to the Section on Epidemiology and Genetics 
where it is classified and analyzed by special methods. 

Major findings : In addition to the findings reported in last year's report, 
an analysis has been made of respiratory distress syndrome in the twins. 
Respiratory distress syndrome occurred in 77 of 1130 livebom twins (or 1 in 
15) and appears to be from 5 to 9 times more frequent than in singletons. 
This increase cannot be entirely accounted for by the higher prematurity rate 
of the twins over singletons. In 46 twin pairs with at least one affected, 
there was a significantly higher concordance rate among MZ than DZ pairs, 
suggesting that genetic factors are of some etiologic importance in this disease. 

An analysis of congenital malfonnations in the twins is also being made. 

Proposed course : To study all twins in terms of a variety of genetic and 
socioeconomic variables, 

159 V 



Serial No. WDS (CF)-63 PR/EG 1175 

Honors and Awards: None 

Publications: Myrianthopoulos, N.C.: A survey of twins in the population of 
a prospective collaborative study. Acta Genet, ^fed. Gemellol . 
19: 15-23, 1970. 

Myrianthopoulos^ N.C.: An epidemiologic survey of twins in a 
large, prospectively studied population. Amer. J» Hum. Genet . 
22: 611-629, 1970. 

IVtyrianthopoulos, N.C.: Respiratory distress syndrome in twins. 
Acta Genet. Med. Gemellol., in press. 



I6OV 



Serial No. -WDS (CF)-63 PR/eG IITT 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 19T1 

Project Title: Genetic and Socioeconomic Factors in Early and Late Fetal 
Death 

Pluvious Serial Number: Same 

Principal Investigator: Dr. N.C, Myrianthopoulos, PRE, NIKDS 

Other Investigators: Esther Jackson, OB, NINDS 

Cooperating Units : None 

Man Years: 

Total: .10 
Professional: .10 
Other: .00 

Project Description : 

Ob jectives : To detennine what effect do genetic and socioeconomic factors 
have on pregnancy wastage and if a distinction can be made etiologically 
between early and late fetal deaths. 

Methodology and major findings : Discriminant analysis of 1,877 early and late 
fetal deaths with respect to kO medical, genetic and socioeconomic variables, 
shows that toxemia, anemia and complications of pregnancy are good discriminants 
in both whites and Negroes; prior pregnancies and prior pregnancy wastage in 
whites only; and socioeconomic status in Negroes only. Knowledge of sex of 
study child improves the power of the discriminant function. 

When the sample is restricted to fetal deaths of mothers who registered in the 
first trimester the results are not appreciably changed. This project is now 
completed. 

Honors and Awards : None 

Publications: None 



161 V 



Serial No. WDS (CF)-63 PR/eG llSij- 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Population Dynamics of Tay-Sachs Disease and other 
Sphingolipidoses 

Previous Serial Number: Same 

Principal Investigator: Dr. N.C, Myrianthopoulos, PRE, NINDS 

Other Investigators : Dr. Stanley Aronson, Brown University 

Cooperating Units : None 

Man Years : 

Total: .15 
Professional: .15 
Other: .00 

Project Description : 

Objectives : This is a continuation of a study to determine whether differ- 
ential fertility favoring the Jewish heterozygote can acco\«it for the 100 fold 
higher frequency of Tay-Sachs disease and the gene responsible for it among 
the Jewish compared with non-Jewish population in the U.S. Evidence for 
selective advantage of the heterozygote was found in the first part of the 
study (Am. Jo Hum. Genet. 18:313-327; I966). 

Proposed coiorse : The plan is to investigate whether or not such advantage 
can be demonstrated in other sphingolipidoses, to study the demography of 
these disorders and to deteimine whether resistance to tuberculosis or some 
other infectious disease confers a selective advantage to the heterozygote. 

Methodology : About ^,000 cases of Jewish immigrant patients who had TB have 
been collected from the records of the American Chronic Disease Center, 
Denver, Colorado. The distribution of places of origin of these patients has 
been compared with that of ancestors of Tay-Sachs disease and a negative rank 
correlation has been found suggesting that the frequency of TB had been low 
in areas of high concentrations of Tay-Sachs disease, and vice versa. A 
more sophisticated analysis is now being perfonned. 



163 V 



Serial No.' NDS (CF)-63 PR/EG 1184 



Honors and Awards: None 
Publications: None 



1614-v 



Serial No. NDS (CF)-65 PR/EG 12T4 
1« Perinatal Research Branch 

2. Section on Epidemiology 

ajid Genetics 

3. Be the s da, Maryland 

PHS-Wm 

Individual Project Report 

July 1, 1970 through June 30;, 1971 

Project Title: Genetic Bases of Neonatal Reflexes 

Previous Serial Numher: Same 

Principal Investigator: Dr. A.F, Naylor, PRB, NINDS 

Other Investigators: Dr. N.C. Myrianthopoiolos, PRB, KINDS 

Cooperating Units : None 

Man Years : 

Total: ,05 
Professional: ,05 
Other: .00 

Project Description : 

Objectives : To investigate the validity of regarding the suck, rooting and 
other neonatal reflexes as genetic entities. 

Major findings : An initial set of cases retrieved for absence of one or more 
of these reflexes was reviewed and seemed to have high frequencies of various 
kinds of trauma whose base line frequencies were unknown. 

Proposed course : To place limits on the frequencies of losses of suck, root- 
ing, palmar grasp, plantar grasp and Moro reflexes because of mutation or 
segregation at gene loci specifically affecting manifestation of these reflexes. 

The completion of the (condensed) Variable Data File makes practical the reacti- 
vation of this project along proper lines. Base populations can be selected 
for general health, especially neurological, and frequencies of isolated absence 
or weakness of single neurological signs can be tested. Active work on this 
project will be undertaken when most current tasks have been carried out. 

Honors and Awards: None 

Publications : None 



165 V 



Serial No. ' EDS (CF)-65 PR/EG 12J6 
lo Perinatal Research Branch 

2, Section on Epidemiology 

and Genetics 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Sequential Aspects of Occurrence of Spontaneous Abortion in 
Family Histories 

Previous Serial Number: Same 

Principal Investigator: Dr. A.F, Naylor, PRB, NIKDS 

Other Investigators: Dr. Dorothy Warburton, College of Physicians and Surgeons 
of Colianbia University 

Cooperating Units : None 

Man Years: 

Total: ,20 
Professional: ,20 
Other: .00 

Project Description : 

Objectives : To relate the risk of spontaneous abortion to maternal age and 
prior reproductive experience. A special point under investigation is whether 
apparent age effects are explicable by a tendency for intrinsic habitual aborters 
to remain in the reproductive population longer in attempts to compensate for 
unsuccessful pregnancies. Also conditional risks have been estimated. 

Proposed course : Although certain portions of the data analysis support a ma- 
ternal aging explanation for part of the trend in abortion risk, a reconsideration 
of the original tab\ilar outputs shows that a very substantial additional parity 
exists. An existing manuscript must be completely redrafted to reflect this 
change in interpretation. 

Honors and Awards: None 

Publications : None 



167 V 



Serial Wo. KDS (CF)-6t PE/eG 15 10 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Bethesda, Maryland 



PHS-NIH 

Individvial Project Report 

July 1, 1970 through Jvne 30, 1971 

Project Title : The Study of Maternal Effects in the Production of 
Congenital Malfoimations 

Previous Serial WumlDer: Same 

Principal Investigator: Dr. W.C, Myrianthopoulos, PRB, NUTOS 

Other Investigators: None 



Cooperating 


Units : 


None 


Man Years: 






Total : 




.10 


Professional: 


.10 


Other: 




.00 


Project Description 


: 


Objectives: 


To deteimin( 



To deteimine the extent to which maternal factors are involved 
in the production of congenital malfoimations, and to single out those mal- 
foimations and conditions of the newborn in which the role of maternal factors, 
genetic and environmental, appears to he deciding. 

Methods employed : The GEN 5-8 was used to obtain a history of outcome of 
prior pregnancies, in families in which half-siblings are present. Study 
pregnancies were also included. 

Ma j or findings : In all, 152 cases were identified and these were screened 
for occurrence of congenital malfoimations and other conditions among 
children by different fathers. Six conditions were found to occur in high 
frequency among half sibs: Rh trouble, convulsions, congenital heart 
disease, club foot, mental retardation and Polydactyly. 

Analysis of the distribution of these abnoimalitie s in first and second sib- 
ships of the same family showed that in the case of seizures and mental re- 
tardation the experience in the first sibship was significantly different 
from that in the second, indicating a genetic etiology; in the case of congenital 
heart defects and club foot the experience seemed to be the same in both 
sibships, indicating that environmental maternal factors predominate. 

169 V 



Serial No,' NDS (CF)-6t PR/EG I5IO 



Additional cases have now became available and a more complete analysis is 
being made. The study is in progress. 

Honors and Awards: None 

Publications : None 



170' 



Serial No. WDS (CF)-6t PR/eG 1514 

1. Perinatal Research Branch 

2, Section on Epidemiology 

ajid Genetics 
3» Bathes da, Marylemd 

PHS-NIH 

Individual Project Report 

July 1, 19T0 through June 30, 1971 

Project Title: Record Linkage of Relatives Registered in the Collaborative 
Study 

Previous Serial Number: Same 

Principal Investigator: Dr. A.F. Nay lor, PRB, NINDS 

Other Investigators: Dr. N.C, Myrianthqpoulos, PRB, NIKDS 

Cooperating Units : None 

Man Years: 

Total: .60 
Professional: ,15 
Other: .45 

Project Description : 

Objectives ; To identify all relatives of gravidae registered in the Collaborative 
Study. 

Methodology : Abstracting of reports of relatives has been completed and the 
infoimation key-punched into card images on magnetic tape. 

Proposed course : In its current foim the file lists, case by case, the NIRDS 
number of reporter and reported relatives and relationship code. The file must 
be reprocessed into pairs so that the relative making the report is retrievable 
when the reported relative is an index case; also transitive pairing — linkage 
of two relatives who are both reported by one woman — must be established. 

Honors and Awards: None 

Publications: None 



171 V 



Serial No. M)S (cr)-6T PR/eG 1515 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Bethesda, Maryland 

PHS-WIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Rh Hemolytic Disease in Negro and White Infants 

Previous Serial Number: Same 

Principal Investigator: Dr. A.F. Naylor, PRB, NINDS 

Other Investigators: None 

Cooperating Units : None 

Man Years: 

Total: .01 
Professional: ,01 
Other: .00 

Project Description : 

Objectives : To confiim a report that high Rh antibody levels have smaller 
morbid effects in Negro than in White babies, althotigh this is not true for 
ABO ajitibodies. 

Major findings : Preliminary and indirect confiimation has been obtained, from 
a small data sample under study in this Section, for reports in the literature 
that high Rh ajitibody titers are not as highly associated with serious morbidity 
in Negroes as in whites. 

Proposed course : The existence of the Variable Data File will make possible 
the easy execution of the required data processing. 

Honors and Awards: None 

Publications : None 



173 V 



Serial No. NDS (CF)-6t PR/eG 1516 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Be the s da, Maryland 

PHS-WIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Size of Placenta in Relation to Mother-Fetus Antigenic 
Difference 

Previous Serial Wumher: Same 

Principal Investigators : Dr. Dorothy Warburton, College of Physicians and 

Surgeons of Columbia University 
Dr. A. P.. Naylor, PRB, WHTOS 

Other Investigators: Dr. G. Nicholas Rogentine, Immunology Branch, NCI 

Dr. Robert Cefalo, Obstetrics, National Naval Medical Center 
Mrs. Lois Dienes, PRB, NINDS 
Mrs. Jean Nemore, PRB, NINDS 

Cooperating Units: Obstetrics Department, National Naval Medical Center 

Man Years : 

Total: 1.20 
Professional: .80 
Other: ,kO 

Project Description : 

Objectives : To investigate the hypothesis published by Billington in 196k, 
that sensitization of the mother during pregnancy against paternal antigens 
leads to non-pathological placental hypertrophy and increased birthweight in 
succeeding pregnancies. 

Major findings : Analysis of a large sample of study data has given convincing 
support to the hypothesis. A paper reporting the evidence has been published. 



Proposed course : The arrangements for a laboratory study, described in the 
19^-70 Annual Report have been active and 25 maternal sera have been tested 
against paternal leucocytes. No attempt at analysis will be made until many 
more cases have accumulated. 

Honors and Awards : None 



r. 

1 



175 V 



Serial Wo. WS (CF)-67 PR/EG I516 

PubU-cations: War-burton, D. and Naylor, A.F. : The effect of parity on 
placental weight and birth weight : An immunological 
phenomenon? A report of the Collaborative Study of Cerebral 
Palsy. Amer. J. Hum. Genet. 23: ^1-5^^, 1971« 



176 V 



Serial No, NDS (CF)-68 PR/eG iGkG 
1, Perinatal Research Branch 
2o Section on Epidemiology 

and Genetics 
3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30^ 1971 

Project Title: Study of the Physical Growth of Children in the Collaborative 
Study 

Previous Serial Number: Same 

Principal Investigator: Dr. Glen S. Bartlett, Case Western Reserve University 

Other Investigators: None 

Cooperating Units: Office of Biometry, NINDS 

This project has been transferred to the jurisdiction of the task force on 
Physical Growth and Development, 



177 V 



Serial No. NDS (CF)-68 PR/eG l6kj 

1, Perinatal Research Branch 

2, Section on Epidemiology 

and Genetics 
3o Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 thro\:igh June 30, 1971 

Project Title: Development and Evaluation of an Index of Reproductive 
Perfoimance for the Puipose of Identifying High Risk 
Pregnancy Suspects 

Previous Serial Number: Same 

Principal Investigator: Dr. Glen S, Bartlett, Case Western Reserve University 

Other Investigators: None 

Cooperating Units : None 

This project has been temporarily discontinued and will be held in abeyance. 



179 V 



Serial No. EDS (cf)-68 PR/eG l6kQ 

1. Peilnatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Farther Investigation of the Socioeconomic Index as a 
Descriptive and Predictive Instrument 

Previous Serial Wumher: Same 

Principal Investigators: Dr. Glen S, Bartlett, Case Western University 

Dr. W.C. Myrianthopoulos, PRE, NBTOS 

Other Investigators : None 

Cooperating Units : None 

Man Years : 

Total: .00 
Professional: .00 
Other: .00 

Project Description : 

Objectives : The socioeconomic status score or socioeconomic index, developed 
by the Bureau of the Census for use with the I960 Census, has been used with 
good results in this Section to describe the Collaborative Study population, 
and by others to describe other populations. This study is designed to ex- 
plore mathematically the three component parts of the index — education and 
occupation of the head of household, and family income — in order to assess 
their relative contributions to the index; to evaluate the ability of the 
index to encompass other socioeconomic variables not used in deriving the 
inde ; and to investigate whether or not the index, in addition to being 
descidptive, can also be used as a reliable predictive instrument. 

Methodology ; Linear discriminant function and multiple regression analysis 
will be used to deteimine appropriate weights to be applied to the component 
factors, and correlation coefficients will be obtained between the index and 
its components, and external variables. 

Current status : A working paper proposing three models has been written. 
Data from first study pregnancy and from the T-year follow-up examination will 
be used to test these models. No further progress has been made in this project. 



181 V 



Honors and Awards : None 
Publications : None 



Serial No. NDS (CF)-68 PR/eg l6k8 



l82^ 



Seri-al No. -NDS (CF)-71 PR/eG I906 

1. Perinatal Research Branch 

2, Section on Bpidemiology 

and Genetics 
3o Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

jTily 1, 19T0 through June 30, 1971 

Project Title: Blood group effects in mother and offspring 

Previous Serial Number: None 

Principal Investigators: Dr. N.C. Myrianthopoulos, PRE, NINDS 

Dr. Be mice Cohen, Johns Hopkins University 

Other Investigators: None 

Cooperating Units: None 

Man Years : 

Total: .10 
Professional: ,10 
Other: .00 

Project Description : 

Objectives : To deteimine the influence, if any, of maternal and/or infant ABO 
and/or Rh types iipon maternal manifestations and fetal-infant- child survivor- 
ship, morbidity and development. 

Proposed course and methodology : A comprehensive investigation will be made 
of the blood group relationships, including possible deleterious consequences 
or beneficial effects in mothers and offspring of various maternal ABO and/or 
Rh types and ABO-Rh combinations both per se, and grouped as potentially 
"compatible" and "incompatible" combinations of mother and conceptus. Maternal 
and offspring effects will be examined in terms of immediate and acute effects 
ajid in terms of long-range effects in mother and offspring, through develop- 
mental milestones at 7 years of age. Comparisons will be made by institution, 
race, age of mother, previous reproductive history, complications of pregnancy, 
n mnrunoglobulin levels, and socioeconomic status. 

Honors and Awards: None 

Publi cati ens : None 



I 



•183 V 



Serial No. NDS (CF)-TI PR/eG I907 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Be the s da, Maryland 

PHS-KIH 

Individual Project Report 

July 1, 1970 through June 30, 19T1 

Project Title: Epidemiologic and genetic study of congenital malfoimations 

Previous Serial Nixnber: None 

Principal Investigator: Dr. N.C, Myrianthopoulos, PRB, NUTDS 

Other Investigators: Dr. J. Drage, PRB, NIKDS 

Dr. C.S. Chung, University of Hawaii 

Cooperating Units : None 

Man Years : 

Total: .20 
Professional: .15 
Other: .05 

Project Description ; 

Objectives : To study the epidemiology of all congenital malfoimations in 
Collaborative Study children at birth ajid one year; to test the hypothesis 
of quasi -continuous distribution as an inheidtance mechanism for most mal- 
foimations; to assess the genetic load due to congenital malfoimations; to 
test the etiologic significance of several medical, genetic and socioeconomic 
factors in the occurrence of selected malfoimations; and to explore more fully 
previo-us findings suggesting a relationship of diabetes in the mother and the 
occurrence of congenital heart disease in the offspring. 

Proposed course emd methodology : The plan is to draw a definitive list of con- 
genital malfoimations and develop a file of all Collaborative Study children 
who had a malfoimatlon between birth and one year of age. Consanguinity data 
will be used to assess the genetic load and aji extension of Falconer's method 
will be employed for the quasi-continuous distribution analysis. A prospective 
analysis will be used to test the etiologic significance of medical, genetic 
and socioeconomic factors. 

Honors and Awards : None 

Publications: None 



185. 



Serial No. KDS (CF)-TI PR/EG I908 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: A chromosomal study of children in the Collaborative Study 

Previous Serial Nimiber: None 

Principal Investigators: Dr. N.C. Myriajithopotilos, PRE, WHTDS 

Dr. H. Lubs, University of Colorado 

Other Investigators: None 

Cooperating Units: Boston Children's Hospital Medical Center, Children's 

Hospital University of Buffalo, University of Tennessee, 
Children's Hospital of Philadelphia and University of 
Oregon Medical School 

Man Years : 

Total: .20 
Professional: «15 
Other: ,05 

Project Description : 

Objectives : To study the epidemiology of chromosomal aberrations in approxi- 
mately 10,000 Study children at age 7 years; and to relate major and minor 
chromosomal deviants to growth, mental and motor development and neurological 
status of these children. 

Proposed course and methodology : A blood sample will be taken from children at 
the five collaborating institutions, during their 7-year examination. Two cells 
will be immediately analyzed; ten cells will be analyzed when chromosomal anom- 
alies are found or when the children have some mental or motor anomaly or a 
congenital malfonnation. All technical work will be done in accordance with 
standardized techniques and new techniques such as fluorescence staining, will 
be employed. All measurements and analysis will be done at Denver with the aid 
of an automatic chromosome analyzer. A second phase of the study will be con- 
cerned with the chromosomal-clinical correlations. 

Honors and Awards: None 

Publications : None 

187 V 



Serial No. EDS (CF)-TI PR/eG 1909 

1, Perinatal Research Branch 

2, Section on Epidemiology 

and Genetics 

3, Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

Jiily 1, 1970 through June 30, 1971 

Project Title: Continuation of study of twins horn in the Collaborative Study 
beyond the age of seven years 

Previous Serial Number: None 

Principal Investigator: Dr. N.C, Myrianthopoulos, PRB, NINDS 

Other Investigators: None 

Cooperating Units: All Collaborating Institutions 

Man Years: 

Total: .10 
Professional: ,05 
Other: .05 

Project Description : 

Objectives : To follow all living twins and their siblings bom in the Collab- 
orative Study to age 15 years in order to study their physical and mental 
development through puberty. 

Proposed course and methodology : All living twins, including unlike-sexed twins, 
will be typed with a broader spectrum of genetic markers to insure utmost ac- 
cxrracy in zygosity and to obtain a distribution of these markers in twins and 
their Study siblings for further studies. The twins and their Study siblings 
will be given ein annual physical examination which will include physical and 
anthropometric measurements, and more extensive examinations with appropidate 
IQ and behavioral tests at ages 12 and 15 years. Investigations will include 
growth and development of genetically identical and environmentally related 
individuals; neurological evaluation, especially in regard to minimal abnor- 
malities; differences in mental development; determination of the occurrence 
of a third type of twin; genetic factors in induction of puberty; and host 
resistance to disease. 

Honors and Awards: None 

Publications: None 



189^ 



I 



Serial No. NDS (CF)-Tl PR/EG 1910 

1. Perinatal Research Branch 

2. Section on Epidemiology 

and Genetics 

3. Bethesda, Maiylemd 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Genetics of Obstetric Variables eind the Role of Maternal 

Factors in the Deteimination of Intelligence and Neurological 
. Perfoimance 

Previous Serial Number: None 

Principal Investigators : Dr. A.F,' Nayldr, PRE, NINDS 

Dr. N.C, MyrianthopoTilos, PRE, NINDS 

Other Investigators : Dr. D. Warburton, College of Physicians and Surgeons of 
Colxmbia University 

Cooperating Units: Department of Human Genetics eind Development, Columbia 
University 

Man Years: 

Total: .50 
Professional: .50 
Other: ,00 

Project Description ; 

Objectives : To analyze the variation of obstetric and gynecological factors 
into heia table and non-heritable components. 

Proposed course : The genetic linkage file necessary for fall realization of 
the objectives, is near but short of completion. Specialized programs necesssiry 
for genetic analyses are being procured from the University of Hawaii but will 
need modification for use on NIH canputers. An expanded version of the Variable 
Data File incorporating needed additional information has been created. This 
expanded file will be used immediately (l) to investigate biases in obstetric 
variation among women reporting relatives in the Study compared to those not 
reporting relatives (2) to investigate obstetric heritability at a "primary" 
level to identify conditions which tend to recur in repeat Study pregnancies. 

Honors and Awards: None 

Publications : None 



I9IV 



Serial No. NDS (CF)-65 PR/P 1278 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Biologic Pattern Data Processing 

Previous Serial Number: Same 

Principal Investigators: Lewis E. Lipkin, M. D. 

Russell A. Kirsch 

Other Investigators: Peter F. Lemkin 
Philip G. Stein 
Stanley E. Shackney, M, D. 

Cooperating Units: Artificial Intelligence Group 
National Bureau of Standards 

Office of Associate Scientific Director 

Clinical Trials 

National Cancer Institute 

PDP-10 Group 

Division of Computer Research and Technology 



Man Years 

Total: 8.2 
Professional: 5.0 
Others: 3.2 

Project Description : 

This project has been extensively reviewed during the past calendar year. 
The first review in June 1970 was by the NCI directorate with uniformly 
favorable comment. The second more formal review in December 1970 by an 
external ad hoc committee, chaired by Dr. Kenneth Earle, Chief of 
Neuropathology of AFIP, concluded: "The committee was unanimous in its 
support of this important project..." 

1. Substantive Projects : There have been three major biologic projects 

begun during the reporting period. These are totally dependent on both 
the concepts and facilities of the emerging image processing facility 
in the Section on Pathology. 



193 V 



Serial No. NDS (CF)-65 PR/P 1278 

a. The Quantitative Characterization of Chroma to lysis : Serial 
histologic sections of hypoglossal neurons of rabbits rendered 
chromatolytic by nerve section one or two weeks prior to sacrifice 
by perfusion fixation and the corresponding contralateral controls 
are selected by the observer for automatic scanning. The tapes so 
generated are displayed on the PDP-lO's 340 display, where under 
the biologist's visual control and by means of a Graf con stylus and 
tablet cells and their components are segregated. The computer is 
providing data on each cell and its components that are measures 

of Nissl distance dispersion and/or loss, nuclear and nucleolar 
swelling, nuclear eccentricity, change in cell size, change in cell 
shape and changes in patterns of distributions of cytoplasmic RNA. 
At this writing, additional subprograms are being written to enable 
us to determine quantitative measures of satellitosis in terms of 
such parameters as the probability of encountering a glial cell as 
a function of distance from the cytoplasmic boundary of the cell 
body. Since the magnetic tapes of the scanned images are preserved, 
these results may be obtained without rescanning. Indeed, as 
additional algorithms are developed, they may be applied to exactly 
the same cell images without further microscopy. 

b. Identification of Marrow and Peripheral Blood Cells in Radio - 
autographs : This ability is one of the remaining requirements for 
specifying the automatic grain counting device requested by the 
National Cancer Institute. A library of scans is now under 
construction, so that following leads such as those provided by 
Mendelsohn and Prewitt, this major requirement for successful 
completion may be met. Of particular interest is the very great 
utility of image masks generable from multiple wavelength scans. 

c. Concurrent DNA Content and Synthesis Rate Determinations in 
Embryonic Neural Tube and Transplanted Lymphomas : The image 
processing facility provides the unique ability to determine not 
only the rate of DNA synthesis in a cell population, but also to 
determine the quantity of DNA/cell in a Feulgen- stained autoradio- 
graph. This ability will provide data which will provide informa- 
tion which hitherto has only been available from laboriously 
examined time-lapse studies and labeled mitosis counts. Material 
presently being examined are transplants of rodent leukemia. In 
process, however, are embryonic neural tissues with the expectation 
of obtaining more quantitative information concerning mantle- 
marginal relationships. 

In addition to the foregoing specific projects, extensive experiments 
involving three-dimensional reconstructions, automatic morphologic 
analysis, etc. were performed. Some indications of their scope is 
given under "4. Computer Analysis and Image Processing Results." 



194 V 



Serial No. NDS (CF)-65 PR/P 1278 

Apparatus Design, Construction and Procurement : Significant design and 
construction attention has gone into improving the scanning microscope, 
while keeping it concurrently usable for productive scanning. Some of 
the modifications have been discovered to be necessary as a consequence 
of having used the system. For example, a new rigid table was 
constructed and procurement was initiated for anti-vibration mounts 
to cure a building vibration induced problem that was discovered during 
the course of scanning at high magnification. In preparing to use the 
scanner, photometric calibration of the photo-multiplier and associated 
circuitry is necessary. To facilitate this, new gain controls were 
constructed and low density filters were obtained which can be used to 
simulate the optical density of biological materials to be scanned, so 
as to facilitate calibration of the scanner over the full optical 
density range. A standard procedure was also adopted for recording on 
the magnetic tape along with an image that has been scanned, the 
corresponding values of the reference beam photomultiplier, so as to 
make completely available for subsequent analysis the information about 
incident illumination necessary to refine the calibration of the 
photometric part of the scanner. For point by point rather than raster 
scanning, the scan axis drivers and associated motor controls necessary 
for driving the scanner mirrors were constructed. Immediately after 
scanning it is often useful to have a crude visual presentation of the 
image as scanned. A beam storage tube display was obtained and 
connected so as to provide both intensity versus x and y type of display 
as well as the previous contouragram form of display to exhibit the 
complete scan output for a single scan. 

During this period the LINC-8 and the PDP-10 computers were connected 
together in a flexible manner to enable the interchange not only of scan 
data but of programs in both directions from the LINC-8 and hence from 
the microscope to the PDP-10 and conversely. The RF08/RS08 disk system 
was connected to the LINC-8 and is currently in operation. An A.B. Dick 
Videojet Line Printer was connected to the LINC-8 and has made it 
possible to produce full resolution displays of scanned images through 
the use of about 16 gray levels of intensity encoded with suitable print 
characters. 

A large disk pack drive installed on the PDP-10 has a five-million word 
capacity and is used exclusively by the image processing project. 

A major effort has been going on in the design and construction of an 
optical bench to provide the capabilities of the present scanner with 
more flexibility for the addition of new scanner options. The basic 
optical bench mount which had previously been obtained had several 
pieces of equipment added to it . A vertical stage to take the place of 
the present optical microscope stage was constructed and installed, as 
was a mount for the use of the monochromator and a mount and changing 
system for mounting microscope objectives on the bench. Investigation 
was initiated for procurement of a new scanner to be used on the optical 
bench and a microscope tube length correction system was designed. 



195V 



Serial No. NDS (CF)-65 PR/P 1278 

Special image measuring devices for use on the optical bench were 
obtained to be used in precise calibration of the optical system when 
it is in operation on the optical bench. The design of a system for 
distance measurement using interferometric techniques on the optical 
bench was begun. 

3. Programs : Many programs have been written during this period for the 
LINC-8 and the PDP-10 which served to facilitate experiments using the 
scanning microscope and to remove the burden from the user of having to 
write detailed programs. Notable among such efforts is the construction 
of successive versions of the MOSS microscope operating supervising 
system which enables complex sequences of scanning and processing on 
the LINC-8 to be initiated by the user with a minimal amount of 
programming. Prior to obtaining special line printer equipment for 
producing images, a program was written to use the NBS Stromberg-Carlson 
Graphic Printer to make digitized printouts of scanned images. This 
served not only the purpose of visual representation of scans, but 
enabled us to gain assurance that the scan data was as intended when 
the microscope was set up. 

During this period the PDP-10 has become the main production device for 
processing scanned data. Largely because of fine cooperation from 
personnel in DCRT, the PDP-10 has proved powerful and has generally 
been satisfactorily available. The total use of the PDP-10 has, at 
various times, consumed over 20 hours of CPU time per month for NBS 
users alone engaged in program writing and development as well as 
production computing. At peak usages, NINDS and NCI users on this 
project have exceeded this total. This total of 40-45 hours of CPU 
time/month can probably be taken as a reasonable projected upper bound 
for computer use during the next year. 

During this period several specialized and some general purpose tools 
have been constructed. A version of the morphological analysis procedure 
for decomposing scanned images that had previously been converted from 
the Q-32 computer to the PDP-10 was modified to use the full resolution 
of the data obtained from the microscope scanner. Another somewhat 
more general version was constructed in the LISP language to provide 
the full generality over all possible scanned images of decomposition 
methods. Of the generalized tools, a systematic effort enabled the 
construction on the PDP-10 system of general purpose image processing 
facility more powerful than the corresponding one previously built on 
the Q-32 computer. This facility is written to operate in the LISP 
language and hence to provide the high level language processing 
capability of LISP with the corresponding efficiency of lower level code 
written either in assembly language or in FORTRAN through the use of a 
FORTRAN-LISP interface written by S. Bryan (DCRT). The list of major 
functions available in the LISP image processing facility is as follows: 

a. A routine for reading whole or partial scans from magnetic tape into 
memory performing optical thresholding on the data. This routine also 
allows the reference photomultiplier data optionally to be read 
instead of the photometric information from the scan data. 

196 V 



Serial No. NDS (CF)-65 PR/P 1278 

b. A routine for variously thresholding image arrays to convert them 
to binary form. 

c. Very high speed non-buffered input/output for swapping images from 
disk to core. Because a typical image involves 16,000 words of 
PDP-10 core storage, it is important to be able to swap such images 
into and out of memory at high speeds to avoid unconscionably large 
amounts of processing time. 

d. A FORTRAN bit processing package to operate within the LISP 
language . 

e. A high speed assembly language program for computing the derivative 
of a scanned image. The derivative data contains information about 
boundaries and gradients and hence is used in morphological analysis 
extensively. 

f . An assembly language program to perform Boolean operations on binary 
arrays considered as masks. 

g. A high speed assembly language program to extract contiguous regions 
(blobs) from binary arrays obtained from images. 

h. A high speed assembly language program for counting the number of 
elements in a binary array. 

i. A FORTRAN routine for circumscribing an extracted object in an 
image with a circumscribing rectangle. 

j. A general purpose statistical routine for taking an image which has 
been previously thresholded into binary form, and then computing 
centers of gravity, mean densities, and various types of moment 
computations, as well as obtaining minimum and maximum values of 
intensity range for an image. 

All the above programs are available at the LISP language level and 
hence can be called with considerable ease by higher-level programs 
without the need for the user to descend to lower level coding. The 
above general and special purpose programs operate in a unique 
environment involving both specialized equipment and users with 
particular kinds of processing problems. Despite the specialized 
nature of this environment, a systematic documentation effort is being 
made to document these programs so as to describe the operations and 
enable their duplication. 



197 V 



Serial No. NDS (CF)-65 PR/P 1278 

Computer Analysis and ImaRe Processing Results : The large amount of 
PDP-10 computer time used during this period is mostly attributable to 
the various analyses that have been run on the computer using scan data 
from various biological sources. We mention below some of these 
results that have been obtained: 

a. Various fluctuations in scanner illumination and line voltages 
result in non-uniformities in an otherwise uniform section of a 
scan. To analyze the degree of non-uniformity, a statistical 
analysis was made on the values of the reference beam on the 
microscope scanner. It was found that this illumination is extremely 
stable, so much so that for a reasonable level of illumination, 
variations in illumination level were small compared to those 
attributable to quantization errors and other sources of scanning 
noise. 

b. In order to test the resolution of the scanner for scanning silver 
grains on blood cells, the Stromberg-Carlson 4020 Display was used 
to resynthesize several scanned images of labeled blood cells. In 
the resynthesized image, the silver grains appeared clearly resolved 
and hence the data necessary for location and recognition can be 
considered to be present in the output of the scanner. 

c. An analysis of a small region from one of these cells containing 
multiple silver grains was made using the morphological analysis 
algorithm. The result was a proper identification of the location 
of silver grains and an indication that if this procedure were used 
over a larger image, the silver grains would properly be identified. 

d. Several experiments were performed using optical serial section data. 
In the first, a specimen with small polystyrene spheres was sectioned 
optically and then the image was reconstructed in three dimensions 
within the computer. Since the spheres were only a few microns in 
diameter, the reconstructed image, rather than being that of a 
sphere, was the diffraction pattern that would be expected from such 
small, largely phase, objects. It was also possible, using the 
three-dimensional reconstruction inside the computer, to rotate the 
image through 90 degrees and hence produce the effects of having 
sectioned perpendicular to the plane of the optical slide. This 
technique can be directly extended to other natural objects and in 
particular can be extended to larger objects than these test 
specimens. 

e. A set of serial sections of two adjacent nerve cells was made which 
exhibited the appropriate low depth-of-f ield for the optics that 
were used and enabled in a resynthesized set of images the easy 
visual identification of the vertical sectioning process. 



198. 



Serial No. NDS (CF)-65 PR/P 1278 

f. An extensive analysis was made of several optical sections of monkey 
vagal nuclei neurons. These analyses consisted of a complete 
morphological decomposition using the morphological analysis algorithm 
along with the determination of optical density statistics for the 
decomposed objects. Using these statistics then, the original 
objects were resynthesized and the computer's reconstruction was 
visually compared with the original scan data. The strong 
correspondence between the original and the resynthesized images 
suggested that the morphological analysis decomposition procedure 
preserves the morphological structure of images for purposes of 
recognition of cells and their components. 

g. A program was written by Bryan (DCRT) to enable manual specification 
of the decomposition procedure analogously to the automatic method 
used in the morphological analyzer. This program is currently being 
worked on to make its operation more precise and more convenient. 
Both versions are being employed particularly in the quantitative 
analysis of chromatolysis. 

Honors and Awards: None 

Publications: Lipkin, L.E.: Resolution, scale change and information 
distortion. In Lipkin, B.S., and Rosenfeld, A. (Eds.): 
Picture Processing and Psychopictorics . New York, N.Y., 
Academic Press, 1970, pp. 203-215. 

Lemkin, P.P.: A patch to focal-w to use the LINC-8 display. 
DECUS Program Library . FOCAL8-58: 1-3, 1969. 

Lemkin, P.F.: OCTMON: Octal monitor for the PDP-8. DECUS 
Program Library . 8-298: 1-7, 1969. 

Stein, P.G.: Image -analyzing microscopes. Anal. Chem. 
42: 103A-105A, 1970. 

Shapiro, H., Biryan, S., Lipkin, L.E«, Stein, P.G., and 
Lemkin, P.P.: Computer aided microspectrophotometry of 
biological specimens. This paper has been accepted for 
publication in Exp. Cell Res. 

Kirsch, R.A.: Computer determination of the constituent 
structure of biological images. This paper has been accepted 
for publication in Comput. Biomed. Res. 



199 V 



Serial No. NDS (CF)-68 PR/P 1650 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Twin Placentation in Relation to Zygosity 

Previous Serial Number: Same 

Principal Investigators: Toshio Fujikura, M. D. 

Luz A. Froehlich, M. D. 

Other Investigators: Ntinos Myrianthopoulos, M. D. 

Cooperating Units: All Collaborating Institutions 

Man Years 

Total: 0.0 
Professional: 0.0 
Others: 0.0 

Project Description : 

Twins totalling 569 pairs were studied in relation to type of twin 
placentation. Separate diamniotic-dichorionic placentas were rarely 
associated with monozygosity. This and the fact that monochorionic 
placentas are all monozygotic make examination of twin placentas 
extremely useful in forecasting zygosity. Relatively heavy infants 
as well as twins with large intrapair birth-weight differences were 
common in the separate diamniotic-dichorionic group, suggesting 
independent intrauterine growth of co-twins. The converse was true in 
twins with fused diamniotic-dichorionic placentas, who also had the lowest 
death rates. Perinatal death rate was the same in whites (14.77o) and 
Negroes (14.37o), highest in male-male twin pairs, but lowest in male-female 
pairs in Negroes and female-female pairs in whites. Compared to singleton 
deaths, the frequency of congenital malformations was not higher in twin 
deaths, but the types of malformations found in monochorionic deaths were 
often multiple and lethal. This study has been completed. 

Honors and Awards: None 

Publications: Fujikura, T. , and Froehlich, L.A.: Twin placentation 
and zygosity. Obstet. Gynec. 37: 34-43, Jan. 1971. 



201 V 



Serial .No. NDS (CF)-69 PR/P 1763 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Kidney Malformations in Fetuses of A x C Line 9935 Rats 

Previous Serial Number: Same 

Principal Investigators: Toshio Fujikura, M. D. 

Other Investigators: None 

Cooperating Units: None 

Man Years 

Total: 0.9 
Professional: 0.5 
Others: 0.4 

Project Description : 

Fifty A X C 9935 strain inbred rat litters were examined near term; 88/315 
fetuses (27.97o) showed renal malformations (renal agenesis and hydroneph- 
rosis). Unilateral renal agenesis occurred more often on the right side 
(11.57o) than the left (3.67o), especially in males. Renal agenesis was 
always associated with absence or hypoplasia of a uterine horn or vas deferens 
and epididymis on the corresponding side. However, the ovaries or testes 
were present and intact. Hydronephrosis was seen more often on the left 
side (11.47o) than the right (3.57o). There was no significant sex difference 
in the frequency of renal agenesis and hydronephrosis. In hydronephrosis 
the ureteropelvic junction was patent, and hydroureter always accompanied 
moderate hydronephrosis. Atresia near the vesicoureteral junction was 
considered as a cause of hydronephrosis. There was a close morphogenet ic 
relation between renal agenesis and hydronephrosis. This study has been 
completed. 

Honors and Awards: None 

Publications: Fujikura, T.: Kidney malformations in fetuses of A x C 
line 9935 rats. Teratology 3: 245-249, Aug. 1970. 



203^ 



Serial No. NDS (CF)-69 PR/P 1764 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Placental Study of Abortion Material 
(Obtained by an induced abortion) 

Previous Serial Number: Same 

Principal Investigators: Toshio Fujikura, M. D. 

Other Investigators: Hideo Nishimura, M. D. 
Kenichiro Ezaki, M. D. 

Cooperating Units: Department of Anatomy 
Kyoto University 
Kyoto , Japan 

Man Years 

Total: 0.1 
Professional: 0.1 
Others: 0.0 

Project Description : 

The placental materials of 114 induced abortions (normal group) and 
87 spontaneous abortions (abnormal group) were histologically compared. 
Degenerative hydropic and necrotic villi were commonly found in the 
chorion laeve of the normal group even in early gestation, but not in the 
chorion frondosum. The sampling site in the chorionic sac was important 
for histological diagnosis. The mean number of chorionic villi in the 
abnormal group was not different from that of the normal group within each 
gestational interval. This indicates that abnormal villous growth may not 
be the primary factor responsible for spontaneous abortion. Up to 14 weeks 
gestation, active syncytial proliferation was present but no substantial 
increase of chorionic villi was found. Beyond this inactive stage the 
villous number increased rapidly and conversely syncytial sprouts decreased 
in numbers. The mechanism of syncytial proliferation was discussed in 
relation to other prenatal conditions. Study completed. 

Honors and Awards: None 

Publications: Fujikura, T., Wishimura,. H., Ezaki, K. : Placental study of 
abortion material (obtained by an induced abortion). 
Amer. J. Obstet. Gynec, in press 



205V 



Serial- No. NDS (CF)-69 PR/P 1765 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: The Interrelationship Between Selected Congenital 
Malformations and Major Pathologic Findings 

Previous Serial Number: Same 

Principal Investigators: Luz A. Froehlich, M. D. 

Toshio Fujikura, M. D. 

Other Investigators: None 

Cooperating Units: All Collaborating Institutions 

Man Years 

Total: 0.05 
Professional: 0.05 
Others: 0.00 

Project Description : 

The incidences of certain variables in the baby, mother and placenta of 
core deaths were compared with deaths having selected congenital malforma- 
tions. Velamentous and marginal cords were more common in single umbilical 
artery deaths compared to core deaths. There was surprisingly little 
association with diabetes, except among the cases with agenesis of the 
kidney. Meconium staining was nearly twice as high in multiple heart 
malformations compared to controls. Hydramnios was common in association 
with anencephaly, and to a lesser extent with spina bifida and hypoplasia 
of lungs. The incidence of toxemia was more than twice as high in 
anencephaly, spina bifida, and agenesis of the kidney compare to controls. 
In addition, retroplacental hemorrhage was twice as high in anencephaly. 
Erythroblastosis was twice as high in accessory spleen but was not found 
in any of the other malformed cases. These correlations will be tested for 
possible significance. The study is being readied for publication. 

Honors and Awards : None 

Publications: None 



207 V 



Serial No. NDS (CF)-69 PR/P 1766 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July I, 1970 through June 30, 1971 

Project Title: Reproductive Ability of the American Negro with Sickling 
and its Public Health Implications 

Previous Serial Number: Same 

Principal Investigators: Luz A. Froehlich, M. D. 

Toshio Fujikura, M. D. 

Other Investigators: None 

Cooperating Units: All Collaborating Institutions 

Man Years 

Total: 0.4 
Professional: 0.4 
Others: 0,0 

Project Description : 

The reproductive performance of 654 sicklers and 1,890 non-sicklers in the 
Collaborative Study was compared. The cumulative fertility rate of mothers 
with sicklemia was the same as that of non-sickling mothers. There was no 
substantial difference in perinatal death rates, birthweight and gestation 
age between the sickling and non-sickling group. Only the infant and child 
death rate was higher in the sickling group. Because of the normal 
reproductive abilities of the sickler, the sickle cell gene may continue 
to propagate in the U.S. Negroes for generations to come. 

The paper has been approved by the Publications Review Board and the NIH 
Review Board. Certain modifications and refinements are felt necessary prior 
to submission for publication. These modifications are strongly dependent 
on the performing of hemoglobin electrophoretic studies on the sicklers, 
to distinguish the homozygotes from the heterozygotes. Such a plan to 
allow uniform testing of all Negroes for sickling and to follow this through 
with hemoglobin electrophoresis on the proven sicklers is being earnestly 
pursued. 

Honors and Awards : None 

Publications: None 



209V 



Serial- No. NDS (CF)-69 PR/P 1769 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: The Significance of Chorioangiomas 

Previous Serial Number: Same 

Principal Investigators: Luz A. Froehlich, M. D. 

Toshio Fujikura, M. D. 

Other Investigators: Pearl Fisher, Ph.D., OB, NINDS 

Cooperating Units: All Collaborating Institutions 

Man Years 

Total: 0.1 
Professional: 0.1 
Others: 0.0 

Project Description : 

Eighty-one cases of chorioangioma in the Collaborative Study were analyzed, 
of which five were in twin pregnancies. Chorioangiomas were more frequent 
in whites (0.337o) than in Negroes (0.207o) and in females (49 cases) more 
than in males (37 cases). Two infants had neonatal thrombocytopenic 
purpura unassociated with skin hemangioma. Congenital anomalies were 
high in twins; in single births, the incidences of several malformations 
were significantly higher than in the general Collaborative Study population. 
An interesting correlation was noted among chorioangioma, skin hemangioma 
and single umbilical artery. In Negroes, in particular, twin rates in 
current and prior pregnancies were high as was the fact that the gravida 
herself was a twin. Acute toxemia was a significant complication in both 
whites and Negroes. This study has been completed. 

The paper was presented at the Teratology Society Meeting in Annapolis, 
Maryland on May 20-22, 1970. 

Honors and Awards: None 

Publications: Froehlich, L.A., Fujikura, T., and Fisher, P.: Chorioangiomas 
and their clinical implications. Obstet. Gynec. 37: 51-59, 
Jan. 1971. 



211v 



Serial No. NDS (CF)-69 PR/P 1772 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Pathologic Effects of Ligation of the Anterior Spinal 
Artery and/or the Great Radicular Artery in Monkeys 

Previous Serial Number: Same 

Principal Investigators: Oscar Aparicio, M. D. 

Other Investigators: Larry C. Fried, M. D. 

Cooperating Units: Surgical Neurology Branch 
Intramural Research 
National Institute of Neurological Diseases and Stroke 

Section on Neuroradiology 

Medical Neurology Branch 

Intramural Research 

National Institute of Neurological Diseases and Stroke 

Man Years 

Total: .05 
Professional: .05 
Others: .00 

Project Description : 

Objectives : The neuropathological evaluation of spinal cords of monkeys 
subjected to vascular ligations, in order to determine the practical 
feasibility of sacrificing any of these vessels if necessary during the 
course of a surgical procedure. 

Methods Employed : The neuropathologic evaluation of the spinal cords by 
means of multiple staining methods, including H&E, Luxol Blue with Cresyl 
Violet, and others, in order to determine the presence, extent and 
location of any lesions due to ischemia. 

Proposed Course : The histopathological evaluation has been completed and 
a publication is being prepared. 

Honors and Awards: None 

Publications: None 



213^ 



Serial No. NDS (CF)-70 PR/P 1858 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Thrombocytopenic Purpura and Placental Hemangioma 

Previous Serial Number: Same 

Principal Investigators: Luz A. Froehlich, M. D. 

Other Investigators: Mary Housler, R. N. 

University of Buffalo 



Cooperating Units: 


None 


Man Years 




Total: 


0.1 


Professional: 


0.1 


Others: 


0.0 


Proiect Description 





A case is presented in which neonatal thrombocytopenic purpura (TP) was 
associated with hemangioma of the placenta. This combination has not been 
reported in the literature, although the combination of TP and skin 
hemangioma is well known. The baby had no skin hemangioma. Other known 
causes of TP in the newborn such as maternal TP and sepsis were not 
present. The mother took a thiazide during the last eight weeks of 
pregnancy. However, the incidence of TP among those whose mothers took 
thiazide (0.017o) was not higher than among those whose mothers did not 
take thiazide (0.017o), indicating that thiazides and TP are not related. 
Study completed. 

Honors and Awards: None 

Publications: Froehlich, L.A. and Housler, M. : Neonatal thrombocytopenia 
and chorangioma. J. Pediat. 78: 5l6-519^ 1971. 



215V 



Serial No. NDS (CF)-70 PR/P 1859 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Mental and Motor Development in Monozygotic 
Co-Twins with Dissimilar Birthweights 

Previous Serial Number: Same 

Principal Investigators: Toshio Fujikura, M. D. 

Luz A. Froehlich, M. D. 

Other Investigators: None 

Cooperating Units: All Collaborating Institutions 

Man Years 

Total: 0.2 
Professional: 0.2 
Others: 0.0 

Project Description : 

Developmental measures in 125 monozygotic twin sets with unequal 
birthweights between co-twins were studied. There were no significant 
differences between co-twins in the Bayley mental and motor scores at 
eight months nor the Stanford-Binet I.Q. at four years. A reportedly 
higher I.Q. for the heavier monozygotic twins was not confirmed in this 
study, even among pairs with large birthweight differences (mean 
differences 26-287o). Although the effects of nutrition on the mental 
development of the fetus are currently of great concern, these data suggest 
that the developing human brain seems to have a strong resistance to 
intrauterine deprivation. 

The future of this manuscript is uncertain. Because interpretation of 
the data is controversial the paper is apparently being shelved until 
data on the seven year I.Q. have been similarly analyzed. 

Honors and Awards: None 

Publications: None 



217 V 



SeriaJ. No. NDS (CF)-71 PR/P 1911 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: A Follow-up of Children with Single Umbilical Artery 

Previous Serial Number: None 

Principal Investigators: Luz A. Froehlich, M. D. 

Other Investigators: Toshio Fujikura, M. D. 
John Churchill, M. D. 

Pearl Fisher, Hi.D. 

Cooperating Units: All Collaborating Institutions 

Man Years 

Total: 0.3 
Professional: 0.2 
Others: 0.1 

Project Description ; 

There are 306 presently living core Study children who were born with 
single umbilical artery (SUA). These are from a total of 355 core SUA 
cases (identified by microscopic examination) of whom 49 (13.8%) have 
died, mostly in utero or during the first week of life. Associated 
congenital malformations were a significant cause of death. The surviving 
cases were compared with surviving controls matched for race, sex, 
institutions, birthweight, gestational age and S-E index. Velamentous 
and marginal insertions of cord were about six times as frequent in SUA. 
There were also slightly more neurologically abnormal cases (7.0%) than 
controls (4.9%). On the whole, however, mean mental and motor and 
four year I.Q. scores, and parameters of physical growth did not distinguish 
between cases and controls. Statistical analysis has been temporarily 
halted because of Dr. Fisher's departure, but all attempts will be made 
to see the paper to completion despite this inconvenience. 

Honors and Awards : None 

Publications: None 



219 V 



Serial No. NDS (CF)-71 PR/P 1912 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: Birthweight in Relation to Renal Glomerular Development 
and Gestational Age in Whites and Negroes 

Previous Serial Number: None 

Principal Investigators: Toshio Fujikura, M. D. 

Luz A. Froehlich, M. D. 

Other Investigators: None 

Cooperating Units: All Collaborating Institutions 

Man Years 



Total: 


0, 


.25 


Professional: 


0. 


.20 


Others: 


0, 


.05 


Proiect Description 







Birthweight, body length and X-ray evaluation of bone deve ''^ypnient continue 

to be widely used as measures of fetal maturity. Howevj^^ these are 

factors of somatic growth and are not necessarily direj,^i^y related to 

growth and maturity of internal viscera. On the conVj^^^-^on that histologic 

examination of an organ would give a more accurate P-Jcture of the maturation 

of internal viscera, a h^istologic evaluation of the Sidneys of 51^ neonatal 

deaths and fresh stillbirths was undertaken. In tl early phases of 

pregnancy, somatic growth appeared to proceed at r gienificantly faster pace 

in the Negro, as evidenced by higher birthweight ^ . ^\^^s race compared to 

whites. However, the developmental rate of vis^j, such as the kidney 

seemed to proceed at similar rates in the two rj ' Racial differences 

were observed in the percent of the presence o' . hroeenesis (PPN) in the 

"small for dates" as well as "large for dates'" /" . f „n«-g in both races 

about half of the "small for dates" infants f ^ -i i pxhibited a nephrogenic 

zone compared to more than 95% of the trul" "■ t'res At the same time 

two to four times as many "large for d ^ f, ^^^^^ts showed nephrogenesis 

as those which were mature both for ^ir' 'weight and gestational age. In 

maternal diabetes more than 807, r ^^fg^^ between 35-37 weeks gestation 

showed nephrogenesis where only r, ,„, . j r^u.^ oaoer is being 

J. J £ ,1- ^. b6.67o were expected, me pcifco. 

readied for publication. "* 

Honors and Awards: None 

Publications: None J 

221V 



\< 



I 



Serial No. NDS (CF)-71 PR/P 1913 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1970 through June 30, 1971 

Project Title: The Clinical Significance of Generalized Petechiae at Birth 

Previous Serial Number: None 

Principal Investigators: Luz A. Froehlich, M. D. 

Jean G. Oliver, M.A. 

Other Investigators: Toshio Fujikura, M. D. 

Jean S. Nemore, R.N., B.A. 

Cooperating Units: Section on Infectious Diseases, PKB, NIKDS 
All Collaborating Institutions 

Man Years 

Total: 0.2 
Professional: 0.2 
Others: 

Project Description : 

In a hand review of charts of some 30 of about 120 living infants diagnosed 
as having had significant skin petechiae at birth, six were found to have 
significant hearing loss, an incidence far greater than expected. Speech 
and behavior problems were also common. Mental retardation was found 
in a mature male along with marked visual impairment. 

On the hypothesis that significant skin petechiae could be accompanied 
by hemorrhages in vital structures such as the brain, inner ear, retina, 
etc., the review of charts of all 120 children will be continued and 
findings compared with those of controls matched for institution, race, 
sex, birthweight and socio-economic index. 

Honors and Awards: None 

Publications: None 



223 V 



Serial Wo. NDS (CF)-70 PR/SLH 1520 

1. Perinatal Research Branch 

2. Section on Speech^ Language and 
Hearing 

3. Bethesda^ Maryland 

PHS-RIH 
Individual Project Report 
July 1, 1970 through June 30^ I97I 

Project Title Explorative Study for the Use of a Speech and Language 

Screening Examination for 3-Year Old Children in the Home 
Situation 

Previous Serial Number: NDS-(CF)-63 PR/BS II67 

Principal Investigator: Dr. Miriam F. Fiedler 

Other Investigator: Dr. Eric H. Lenneberg, Cornell University 

Cooperating Units: Children's Medical Center, Boston, Massachusetts 

Section on Behavioral Sciences, PRB, NINDS 
Section on Pediatric-Neujrology, PRB, NIWDS 

Man Years: 

Total: .5 

Professional: .2 

Other : . 3 

Project Description : 

The study concerns children with non-normal speech identified at age three 
years by means of interviews with the mothers of Perinatal Project children 
in Boston. Findings were confirmed by subsequent speech, language and 
hearing examinations. The speech data were considered in relation to 
perinatal findings as an initial study, and, in a second study, with regard 
to outcome at age seven years in the light of psychological and neurological 
examinations. Most of the children identified as abnormal or suspect at 
age three years were found to be similarly regarded by other measures at 
age seven years. 

Part of the study was presented by Dr. Fiedler as a paper delivered at the 
Tri-City Meeting of the Obstetrical Society in Boston on May 16, I967, 
\nider the title "Delayed Speech Development in Children at 3 Years of Age 
Related to Peri and Postnatal Findings". The study has been completed. 

Honors and Awards: None 

Publications: 

Fiedler, M. F. , and Lenneberg, E. H. : Explorative Study for the Use of a 
Speech and Language Screening Examination for 3-Year Old Children in the Home 
Situation. Pediatrics . In press. 

225v 



Annual Report 

Associate Director's Report 

July 1, 1970 through June 30, 1971 

Extramural Programs 

National Institute of Neurological Diseases and Stroke 

A detailed summary of the NINDS extramural research grant effort and training 
accomplishments in Fiscal Year 1971 are provided in the reports of the Research 
Grant and Training Branches; organizational progress including program reporting 
activities are included in the report of the Administrative Officer. In broad 
terms, the highlights of NINDS Extramural Programs for Fiscal Year 1971 have 
been characterized by: 

A. A CONTINUED REDUCTION IN TRAINING ACTIVITIES : With a "hold the line" 
philosophy in the training activities of the Institute, 10 additional grant 
supported training programs have had to be discontinued. This was necessitated 
by the continuing increase in funding requirements of active programs despite 
an across-the-board "negotiated cut" for the third successive year, this year 
at the 157o level. The plan to stabilize the number of new RCDA awards each 
year at 20 is reaching fruition; if there are no further reductions in this 
program area in the future, the fiscal requirements for this minimal level of 
activity will be reached in Fiscal Year 1975. Fiscal Year 1971 will end with 
$8.5 million approved training grant and traineeship applications unfunded and 
$1.5 million approved fellowship applications unfunded. The President's Budget 
for Fiscal Year 1972 will require further reductions in NINDS training activi- 
ties. This will have particularly unfavorable effects on the recruitment and 
training of personnel for careers in teaching and research in neurology, child 
neurology, neurological surgery and otolaryngology. In addition, desperately 
needed new training programs in the basic science areas of neurochemistry, 
neurophysiology and neurobiology will not be activated. 

B. A TEMPORARY STABILIZATION OF THE LEVEL OF RESEARCH GRANT ACTIVITY : An 
increase of $5 million in research grant funds in Fiscal Year 1971 permitted 
the Institute to maintain its research grant activities at approximately the 
same level as in Fiscal Year 1970. The additional funds were utilized to help 
offset a 77c increase in cost-of-living; to provide for a reduction to 107o of 
the previous "across-the-board" budget negotiation level of 157=,; to meet a 
peak in the triannual level of previously committed applications; and to 
initiate feasibility studies for the establishment of acute spinal cord injury 
research centers. The President's Budget for Fiscal Year 1972 reduces the 
research grant activity by over a half-million dollars. This reduction will 
interfere seriously with the Institute's plan to further reduce across-the- 
board negotiations to the 57= level, to meet the continuing increase in research 
costs, and to provide additional emphasis on selected high priority research 
programs. 

C. RESEARCH AREAS OF SPECIAL EMPHASIS : 

(1) Stroke : The Institute's principal instrument for research grant 
support of stroke research is its program of stroke clinical research centers, 
eighteen of which are now active. These are centers of coordinated clinical 
and basic research focused on problems such as local and regional cerebral 
blood flow, cerebral hypoxia, cerebral edema, cerebral arterial thrombosis. 



hemorrhage and spasm, stroke epidemiology, the pathophj'siology of the transient 
ischemic episode, etc. In the area of cooperative research; The cerebral 
aneurysm study has been completed with significant results; the hypertension- 
stroke study is in its final phases; plans have been completed for the initia- 
tion of a study to evaluate the efficacy of amino-caporic acid in the treatment 
of the acute stages of intracerebral hemorrhage. As a result of the activity 
of the Joint Council Subcommittee on Cerebrovascular Disease (NINDS and 
NHLI) , plans have been developed for a program of coordinated research on the 
problem of acute stroke care (nervous system monitoring and acute medical- 
surgical intervention) through the establishment of acute stroke research units; 
this activity can be initiated if the additional funds required are made avail- 
able for stroke research. 

(2) Trauma To The Central Nervous System : In 1968, the NINDS initiated 
its program of Head Injury Clinical Research Centers by means of specialized 
research center awards to plan and develop these activities. In Fiscal Years 

1971 and 1972, the pilot centers will compete for future support in the cate- 
gorical clinical center program. In order to provide for both a progress report 
on research accomplishments to date in this area and to establish specific 
scientific merit criteria for review of applications, the Institute has organ- 
ized a Head Injury Research Workshop. This Workshop will be held in February 

1972 and will include scientists from each of the 6 active head injury research 
centers and selected scientists from other laboratories. 

In the area of acute spinal cord trauma, fifteen applications for feasibility 
studies have been received. These applications to develop the scientific, 
medical, community and organizational feasibility of establishing up to 4 
acute spinal cord injury clinical research centers in the U.S. will undergo 
review during the summer of 1971 and receive final Council recommendation in 
September 1971. Funds for the feasibility studies have been set aside. A 
minimum of $4 million will be required to launch the 4 center activity in late 
Fiscal Year 1972 or early Fiscal Year 1973. 

(3) Communicative Disorders (Hearing; Speech; Language): A Task Force of 
the NINDS Communicative Disorders Program Project Review Committee has completed 
its report on opportunities for research in these areas. The report documents 
the several research problems of high priority and opportunity which justify 
increased Institute effort; included are the biological and medical effects 

of noise on man; chronic and episodic vertigo; genetics and deafness; pre-school 
deafness; hearing loss and chronic secretory otitis media; and traumatic neural 
impairment and language. The documents supporting these specific program area 
needs will serve as a basis for program planning and development in FY 1972. 

(4) Parkinsonism; L-DOPA; Neurotransmitters : The availability of L-DOPA 
and its acceptance as the therapy of choice for moderate to severe Parkinsonism 
has redirected and accelerated the entire area of research on movement disorders, 
neurotransmitters and the neural and neuromuscular synapse. L-DOPA potentiators 
are now in phase 2 and phase 3 study; L-DOPA analogues, dopamine-o-mimetic 
drugs and antichoreic agents are now potentially available; the delicate balance 
of protagonists and antagonists of synaptic transmission is being described; 

and the metabolic pathways of neurotransmitter saturation therapy are being 
unraveled. The four research centers in this area supported by the Institute 



2 w 



are the leaders in these developments; the additional $3 million in research 
projects provides for an independent but complementary research effort. 

D, PROGRAM REPORTING : With the completion of the conversion of our routine 
data storage and retrieval system to tape and the development of programmed 
search procedures, for the first time a comprehensive search and reporting 
system is now in effect. Staff scientist administrators responsible for 
program- area activities now have regular reports available to them in both 
the research grant and training grant programs; similar reporting will be 
available in the fellowship programs in Fiscal Year 1972. Computer data 
also are being used regularly for fiscal monitoring and reporting. In 
Fiscal Year 1972, pilot programs will be developed and tested for the 
reporting of scientific status and accomplishments. 



3w 



Annual Report 

Administrative Report 

July 1, 1970 through June 30, 1971 

Extramural Programs 

National Institute of Neurological Diseases and Stroke 

Extramural Programs found the prohlem of diminishing staff particularly 
difficult during Fiscal Year 1971- During a year when position strength was 
more than 15^ below normal operating levels and the program data maintenance 
and analysis organization was "being expanded to meet increasing demands 
against it, the normal ability to reassign support personnel to cover absences 
or heavy load demands was most difficult. The sacrifice of certain central 
service positions has caused a further drain on the time and effort of exist- 
ing clerical staff. Mail and messenger service and local duplication have 
all become do-it-yourself tasks. The sum of this was that certain minimum 
compromises with total efficiency had to be made. 

The Extramural Programs participated with other Institute program areas 
in two Equal Employment Opportunity conferences during FY 1971- These con- 
ferences were held to identify specific problems and attitudes that foster 
inequitable job situations and to develop an Institute commitment to improve 
equal opportunity. An affirmative action plan has been developed and is being 
implemented at this time. 

The Data Analysis and Reports Unit has completed its conversion from 
manual records to computer files. This new data file has begun to provide 
Extramural Program's management with an information capability that enhances 
management planning, analysis, and operations and improves projections of 
future needs . 

The full impact of the concurrent project period ruling was felt in grants 
management during FY 1971- A significant amount of additional grant support 
was permitted by this method of allowing the use of unexpended grant funds 
during an extension of the project period. 

The administration of the NINDS Extramural Programs budget was made diffi- 
cult due to the late release of funds with the appropriation being signed on 
January 11 and the apportionment approved by the Office of Management and 
Budget on March 1, 1971. This late release of funds resulted in the delay of 
the award of many new as well as competing renewal Research Grants until the 
fourth quarter of the fiscal year. As in the past, reserves were withheld 
from the appropriation in the amount of $2, 957 > 000 for Research Grants and 
$672,000 for Graduate Training Grants, a total of $3,629,000. Of this amount 
withheld from grants, $1,307,000 was allocated to NINDS direct operations for 
pay raises, and the balance of $2,322,000 to other Institutes of N.I.H. and 
the DHEW for pay raises. However the amounts released this year were increas- 
ed over the President's Budget by $U, 211, 000 for Research Grants and $187,000 
for Fellowships. There was no change in Training Grants. Of the increase 
for Research Grants, $3,522,000 was for research projects and centers and 
$689,000 for General Research Support Grants. Even with these increases, the 
level of unfunded approvals continued to climb upward as a result of an in- 



5 w 



creased number of approvals, the average cost of grants increasing coupled 
with a lesser amount of negotiated reduction. The amounts of unfunded approv- 
als were $15,300,000 for Research Grants, $8,500,000 for Training Grants, and 
$1,^50,000 for Fellowships. To assure the most efficient use of available 
funds, the following steps were taken. 

1. Research Grants were negotiated do^mward in an amount averaging 10^, re- 
sulting in savings of approximately $U.O million which resulted in the award 
of "^V^o more new and competing renewal grants than would have been possible 
without negotiations. 

2 . Graduate Training Grants were negotiated downward an average of 15*5^ which 
made possible the award of twice as many competing grants as would have been 
awarded without negotiations. 

3- Additional savings were realized for Research Grants and Graduate Training 
Grants by utilizing the concurrent project concept. This means that competing 
renewal grants were reduced by the amount of funds remaining in the previous 
project period and extended up to a year so that the grantee could use these 
unexpended funds during the new project period. 



6 w 



Annual Report 
July 1, 19T0, through June 30, 1971 
Research Grants Branch 
National Institute of Neurological Diseases and Stroke 

Introduction 

The "brain is by far the most intricate^ sensitive and 
versatile organ in the body. As a result^ it has been the subject of 
ejctensive study and research for centuries. However, it has yielded 
only slowly to scientific exploration because of its complexity and 
because of its relative inaccessibility due to being enclosed in the 
skull and due to the blood-brain barrier which separates the brain 
metabolic ally in many respects from the rest of the body. Nevertheless, 
research is gradually bringing a greater understanding of how the 
10 to 13 billion individual nerve cells in the brain, together with the 
additional billions comprising the nervous system, work together to 
make the human body an effective and coordinated living organism. 

The economic burden of the neurological and sensory diseases 
amounts to billions of dollars each year, with immeasurable human 
suffering. Although the human spirit can often adjust to the effects 
of physical disability, even the greatest courage may be broken by the 
devastating consequences of brain injtiry or disease which may continue 
or exacerbate during the remainder of a person's life. 

More than 200 disorders are known to afflict the brain, 
sense organs, nervous system and neuromuscular apparatus, the most 
familiar of which are stroke, head and spinal cord injury, epilepsy, 
cerebral palsy, aphasia, multiple sclerosis, muscular dystrophy, 
parkinsonism, brain tumors, and otosclerosis. These diseases lead to 
paralysis, loss of speech, paraplegia and deafness, and are among the 
major causes of death and permanent disability in the United States. 

The research grant programs of the National Institute of 
Weiu-ological Diseases and Stroke include research projects, research 
program projects, cerebrovascular and commimicative disorder outpatient 
clinical research projects, clinical research centers, and grants to 
establish cerebrovascular and head injury clinical research centers. 
The objectives of these programs are the identification, stimulation, 
and support of important research problems related to the diagnosis, 
treatment, and prevention of disorders such as those mentioned above. 

For many years the question of whether hypertension played a 
major role in the development of stroke has been controversial. About 
one year ago the Veterans Administration reported a study indicating 
that antihypertensive therapy had a marked protective affect on stroke 
morbidity and mortality. This whole question was then reviewed by the 
Joint Council Subcommittee on Cerebrovascular Disease of the NHLI and 
NINDS and by the NAIJDS Council. Both groups recommended that further 



7 w 



steps be taken as expeditiously and aggressively as possible to implement 
the application of antihypertensive therapy on commimity bases. Since 
this problem is now beyond the clinical research stage and is in the area 
of commimity application, this recommendation was referred to the Regional 
Medical Programs Service which was urged to give it a high priority. 

Injuries to the spinal cord are occurring with increasing 
frequency. Once the spinal cord degenerates in the area of injiiry, 
paralysis always develops. In March 1971^ the HANDS Coimcil recommended 
approval of plans to support a few Acute Spinal Cord Injury Centers. At 
the first stage of development, funds would be provided only to plan the 
requirements and test the feasibility of a few centers for acute spinal 
cord injury. Each Center's plan would include investigation, development 
and evaluation of improved methods of emergency treatment, rapid trans- 
portation, diagnostic techniques, medical- surgical repair, and the train- 
ing of required professional, scientific and technical personnel. Ultimately, 
it is expected that a fully developed Center would contribute important 
information on the prevention of degeneration of the spinal cord and on 
the restoration of spinal cord function. Close liaison with other 
government and non-government agencies with responsibilities in this 
area is being maintained. 

The importance of improved stroke acute care is obvious since 
70 to 80 percent of the mortality occurs in the first ten days. Also, 
the consequences of not recognizing a progression or extension of the 
infarct may be catastrophic. After extended review of this problem, the 
Joint Council Subcommittee on Cerebrovascular Disease, KHLI-WIKDS, agreed 
that the present information about stroke acute care is exrtremely limited 
and that the individual efforts on the part of investigators, singly 
and in teams, to push forward with the problem have met with only limited 
success to date. In view of these considerations, the NANDS Council in 
March 1971 approved the organization of a Problem Commission on Stroke 
Acute Care Research. The Commission will consist of 10-12 experts in 
this and closely related areas and will be closely linked to the present 
Stroke Clinical Research Centers. It will be the responsibility of the 
Commission to (l) develop the strategy required to explore the problem 
(e.g., identify the neurological variables requiring attention and 
possible human and mechanical monitoring as a basis for intervention); 
(2) describe the specifications of the required organization, software 
and hardware; and (3) assume the responsibility for testing and evaluating 
these specifications in their own Stroke Clinical Research Units. 

This was the third year in which every research grant, competing 
and committed, was subject to a negotiated reduction. All grants were 
negotiated downward by an amount which would hopefully still allow the 
work to proceed, although productivity doubtless was reduced even more 
than in the previous two years because costs have increased. As a result 
of this arrangement, however, over 50^0 more f\inds were available for 
competing applications. Even so, approximately half of the approved 
projects could not be supported. The Institute has been advised that 
grants may be reduced by only a minimal amount {%) next year. In effect 



this will amount to a reduction of about $2.5 million or 25 percent in the 
fluids available for competing applications. Two years ago^ the National 
Eye Institute was organized and all of the activities related to the 
visual system and disorders of vision were transferred out of WIKDS. This 
took more than 20 percent of the grants, applications and funds. Althoiigh 
there has been little increase in the f\xnds available, it has been possible 
to maintain about the same number of projects by reducing each grant as 
described above. That is, last year 1,267 research grants were supported 
at a total cost of $U8.8 million. This year 1,256 grants were supported 
at a cost of $53.6 million. 

Within one year after the formation of the National Eye Institute, 
the number of applications to IttRDS was just as large as it was before 
the operation of the two. For example, in March 1970, the RAKDS Council 
reviewed about 375 applications. In June 1971 the Council reviewed about 
^50 applications, a 20 percent increase over the average for the previous 
year. This continued and dramatic increase in the number of requests is 
due primarily to the effectiveness of the research training programs of 
this Institute in which the output of fully trained investigators has 
only in recent years reached its full potential. 

There were two replacements in the professional staff of the 
Research Grants Branch since last year. One was due to a retirement and 
the other was due to a resignation. One of the new staff members is 
Executive Secretary of Program Project Committee A and the other is 
responsible for research grants in the areas of pharmacology, medicinal 
chemistry, and toxicology. Both have proved to be mature and competent 
scientists and are becoming expert administrators. Therefore, the Branch 
continued to enjoy the services of an active and experienced staff. 

The numbers of grants and amounts of funds in the various 
disorder categories are shown in Appendix A. 



Cerebrovascular Disorders 

In FY 1971 support for research in cerebrovascular disease was 
at the level of $5*6 million, representing 63 research grants and I9 
clinical research centers. 

Studies directed toward cerebral blood flow continue to be of 
prime importance in research on cerebrovascular disorders. Some 
investigators direct their efforts to such facets as development and 
refinement of methodology and techniques for measurement of cerebral 
blood flow at a regional and focal level, others to problems of regulation, 
others to correlative studies of cerebral blood flow with other parameters 
which might be used as predictive studies in the course of a stroke. 

In one clinical research center, the relationship of cerebral 
blood flow (CBF) to pH in cerebral venous blood and in cerebral cortex 
is being explored. Dogs were lightly anesthetized, paralyzed and 
ventilated with oxygen. CBF was meas\u:"ed by an electromagnetic flowmeter 
in a shunt between the torcular and the superior vena cava. Cerebral 
venous blood pH, p02^ and pC02 were measured continuously in a flow- 
through within the shunt. The same parameters were measured on the surface 
of the cerebral cortex by the application of flat surface counterbalanced 
electrodes on the exposed brain through a parietal craniotomy. The brain 
surface was protected by a shallow pool of warmed artificial cerebrospinal 
fluid. Simple seizures were produced by electroconvulsive stimuli or by 
pentylenetetrazol loV. and recorded by electroencephalographic monitor. 
In eight dogs so studied, CBF increased immediately with the onset of 
each seizure. Both brain and cerebral venous p02 increased consistently 
with seizures in well ventilated animals, but fell when seizures were 
accompanied by apnea. Brain pH showed little change initially during 
the acute phase of cerebral hyperemia. Thereafter brain pH showed 
variable changes. It decreased by O.IO-O.I5 units in most observations, 
but in several experiments a primary increase in pH was found. In the 
post-ictal phase brain pH was consistently increased above baseline values 
at a time when CBF was still elevated. In the post-ictal phase of 
impaired brains, autoregulation increases, induced by hypertension if CBF 
was accompanied by increases in brain pOg and pH, and decreases in pCOg. 

The lack of consistent correlation between CBF and brain pH 
during and after seizures suggests that factors other than hydrogen ion 
activity play a role in regulation of cerebral vascular resistance. Such 
factors, whether neurogenic or metabolic, serve the important function of 
augmenting CBF to meet the brain's increased oxidative metabolism during 
seizures. Further studies are continuing to define mechanisms of 
functional hyperemia in brain. 

Another study cites experimental evidence that microparticles 
present in blood during cardiopulmonary bypass cause a reduction in 
cerebral blood flow and metabolism during this procedure. The extent and 
distribution of ischemic cell change in dog brains following prolonged 
cardiopulmonary bypass are being investigated. Six animals have undergone 



10 w 



thoracotomy under anesthesia and four were maintained on complete bypass 
for four ho\:irs under conditions of normal temperature^ arterial pressure, 
"blood gas tensions, and pH. T\<ro control animals were maintained \inder 
identical conditions, "but without bypass. Thereafter, perfusion with 
saline followed by perf us ion-fixation with a formaldehyde-acetic acid- 
methanol mixt-ure was continued for at least 20 minutes at perfusion 
press-ures of 100-110 mg Hg. Following an additional period of immersion 
fixation, 20 micron sections were prepared from paraffin-embedded blocks 
at 6 mm intervals throughout the cerebral hemispheres, cerebellum and 
brain stem, and were stained by several critical staining methods. 

The four experimental animals studied following bypass showed 
early ischemic cell changes by the criteria of Brierley and Brown for 
light microscopy. These changes were non-focal and present in neiirons of 
layers "VI and V of the cerebral cortex, in thalamic nuclei, and in 
Itirkinje cells in the cerebellum.. Although there was no absolute regional 
localization, ischemic changes were more apparent within the parietal 
cortex than in other cortical areas. While occasionally small and medium 
sized arterioles were occluded by aggregates of red blood cells and 
amorphous PTAH positive material, there were no occlusive lesions of 
larger sized arterial vessels. Small punctate parenchymal hemorrhages 
were present in two specimens, and two instances of intraventricular 
bleeding, apparently originating in or near the choroid plexus, were 
found. Control sections were histologically normal. Continuation of 
these studies is planned, including microangiographic radiography of thick 
sections of brain following perfusion with colloidal barium. These studies 
are to be correlated with direct measiorements of microembolic material in 
blood during bypass by sonar detector methods. Also, the effectiveness 
of micropore filtration in preventing such changes will be tested. 

A serial circular angiotomographic device has been constructed 
and tested extensively on dogs during angiography. It has been possible 
to demonstrate a diffuse "brain blush" in tomographic sections 1 cm. 
thick. Extensive physical tests have been conducted to establish the 
relation of thickness of cut to tomographic angle and the relation of 
speed of rotation to resolution. The results of animal studies were so 
encoiuraging that the unit was moved to the clinical area and patient 
studies have been initiated. Fifteen studies have been carried out on 
patients. In two cases the angiotomogram gave diagnostic information not 
obtained from conventional films. Continued patient studies will be 
carried out to evaluate the unit for its clinical value. 

In the same radiology group, a new monitoring system for use 
dioring cerebral angiography is being evaluated. The system incorporates 
an industrial transducer and alarm system, and has proved to be an added 
safety factor during angiography. It indicates blood clots in the needles 
or catheter, sub-intimal position of the needle tip and changes of blood 
pressure or pulse rate. A unique feature of the device is that the trans- 
ducer may be left connected during the injection of contrast media. The 
part of the pressure tracing immediately after the injection of contrast 
media is of considerable interest. 



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In another clinical research center^ several members of the 
group are continuing studies on newly developed techniques for measuring 
hemispheric blood flow and metabolism in man. They have shown a bilateral 
reduction in hemispheric blood flow and in metabolism in unilateral stroke 
due to diaschisis. They have found that cerebral acetoacetate and alpha- 
butyrate metabolism is not increased in the infarcted hemisphere. The 
group now plans to direct their attention to the phosphate-oxygen ratio 
which may define whether or not uncoupling is present in the human brain 
following cerebral infarction. 

Another group of investigators is studying new methods for 
measuring regional cerebral blood flow using the Anger Camera and a tape 
storage and retrieval system following the intracarotid injection of 
radioactive xenon and technecium. These methods provide greater resolution 
for regional measurements than previously available, and the play back 
system permits measurement of up to 100 different areas of brain with a 
single injection. Evidence is being provided of definitive and diagnostic 
patterns of abnormal flow and changes in blood volume and blood brain 
barrier in intracranial hematoma and brain tumor as well as in patients 
with occlusive cerebral vascular disease and infarction. 

Another group has induced experimental subarachnoid hemorrhage 
by the injection of fresh blood into the subarachnoid space. This 
produced a spasm of the cerebral vessels with a marked reduction of 
cerebral blood flow on the release of serotonin. The relation of this 
to ruptured aneiirysm and spasm in hiunan subjects is under active investi- 
gation and methods of treatment are being considered. 

In another set of experiments, regional cerebral blood flow is 
being measured by implanted electrodes before and after occlusion of the 
middle cerebral artery and the effects of various forms of treatment on 
regional blood flow are under study. These studies are being combined 
with measurements of regional cerebral blood flow using the radioactive 
antipyrine method originally developed by Landau, Sokoloff, et a], as 
modified by Reivich and Isaacs. In the human laboratory, using the 
hemispheric blood flow method and the regional xenon techniques, the 
therapeutic usefulness of glycerol has been shown, which reduces brain 
swelling and increases blood flow in the area of cerebral edema. 

Therapeutic trials are also ongoing on a double blind basis 
with two different dri;igs. One is Hexobendine, a potent cerebrovasodilator. 
Investigations on man and in animals have sho-vm that it increases 
cerebral blood flow. The drug has been given to 11 patients intra- 
venously, and it has been observed that five improved, four showed no 
change and two died of unrelated causes. Wo toxic effects have been 
noted. These studies are being extended to evaluation of therapy in 
cases of acute and chronic cerebrovascular disease. Additionally, drug 
studies with Cyclandelate are now being initiated. 

Researchers in another clinical research center have completed 
a study in animals of the effect of change in blood pressure on blood 
flow through cerebral cortex made acutely Ischemic by occlusion of a 

12W 



middle cerebral artery. The effect of CO2 inhalation on blood flow 
through cortex which was either acutely or chronically ischemic was 
studied in animals and revealed the loss of autoregulation in ischemic 
cortex. The auto-radiographic technique for the measurement of regional 
cerebral blood flow in animals was used in the study of the effects of 
occlusion of a middle cerebral artery on blood flow and various regions 
of the brain correlated with changes in surface vessels. It was then 
possible to get quantitative estimates of flow in portions of the brain 
remote to those areas being visualized by the investigator. A study was 
begun of the effects of stimulation of the sympathetic nerves in the neck 
on blood flow through the cerebral cortex and on the diameter of s\irface 
blood vessels in animals. A pilot study has been initiated in hiMians of 
the measurement of cerebral blood flow by inhalation of xenon-133 and 
has so far shown that the technique is useful but needs further evaluation, 
particularly in patients with cerebrovascular and other neurological 
disorders. This evaluation study is continuing. 

Another group of investigators at this research center is 
pursuing the development of clip grafts for aneurysms in small vessel surgery 
and is beginning to apply the techniques in human subjects. Among personnel 
of various stroke research centers where carotid endarterectomy is 
performed, there continues to be active debate concerning the various 
methods for keeping morbidity and mortality at the lowest possible levels. 
The ability to rapidly and safely evaluate the changing pattern of regional 
cerebral blood flow prior to, during and following endarterectomy would 
be of potential value in making clinical correlations concerning surgical 
technique, pre-operative neurological findings, including the results of 
arteriography and the post-operative neurological state of the patient. 

Another research group are delineating the temporal profile of 
acute progressing cerebral infarction in the carotid system and of acute 
progressing infarction in the vertebral basilar system. This is a problem 
of practical importance since there are some patients who develop severe 
worsening of clinical evidences of focal cerebral ischemia some hours 
after a cerebral infarct is thought to have been "clinically completed." 
The occurrence of this delayed worsening suggests certain pathophysiologic 
mechanisms which might be subject to prevention by appropriate therapy. 

Research to develop a method for measirrement of regional cerebral 
glucose metabolism has been pursued at another research center. To do 
this, the characteristics of a tracer for the proposed technique was 
thought to be one which was taken up by the brain at a rate proportional 
to that of glucose and whose metabolic products remained in the tissue 
under study. The use of Cl^-2-deoxyglucose was selected for study and 
experiments were conducted to establish a mathematical model and to 
substantiate the findings by actual experiments in dogs. Favorable results 
have been obtained to date, and further development to refine the technique 
is being pursued. It is anticipated that this research will make possible 
(1) a means of studying regional CBF simultaneously in vivo, (2) will also 
make possible a direct assessment of the role of regional metabolism in 
the control of cerebral circulation, and (3) finally when validated it 
will make possible the extension to humian studies, by the use of Cll-2- 

13 w 



deoxyglucose which can be detected with external detectors. 

The investigator has also directed his efforts to an evaluation 
of rapid diagnostic methods in extracranial cerebrovascular disease. Four 
rapid diagnostic tests have been examined (direct thermometry^ ophthalmo- 
dynamometry, bruit and pulse assessment) to discriminate for the presence 
or absence of significant extracranial cerebrovascular disease. Utilized 
separately^ thermometry, ophthalmodynamometry and the examination for a bruit 
will find over 75^ to 85^ of cases. Pulse assessment alone reveals only kO'fo 
of significant lesions. The combination of neck examination for the presence 
of bruits or pulse deficit alone will discover 93^ of the vascular lesions. 
In this investigator's present series, lOO/o of the cases were found by including 
either direct thermometry or ophthalmodynamometry in combination with the 
aforementioned two tests. 

Ophthalmodynamometry accurately located the site of the major 
lesion in more than 9^'fo of the cases. The direct thermometry technique 
scored below kO'fo in lesion localization. It is suggested that the thermom- 
etry technique, because of its ease of performance, be utilized in combina- 
tion with neck examination as the preliminary step in assessing patients 
with suspected extracranial cerebrovascular disease. Ophthalmodynamometry 
should then be used for more accura'te localization of the lesion prior to 
angiography. 

An understanding of the normal control mechanisms of the cerebral 
circulation is essential to any investigation of the derangement present 
in various abnormal conditions. This is especially important in the study 
of cerebrovascular disease. Knowledge of the important regulating factors 
in both normal and pathologic states may lead to concepts that will be of 
use in preventing or correcting abnormalities of the cerebral circulatory 
system. The relationship between regional cerebral metabolism and regional 
cerebral blood flow is one important aspect of the control system which 
has not yet been able to be directly studied. The development of a method 
for measuring regional cerebral glucose metabolism in vivo should be of 
significance in this regard. 

Considerable interest has developed in studies of cerebral 
vasospasm. One research group reports progress in determining the actual 
effect on regional blood flow of angiographic experimental cerebral vaso- 
spasm. Vasospasm was produced both by vessel puncture (as recommended by 
this group) and by subarachnoid injection of blood (as recommended by 
others). It was found that, despite apparent significant cerebral vaso- 
spasm (vessel caliber reduced to l/2 of control), there was still no 
statistically significant reduction in cerebral blood flow. Only when 
the vasospasm became intense, that is the cerebral arteries appeared to 
be 1/3 or less of the original caliber, was there reduction in cerebral 
blood flow. When this critical point was reached, the cerebral blood flow 
dropped dramatically. In a series of 25 monkeys in which subarachnoid 
hemorrhage was produced by puncture of internal carotid artery, virtually 
all showed a precipitous drop in cerebral blood flow determinations made 
within 1/2 hour of puncture. Cerebral blood flow determinations rapidly 
stabilized and, quite -unexplainedly, increased s.lightly above control 
from the first to the fifth hour after hemorrhage. Following this, 
cerebral blood flow would usually stabilize despite the presence of mild 

Ikv 



to moderate spasm on the arteriograms. If severe vasospasm developed^ 
angiography would demonstrate marked arterial constriction. CBF values 
at this time were often half of the control value. This second phase of 
spasm (probahly part of the biphasic response described by Brawley) would 
persist throughout duration of the survival of the animals. Upon the 
injection of blood into the chiasmatic cistern, inmiediate angiographic 
spasm was noted, associated with a significant drop in cerebral blood flow. 
The spasm would never become secondarily severe, though some degree of 
spasm was seen throughout the experiment. Marked reduction in CBF and this 
characteristic second stage of spasm are not seen with subarachnoid blood 
injections. 

It is the opinion of this investigator that both rupture of an 
intracranial vessel and subarachnoid injection of blood are capable of 
producing an immediate transient angiographic spasm which does reduce CHF. 
Apparently, the puncture technique is more likely to produce the character- 
istic second phase of spasm which is probably the clinically significant 
phenomenon. 

Concomitant experiments carried out by this researcher related 
to the determination of stability of regional CBF determinations by the 
radioisotope method in experimental animals for long durations. A series 
of long d-uration regional blood flow experiments (over 36 hours) were 
carried out in ten animals. It was found that: (l) RBF determinations 
within the first hour following the preparation fluctuate and are invalid, 
and (2) the coefficient of variation among the determinations increases 
with the d\iration of the experiment, within the first 5-6 hours it being 
only 3-^io, but after 20 hours it may be as high as ik^o. The practicality 
of this technique for inducing cerebral vasospasm may be open to question 
since the technique of this investigator produced severe reduction in 
blood flow in only about I/3 of the preparations, but not at all in 
preparations produced by techniques described by others. 

Another phase of this investigator's studies consisted of the 
application of physical forces to overcome the effects of cerebral vaso- 
spasm. This researcher has been able to reliably maintain regional cerebral 
blood flow above control levels in normal autoregulating brains by inter- 
mittently occluding the descending aorta with an inflatable balloon passed 
up throLigh the femoral artery. He has determined the appropriate intervals 
of inflation and deflation which allow for adequate renal perfusion, yet 
can increase CBF significantly. The assumption is that in the presence of 
cerebral vasospasm, increased perfusion pressure for long intervals may 
adequately nourish the brain until the spasm spontaneously subsides. The 
technique of intermittent aortic occlusion was performed in 35 dogs and 
25 monkeys without pathologic alterations. Intermittent aortic occlusion 
in the presence of spasm has been attempted. It has been shown that it is 
capable of perfusing past severely constructed vessels used in the 
experiment described. In this experiment there was intense spasm of the 
middle cerebral artery, which barely visualized angoigraphically, associated 
with a marked reduction in CBF records. On institution of intermittent 
aortic occlusion, CBF values returned to near normal levels. The results 
appear encouraging even though there are many pi-oblems yet to be solved. 

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These include cerebral edema which can result in certain instances when the 
brain is so hyper-perfused. Additionally^ certain animals develop 
alterations in blood pressure after long intervals with intermittent aortic 
occlusion. The importance of these studies is heavily based on the pre- 
sumption that a model of cerebral vasospasm, which behaves in a manner like 
zhe human vasospasm phenomenon, is necessary before any research in this 
area can be continued. The significance of this model as it may apply to 
problems of cerebral blood flow in cerebrovascular disease is not certain. 
Fiorther studies are needed. 



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Trauma (Head Injury) 

Since trauma is the number one killer of man while he is in the 
priTie of life^ it is only natiiral that a major aspect of trauma^ head 
injury^ should receive special attention from the KINDS. During 1971, 
approximately JO research grants representing $5,228^000 supported 
investigators working in medical and scientific areas directly or indirectly 
related to central nervous system trauma, particiilarly head injury. More 
recently, this Institute has also begun to expand its support of research 
on acute trauma to the spinal cord. 

In general, the care and study of the seriously injured head 
trauma patient has had as its primary concern the prevention of brain 
swelling and therefore a search for the mechanisms that trigger the onset 
of brain edema. Recent work in one of the head injury centers demonstrated 
a marked diminution in arteriolar calibre (720 micra) in the injured area 
following controlled impact trauma to the intact monkey skull. One day 
after trauma the microangiograms showed a return toward normal in the 
calibre of the larger vessels and an increase in tortuosity of these vessels 
with a continuing diminution of capillary filling. Histologically, the 
most significant feature was the presence of cerebral edema which became 
apparent one day after injury, the edema being present in both the cortex 
and the white matter. The brain swelling was well established between the 
first and third days when the caliber of the arterioles was larger than 
that noted in the immediate post traumatic period. This was also the 
period when the capillary filling was abnormal. 

Workers in another head injury center also have been investigating 
cerebral vasomotor paralysis following artificial concussion in animals 
and during the decompression which usually occurs in patients following 
brain tumor extirpation. In the latter case, patients who had had prolonged 
retraction of the brain during removal of a deep seated tiimor or clipping 
of an aneurysm, had an intracranial pressure which was extremely sensitive 
to changes in respiration. These patients were maintained on respiratory 
assistance immediately following surgery and, quite often, removal of 
the respirator was followed within two minutes by an increase in intra- 
cranial pressiire of 30-50 mm Hg. The pressure promptly fell when respiratory 
assistance was reinstituted. Several continued to decompensate and 
ultimately the responsiveness of intracranial pressure to respiratory 
control disappeared. These investigators attributed the continued rise 
in Intracranial pressure to brain swelling and the lack of response to 
changes in arterial p02 and pC02 to paralysis of the vasomotor tone of the 
cerebral vessels. 

This concept of cerebral vasomotor paralysis was developed in 
monkey experiments in which pressirre waves were first identified and 
studied. A series of moderately high intracranial pressure waves that had 
no effect on the animal's EEG was often followed by a "terminal pressure 
wave, " at which time the intracranial press\ire rose to the level of the 
blood pressure, the EEG became isoelectric, and the animal passed rapidly 
into shock unless fluid was rapidly withdrawn from the pressure inducing 
balloon. 

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In another series of experiments the relationship of cerebral 
blood flow (CBF) and intracranial pressure (IP) was investigated. During 
the control period^ when the arterial pressure was elevated with 
norepinephrine, the CBF remained essentially constant. After a series of 
balloon induced intracranial pressixre waves, norepinephrine was injected 
again. The same increase in blood pressure now produced a greater rise 
in CEF, leading these investigators to postulate that the vascular auto- 
regulatory mechanism had been damaged. As with the patients discussed 
above, the response of the cerebral arterioles to hypercapnia and hyper- 
ventilation also disappeared. In the terminal stage, when intracranial 
and arterial pressures were equal, cortical electrical activity had 
disappeared, spontaneous breathing had ceased, and rapid evacuation of the 
balloon produced a drop in IP to normal. This sudden expansion of the brain 
following balloon deflation was attributed to rapid filling of flaccid 
cerebral arterioles (vasomotor paralysis) and it was suggested that the 
increase in brain volume was due to the rapid expansion of cerebral vessels, 
cerebral edema, or a combination of the two processes. 

Attempts to control and reverse the trend of progressive brain 
edema in the head injured patient have involved the energies of scientific 
personnel in a number of head injury centers. The efficacy of using 
diuretics to successfully treat brain edema following head trauma has been 
a subject of major concern because the results have had a wide range of 
variability. Precisely controlled studies by one head injury center have 
demonstrated that the brain swelling which follows experimental brain trauma 
in animals is not reversed by steroid (dexamethasone) therapy. In terms 
of human disease, this would certainly mean that, in those conditions 
involving cerebral edema in which there is associated significant necrosis 
and hemorrhage, the edema would not be expected to respond to steroid 
therapy. The possible effect of steroids on inflammation and thus on 
edema has not, however, been entirely ruled out. 

Long-term studies on seizure incidence in children (birth to 
lif years) following injury has shown that 10 percent of the head injured 
children had at least one seizure during the first 2k hours after injury. 
The incidence of cases with seizures dropped to 3 percent by the end of the 
first year and approximately 10 percent of the children without seizures 
(at the time of injury) developed EEG abnormalities, including spike foci 
within 2-k years after injury. In addition, the head-injured child had 
an increased incidence of negative behavioral traits, compared with their 
pre-injury incidence. Among the major symptoms were hyperkinesis, 
aggression, irritability and poor control of affect So 

The effects of brain damage on performance and physiological 
responses have been studied by another group of scientific investigators 
who found that brain-injured patients have slower reaction times, diminished 
alpha blocking, and disturbed orienting responses to auditory stimuli. In 
addition, cold pressor stimulation to the brain-injured patient results in 
depressed plethysmographic rebounds, suggesting differently tuned vascular 
responsivity. 



18 



Another area of major concern^ and therefore major research 
effort^ is regeneration of nerve tissue following central nervous system 
trauma. In general most of the research models have employed the spinal 
cord for study but one group has concerned itself with investigating 
regeneration in the pyramidal tract. In man^ damage of the pyramidal tract 
produces a total hemiplegia^ but these patients generally recover during 
the following eight months with excellent control of the involved face, 
arm and leg without developing the "pyramidal syndrome, " except for the sign 
of Babinski. Parallel studies in monkeys revealed that the recovery of 
motor function was not as complete as that recorded in man. These differing 
results suggest that in man, as opposed to the monkey, there are at least 
three motor systems; the first of which is the one mediated by the cortico- 
spinal tract at the pyramidal level; the second being a corticospinal 
pontine system; and the third, a system whose cortical origins and paths 
are unknown. 

An investigation concerning the origin and activity of cells 
invading the damaged animal nervous tissue (spinal cord) has revealed that 
they are mononuclear leukocytes. Large numbers of these cells enter the 
white matter of the cord at all levels following root transection. Some 
of these cells underwent cell division after entering the nervous tissue, 
as evidence by the labeled mitotic figures. In addition, increased DM 
synthesis was recorded in cells adjacent to the nerve cell bodies, both 
in the cord and in the dorsal root ganglia. 

Even more exciting are results of studies carried out in 
another laboratory on regeneration following spinal cord damage. Axons 
at the proximal end of the animal spinal cord following transection were 
found to produce growth cones and characteristic new bouton connections on 
axons and dendrites. This is the first positive data that mammalian 
central nervous system neurones have the capacity to regenerate. 
Additional work on regeneration in damaged animal spinal cord by another 
group of investigators has demonstrated that one week following cord damage 
the area of insult becomes filled with Gitter cells. At two weeks, axons 
begin to appear beneath the pia, following the s\irface of capillaries, and 
at seven weeks the area of destruction is filled with a heavy growth of 
myelinated axons. Electronmicroscopy has shown that the cells giving origin 
to the myelin are both Schwann cells and glial cells. The premise is that 
the axons regenerating and encased by a peripheral type of myelin grow in 
from the dorsal spinal nerve roots and blood vessels of the st-umps, while 
those possessing a central type of myelin grow out from the spinal cord 
stumps beyond the zone of damage. 

That cultured embryonic spinal ganglion cells can form functional 

synapses has been an area of endeavor in which considerable effori. has been 

devoted by one group of investigators. They have shown that "typical 

synapses" are formed, even though they lack other innervation territories. 

The newly formed dendrites have functional meaning as recipients of input 

rather than fortuitous efforts as abortive gro^rth. Intracellular records 

of these ganglion cell cultures have been obtained, providing evidence 

of synaptic interaction. Nerve growth factor (NGF) was employed in the 

tissue culture medium and future studies are planned to answer the question 

of how crucial this material is and if so, is it necessary for the formation 

of functional synapses. 

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OTHER NEUROLOGICAL DISORDERS 
Sclerosing Disorders - General 

Studies relating to sclerosing disorders in general continue to focus 
;jrimarily on detailed investigations of lipids and myelin. The lipid studies 
pertain to central nervous system lipids , lipid metabolism in normal and 
pathological states, the relationship between brain lipids and electrolytes, 
and a biochemical genetic study of bacterial lipids, phospholipids are the 
subject of study with regard to their physical state, biological activity in 
general and nerve-muscle function in particular, together with their metabolism 
in membrane synthesis. The biochemistry of glycolipids, fatty acids, long 
chain bases, inositol and phosphoinositides, along with the mechanism of ion 
transport in nerve membrane and studies on subcellular fractions of developing 
brain, continues in the service of a better understanding of general sclerosing 
disorders. Myelin studies continue to emphasize structure, function, metabolism, 
pathology and comparative studies, together with its formation and relation 
to the neuron, its abnormal composition in plasma membranes and the inter- 
actions of phosphoinositides in myelin itself. 

Multiple Sclerosis 

Multiple sclerosis is a disorder of the nervous system characterized by the 
presence of tiny sclerotic areas or plaques of degenerating or destroyed 
myelin in the white matter of the brain and spinal cord. Its symptoms are 
referable to disruption of nerve impulses in the affected fiber tracts. Its 
etiology remains to be determined and treatment is only symptomatic. 

A current investigation is aimed at the etiology of multiple sclerosis and 
the mechanism involved in its development. The work is directed at the 
identification of infectious agents, measles virus in particular, that may be 
responsible for the signs and symptoms of demyelinating diseases appearing 
years after the initial illness. It is postulated that the measles virus in 
some form may persist and provide active Immunity throughout life for most 
people but on rare occasions may produce demyelination and symptoms commonly 
associated with central nervous system disease, behaving generally in the 
manner of the "slow" viruses. Evidence is now available which indicates that 
the measles virus may be responsible for the pathologic changes known to be 
present in various forms of measles encephalitis, a world-wide disease witn a 
geographic distribution similar to that of multiple sclerosis, since its 
incidence tends to be low in tropical areas and high in the more temperate 
zones . 

The discovery of inclusion bodies in the cytoplasm and nuclei of cells and the 
formation of giant cells in areas of demyelination strongly suggest that the 
measles virus is related to encephalitic manifestations. The hallmark. of the 
virus has been found in individuals suffering from degenerative brain disease 
years after childhood measles. Further, several patients with a classic form 
of multiple sclerosis have shown inclusion bodies and giant cells similar to 
those seen in acute and chronic measles encephalitis. Continuing attempts 
will be made to recover active measles virus by co- cultivation with Hela and 
human embryonic kidney cells. • ■ 

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Amyotrophic Lateral Sclerosis 

Amyotrophic lateral sclerosis is a neurological disorder of unknown etiology 
with an incidence of 8,000 to 10,000 patients in the United States. It is 
characterized initially "by muscle weakness and wasting which is brought 
about by degeneration of motor cells in the brain and spinal cord. Treatment 

is symptomatic only. 

A new study is in progress to develop an experimental model for amyotrophic 
lateral sclerosis by immunomanipulation of mice infected with mouse encephalo- 
myelitis virus which causes mouse poliomyelitis. 

This study arose from suggestive evidence that certain progressive neurological 
diseases of later life may be caused by a reactivated virus which seemed to 
have been inactivated effectively many years earlier. Particularly intriguing 
are some cases of amyotrophic lateral sclerosis which have appeared many years 
subsequent to recovery from antecedent poliomyelitis. A not unreasonable 
conjecture with regard to mechanism might be that the poliovirus has been 
reactivated. Similar cases can be made for paralysis agitans, multiple 
sclerosis and possibly Jakob- Creutzfeldt disease and parkinsonism. 

The main objective of the program will be to develop a model system for the 
reactivation of a central nervous system virus which can be applied to human 
amyotrophic lateral sclerosis. The approach will be to infect animals with 
mouse poliovirus so they contract the disease but ultimately recover. Follow- 
ing recovery, the immunologic competence of the animals will be compromised 
by various means with the expectation that latent viruses will manifest them- 
selves under these circumstances. 

Experimental Allergic Encephalitis 

Experimental allergic encephalitis is an artificial, abnormally induced, 
auto- immune disease in which the immune defenses of the organism are 
manipulated and directed against its .own tissues. It has been used as a model 
system for the study of immunological factors implicated in central nervous 
system disease. Demyelinating diseases bearing resemblance to miiltiple 
sclerosis may be induced by this means. 

Serial measurement of cerebrospinal fluid beta- glucuronidase has been found 
to be a sensitive method for detecting experimental allergic encephalitis and 
for monitoring the course of the disease in the rabbit. Elevated levels have 
been found only with clinical or histological evidence of the disease. Control 
studies indicate that elevated cerebrospinal fluid enzyme does not resilLt from 
induced serum increases of beta- glucuronidase levels. The suspicion is that 
the increased cerebrospinal fluid enzyme levels in active experimental allergic 
encephalitis reflect release of enzyme from lymphoid and other inflammatory 
cells contributing to the encephalitic lesion. There are implications here 
for human neurological disease states. Continuing studies indicate that cats 
can develop experimental allergic encephalitis providing an elaborate 
sensitization procedure is used, but so far, the disease has been induced in 
a high proportion of animals only after placing them on a thiamine deficient 
diet. 



21 w 



It was found that experimental allergic encephalitis could be inhibited in 
rats by cyclophosphamide when given in daily doses beginning as early as the 
first day of paralysis or as late as the fifth day of advanced paralytic 
signs. It caused reversal of clinical signs and disappearance of lesions in 
two thirds of the animals. Fully recovered animals showed a prompt relapse 
when cyclophosphamide was discontinued. Continued remission could be obtained 
with continued treatment. 

Continuing investigation will determine the relative roles of different cell 
types, subcellular factors and circulating antibodies in the development of 
this auto- immune disease. Mechanisms underlying the inhibitory and therapeutic 
effects of cyclophosphamide will be further investigated as will the apparent 
ability of thiamine deficiency to enhance the susceptibility of cats to the 
disease. 

Experiments are being devised to discover the phylogenetic capacity to 
develop auto immune disease and autoantibody along with che role of the thymus 
and other central lymphoid tissue in the interest of preventing and treating 
experimental auto- immune disorders. 

Parkinsonism 



Parkinson's disease is a progressive neurological disorder of unknown cause 
affecting certain brain areas involved in the control and regulation of 
movement. Its onset may be incidious and its progression may be almost 
imperceptible ;, but is ultimately characterized by tremor, rigidity and bent 
posture. Standard treatment in the past consisted of physical therapy, a 
variety of driigs, and surgery. A highly effective form of replacement therapy 
making use of large oral doses of L-Dopa or thalamic surgery are now the 
treatments of choice. 

A third center for research in parkinsonism has recently come into existence 
and will be concerned with a correlated approach involving biochemical, 
ultrastructural and neurophysiological methods in the same systems to elucidate 
the role of catecholamines in the basal ganglia. The individual projects will 
be concerned with anatomical studies of biogenic amines in the primate brain, 
neuromelanin, neurophysiology of the substantia nigracaudate nucleus inter- 
actions, blood-brain barrier to L-Dopa, amine metabolism, and motor potentials 
in parkinsonism. 

The anatomical studies of aminergic systems in primate brain that may be 
related to the effects of L-Dopa administration and nigro- striatal pathology 
will deal with catecholamine bodies in the substantia nigra and regions rich 
in catecholamine terminals. They will also include mapping of nigro striatal 
dopaminergic pathways in the primate. Experiments are planned which will 
examine in detail the modulatary effect of the substantia nigra on the 
cortico-striato- thalamic activity of the brain using both anatomical and 
physiological techniques. Studies are also designed to elucidate the mechanism 
of entry of Dopa or its metabolites into the central nervous system across the 
blood-brain barrier and possibly across the choroid plexus. There are 
suggestions that a central mechanism, not located in the motor cortex, develops 
during learning of skilled movements which monitors and controls subsequent 



22 w 



vol\intary execution of specific motor acts. Selective inhibition of afferent 
stimuli is essential to this mechansim. Interference with this system can 
lead to slow behavior not unlike that seen in Parkinson's disease. It is 
proposed to examine these hypotheses in animals and in Parkinson's disease 
patients before and after L-Dopa therapy. 

Work is in progress now to determine the effects of Dopa decarboxylase 
inhibitors alone and in combination with monoamine oxidase inhibitors on the 
amount of systemically administered L-Dopa that gets into the brain across 
the blood-brain barrier and to explore dimethyl sulfoxide as a transport 
carrier of Dopa molecules. 

Finally, there is a program for the long-term follow-up of patients with 
parkinsonism receiving L-Dopa and for investigations of L-Dcpa in selected 
disorders other than Parkinson's disease. These studies are to be supplemented 
with peripheral metabolic inhibitors and centrally acting monoamine oxidase 
inhibitors when synthesized. Evaluation of dopamine-like substances, substances 
which change the sensitivity of the central nervous system to L-Dopa, and the 
role of serotonin as related to the involuntary movements induced by L-Dopa 
will be included in these studies. 

Neuromuscular Disorders 

The most prominent neuromuscular diseases are myasthenia gravis (MJ) and the 
muscular dystrophies (MD) which have prevalence rates of h and 6 per 100,000 
respectively. While MJ has been recognized as a clinical entity since the 
seventeenth century, the dystrophies constitute a variety of clinical 
conditions. New dystrophic variants in both animals and humans are described 
yearly and their classification constitutes a formidable neurological problem. 
Myasthenia gravis has been related to impaired transmission through the 
myoneural junction while the dystrophies are variously correlated with 
deficiencies in muscle, nerve, intermediary metabolism, or the myoneural 
junction. 

MG is characterized by progressive impairment of neuromuscular transmission 
with continuous exercise. Research in recent years has demonstrated that a 
diminished quantity of acetylcholine is liberated from nerve terminals, 
preventing adequate depolarization of the muscle membrane. Current observations 
suggest that a decreased amount of the neurohumor is synthesized at the nerve 
terminals, as judged by the lower activity of choline acetyltransterase in 
muscle biopsies from M5 patients. It is not known whether the diminution of 
enzyme activity results from the presence of an endogenous inhibitor or from 
more primary structural or metabolic factors. 

Treatment, for the most part, consists of the administration of anticholinesterase 
drugs which act to increase the effective concentration of acetylcholine at 
the endplate. However, in a significant number of patients, marked improvement 
may be obtained by the chronic administration of ACTH or prednisone. The 
basis for the efficacy of these latter drugs is not clear. Finally, the 
surgical removal of a thymoma in a patient with M} often leads to marked 
improvement. Curiously, the incidence of thymomas in MJ is approximately 15'/o 
or about five thousand times more frequent than in control patients. 

23 w 



Recent studies continue to confirm the possibility that MG is an autoimmune 
disease. The serum of any patients contain antibodies against their own 
muscle, thymus, thyroid or other tissue. No antibodies have been identified 
as yet as the myoneural jionction, and the presence of antimuscle antibodies 
doeo not necessarily correlate with the course of the patients illness. 
However, it has been reported that the serum of MG patients inhibits the 
synthesis of acetylcholine by frog or rat brain. 

Research efforts in the area of muscular dystrophy follow well-established 
approaches. Substances which may alter the dystrophic condition are being 
tested in humans, in animal models, and in tissue culture. Recently, it 
has been reported that safflower oil reverses the hereditary muscular 
distrophy of chickens. The testing of components of the oil as well as 
structurally related analogues is currently in progress. Methods for the 
detection of carriers of X-linked Duchenne muscular dystrophy continue to 
improve. Approximately lOPJo of known female carriers have elevated levels of 
serum creatine phosphatase. For the remaining 30fo who have normal serum 
values, light and electron microscopic evaluation of biopsy material may prove 
helpful. Myopathic changes in the muscle of carriers may be extensive, 
minimal or absent. 

The properties of dystrophic muscle continue to be elucidated. In tissue 
culture, muscle cells from dystrophic chickens differ in rate of fiber growth, 
rate of development of enzyme activity and strength of spontaneous contrations. 
It has been shown that muscle fibers of dystrophic mice have significantly 
lower resting potentials. The decrease for fast fibers is approximately 
twice as large as that for slow fibers. Finally, it may be assumed that 
present research on the molecular events in contraction and on the chemistry 
of cell and organelle membranes will soon include dystrophic muscle. 

Bioengineering Studies of Muscle Dynamics 

During the past decade there was a surge in activity initiated by the joint 
efforts of engineering, physical, biological, and medical scientists to 
develop suitable methods for the prevention, treatment, and management of 
disease. 

The KINDS has played a catalytic role in this activity by providing research 
grant support during the developing years of biomedical engineering, a field 
which deals with the interaction between the engineering sciences in biology 
and medicine. The report of the research activities of such a multidisciplinary' 
team is presented below. 

This group of investigators includes engineers with considerable experience in 
experimental mechanics and in developing mathematical models; biophysicists 
knowledgeable in systems analysis; biomedical scientists well qualified in the 
field of nerve and muscle physiology; and a surgeon whose major research 
interest is muscular- skeletal physiology and orthopedic surgery. 

The long-term goal of their research program is to determine what differences 
exist between the mechanical and control behavior of the limbs of normal 
subjects and those afflicted with various known neuromuscular disorders. The 
investigators' working hypothesis is that the neuromuscular skeletal system 

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can be represented by a mechanical plus control' system model of muscle dynamics. 
In disease- or disability afflicted subjects part of this system, depending 
on the disease, will be functioning abnormally or not at all. 

The model has undergone many modifications as experimental data have been 
collected to check its various details. Since the system under analysis is 
complete, the mathematical determination of the model constants is not a 
simple process. A digital computer program has therefore been written to 
process the experimental data and systematically determine the model parameters 
from them. The program modifies parameter values in order to minimize the 
error with which the model can predict the experimental data submitted to it. 
The mathematical scheme used in this program, called an identification scheme, 
has been treated extensively in control theory. The model is mathematically 
non-linear, however, so that the development of the scheme has not been a 
simple straight- forward procedure. The principal part of the successful 
reduction system the investigators are now using is a non-linear simplex 
method. 

On the practical level, it appears that quantitative measures of neuromuscular 
function in human neuromuscular disorders are almost non-existant. For example, 
in connection with spasticity, it was pointed out a year ago that an objective, 
accurately reproducible and easily applied test for muscle spasticity has yet 
to be devised. 

This group has developed control systems and mechanical models in the 
relatively neglected field of muscle dynamics. The first stage of the study 
was concerned with correcting, improving, and expanding conventional analyses 
with particular regard to the theoretical effects of heat production on the 
parameters of the model. Movement of the forearm around the elbow was studied 
for this purpose. Movements of the forearm against various types of loading 
in response to sensory signals were monitored both by measuring displacement 
parameters and by electromyography. The study was extended to other muscle 
systems of the body. 

From the dynamic models of the system, the investigators hope to be able to 
localize the malfunctions, and use the quantitative information obtained as a 
basis for improved diagnosis and therapy. 

The approach to this problem has two major components. First, the quantitative 
approaches of engineering--especially in the areas of human dynamic structural 
analysis, solid mechanics, and control theory--can now be applied to complex 
non-linear biological systems such as human muscle. Second, such an engineering 
analysis makes it possible to derive quantitative parameters. 

To achieve this goal, the problem was attacked in four basic steps, three of 
which are now well in hand. In the analytical and modelling phases of the 
study, progress has occurred in three directions; analysis, model construction, 
and parameter evaluation. In the analytical work, the investigators combined 
current physiological understanding of muscle constitution and behavior with 
continuum mechanics. They have thus established a useable quantitative model 
for forearm movements including limb dynamics, muscle mechanics, and neural 
control. It provides numerical measures of both neuromuscular parameters and 

25 w 



muscle tensions. A series of simple non- invasive tests provides a normal 
data base. 

Three types of studies designed to evaluate the parameters in the model have 
been performed on human subjects. The first, a series of static tests, 
demonstrates on the basis of the principles of structural mechanics that 
averaged electromyogram amplitude is directly related to muscle contractile 
force. This series of tests also provides numerical values for a number of 
parameters in the model and for the forces in the individual muscles. The 
second study, consisting of a set of constant angiilar velocity tests, 
independently evaluates these same parameters as well as a damping coefficient. 
The results are in agreement with those of the first study, and in addition 
show that damping is small. The third study, a series of "quick- release" 
tests, serves as an evaluation of the control parameters in the model. The 
results indicate that direct positional control is much more important for 
present experimental conditions than velocity control. 

A model that includes central nervous system characteristics, local thermal 
effects, wave propagation effects, and the dynamics of the skeletal frame 
would be of considerable value to those physiologists, biophysicists, and 
engineers who are working in the field of control theory and system performance 
in other biological systems. This effort to provide a systematic account of 
how muscle systems function in situ shoiild contribute basic information of 
ultimate value to many medical specialties. 

The investigators plan to continue this line of research to refine both the 
structure and numerical constants in their model for the system so that it 
represents the physiological condition more accurately. 

At this time a complete testing and evaluation facility is already functioning. 
Human subjects are being studies on a regular basis. The results obtained to 
date indicate that numerical values for parameters, separately identifying 
muscle properties and nervous system control properties, have been calculated 
from measurements during a very simple (for the subject) dynamical test. This 
means that changes in individual parameters in the neuromusclar systems of human 
subjects can be monitored quantitatively. Such a quantitative technique is 
much needed, not only for differential diagnosis, but also for monitoring 
therapy. 

In addition, studies of patients with a well characterized disability, e.g., 

muscular dystrophy, are planned to determine the clinical usefulness of the 

model, thus leading to improved quantitative tools for diagnosis and clinical 
studies . 

Convulsive Disorders 

Patients who suffer from convulsive disorders often conceal this fact because 
it can adversely affect their social and economic status and their insurability. 
It is therefore difficult to estimate numbers of persons affected. Their 
number is estimated between 2 and k million. Conservatively, loss of earning 
capacity caused by this disease, costs the Nation more than a billion dollars 
a year. Much of this' would be unnecessary if tlae general public, by better 



26w 



understanding of the disorders ^ would make available to victims the social, 
educational, and employment opportunities now denied them largely because of 
ignorance and fear. Gnerally, persons with epilepsy have average or bex.ter- 
than- average intelligence and good earning potential. A majority of them can 
live normal lives because their convulsive disorders can be controlled with 
anticonvulsant drugs. 

Epilepsy is a disease characterized by one or more of the following symptoms: 
(l) paroxysmally recurring impairment or loss of consciousness; (2) involuntary 
excess or cessation of muscle movements; (3) psychic or sensory disturbances; 
(k) perturbation of the autonomic nervous system. On the basis of origin, 
duration of seizures , clinical and electroencephalographic evidences, epilepsy 
is classified into four subdivisions. 

In epilepsy known as "Grand Mai," in more modern terminology, "major motor 
seizures," the attack is most violent and convulsions last longer. The 
afflicted person may bite his tongue and invariably loses control of his 
faculties. If the area or areas of the brain can be located from which these 
abnormal electrical discharges emanate, the affliction is termed "Focal" or 
Jacksonian epilepsy. In "Petit Mai," a disorder of the young, the seizures 
are of short duration, and attacks occur more frequently. Seizures are 
characterized by myoclonic jerks and they generally start in the extremities 
and/or in one corner of the mouth. The affected part trembles violently. The 
trembling movement moves upwards. It either ends up in a minor conviolsion or 
the individual loses consciousness in the same way he does in grand mal. 
"Psychomotor" epilepsy is caused by discharging neurons which exert their 
influence upon the mental processes as well as upon the muscle movements. A 
patient exhibits adversive or torsion movements, dreadful fear, flinging of 
arms aimlessly, smacking of lips, and other incoherent physical and mental 
behaviors. The last type is called "Autonomic" epilepsy. A person suffering 
from it experiences a sharp, acute, sudden pain in the stomach. Hypertension, 
perspiration and other visceral disorders are the common symptoms. 

Epilepsy may be caused by several factors, such as brain damage, presence 
of a scar caused by a wound or an injury, drugs, congenital malformation, 
nutritional deficiences, metabolic abnormalities, fever (in infants), infectious 
diseases (encephalitis, meningitis), brain tumors and abscesses. In a recent 
study of the long term effects of administering a diet rich in median chain 
triglyceride (MCT) on seizure control in children indicated that a diet 
deriving 60 percent of its calories from MCT, when administered to a group of 
children with a variety of seizures refractory to conventional modes of therapy, 
was effective in controlling seizures, particularly myoclonic and akinetic types. 
A less sustained control was obtained in some older children with psychomotor, 
focal motor and grand mal convulsions. These studies provided very little 
information regarding the mechanism of the effect of the diet on seizure 
control. They only indicated that the diet produces hyperketonemia but no 
metabolic acidosis. 

The role that heredity plays is as yet undetermined. If one twin develops 
epilepsy, the other one is likely to develop it also. However, patients with 
this disease do not necessarily have epileptogenic offspring. Comparative 
intelligence evaluations and electroencephalogrfim studies show that patients 

27 w 



with familial idiopathic epilepsy have lower intelligence quotients than their 
non-epileptic relatives. It is estimated that a significant portion of the 
U. S. population is epileptic with seizures that are symptomatic of a primary 
disorder or of unknown idiopathic etiology. Evidence has been developed 
indicating that at least a portion of the idiopathic epilepsy observed in man 
has a genetic basis, although the specific mode of inheritance is not well 
understood. Experimentally, using Beagle dogs with spontaneous, recurrent 
convulsive seizures similar to those observed in man, evidence is being provided 
indicating a strong genetic basis for conviolsive episodes. Based upon the 
comparative studies of spike- and- wave EEG train in groups of patients suffer- 
ing from febrile convulsions, focal EEG anomaly, or focal epilepsy, and in a 
sample of normal people, it is suggested that several of the epilepsies might 
liave a common genetic factor in their etiology. A number of investigators 
have reported genetic involvement in other forms of epilepsy. These include 
photogenic epilepsy, idiopathic epilepsy, benign familial neonatal convulsions 
and partial epilepsy. 

Although the exact mode of action of mechanisms underlying epilepsy is largely 
unknown, the experimental evidence based on electroencephalographic studies 
indicates that the neurons in the affected area build up a supply of electrical 
energy through their metabolic action. By repetitive activity of charge and 
discharge, the cells become overactive and start firing in synchrony. The 
firing pattern may spread to other areas of the brain resulting in an epileptic 
seizure. When the neurons start firing again in dysharmony, the seizure is 
over. Brain waves so monitored give no indication of the individual's 
intelligence, thoughts, or his mental health. However, they provide strong 
clues as to whether or not a person has epilepsy. An EEG recorded during a 
seizure is likely to show unusually high bursts of energy release. The 
pattern indicates the type of seizure an individual has suffered. 

One of the most versatile electrical instruments used in the detection of 
electrical discharges from the nerve cells is an electroencephalograph (EEG) 
which picks up electrical discharges from the brain cells, amplifies them and 
records the impulses in the form of graphs. With its help, various areas of 
the brain have been scanned and, using microelectrodes, electrical energy 
discharge patterns in groups of cells or a single cell, both in vivo and 
in vitro, can be recorded. 

In order to understand the seizure mechanisms, epilepsy has been induced 
experimentally in laboratory animals. The methods of induction can be divided 
into four main groups. Methods based on electrically- induced seizures include 
generalized stimulation of the intact animal to produce either threshold 
(clonic) seizures or maximal (flexor-extensor) seizures, and local stimulation 
of selected areas of the brain or spinal cord. Methods based on the 
administration of chemicals induce seizures similar to the ones indicated for ' 
electrical stimiolation. Although chemicals, such as strychnine, picrotoxin, 
methionine sulfoximine, and thiosemicarbazide, have limited value in inducing 
seizures for evaluation of anti-convilsant drugs, they nevertheless provide 
valuable tools for the study of seizure induction. The most important 
chemicals used for producing seizures are convulsant drugs and irritant agents, i 
For example, irritant agents, such as penicillin, applied to the cerebral 
cortex or injected into various brain structures, result in an acute irritable 



28w 



focus, impulses from which may rapidly spread to other areas of the brain and 
produce generalized seizures. Included in this section are the methods for 
producing chronic epileptogenic animals by inducing various forms of brain 
injury. Evidence linking acetylcholine to synaptic transmission in the 
central nervous system and especially to cortical arousal systems, plus the 
evidence that seizure activity can at times result from endogenously produced 
acetylcholine, has led several investigators to focus their attention on this 
agent with a hope to elucidate more general principles of epileptogenesis. 
The injection of alumina cream in the brain of monkeys appears to produce a 
reliable model which resembles human epilepsy. Investigations with iron 
hydroxide, magnesium hydroxide, zinc oxide, chromium oxide and mixed earth 
oxides showed that all these oxides or hydroxides are without epileptogenic 
effect when applied to the motor cortex of the monkey. On the other hand, 
nickel and antimony proved most epileptogenic only when implanted as pellets 
into the motor cortex. Mild effects were observed with bismuth, zirconium, 
tin, titanium, molybdenum and tungsten. Twenty- three more metallic powders 
have been tested on the sensorimotor cortex of monkeys. Some have shown very 
severe effects, while others remained ineffective. A third technique used in 
inducing experimental seizures is based on sensory stimiolation, such as 
elicitation of audiogenic seizures in susceptible mice and rats and photic 
seizures in rabbits and baboons. Interest has recently been directed to 
baboons which show a high incidence of spontaneous photic seizures comparable 
to those observed in photic- sensitive epileptic patients. Lastly, seizures 
may be induced by metabolic deviations, including water and electrolyte 
alterations, CO2 withdrawal, hyperthermia, endocrine ablation, or hormone 
administration. The aim of these studies is to elucidate the neural mechanism 
involved in the transition of a localized lesion in the brain to an epilepto- 
genic zone. This type of experimental approach has contributed new methods 
of inducing chronic epilepsy in animals which provides an important new tool 
for the study of this disease. By applying EEG techniques to these experi- 
mentally induced epileptic animals, the efficacy of new drugs is being tested, 
epileptic loci (foci) have been determined, and new and improved surgical 
techniques for therapeutic application are being developed. 

In a recent classification of the epilepsies, seizures elicited in the newborn 
have been described as showing partial discharges, susceptible to change from 
time to time, in both morphology and topography, from area to area and some- 
times from one side to the other. In terms of epilepsy the maturing nervous 
system acquires with age, an augmented capacity for generalized seizures, an 
increase in the frequency, duration and spread of after discharges, and an 
overall reduction of seizure thresholds. The developing nervous system has 
thus been shown to be in an advantageous position in terms of protection from 
convulsive disorders. Using developing brain as an experimental model, 
greater insight into the pathophysiological mechanisms involved in the production 
of seizures is currently being investigated. 

One of the experimental leads to the study of the ontogenesis of seizure 
mechanisms is based on the potassium shift across the nerve membrane. A 
large increase in the radioactive potassium surface efflux prior to, and 
during the early stages of seizures elicited by metrazol and electrical 
stimulation of both the hippocampus and neocortex of cats and rats, was 
observed. Using this as supportive evidence for the potassium accumulation 

29 ^ 



hypothesis, a model for the regenerative, all-or-none aspects of the initiation 
of seizure activity was proposed. Added strength was gained by a recent 
observation that Dilantin ( diphenylhydantoin) , one of the most widely used 
antiseiziire drugs, despite its great clinical utility, has thus far not been 
shown to have any specific effect on the nervous system other than that of 
antagonizing seizures. For this reason it has been of little value in under- 
standing the pathophysiology of epilepsy. However, in later experiments 
Dilantin was found to stimulate the rate of potassium uptake in an isolated 
lobster leg nerve preparation. This effect was completely abolished by 2, k, 
di- nit ro phenol. These observations, together with the finding that Dilantin 
was without effect in changing the potassium efflux rate constant, suggested 
that Dilantin may be stimulating the active uptake of potassium. The idea 
that Dilantin exerts its antiseizure effect by stimulating the so called 
Na-K pump is supportive of the hypothesis that accumulation of potassium 
contributes to the seizure induction. By stimulating the active uptake of 
extracellular potassium, Dilantin stabilizes the nervous system against 
excessive changes in potassium which, according to this hypothesis, is an 
effect that would be expected to diminish the likelihood for seizure activity. 

Several physio- chemical differences between normal and epileptogenic brain 
tissues have been found. For example, tissue from the area giving rise to 
abnormal electric discharge did not bind as much acetylcholine as did the 
normal tissue. This metabolite has been shown to occur in high quantities in 
epileptic regions of the brain, which suggests that, when this substance 
accumulates, seizures result. Furthermore, the widely accepted idea that 
acetylcholine functions primarily as an excitatory transmitter in the central 
nervous system is supported by the fact that intracarotid, intraventricular or 
intracisternal injections, or the local application of this chemical to various 
exposed areas of the brain provokes grand mal seizures. This observation is 
further supported by the fact that inhibition of acetylcholinesterase led to 
the same effects on seizure threshold as did the exogenously applied 
acetylcholine. On the other hand, norepinephrine decreases seizure suscepti- 
bility and dopamine and serotonin appear to have anticonvulsant activity. 
Gamma- aminobutyrate (GABA) is of special interest in research on epilepsy 
because of abundant evidence that it may function as an Inhibitory transmitter. 
Applied topically to the exposed cerebral cortex or injected into the carotid 
artery or the brain ventricles, it elevates thresholds for both chemically 
and electrically induced seizures. In this connection, a deficiency of vitamin 
B5 (pyridoxine) has induced seizures by producing a deficiency of GABA. Also, 
a number of known convulsant toxins appear to interfere with the use of GABA 
in the body. Disturbance of electrolyte balance may be related to the common 
convulsion during fever in infancy. Information about the cause and correction 
of such disturbances may be applied to reduce the severity of recurrent 
epileptic attacks and to reduce the possibility of permanent brain injury 
which may be caused by such episodes. 

Clinical solutions to the problem of epilepsy are concerned primarily with 
medical and surgical approaches. Before surgery is performed, it is necessary 
to ascertain that the seizures originate wholly or in part from an area of 
the temporal lobes that can be safely removed without causing serious neuro- 
logical damage, the seizures occur frequently and are incapacitating, and 
presently available drugs are ineffective in controlling the seizures. Also, 



30w 



the surgery must be accomplished so as to avoid creating a secondary scar 

which may in turn cause recurrence of the seizures. Recently, it has been 

found that barbituate and thiopental used in surgery can pinpoint areas of 

the diseased brain tissue responsible for epileptic seizures. Locating this 

tissue precisely permits it to be removed surgically. 

Several investigators have confirmed the observation that sexual disturbances 
in man occur frequently in temporal lobe epilepsy and less so in other varieties 
of the disorder. The exception to this is the relatively rare instance in 
which ant iconviils ants used in average therapeutic doses impair libido. It is 
widely held that anticonvulsants are often responsible for impotence. This 
may happen if the therapy is excessive. The majority of the patients with 
temporal lobe epilepsy and with hyposexuality regained their normal sexuality 
after operation. Attempts have been made to locate the precise region of the 
temporal lobe which may be disordered in the impotent patients. Several 
investigations have presented clinical evidence implicating the rhinencephalic 
structures and their connections in sexual disorders, and have produced 
convincing operative evidence that anterior temporal lobectomy may have a 
beneficial effect on sexual functions. 

Hypothermia (90 - 92 F) as a tool in therapy of patients with acute cerebral 
lesions, such as cerebral contusions, cranial hemorrhage, tumors, and cardiac 
arrest has been tried. Patients that were in status epilepticus, or had 
intermittent seizures which could not be controlled by medication were 
completely relieved of attacks or at least were benefited by cooling. Local 
cooling of the epileptic brain resulted in the suppression of spiking activity 
which also facilitated its response to anticonvulsant drugs. From this study 
it was concluded that hypothermia is another treatment for problems of status 
epilepticus where drugs are not effective or are contraindicated. Wo 
complications were observed that could be attributed to the hypothermia. 

The treatment of epilepsy has depended to a major extent on drugs. One of the 
earliest medications was a sedative called bromide. This was followed by 
another sedative, phenobarbital, which worked better, but caused drowsiness 
in some cases. About twenty years ago diphenlhydantoin (Dilantin) was 
introduced in the treatment of epilepsy. It has been widely used and has 
proved to be a valuable anticonvulsant drug. However, it was soon recognized 
that ataxia sometimes occurred as a complication of therapy with this drug. 
The ataxia may develop rapidly over a period of a few days, or insidiously 
over weeks or even months. One unexplained feature about this ataxia is the 
fact that occasionally a patient will rapidly develop ataxia in spite of 
having taken the same dose of Dilantin for several years. It has been thought 
that ataxia is a benign symptom only requiring a reduction in dosage or 
occasionally withdrawing of the drug. It was, however, later conclusively 
shown that cats subjected to Dilantin medication had developed severe loss of 
Purkinje cells in the cerebellum and cystic gliosis of the cerebellar white 
matter. A number of cases of permanent damage to the cerebellum, apparently 
due to this drug, have also been reported. 

Further investigation on epileptic patients who had suffered Dilantin 
intoxication indicated a high level of CSF protein levels during the period 
of intoxication. All the patients improved when the drug was withdrawn, but 



31^ 



abnormal signs occasionally persisted for several months. Inhibition of the 
NA, K, Mg, ATPase enzyme system by Dilantin was observed which may be an 
important factor in causing neuronal damage. It is suggested that Dilantin 
intoxication may not be such a benign complication as was previously thought, 
but this still does not alter the fact that Dilantin is probably the most 
effective single drug presently available in the management of epilepsy. The 
need of potent drugs of low toxicity is still paramoimt, especially in 
psychomotor epilepsy. The use of Dilantin, phenobarbital, primidone, and 
mephenytoin (Mesantoin) has left a relatively large group of inadequately 
responding patients. Recently, considerable interest has been centered on 
the use of benzodiazepines in the treatment of epilepsy. Diazepam (Valium) 
I.V. has been proved to be effective in the interruption of epileptic seizures, 
particularly Status Epilepticus. Similarly, chlordiazepoxide (Librium) and 
especially nitrozepam (Mogadon) have been shown to be useful in prophylactic 
treatment. 

Oxazepam (Serax) which is a metabolite of diazepam is easily absorbed from the 
intestines and is excreted quantitatively, or nearly so, as the glucuronide. 
Oxazepam has been widely used as an ataraxic because of the large safety 
ratio and the infrequent occurence of side effects (drowsiness, ataxia, skin 
rash, headache, etc.). Clinical trials made it clear that Serax is a potent 
drug in the treatment of psychomotor epilepsy and is of low toxicity compared 
to Dilantin and other anticonvulsant agents. The effect is seen not only in 
the reduction of seiziire frequency, but also in the KEGs. The fact that his 
compound does not interact with Dilantin metabolism, facilitates its use in 
combination with Dilantin. 

In order to establish relationships between biological activity and molecular 
structure, studies involving X-ray crystal structures are being undertaken on 
anticonvulsant drugs used in the treatment of grand mal epilepsy. Recent 
structural determinations of the clinically important anticonvulsants dmgs, 
Dilantin and Valium, have shown that although these dn;igs are chemically 
unrelated, conformationally the two drugs have many similar features. It thus 
appears that their efficacies against grand mal epilepsy are primarily due to 
stereochemical features. Similarly the drugs, procyclidine and triexyphenidyl, 
newly important anticonvulsants, are being investigated by X-ray diffraction 
methods and their molecular structure is being compared in detail with Dilantin 
and Valium. Such biochemical and biophysical tools have provided new avenues 
for the synthesis of more efficacious and less toxic anticonvulsants. 

Infectious Diseases 

Infectious diseases of the nervous system include many types of illness caused 
by, or communicated by parasites, such as bacteria, protozoa, fungi or viruses. 
Studies of infectious diseases are primarily concerned with epidemiology and 
etiology of causative agents, their mode of transmission, host relationships, 
and possibly ways to control their propagation. 

Methods 'most commonly employed for recognizing the presence of viral agents in 
cell cultures include; observations for cytopathic effects, hemaglutination, 
hemadsorption, and interference, fluorescence and electron microscopies. 
Immuno- fluorescent techniques are being used in diagnosing and studying brain 



32v,' 



inflammation due to viruses. Other studies are concerned with experimental 
encephalitis, the epidemiology of Eastern equine encephalitis, the effects of 
parasites on the nervous system, testing vaccines for protection against 
arboviruses, and the possible role of viruses in acute neruological syndromes 
in children. 

Criteria of the responses of the experimental animal to viral infection are 
established by careful monitoring of the physiologic behavior of the CNS 
following inoculation with tissue extracts and whole cells derived from brains 
of patients with chronic encephalopathies. The early events of the experimental 
disease in animals have been detected by electroencephalography because the 
EEG record becomes abnormal before any other signs of diseases appear. These 
techniques are being perfected for clinical application. However, at present 
it permits the selection of diseased animals for study during initial phases 
of the pathogenic process. Before drawing any definitive conclusions about the 
nature of the disease produced, especially about its relationship to the 
original human disease, it is well to remember that a CNS disease produced in 
animals may not be identical with the diseases of the patient from whom the 
tissue was obtained. A dramatic example of species difference in host response 
is the effect of simian virus B. In man it induces a disease that is severe, 
and usually fatal, whereas for monkeys this virus is innocuous. 

A considerable amount of work is being done on (l) after effects of infections 
during pregnancy, where they may result in the offspring's brain damage and 
mental retardation; (2) analysis of viral polioencephalomyelitis in animals; 
(3) experimental measles encephalomyelitis; (k) mode and spread of a variety 
of neurotropic agents; and (5) so-called "slow virus" diseases of the nervous 
system. Several years ago a viral polioencephalomyelitis was identified in 
pigs. The virus has now been isolated from several different organs and has 
been shown to be an enterovirus quite unrelated to many other known viruses. 
All isolates but one produced polioencephalomyelitis in germ free pigs 
indistinguishable from naturally occurring infections. 

Recently an agent causing paralysis of the CNS in rats has been isolated and 
is called hemorrhagic encephalitis of rats (HER). It produces acutely lethal 
encephalomyelitis when injected into suckling rats, including severe hemorrhagic 
lesions of the brain and spinal cord. 

Subacute sclerosing panencephalitis (SSPE) is a degenerative neurologic 
disease of children and young adults. It is characterized by progressive 
mental and motor deterioration, myoclonic jerks, and coma. The patients 
become severely emaciated and die from intercurrent infections. The diagnosis 
established during the incipient stages often shows a personality disorder or 
mental retardation. At that time, the EEG shows slowing and dysrhythmia. 
However, high amplitude, low frequency synchronous waves do not develop until 
the patient exhibits myoclonic jerking. Spinal fluid proteins and cell counts 
remain normal or increase slightly during the entire course of the disease. 
Transmission of encephalomyelitis from humans to animals and further from 
animal to animals, producing symiptoms typical of SSPE in the animal, has 
provided an important new lead in isolating and understanding the causative 
agent in SSPE. During the last few years, evidence of a relationship between 
SSPE and measles virus has been established. With the help of electron 



33^ 



microscopy, tubular structures have been seen in the brain with SSPE which 
resembled the nucleocapsid.es of measles virus. Further investigation showed 
a high or rising titer of measles antibody in serum, measles antibody in 
cerebrospinal fluid, and measles viral antigen in the brain of SSPE patients. 
On the basis of cytopathology, filtration, and serology, measles virus has been 
isolated from patients with SSPE. These patients had no history of clinical 
measles, but had received live measles virus vaccine; some had rubeola once 
in their life time. Several characteristics common to both measles virus and 
SSPE have been foiuid. They include antigenic properties, host- range, 
cytopathogenic effects, temperature sensitivity, thermal inactivation and 
interferon production. Although basic similarities have been established, 
still unanswered are the questions of how and why the virus persists during 
the long period after the patient has recovered from measles and before he 
develops SSPE. Is the virus different in some way from regular rubeola? Will 
measles vaccine protect against SSPE? Is the isolated paramyxovirus a neuro- 
adapted strain of measles virus or a distinct but antigenically related virus? 
Is a second "helper" or "co-virus" necessary to produce disease? If two 
etiologic agents exist, how do they interact? How does virus reach the brain 
and spread within the CNS? How is it maintained for years if indeed it is 
measles virus? Are the unusually high serum measles antibody levels noted in 
most SSPE patients in response to viral antigen in brain and how is the virus 
protected from antibody? Does interferon play a role in limiting the disease? 
Is it possible to reactivate a latent or defective virus in the CWS? 

Measles encephalitis is a disease characterized pathologically by demyelination. 
Evidence that the measles virus is directly responsible for the brain damaging 
effects has been inconclusive. This consisted primarily of the demonstration 
of cytoplasmic and nuclear inclusions, and small giant cells in brain material 
from fatal cases in various stages of the disease. Presently, research is 
directed towards the etiology of miiltiple sclerosis (MS) and the mechanism 
involved in its development. It seems likely that the measles virus is 
responsible for an acute demyelinating disease, known as "measles meningoence- 
phalitis." It is postulated that MS may be a "slow" infection of the nervous 
system by measles virus, i.e., that post-measles encephalitis, SSPE and MS 
are different temporal manifestations of measles. It has been reported that 
MS patients have significantly higher serum measles antibody titers than 
patients with other neurological diseases or normal individuals. These are 
the only observations that provide a possible link between measles and MS at 
the present time. 

The concept of slow viruses was originated by Sigurdsson in the year 195^ who 
recognized in Icelandic sheep a special category of diseases with prolonged 
incubation period, slow in clinical course, and restricted to a single organ 
and species. Rlda (or scrapie), a chronic neiirological disease, and maedi, a 
chronic pneumonia, were included. Their relevance to human diseases stems from 
the similarity of sheep scrapie to human kuru. Kuru and Creutzfeldt- Jakob 
Syndrome are two chronic human neurologic diseases. Kuru is a fatal neurologic 
illness restricted to the Fore people of New Guinea. It has been transmitted 
to chimpanzees. The transmissible agents involved have not been characterized 
and studies are limited because a susceptible small laboratory animal or tissue 
culture system has not been found. 



31^ w 



Electron microscopy has revealed particles similar to viruses of the papova 
group in the brain from patients with progressive multifocal leucoencephalopathy. 
No evidence has been presented on the transmissibility of this disease. Visna 
virus is found in sheep and is also capable of rapid growth in tissue culture, 
but pathogenesis in vivo is slow. Aleutian disease of mink is also considered 
a slow virus . 

Of all the slow viruses implicated in acute, subacute and chronic neuropathies, 
scrapie is the only virus studied extensively because of the relative speed 
and economy of using mice. Reliable criteria of its induction have been 
established. Mice in advanced scrapie, when suspended by the tail, character- 
istically clasp their hind legs together, whereas normal mice splay theirs 
outwards. During uhe developmental stages, mice exhibit a sequence of 
responses reflecting progressive central nervous system damage before the 
clasping reaction is manifested. These studies have also generated many of 
the seeming paradoxes of scrapie virus, such as its size relative to its 
molecular weight and apparent absence of any nucleic acid. Hypotheses 
explaining the special features of the scrapie agent include persisting tolerated 
infection; defective virus; special auto- immune response; and a replicating 
polypeptide, polysaccharide or membrane defect. Directly or indirectly these 
concepts have provided rational bases for research. For example, if scrapie 
is a defective virus, Sendai virus may be acting as a helper in the accelerated 
course observed. If scrapie represents a membrane defect, it may be visible 
by scanning electron microscopy. Scrapie virus has been successfully grown 
in brain cell cultures of sheep, mouse, and mink. All workers have observed 
differences in vigor and appearance between infected and normal brain cells 
in vitro. Biochemical differences associated with scrapie have also been 
reported. Increases in enzyme activity, such as glucosaminidase, offer the 
possibility of quantitative assay using florescent- tagged enzyme substrates 
and detecting the split produced with ultraviolet microscopy. Mice inoculated 
with scrapie virus showed increased DNA metabolism. Inhibitors of DNA 
metabolism ( idoxuridine and hydroxyurea) did not alter the course of disease. 
Interferon has also been studied in scrapie. Extrinsic interferon, injected 
peripherally, had no effect. Other studies showed that scrapie neither 
induced interferon nor prevented its induction by viruses. Studies with 
poliomyelitis and a crude predecessor of statolon, an interferon inducer, 
showed that peripheral treatment was active against peripherally inoculated 
polio virus, but not against polio virus inoculated intracerebrally unless 
treatment was also given intracerebrally. Experiments currently in progress 
indicate that scrapie can be altered in vivo . Test materials have been 
selected for their relation to interferon, immunity, DNA metabolism, and 
antiviral or other therapeutic activity. Wo effect on survival was noted with 
adenine arabinoside, started the date before scrapie inoculation and continued 
throughout the course of disease. Tentative findings indicate that short-term 
amantadine treatment late in the disease did not reverse its neurologic effect. 
The results of cyclophoshamide treatment varied depending on the time of 
initiation. Recently, scrapie virus has been found in sheep brain inoculated 
with SSPE and IVIS infected brain tissue. Also, scrapie-like illness in mice 
inoculated with MS brain has been observed. Acceptance of the importance of 
these reports awaits clear evidence that an agent responsible for the disease 
in sheep and mice had its origin in the MS tissue and not in the experimental 
animals or their surroundings. 



35" 



Current studies are directed towards the development of new methods of evaluating j 
scrapie in vivo and in vitro , increased understanding of the disease and 
etiologic~aient associated with scrapie, its related neurologic disorders, 
and the means of altering them. 

Myxoviruses are medium sized, ether sensitive RNA viruses, which in man have 
the common effect of causing respiratory disease. Although they are not 
regarded as "neurotrophic" viruses in man, mumps is the most common virus 
causing central nervous system infection in rodents. In these studies ( 

occasional neurons are infected by the unadapted strain, and repeated passage 
has yielded a strain which infects parenchymal cells of the brain, causing 
acute encephalitis. However, mumps virus in hamsters has been shown to cause 
a clinically inapparent infection limited largely to ependymal cells and neurons, 
resulting in acute encephalitis. 

Hydrocephalus may be caused by infectious agents contracted prior to birth. 
Suckling hamsters infected by mumps virus developed a narrowing of the 
aqueduct of Sylvius with subsequent hydrocephalus. The narrowing of this 
canal, responsible for draining the ventricles of the brain, is the most common 
cause of hydrocephalus in man. 

The study was conducted with experimental models to determine the histopatho- 
logical changes in the brain, aqueductal narrowing, and hydrocephalus induction. 
The signs of clinical disease developed only after the resolution of the acute 
infection (about fourteen days), during which the lining cells of the aqueduct 
had been almost destroyed. Of the suckling hamsters inoculated with mumps 
virus, ninety-five percent developed clinical hydrocephalus. These and other 
histological changes were shown to be specifically related to the mumps virus 
infection. 

Although studies of experimental infections indicate that viruses may reach 
the nervous system by various routes, studies of naturally occurring infections 
indicate that the major pathway of virus spread to the nervous system is by the 
hematogenous route. The various humoral and cellular defenses which act as 
deterrents to limit the occurrence, intensity and duration of viremia, as well 
as subsequent spread to the nervous system, constitute a physiologic blood- 
brain barrier which must be overcome if infection of the nervous system is to 
occur. It has been shown that herpes simplex virus gains access to the central 
nervous system by the hematogenous route. Since blood contains both phagocytic 
and immuno- competent cells, the extent and duration of hematogenous dissemination 
of virus depends upon the outcome of interactions between host defenses and 
virus. The characteristics of herpes simplex virus are: (l) it is a DNA virus 
whose replication is susceptible cells is generally associated with formation 
of easily demonstrable intranuclear inclusions; (2) it is associated with viremiaj 
and central nervous system disease in both man. and animals; (3) it can cause 
acute and subclinical infections; (k) it may persist in the tissue in occult 
form for prolonged periods, giving rise to exacerbations of disease following 
a variety of stimuli and; (5) it has been associated with chronic and recurrent 
encephalitis in the rabbit, suggesting that it may also be responsible for 
such illness in man. From these studies it has been ascertained that human 
white cells, specifically lymphocytes will support the replication of herpes 
simplex virus j^ vitro. This was the first DNA virus to be propagated in 

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human leukocytes. Viral replication failed to -occur in the absence of 
phytohemagglutinin, suggesting that lymphocytes must be in a particular growth 
phase to be susceptible to infection with this agent. 

Woi-K with the Sindbis encephalitis virus in mice demonstrated a precipitous 
development of resistence during the second week of life, due apparently to 
a limitation in the spread of infection. Wo non-specific viral inhibitors 
could be detected. Studies on mumps vims encephalitis in hamsters showed 
that disease and death resulted from the infection of neurons which remained 
morphologically normal. It is suggested that this material may be useful in 
investigating the "slow" viruses which seem to act in a similar way. 

In Schilder's encephaloclastic sclerosis, virus-like particles were identified 
by electron microscopy which are distinctly different from those observed in 
the case with herpes simplex encephalomyelitis , but are similar to those that 
occur in subacute sclerosing leukoencephalitis. The nature of these virus-like 
particles is being investigated. 

In 1958 there appeared the first report that free-living ameba could produce 
fatal meningoencephalitis in animals. More recently a death of a boy was 
reported who died five days after initiating the symptoms now being recognized 
as typical of amebic nemingoencephalitis (AME). It is believed that the 
invading ameba, Maegleria gruberi , enters the mouth and nose, possibly during 
swimming. The victim soon suffers flu-like symptoms followed by rapid 
deterioration and death. Unlike most of the parasites that live with man, 
Naegleria, as a free-living organism, requires adequate oxygen. This may 
explain why it has a great affinity for the oxygen- rich capillaries of lung 
and brain tissue. The disease appears to be fatal and is without response to 
anti- amebic drug therapy. Virus-like particles measuring about 100 mm in 
diameter have been observed in some strains of these invading pathogens and 
their morphological characteristics have been recorded. Preliminary experi- 
ments indicate that mice are killed by introducing axenic EG ameba either 
intracranially or interanasally. Other studies involving isolation, purifica- 
tion, characterization of the active agent are currently underway. 

In 19^3 and 19^4 a virus was recovered from mosquitoes in the San Joaquin 
Valley of California. Until I965 virtually nothing was known about its 
disease- causing potential in humans when the recovery of a representative 
strain of this virus was reported from the brain of a child with a fatal 
illness. It was apparent from serologic studies that many patients examined 
were infected with California encephalitis virus coincident with rather severe 
illnesses. These patients were studie very carefully to exclude other known — 
causes of viral CMS diseases. All patients had encephalitic signs and symptoms 
different from those due to enteroviruses. In contrast to enteroviral illnesses, 
none of the patients in this study had a biphasic illness, none were associated 
with upper respiratory of gastroenteric signs or symptoms, and no illnesses 
occurred in siblings or other family contacts of the patients. Moreover, 
seizures, coma, and disorientation were more frequent in the patients in this 
study in comparison with children with CMS infection due to enteroviruses. 
Behavioral correlaries of patients suffering with this virus showed difficulty 
in receiving consistently meaningful, basic visual and auditory perceptual 
information. They had little or no difficulty in the higher language functions. 

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The personality of these children seemed to fit the "organic hyperkinetic 
syndrome." The result is a predictable behavior pattern seemingly associated 
with specific learning disorder. 

Tumor C1300 has been in serial transplantation since 19^0 at the Jackson 
Laboratory, Bar Harbor, Maine. This tumor arose spontaneously from the 
region of the spinal cord of a albino mouse and was diagnosed tentatively as 
neuroblastoma . Until recently no definite proof existed regarding its 
neuronal characteristics. One of the characteristic enzymes of nerve cells, 
tyrosine hydroxylase, is present in tumor C1300, confirming the diagnosis of 
neuroblastoma. Recehtly it has been demonstrated that cell-free extract (CFE) 
of neuroblastoma tumor induces a similar neoplasm in the dermis, mesentery, 
spinal cord and sympathetic chain after subcutaneous injection. A total of 
70 percent of CFE- injected mice developed neuroblastoma tumors. Like tissue- 
transplanted tumors, CFE- induced tumors possess a similar amount of tyrosine 
hydroxylase and, in vitro, differentiate into neuron-like structures. Electron 
microscopic studies show that virus-like particles are present in CFE- and tissue- 
induced neuroblastoma. The viruses are doughnut- shaped and appear to bud from 
endoplasmic reticulum. Whether these viruses are causative agents in producing 
neuroblastoma by CFE remains to be ascertained. The following studies using 
partially- purified neuroblastoma viruses are in progress: (l) viral etiology 
of neuroblastoma; (2) induction of neuroblastoma using different routes of 
injection; (3) induction of neuroblastoma in different species; (k) deomonstra- 
tion of the type of virus, DWA vs. RNA; (5) transformation study ±n vitro using 
different mammalian cell-lines; (6) selection of a cell-line which sustains 
viral multiplication without transformation; (7) exploration of the presence 
of viruses in human neuroblastoma; and (8) the induction of tumor using cell- 
free extract of glioma, medulloblastoma, astrocytoma and neurofibromatosis. 
The animals and human tumors will be used for this purpose. 



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RIMAMENTAL RESEARCH M THE NEUROLOGICAL SCIENCES 

Introduction 

The great importance of fundamental studies on neurological, sensory, and 
communicative components is frequently overlooked because they may have little 
or no immediate clinical application. However, the history of science shovs 
clearly that improvements in the treatment of disorders are largely dependent 
on sound basic research. 

It is generally accepted that the membrane around a nerve fiber plays an 
essential role in the transmission of the nerve impulse along the fiber. 
However, relatively little is known about the structure and mechanism of action 
of the nerve membrane or of living membranes in general. There is no doubt 
that research on membrane structure will contribute important information about 
how the nerve impulse is transmitted along the nerve fiber . There is a 
substance known as the nerve growth factor, the exact composition and source 
of which is still uncertain, which has a remarkable stimulatory effect on the 
growth of nerve fibers. The ultimate importance of the factor is still to be 
determined, but it surely will have important functions in nerve regeneration 
and it may well turn out to be useful in healing lesions and restoring function 
in the brain and spinal cord. 

There are several anti-convulsant drugs that are relatively effective in 
controlling seizures in a good proportion of epileptic patients. Some of these 
drugs are quite unrelated chemically and there is no explanation why they are 
effective and why closely similar compounds are entirely ineffective. It has 
been suggested that the efficacy of a drug may depend upon a highly specific 
molecular structirre rather than on the chemical composition of the molecule. 
Therefore, studies on molecular structure are now highly Important and involve 
such sophisticated techniques as x-ray crystallography, electron paramagnetic 
resonance spectroscopy, and nuclear magnetic resonance spectroscopy. On the 
basis of such highly specialized information on the atomic interrelationships 
and the electron activity in the effective molecules, it will be possible to 
find or synthesize other molecules that are more effective in the treatment of 
disease. It may also lead to an understanding of how the present drugs work, 
which often is completely unknown now. 

One of the great \xnknowns in neurophysiology has been the exact mechanism 
by which the nerve Impulse is transmitted across the synapse from one nerve 
fiber to the next or from a nerve to a muscle or other end organ. In 190^4- it 
was suggested that some nerves accomplished this by the liberation of epinephrine. 
In 1914 another investigator si;iggested that acetylcholine was the synaptic 
transmitter in some cases. These postulates were finally proven in I92I by the 
classical experiment in which two frog hearts were connected by a small glass 
tube. When the beat of one heart was slowed by stimulating the vagus nerve, 
the beat of the other heart also slowed a few seconds later, demonstrating 
conclusively that some chemical was produced by the nerve in the first heart 
which was carried in the perfusion fluid and produced a similar effect in the 
completely separate second heart. It is now known that a number of compounds 
(acetylcholine, noradrenaline, dopamine, 5-hydroxytryptamlne, gamma-amlnobutyric 
acid, glutamic acid, glycine) may act as synaptic transmitters. However, the 
mechanism by which the transmitter may be produced and act very rapidly 

39 w 



(many times per second) and how the receptor nerve receives the stimulations 
is still largely unknown. As mentioned at the beginning of this Report, there 
are 10 to 13 billion nerve cells in the brain alone. Each then coimects with 
from two to several other nerve cells. Therefore, the number of synapses is 
almosc infinite and research on synaptic transmission is of the utmost 
importance. Also, this is a good example of why the application of basic 
research must be a time consuming process. The first evidence of the chemical 
transmission across synapses was obtained more than 65 years ago. However, 
the development of the area, even to its present imresolved state, had to 
await the discovery of highly sophisticated techniques such as electron 
microscopy and methods of chemical analysis sensitive enough to detect literally 
only a few molecules of a specific compound. 

The Nerve Membrane 

One team of investigators has used spin labels (paramagnetic free radical 
probes) to detect inside -out side conformational transitions of specific spin- 
labeled lipids and proteins in biological membranes. A vai'iety of spin labels 
have been synthesized and they are designed so they may be directed toward 
a special region of the membrane or may serve as analogues of natural membrane 
components. It was found that in membranes, particularly nerve and erythrocyte, 
the motion of a series of spin-labeled fatty acid labels leads to a quantitative 
estimate of the amounts of gauche and trans conformations about carbon-carbon 
bonds of the fatty acid chains of phospholipids. Analysis of the magnetic 
resonance spectra showed that the preferred orientation of the amphiphilic 
labels I and II in nerve fibers was that in which the".long molecular axis is 
perpendicular to the membrane surface. The use of spin-labels in nerves has 
led to the development of a new theory of anesthesia. The magnetic resonance 
spectra of lipid labels have revealed the fluidization of membranes by local 
anesthetics and the tightening of the membrane in the presence of cholesterol 
and antibiotics. ATPase of the cerebral cortex cell membranes is presumably 
part of the active sodium potassium pump of the membranes. The fact that 
biogenic amines such as serotonin and norepinephrine affect enzyme activity 
and that the ATPase is foimd primarily in the synaptic region suggests that, 
in addition to its function in maintaining sodium and potassium gradients, 
it may be involved in nerve impulse initiation. 

Another project is working toward providing a mathematical physical- 
chemical theory of the activity of the axonal membrane and thereby to correlate 
observed membrane phenomena and predict new phenomena. Further objectives are 
to provide a theory of nerve activation by transmitter substances, to assist 
in understanding the action of phannacological agents, and possibly to explain 
cyclical nerve cell activity such as is detected by the EEG. The project is 
designed to show that the excitability of membranes results from classical 
physical -chemical forces, and does not depend upon any unusual specialized 
structures, such as specific ion species channels, or special field- sensitive 
gates. Some years ago it was shown elsewhere that the activity cycle consisted 
first of all of a large increase in the inward Na+ current followed by an 
increase in outward K+ flow. However, no attempt was made to provide a 
theoretical foundation for the empirical eqimtion obtained since this was felt 
to be beyond the available physical knowledge at this time. 



40w 



The problem is two-fold; the calculation of the steady state ion dis- 
tribution, and the transient. The first requires the solution of the total 
differential equation two -boundary problem, and the second, the PDE two- 
value boundary problem. The total DE solution gives the steady state, from 
which the activity cycle commences, and the PDE solution gives the activity 
cycle. Both problems were much more complex than the investigators anticipated. 
Much of the effort during the first two years was required to simply develop 
the necessary numerical methods. 

In this "model" of an excitable membrane, it has been shown that the 
important factors are the height of the potential barriers for the ions to 
traverse the two interfaces, the effect of the local electric field on these 
barriers, and the blocking effect of absorbed CA++. A high negative charge 
concentration near the outer surface of the membrane was found to be of 
importance in giving the local field necessary at the interface. An interest- 
ing and suggestive finding was the existence of oscillatory currents following 
a disturbance. Their magnitude depends on factors yet to be investigated. 
However, it appears that some clue to the source of EEG rhythms may be found. 

Nerve Growth Factor 

More than two decades ago it was observed that a crude extract of mouse 
submaxillary gland coiold produce an explosive growth when applied to cultured 
ganglion cells. Several years later a protein was isolated and purified which 
possessed the characteristics of nerve growth factor (NGP). It was deteimiined 
that the growth promoting factor was present in lower concentrations in a 
number of other tissues, and that the extract of mouse salivary gland contained 
a number of proteins other than NGP. Most notable among the latter is a 
protein that specifically stimulates the growth of epithelial cells. Still 
other growth factors may be identified in the future. The chemical and 
biological inter -relationships among the various substahces isolated from the 
mouse submaxillary gland are both complex and uncertain since chemical con- 
stitution and biological activity are closely dependent on the isolation 
procedures employed. However, it is now generally agreed that the simplest 
NGP protein extractable from the gland is relatively small with a molecular 
weight of 25,000 and is very basic. A larger species with NGP activity is a 
combination of this basic protein with two other types of subunits, one of 
which is an esteropeptidase. Whether or not other preparations with NGP activity 
are actually distinct from these two, has yet to be determined. In any case 
biological activity as reflected by neurite growth in tissue culture can be ob- 
tained with most preparations at a concentration of 10"T grams /mililiter. 

The mechanism of NGP activity is being pursued intensively in a number of 
laboratories utilizing modern techniques of biochemistry and cell physiology. 
We may soon know whether the NGP protein acts on the nerve membrane or moves 
through it, how cellular synthesis of nucleic acid and protein is initiated by 
NGP and how NGP inactivates the mechanisms which control cell size, shape and 
volume. As may be expected a number or laboratories are investigating the 
effects of NGP on regeneration in the nervous system. 

Structural Studies of Drugs 

By methods of single crystal structure determination, using X-ray and 

i^iw 



neutron diffraction^ one project is studying the structural details of the 
■way in which barbiturate molecules associate with each other and with molecular 
species of biological systems^ such as water, peptides and lipids. The use 
of barbit\rrates in neurological disorders is well known. It has been assumed 
that for effective action a barbiturate, initially in an aqueous medium, 
must be capable of associating with the lipids of a biological membrane to 
modify the normal metabolic cellular processes such as membrane transport. A 
related possibility is that the barbiturate may directly inhibit the complex 
enzymatic processes of oxidative phosphorylation. It is now believed that the 
multi-enzyme system in electron transport is effectively blocked by barbiturate 
in the area of a flavoprotein. 

The detailed crystal structures of some 20 barbiturate derivatives have 
been determined and tested in chemical and/or biological systems. It was 
found that the barbiturate ring is planar, apart from slight distortions 
depending on the crystal environment. Exceptions are found in the "half chair" 
conformation of 5^ 5-<3-isubstituted barbiturates in which at least one 
substituent is hydroxyl. With one exception (alloxan), all potentially hydrogen 
bonding hydrogen atoms were found to be hydrogen bonded. 

There was a strong preference for barbiturate imine groups to hydrogen- 
bond with carbonyl groups of other barbiturate rings rather than with C(5) sub- 
stituent groups or water molecules. Almost all barbitiirate structures thus 
exhibited a one- or two-dimensional framework of NH. ..0-C hydrogen bonded 
barbiturate rings which appears to be the dominant crystal structure determin- 
ing activity. Although the nature of the C(5) substituents and the possible 
presence of water of crystallization determine the manner of ring hydrogen 
bonding to form this framework, their own hydrogen bonding requirements appeared 
to be of secondary importance. The latter may frequently be of the "bent" or 
"bifurcated" type, indicating relatively weak hydrogen bonding interactions. 

The overall aim of another project is to determine the conformations of 
acetylcholine (ACh) when it becomes attached to its nicotinic and its muscarinic 
receptors. The approach was to synthesize analogues of ACh in which a degree 
of rigidity was introduced into the amino alcohol portion of the ester. 
Cyclopropane and cyclobutane rings were used to impart the desired rigidity in 
the ACh analogues and congeners. Free rotation was thus prevented, and the 
number of possible conformers was greatly reduced. The overall shape of the 
molecules and pertinent ranges of interatomic distances could then be estimated. 
This work is based on the premise that the conformation and moleciolar shape of 
ACh are critical factors in its actions at the two types of cholinergic 
receptors. 

Some 50 analogues of ACh have been synthesized, particularly those in 
which the amino alcohol portion is a part of a cyclopropane or a cyclobutajie 
ring system. Each of the enantiomers of trans-2-acetoxycyclopropyltrimethylam- 
monium iodide has been prepared. The absolute configurations have been 
determined and have been correlated with biological data. A new approach to 
the sjmthesis of c is -2 -acetoxycyclopropyltr imethylammoniuin has been developed 
which Hiay permit higher yields of the compound. 

In the studies on 'the cyclobutane series, cyclobutane carboxamides have 

42w 



been converted to cyclobutylamines with lead tetraacetate. 2-Substituted 
cyclobutylmethyi ketones have been prepared from 2-substituted cyclobutane- 
carbonyl chlorides, and these ketones have been shown to undergo a facile 
Baeyer-Villiger reaction to give high yields of cyclobutyl acetates. 

Despite its relative chemical simplicity and its importance as a neuro- 
transmitter, the mode of action of ACh is still unknown. A logical hypothesis 
is that the flexible ACh molecule assumes different molecular shapes and 
different interatomic distances at the nicotinic and the muscarine reptors, 
conditions which are imposed on the ACh molecule by the chemical and steric 
nature of the receptor area. With the synthesis and testing of ACh analogues, 
this project is trying to clarify the mode of stereo-chemical attachment of 
ACh at its nicotinic and muscorinic receptor sites and in this way truly explain 
the mechanism of action of ACh. 

The Synapse and Neixrotransmitters 

The identification of synaptic transmitter compounds and the mechanisms 
by which lobster nerve cells regulate their accumiilation has been studied 
extensively in one laboratory. Early work showed that gamma-aminobutyric acid 
(gam) was the inhibitory transmitter at the neuromuscular junction, that it 
was the most active inhibitory substance present in nerve extracts, and was the 
only physiologically active compound selectively localized in inhibitory 
neurons. Glutamate was the only excitatory substance found. 

It has now been shown that GABA is actually released when an inhibitory 
neiiron is stimulated and the amount released is proportional to the number of 
stimuli applied. Like the release of chemical transmitters at other synapeses, 
it depended on the presence of calcium in the bathing fluid. Recently, it has 
been possible to localize the site of GABA uptake in lobster nerve-muscle 
preparations. Certain aldehyde fixatives will covalently bond amino acids to 
tissue components. With this method, using h3 labeled GABA, the site of uptake 
was shown to be the connective tissue cells and Schwann cells that surround 
the nerve -muscle preparation. 

Using single excitatory and inhibitory axons, enzymic studies showed that 
the pathway of GABA metabolism in the lobster was identicaJ. to that in the 
mammalian nervous system. The substrates, glutamate and alpha -ketoglutarate, 
were present in the same concentrations in excitatory and inhibitory axons, 
and transaminase activity was similar in both kinds of cells. Only decarboxylase 
(and GAEA) was different, with about 100 times as much activity in the inhibitory 
axon. It was found that high concentrations of GABA could inhibit the decar- 
bo:cylase. At the normal concentrations of GABA and glutamate in inhibitory 
axons, the synthesis of GABA approximately balanced its destruction. This led 
to a new mechanism to explain the selective accumulation of GABA in inhibitory 
axons, with the decarboxylase being the key to accumulation and the GABA 
inhibition of the decarboxylase the key to the final level to which GABA 
accumulated. 

In recent years, glutamate has been thought to be the excitatory trans- 
mitter compound. After a prolonged series of experiments, it was possible to 
demonstrate a small (lO^) release of glutamate after excitatory nerve 



l^y 



stimulation. The lack of adequate methods made 'it impractical to try to 
obtain the necessary controls of calcium dependence on release or the pro- 
portionality between frequency of stimulation and amount of glutamate released. 
Quantitative comparisons of glutamate in excitatory axon extracts and the amount 
of excitatory activity in glutamate units in the extracts showed that there is 
about 2 to 3 times more activity than glutamate. After much work, it was 
shown that most of the excess activity could be accounted for as asparate. 
Asparate alone is relatively ineffective^ but it can add to the excitatory 
effect of glutamate^ causing an apparent potentiation of the glutamate res- 
ponse. 

Studies on the post-tetanic potentiation (PTP) and post-tetanic repetition 
(PTR) are particularly important because these responses disclose the activity 
of motor nerve terminals. By this mechanism, another laboratory has studied 
the effects of many compounds on the soleus motor nerve terminals (MNTS) in vivo . 
Using increasing doses of d-tubocurarine (dTC), it was fo\md that PTR and 
PTP were progressively suppressed and finally obliterated. It is impressive 
that PTR and PTP were abolished by a curare (dTC) dose that had only a minimal 
effect on neuromuscular transmission. This showed that the MIITS were a highly 
sensitive locus of dTC action, but that this in itself was not the source of 
the neuromuscular blocking action of curare. Nevertheless, this effect suggests 
that the pre-jimctional structure may still be important in the action of de- 
polarizing neuromuscular blocking drugs. 

Because of its importance in neuromuscular studies, Mg was investigated. 
It was found that Mg also depressed the PTP and PTR, This demonstrates that 
the action of llg is on the MNTS. The pre- junctional effect of Mg, like the 
dTC action, preceded the effect on transmission. Also, it was found that 
transmission block and depression of directly stimulated muscle coincided. 
Thus, Mg depressed both muscle and MNTS, whereas dTC acted selectively on MNTS. 

The local anesthetics, procaine, lidocaine, tetracaine, and debucaine had 
been shown by other investigators to suppress PTP and PTR. Studies on this 
project showed that the respective potencies of these drx;igs to depress MNTS 
correlate exactly with the order of their condution blocking action on 
peripheral nerve. This result is important because analysis of procaine action 
on neuromuscular transmission by in vitro quantal methods by other workers had 
indicated that the major action is curariform on the post-junctional membrane. 
No doubt these drugs, like Mg, will affect excitable tissues non-selectively, 
but at the neuromuscular junction the first effect, as dose is increased, -^ is 
the paralysis of MTS. 

Diphenylhydantoin (DPH), a drug used in epilepsy, was known to suppress 
PTP of the monosynaptic reflex pathway. Therefore, its effect was studied on 
the hyperpolarization of MNTS that follows high frequency stim\£Lation. Anti- 
conviasant doses were found to suppress MNTS PTR and consequently the muscle 
PTP. Apparently, DPH opposes the hyperpolarization of MNTS just as it does in 
the primary afferent endings in the 2N pathway. The DPH action on post-tetanic 
hyperpolarization indicates that the drug selectively suppresses after-potential 
production in the terminals. 

All of this work has been done in vivo and, therefore, the results are 



kk 



unquestionably relevant to clinical neuropharmacology. AlsO;, the actions 
of the various drugs classically associated with modification of neuromuscular 
function have been seen in a new light and new understanding of ^their action 
has been developed. 

With the use of electron microscopy and semi-quantitative microscopic 
spectrofluorometry, a team of investigators has been studying the function 
of neurohumors such as serotonin (5-HT)^ nor-epinephrine (NE) and acetylcholine 
(ACh). It was possible to demonstrate a relationship between the dense-core, 
or granular, nerve ending vesicles and a storage form of NE in the rat vas 
deferens. Vas deferens depleted of endogenous WE were exposed to WE or to 
one of its precursors, DOPA or dopamine, in the presence of an inhibitor of the 
conversion of precursors to WE. Only WE specifically restored the proportion 
of granular vesicles to nearly normal levels. Thus, the granular vesicles were 
identified as the intra-vesicular compartment of WE storage. In the presence of 
an inhibitor of monoamine oxidase iproniazid, exogenous WE accumulated in the 
extravesicular compartment, but did not enter the granular vesicles. Under 
these circumstances, the contractions of the vas deferens usually produced by 
stimulation of the nerve were markedly inhibited, presumably due to excessive 
NE in the extravesicular compartment. It is clear from these studies that the 
distribution of WE can be identified in its storage sites at the subcellular 
level and that the subcellular localization of the amine influences physiolog- 
ical and pharmacological effects. 

This group has also studied neurohumors in the rat brain. They showed 
that gamma-hydroxybuturic acid lactone (GBL) induced anesthesia, but had no 
influence on 5-HT and gamma -aminobytyric acid (GABA) levels in the brain, and 
iTke all central nervous depressants, produced an increase in brain ACh. In 
regard to effects on brain catecholamines, neither gamma-hydroxybutyric acid 
(GHB) nor GBL altered brain NE levels, but they did selectively increase 
brain dopamine (M) levels, occasior-ally as much as 100^. In addition 
gamma -methylpara tyro sine, which blocks the synthesis of both NE and DA, caused 
about 100^ increase in the sleep time of animals given GHB or GBL. This 
indicates that DA may be necessary for the arousal of an animal from a sleeping 
state. The increase in DA during GHB-induced sleep may be due to a reduced 
utilization of DA or to a compensatory activation of synthesis as a resiolt of 
suppression of dopamine receptors by the GHB. Thus, a study of the ultramicro- 
scopic localization of neurotransmitters may make important contributions to 
knowledge of sleep mechanisms, a problem of utmost importance in itself. 



1^.5w 



COMMUNICATIVE SCIENCES 

In communication, language is the carrier of the vast amount of what 
we call culture. Knowledge of the past, techniques of food getting, science, 
and social rituals are all carried in language. The social psychologist tells 
us that language is important first as it relates to communication and secondly 
as it functions in the socialization of the individual, particularly in the 
development of his personality. Moreover, it carries for the person the social 
definitions of situations, the world of discoiirse and the whole range of cul- 
ture content which impinges upon and shapes the individual. The National 
Institute of Neurological Diseases and Stroke is pleased to report many 
important advances in knowledge in the Communicative Sciences this past year. 

Sensory Processes 

A physiological and psychophysical investigation in three sense modalities: 
audition, touch and vision, complemented by analytical procedures involving 
mathematics, electrical network modeling and digital computers, has extended 
the analogy between auditory and tactile psychophysical characteristics to 
cover several aspects in the frequency, time and space domain at threshold, 
as well as at suprathreshold levels. The generalized functional relationship 
between the firing rate produced by sensory reception and stimulus intensity, 
which was discovered in the previous year and which seems to hold indepen- 
dent of sense modality and animal species, was further documented with the 
help of neurophysiological experiments on the Limulus lateral eye and the 
auditory organ of Southern Army Worm Moths. A theorem resulting from the 
mathematical formation of the relationship was confirmed by studying summation 
between sinusoid and a band limited random noise. In audition measurements 
of central masking characteristics have continued, and the psychophysiological 
theory of central masking developed in the previous year was further validated. 
In electrophysiological investigations of the auditory receptors of the 
Southern Army Worm Moth, it was possible to infer the principal site of mechan- 
ical stimulation. In tactile studies, it could be demonstrated that the slope 
of the subjective intensity functions is negatively correlated with the thresh- 
hold intensity and the density of innervation. Sensory studies continued to 
focus on the Limulus lateral eye. The neurophysiological recordings from 
single receptor units have brought out a nonlinear interaction between 
excitatory and inhibitory processes and have made it necessary to introduce a 
nonlinear correction factor into the Hartline-Eatliff equations. 

Another investigator, attempting to determine the extent to which 
evidence for the Stevens power law shows itself in the recorded potentials 
from sensory cells, determined that in numerous sense modalities the neuro- 
electric potentials can be shown to grow as a power function of stimulus 
intensity. The power function exponents for two modalities, vibration and 
hearing, have exhibited an interesting dependence on frequency. 

Auditory Masking 

In attempting to identify differences between sinusoid and noise masking. 



k6^ 



an investigator developed a new hypothesis which accounts for some of the 
differences. The observer;, faced with the problem of trying to detect a 
weak sinusoid in some masking stimixlus, searches the frequency spectrum and 
listens primarily at the region in frequency where the signal - to - 
masker ratio is most favorable. In detecting a sinusoid in a white noise 
spectrum^ there is no problem. The maximum signal - to - noise ratio occurs 
at the signal frequency^ and asserting that the observer listens primarily 
at that frequency^, is simply the statement of the critical band hypothesis.. 
Detecting a gated sinusoid in a continuous sinusoidal masker is a different 
problem. The mere fact that a signal is turned on and off implies presence 
of energy in regions of the spectrum other than the signal frequency. The 
amount of this splattered energy depends on the exact shaping employed in 
the process of gating the signal. The ratio of signal - to - masker energy 
depends both on how the signal is gated and how much the masker is attenu- 
ated by the observer's auditory filter. The critical item is not the rise 
time of the gating filter, but the off -band attenuation rate of the auditory 
filter. 

Vestibular Mechanisms 

One group of researchers developed an analog equivalent electrical 
circuit of the cochlea that follows the principles of the electromechanical 
hypothesis for the function of the organ of Corti. The steady-state 
parameters of this heuristic electrical model were calculated from data 
about the impedance and potential differences that exist across the walls 
of the cochlear partition. The behavior of the model is such that it replicates 
many observations obtained during experiments on the cochleas of guinea pigs. 
The model affords a qualitative as well as quantitative approach to the study 
of the direction, magnitude, and source of electric currents flowing through the 
inner ear dirring acoustic stimixlation. A series of experimental investigations 
on optokinetic nystagmus revealed that the oculomotor responses to optokinetic 
stimulation of rabbits, cats and man were qualitatively similar when studied 
by electronystagmographic methods. There were, however, some noticeable 
quantitative differences. The frequency of nystagmus increased as the velocity 
of the stimulus was made greater. For comparable velocities, the frequency of 
the nystagmus in the rabbit is smaller than that of the cat and of the "stare" 
nystagmus in man, but is larger than that of the "look" nystagmus in man. The 
greater effectiveness of the optokinetic stimulus when presented monocularly 
and moving in the usual field from lateral to medial direction depends on the 
existence of a large number of ganglion cells in the retina that are pre- 
ferentially stimulated by visual targets moving in that direction. 

Another group of investigators described the discharge characteristics 
of peripheral vestibular neurons innervating each of the semicircular 
canals in the squirrel monkey. The average resting discharge was about 90 
spikes/second. All neurons responded to angular accelerations in one direction 
with an increase in the discharge and to oppositely directed accelerations 
with a decrease. The response was always consistent with the morphological 
polarization of the hair cells. Many units did not adapt and their response to 
constant accelerations resembled the response predicted by the torsion-pendiilum 
model. This and two other studies on the physiology of peripheral neurons 
innervating the otolithic organs of the squirrel ;3Jonkey produced findings that 



reinforced to a certain extent the current theory on the mechanics of the 
semicircular canals (overdamped torsion-pendixlum model) and also provided a 
basis supporting various observations in m.an and animals^ which deviated or 
are not predicted from theoretical considerations. Further, these studies 
may help to interpret the response of semicircular canals when exposed to 
rotatory or caloric stimiolation as practiced in the clinic. 

Cochlear Mechanisms 

The cochlear potentials (coclilear microphonics, summating potential and 
endocochlear potential) as recorded inside the cochlear duct showed much 
less variability than when these potentials were recorded by one investigator 
in the perilymphatic spaces. The magnitude of the cochlear microphonics 
recorded in the same place in the cochlear duct showed a range of less than 
+ 3 d^. around the mean. At the same time the magnitude of the cochlear 
microphonic for different species varied as a function of frequency in a 
manner that is different in different animals. Cats showed the greatest 
resolution in their tuning curve. The summating potential behaved similarly 
among different animals of the same species, but also showed quantitative 
differences among species. 

Another investigator working on cochlear hair cell metabolism and 
cochlear potentials found that scala tympani resting pressirre in the 
anesthetized guinea pig was usually slightly higher than atmospheric pressirre, 
the greatest value measured being 10 mm Hg. with an average of 6 mm Hg. The 
scala typmani pressure rose markedly during breathing of low oxygen concen- 
trations and the scala vestibuli pressure changed in the -same direction as 
scala tympani pressure diiring anoxemia. Finally, the scala tympani pressiire 
during anoxemia followed a rise of cerebral spinal fluid pressure. It was 
also noted that intravenously administered dinitrophenol caused an increase of 
cochlear endolymphatic dc potential, in some cases 15 to 20 percent. 

Inner Ear 

A group involved in microscopic studies of the inner ear reported 
pathological changes produced by age, disease, drugs, and noise. They found 
that interest tends to focus more and more on tissues, such as the stria 
vascularis and spiral ligament, that provide by their metabolic activity the 
special homeostatic controls that the organ of corti requires for its func- 
tioning, and the blood vessels supplying those tissues. The degenerative changes 
that occur in Corti 's organ itself tend to follow a more or less stereotyped 
pattern, regardless of their cause, and the sequence of changes, especially as 
they affect the hair cells, is now well established. In guinea pigs treated 
with Kanamycin they found there is an increase in the niimber of avascular channels 
of the spiral ligament above Reissner's membrane, indicating a loss of \ 

capillaries. Kanamycin causes not only loss of capillaries but changes in the 
cells of the stria, especially the intermediate or chromophobe cells. Exposure 
to white noise for 6 to 30 hours at 120 dB. produces capillary vasoconstriction 
in the spiral ligament and in the vas spiral system, as well as vacuolization 
of Reissner's membrane. The hypothesis is that capillary vasoconstriction 
is the primary result of noise exposure associated with the temporary elevation 
of auditory threshold. Similar vascular changes are caused by quinine and 



k&v 



salicylates. If long continued, the ischemia causes hair cell loss and per- 
manent threshold shifts. 

Another group engaged in the chronological development and accumulation 
of acid mucopolysaccharide and various enzymes in the endochrondrol model 
Gi rodent temporal bones spent this year working out techniques which will 
allow them to assess more critically acid mucopolysaccharides, phosphatases, 
phosphorylase and non-specific esterase in the otic capsule. The specific 
activity of these enzymes are also being investigated with controlled chemical 
procedures including colorimetric determination of the various enzyme systems. 

Hearing Disorders 

Investigators concerned with determining the characteristics of very 
low frequency auditory sensitivity observed that the rate at which auditory 
sensitivity (as determined by microphonic potentials) decreases with the 
decrease of stimulus frequency below approximately 100 Hz is apparently species 
dependent. A combined anatomical and electrophysiological study was completed on 
four species: cat, guinea pig, chinchilla, and kangaroo rat. It was demonstrated 
that the sensitivity change below 100 Hz is 12 dB. /octave for cat and chinchilla, 
while it is only 6 dB. /octave for guinea pig and kangaroo rat. Concomitant low 
frequency cochlear microphonic phase differences were also seen. They have 
shown that it is the size of the heliotrema that is primarily responsible for 
the differences. The area of this opening is approximately ten times greater 
in the cat and chinchilla than in the other two species. 

Comparative Hearing 

A group of investigators working on the problem of sound conduction in the 
ear foimd that some problems could be solved by studying certain species of 
lizards. Previously, they found that some chameleons, which lack an external 
ear opening and lack the tympanic membrane, hear rather well. Some possess a 
substitute tympanic membrane in the form of a thin plate of bone (a modified 
part of the pteryoid bone). Recently, two additional species have been in- 
vestigated; one ( Chamaeleo dilepis ) that has this substitute mechanism, and 
another ( Chamaeleo ellioti ) that lacks it. The hearing was found to be 
significantly better in C. dilepis . A second form of substitute for the 
tympanic membrane was encountered in Cophosourus texona . It consists of an 
air sac within the middle ear cavity with which the columella is in contact. 
This ear performs remarkable well in sound reception. A problem of central 
interest is the manner of stim\ilation of the hair cells, and the relative 
effectiveness of different structures and modes of operation. In a study of the 
basilar papilla of the crocodilian, Caniman crocodilus , researchers found 
that the papilla differs from that known in other reptiles and in vertebrates 
in general. The general morphology of the papilla bears some resemblances 
to that in birds, but the structure of the short hair cells and supporting 
cells appears to be unique among vertebrates in which the fine structure of the 
ear is known. The Chamaeleontid shows papillar structure suggestive of a 
cytologically unspecialized condition, somewhat reminiscent of that in tiortles. 



49W 



Echolocation 

Investigators are pursuing the design of echolocation in hats and the 
processing of acoustic information hy the bat brain by analyses of the 
"sonar" output of several genera of hats - Hyposideros , Mo r mo ops , 
Chilonycteris , and others; some during goal directed flight and electro- 
physiological recording from the cochlea, auditory nerve, and inferior 
colliculus. Bats using pulses of long duration, Chilonycteris parnellii and 
Rhinolophic , sire now kno-^m to use pulse duration, in effect, to separate two 
channels of information arriving via echo. The constant frequency portion 
is processed for some 25 to 40 msec, and presumably yields velocity information 
via Doppler shift while FM sweeps are actively held off until later to be 
processed separately, probably for position determination. The loltimate goal 
will be to understand how signals of given pitch, loudness, and timing can 
be processed by the brain into a form that reveals the position, velocity and 
the nature of objects in the surroundings. 

Another group working in tissue transmission found that a topography of 
sound transmission velocities exists in the porpoise forehead that causes 
a focussing of sound within the fatty melon, either upon transmission or 
reception. Sounds transmitted into the fatty melon tend to be focussed very 
strongly into the right nasal plug lip. In general, velocities are slower 
than in either fresh or sea water except near the plug. In the sperm whale, the 
entire spermaceti organ is filled with waxy oil that transmits much faster than 
sea water and which is probably the horaologue of the area adjacent to the 
right nasal plug. 

Noise Induced Deafness 

Interaural alternated speech with and without intervening noise a.nd 
interrupted speech was presented to subjects with normal and impaired hearing. 
The results were found by a group of researchers to indicate little effect on 
speech discrimination among normal listeners for interaural alternated speech 
until intervening noise is introduced. Although there were marked individual 
differences in the presence of intervening noise, the difficiilty in discrim- 
inating speech increases as the rate of alternation increases. For subjects 
with impaired hearing, the trend indicates that they perform all tasks more 
poorly than normals even though there may be little improvement of speech 
discrimination for continuous speech. Normal and hard of hearing subjects 
make equal loudness balance judgements with various bands of noise at the 
same and different center frequencies. The normal subjects sum loudness over 
a much narrower band width than do subjects with cochlear involvement. In a 
study on perception of complex auditory stim-uli by the deaf it was found that 
Goarticulation of the consonants and vowels of speech produces transitions in the 
frequency location of the vowel formants. The results suggest that sensorineural 
damage does not necessarily impair a persons ability to use brief, small trans- 
itions in formant frequency as cues for discrimination. It does seem to reduce 
the ability to find cues when they occur in a speech-like environment. 

Speech 

A group of researchers has been investigatirg the efficiency of function 



50 w 



of the reinnervated canine larynx and on in vitro and in vivo tests of their 
antilymphocyte serum. In addition, they have pursued other means of improving 
the chances of success after transplantation of the canine larynx by removing 
the offending cell, i.e., the small lymphocyte, and of accelerating rein- 
nervation. They axe making a serious attempt to tissue-type their dogs before 
transplantation of the larynx. Heretofore, no effort vas made to match animals 
in regard to histocompatibility of antigens. They are using antilymphocyte 
serum (ALS) as the immunosuppressive agent of choice in their laryngeal 
transplantation. The problem of voice function of a transplanted larynx is 
closely related to that of voice function of a reinnervated larynx. 

Another group found that in humans, when background talk reaches a level 
where it is just mildly disruptive to intelligibility for the norm.al hearer, 
it can be a serious masker for the individual with sensorineural hearing loss. 
They showed that competing speech causes a pathological increment in masking, 
and suggest that the traditional measurements of hearing; loss, such as thresh- 
hold shift and discrimination loss, as defined by reduced intelligibility 
in quiet, should be supplemented with specification of the increase in the 
masking efficiency of competing speech and other background sounds. In a 
test of 2U subjects, measuring nonaurally discrimination of monosyllables 
against competing sentences, findings showed that a third dimension of handi- 
cap is imposed by sensorineural pathology. Such pathology not only changes 
threshold and impairs intelligibility in quiet, but also disturbes the ability 
to resist masking in complex environments containing background noise, 
particularly speech. 

Olfactory Communication 

A team investigated the olfactory connections of the limbic system and hy- 
pothalamus in the rodent. The tertiary olfactory contribution to a myelinated 
bundle in the lateral hypothalamus previously identified in the rat has now been 
found in both hamster and mouse. In all three species, lesions which Include 
the olfactory tubercle cause degeneration of fibers which travel in the far 
lateral region of the medial forebrain bundle to the posterior hypothalamus 
border of the midbrain. In the rat, many regions of the olfactory cortex project 
to the lateral preoptic area, mediodorsal nucleus of the thalmus, and lateral 
hypothalamus. Other secondary olfactory zones (medial and cortical amygdaloid 
nuclei) do not show this pattern of projection. Lesions of the medial nucleus 
cause degeneration of fibers reaching the bed nucleus of the stria terminalis 
through its post commissural component. Other work included electrophysiological 
evaluation on hormone effects with urethane-anesthetized male rats. Comparing 
recorded results from nonnal and castrated male rats to study the effects of 
testicular androgens on the activity of neurons in the preoptic area, a 
hypothalamic region which receives a strong olfactory input, it was found that 
some units in the preoptic region reliably change their ongoing discharge rate 
when the cortical EEG shows sudden transitions between activation and synchrony 
characteristic of the urethanized rat preparation. Many more neurons in the 
normal male than in the castrate show this correlation with the EEG. Preoptic 
units which increase their discharge rate during EEG activation tend to be 
those which show excitatory responses to odors and tend to change activity be- 
fore the EEG change in transition from activationary to synchrony but not vice 
versa. 



51" 



Sonic Booms 

Sonic booms may damage the apical turn of the cochlea causing weakening 
which in time leads to hearing loss according to another team of investigators. 
Guinea pigs were exposed to 1,000 bursts of 130 dB. lasting 2., h, 5, or 125 
msec, and were then retested for the Preyer reflex. Histological examinations 
revealed damage to hair cells of the apical turn of all test animals. 

Otitis Media 

Incidence of deafness, otitis media, and perforation of the eardrum in 
Guam populations is the highest reported anywhere. In school children hearing 
loss was found to be four times that in most Worth American cities. Selective 
Service examinations of young men in Guam showed similar incidence rates. Per- 
foration of the eardrum accounted for 10^ of Guam young men being disqualified 
for military service as compared with .Olfo of the Selective Service population 
of the U.S. 

Rubella 

Another research team reported that many pre-school children may have had 
congenital rubella and are likely to remain undiagnosed until a specific com- 
plaint develops, such as a language or hearing defect. The findings indicate 
that such conditions are recognized too often only after severe academic 
retardation and emotional suffering have occurred. Therefore, they suggest a two 
-part screening program as a model for identifying children with unrecognized 
communication problems so that treatment or special training can begin as early as 
possible. This team cooperated with Montgomery County Hesilth Department in a 
program in which I36 children were located by radio, TV, newspaper notices and 
letters to the locai medical society. Children with possible communication 
disorders were given a clinical examination which included evaluations of growth, 
speech, language, hearing, vision, and auditory memory. Blood specimens were 
obtained from 11 i+ children and rubella hemogglutination inhibition (H. I. ) 
antibody titrations were performed on the sera. 

Aphasia 

In an out-patient clinical research facility for aphasic involvements 
of children, studies on visual sequencing performance demonstrated that 
aphasic children are generally inferior to normal children on various types 
of sequencing tasks. A modality study by the same group of investigators has 
produced preliminary data which indicates that aphasic children make more errors 
in judging non-equivalent forms than equivalent form.s, and they also had longer 
latencies for an intramodal haptic condition than for an intramodal vis\ial i 
condition. Another continuing study in this out-patient facility is a series ' 
of directionality investigations progressing from gross visual forms to 
discrimination of letters such as p and b, b and d. Findings show that aphasic 
children who failed directional discrimination on the initial pre-test, were 
able to complete this task after they had completed the program. Studies are 
continuing to determine which of the tasks in the progression of discrimination 
difficulty are essential for improved performance. 



52W 



Another group of investigators has been working in a multidisciplinary 
effort on aphasia and related problems of "brain function. They have reported 
that diagnosis and lesion site have a predictable relationship to pattern 
of work comprehension and production, with respect to semantic class, word 
frequency, and picturability. Scaling of syntactic tasks reflecting levels of 
agrammatism is currently in progress by this group. In this area of non- 
linguistic ne-uropsychological research, these investigators have presented 
evidence that the left parietal lobe is found to be more critical than the 
ritjht in cross-modal transfer, but less critical for short term visual memory. 
Neirroanatomical studies have suggested an anatomical basis for cerebral 
dominance based on the larger average size on the left than on the right planum 
temporal in man. This same team of investigators has demonstrated that aphasic 
patients not only vary in their ability to comprehend what they hear, but do so 
according to at least four distinct patterns of auditory comprehension. They 
showed that five diagnostic classes of aphasic patients differed in these 
factors: (l) breadth of vocabulary; (2) auditory sequential pointing -span; 
(3) comprehension of directional prepositions; (k) recognition of correct 
grammatical usage of prepositions. 



53w 



AFFEEDIX A 
RESEARCH GRANTS AWARDED IN FY 19T1 BY DISORDER CATEGORY 
(Dollars in Tliousands) 

DISORDER CATEGORY N0_; AMOUNT jo of $ 

TOTAL ALL DISORDERS I256 $53,6U5 100.0 

1. NEUROLOGICAL DISORDERS 

A. Neurological Disorders of 172 $ 6,200 11.6 
Early Life 

B. Neurological Disorders of Aging 59 2,810 5.2 

C. Cerebrovascular Disorders 80 5^350 10.0 

D. Epilepsy and Related Paroxysmal 52 2,500 h.G 
Disorders 

E. Sclerosing Disorders 63 2, 36O 4.U 

F. Muscular and Neuromuscular Disorders 1^^ ^,690 8.7 

G. Infectious Diseases 10 25O O.5 



H. Trauma and Injury 68 3,120 5.1 

J. Tumors of Nervous System 26 590 1.1 

M. Neuroendocrine Studies 86 3;,050 5.7 

W. Neural Aspects of Learning and ^3 1^690 3.2 
Behavior 

F. Nervous System Studies - li<-0 5,0U0 9.h 
Normal Function 

TOTAL - NEUROLOGICAL DISORDERS 9il3 $ 37,650 70.2 



59 


2,810 


80 


5,350 


52 


2,500 


63 


2,360 


ihk 


i+,690 


10 


250 


68 


3.120 


26 


590 


86 


3,050 


^3 


1,690 


li^O 


5,0U0 


9^3 


$ 37,650 



52j.w 



APPEEDIX A 
RESEARCH GRANTS AWARDED IR FY 1971 BY DISORDER CATEGORY (Contd.) 
(Dollars in Thousands) 
DISORDERS CATEGORY NO. ^AMOUNT jo of $ 



2. SENSORY AND PERCEPTUAL DISORDERS 

A, Disorders of Hearing and 
Equilibrium 

B. Disorders of Speech and Other 
Higher CWS Functions 



li+5 
36 



C. Disorders of Other Senses IO8 

TOTAL - SENSORY & PERCEPTUAL DISORDERS 289 



$ 6,605 

1,890 

3.750 
12, 2U5 



12.3 
3.5 

7.0 

22.8 



3. MULTI-CATEGORICAL 



20 $ 3, 660 



6.8 



h, COKFERENCES 



90 



0.2 



55w 



ANNUAL REPORT 
July 1, 1970 through June 30, 1971 

Extramural Programs 

Training Grants and Awards Branch 

National Institute of Neurological Diseases and Stroke 

The primary aim of the Training Grants and Awards Branch is the specialized 
training of skilled professional and scientific personnel for careers in the 
research and teaching aspects of the prevention, diagnosis, and treatment 
of neurological and communicative impairments. To accomplish the Institute's 
objective, two general types of awards are made, training grants and fellow- 
ships. Included in the Training Grants category are (1) awards to superior 
training institutions; (2) awards to institutions which wish to develop 
programs or strengthen existing weak programs; (3) Special Traineeships ; and 
(4) Teacher-Investigator Special Traineeships. Included in the Fellowships 
category are (1) Postdoctoral Fellowships; (2) Research Career Development 
Awards; and (3) Research Career Awards. The methods of applying for support 
and the review of applications vary among the types of programs. Institutions 
apply for Training Grants and Developmental Training Grants; institutions 
also apply for Research Career Development Awards, except they apply for 
these awards on behalf of a specific candidate. (New Research Career Awards 
are no longer being made.) Special Traineeships, Teacher-Investigator 
Special Traineeships, and Postdoctoral Fellowships, are awards made directly 
to individuals. 

To support the NINDS training activities in FY' 71, the House-Senate Conference 
Committee recommended an allowance of $17,754 million which became the 
appropriation. However, only $17,082 million were allocated to the Institute 
which provided $14.3 million for Training Grants and Special Traineeships 
and $2,782 million for RCDA' s and Postdoctoral Fellowships. The training 
grant funds enabled the Institute to support 219 training programs and 
201 special trainees; the fellowship funds were used to continue 12 Research 
Career Awards and to award 76 Research Career Development Awards and 91 
postdoctoral fellowships. 

In FY' 71, the Institute made training grant awards to 55 institutions that 
submitted renewal applications and to 9 institutions that submitted new 
applications. In view of the large number (27) of renewal applications that 
were not funded the previous year and the fact that no new awards were made, 
it was anticipated that a number of institutions would submit amended 
applications. In this regard, of the 55 renewal applications that were 
funded, 11 were amended applications and of the 9 new applications that 
were funded, two were amended applications. Thus, it was advantageous to 13 
program directors who had previously approved programs to submit amended 
applications. Four other amended renewal applications were recommended for 
approval but again were not funded because the merit ratings they received 
were too low. 

Following the March meeting of the Council, when recommendations were made 
concerning the funding of new and renewal applications, ten programs which 

57w 



had been supported for a number of years had merit ratings too low to fund. 
These have been either already terminated or have been awarded phase-out 
support. Eight of these programs were in neurology and two were in communica- 
tive disorders. In FY' 68, the latest year for which information is available, 
these ten programs provided full support to 36 trainees and partial support 
to 20 trainees. In addition, 16 individuals were supported for a two or 
three month period in order to give them an exposure to neurology or the 
communicative disorders. The following table identifies the institutions 
which were recommended for approval in Fy'71 but not funded and it indicates 
the number of trainees in FY' 68 who received full, partial, or short-term 
stipend support from grant funds: 



Grant Number Institution 

TOl NS 5049 Northwestern University 
TOl NS 5099 University of Miami 
TOl NS 5109 State University of New 

York, Downs tate 
TOl NS 5166 Wayne State 
TOl NS 5182 Mayo Foundation 
TOl NS 5201 University of Miami 
TOl NS 5262 University of Illinois 5 
TOl NS 5409 University of North 

Carolina 
TOl NS 5573 Mt. Sinai School of 

Medicine 1 

TOl NS 5574 Purdue University 6 



Trainees Stipended 



Full 


Partial 


Short-term 


4 


1 


2 


- 


14 


- 


_ 


_ 


2 


6 


- 


2 


13 


4 


8 


1 


- 


- 



36 20 16 

RESEARCH CAREER DEVELOPMENT AWARDS 

Having made no new or renewal Research Career Development Awards in FY' 70, 
FY' 71 was a banner year for this program. Of 36 applications recommended 
for approval, 22 were funded. With only one exception, these 22 applications 
were all new; nine were amended applications submitted in behalf of individuals 
for whom support was sought the previous year. 

In evaluating applications for renewal support, grantee institutions were 
asked to provide a special justification for needing additional support 
for the candidate. In addition, they were asked to (1) indicate their 
permanent plans for the candidates; (2) state specific accomplishments of 
the awardees during the initial award period; and (3) signify the need for 
additional periods of training and supervised experiences. For the most 
part, the institutions did not make strong cases for needing additional 
support and the general philosophy of both the training committees and the 
Council was to not recommend continued support for individuals who had 
already received five years of RCDA support unless special justification 
was provided. These candidates were considered independent investigators 
and as such they had achieved the objective for which the award was initially 
made . 



58 w 



New guidelines adopted this year, governing the Research Career Development 
Award program, provide that any application terminating after June 30, 1972, 
will not be eligible for renewal support. Thus, the RCDA becomes a single 
five-year award with no possiblity of being renewed. Another change concerns 
the evaluation of the applications. The initial review will be made by the 
appropriate DRG Study Section. The National Advisory Council has requested 
that all applications also be reviewed by an NINDS training review committee. 

TEACHER-INVESTIGATOR SPECIAL TRAINEESHIP PROGRAM 

FY' 71 was the second year of the Teacher-Investigator Special Traineeship 
program. This program aims to recruit and prepare future teacher-investigators 
of the highest caliber for academic careers in disciplines or areas of the 
neurological, neurosensory, or communicative disorders. The award is the 
most competitive and prestigious training and development award made by the 
Institute to an individual. 

During FY' 71, 19 teacher-investigator applications were submitted. Fifteen 
of the applicants were interested in the neurological sciences and four were 
interested in communicative disorders. Eleven applicants were invited to 
Bethesda on February 27 to be interviewed by an NINDS Special Ad Hoc 
Interview Committee. 

The following individuals were selected to receive Teacher-Investigator Awards: 



AWARDEE 



SPONSOR AND 
INSTITUTION 



DISCIPLINE 



Irving K. Arenberg, M.D. 



Richard Torack, M.D. 
Professor, Pathology and 
Anatomy 
Department of Pathology 
Washington University 
School of Medicine 
St. Louis, Missouri 



Oto- 

neuropathology 



Ira B. Black, M.D. 



Fred Plxjm, M.D. Neurobiology 

Professor and Chairman 

Department of Neurology 

Cornell University Medical College 

New York, New York 



Mary A. Guggenheim, M.D. 



C. Henry Kempe, M.D. Neurology- 

Professor and Chairman Virology 

Department of Pediatrics 

and 
James Austin, M.D. 
Professor and Chairman 
Department of Neurology 
University of Colorado Medical Center 
Denver, Colorado 



59 w 



Mark E. Molliver, M.D. 



Donald J. Woodward, Ph.D. 



David Bodian, M.D. 
Professor of Anatomy 

and 
Guy M. McKhann, M.D. 
Professor of Neurology 
Johns Hopkins University 
School of Medicine 
Baltimore, Maryland 

Paul Horowicz, Ph.D. 
Chairman, Department of 

Physiology 
University of Rochester 
School of Medicine 
Rochester, New York 



Child Neurology 

Neuroanatomy 



Developmental 
Neurobiology 



D. C. UNIVERSITY CONSORTIUM: ACADEMIC NEUROSURGERY 

During FY' 71, the staff held meetings with neurosurgery representatives of 
the D. C, universities and other neurosurgeons in an attempt to stimulate 
the development of a Ph.D. postdoctoral program in neurosurgery as a 
multi-university Consortium undertaking. The organizational unit for the 
program would be the D. C. Consortium and the objective of the program would 
be to develop a program in the Washington area utilizing several of the 
local facilities to train neurosurgeons for careers in research and academic 
medicine. The meetings were attended by the following individuals: 

Dr. Jesse B. Barber, Jr. 
Howard University 

Dr. Calvin Early 

National Naval Medical Center 

Dr. Ludwig Kempe 

Walter Reed Army Medical Center 

Dr. John Luessenhop 
Georgetown University 

Dr. Hugo V. Rizzoli 

George Washington University 

Dr» John M. Van Buren 
National Institute of Neurological 
Diseases and Stroke 

The advantage of utilizing the Consortium is that the administrative unit 
already exists and any facility in the Washington area could be utilized 
to provide for a training program which would be unique to the specific 
needs of the trainees. 



60 1 



It is anticipated that a formal application requesting support for this new 
program will be submitted prior to October 1, 1971 

FRONTIERS IN RESEARCH IN TEACHING 
IN NEUROSCIENCE: A MINORITY TRAINING PROGRAM 

In FY' 71, the Institute made an award to the Marine Biological Laboratory, 
Woods Hole, to advance training in the neurosciences primarily for minority 
group individuals at the postdoctoral level who are seeking opportunities 
for further experience in research and who desire to strengthen their 
capacities as teachers of future neuro-scientists. The program is designed 
to enable trainees to carry out a personalized program in one or more areas 
of neurobiology. 

An award to the MBL provided support for approximately 6 trainees. Although 
these individuals would be selected on a competitive basis, it was agreed 
that for this pilot effort the program would be experimental in design and 
it would be generally publicized with specific publicity aimed at institutions 
that are composed mainly of minority ethnic groups. 

Should the project be successful during the pilot period, the Marine 
Biological Laboratory would submit an application for a similar grant 
for long term support. 

STATUS OF OTOLARYNGOLOGY TRAINING PROGRAMS IN THE UNITED STATES - 1969 

During FY' 71 a report was prepared by the Committee on Education, Society 
of University Otolaryngologists, on the status of otolaryngology training 
programs in the United States in 1969. The data for this report were 
collected by Dr. Dean Lierle and the study was supported by an NINDS grant. 

The survey included 103 programs which were approved by the American Medical 
Association and the Review Committee for Otolaryngology. Sixty-nine programs 
were in university teaching centers; of these, 36 were independent departments 
while 33 were sections of general surgery. Thirty-four were in government 
or private institutions. 

The report calls attention to the critical manpower situation. In 1969, 
there were 253 first-year residency positions whereas there was a need 
for a minimum of 500 such positions. There should be one certified 
otolaryngologist for every 40,000 people in the United States and at 
least 500 residency positions should be available to develop adequate 
health care for the public, and provide the teacher-investigators required 
for academic positions. 

The formation of new medical schools (eight in 1969) added a few residency 
positions, but not nearly enough to keep pace with increased needs. The 
manpower situation cannot improve unless there is an expansion of many of 
the residency programs. In addition to the lack of professional staff there 
is also a lack of technicians in otolaryngology. The report points out that 
paramedical personnel, properly trained, could be of value in lessening the 
workload of faculty and residents. There appeared to be a fairly good supply 
of audiologists and speech pathologists. 

6iw 



In the 103 institutions surveyed, a minimum of 150 more full-time teachers 
are needed to improve the quality of teaching and properly supervise the 
clinical and research areas. It is practically impossible for one full- 
time staff member to attempt to administer, teach, and conduct research, 
even in the smallest department of otolaryngology. 

The report concludes b)' making the following recommendations to alleviate 
the manpower situation and improve the quality of otolaryngology training 
in the United States: 

1. Increase the number of beds, the budgets and the faculty, in 
otolaryngology, particulary in smaller departments, thus providing more 
residency positions and improving the quality of teaching. 

2. Utilize many available, competent young private practitioners in 
areas adjacent to medical school and hospitals. These young men could 
contribute greatly as part-time teachers and should be well compensated 
by the institutions. 

3. Train more paramedical personnel in otolaryngology to help 
relieve the work load of residents and staff. 

4. Make all departments and divisions of otolarjmgology independent, 
particularly for budgets, in-patient beds, staff, access to the dean's 
office, and membership on medical school committees. 

5. Require clinical clerkships for undergraduate medical students 
for a minimum period of two weeks; also, time should be provided for 
electives in otolaryngology. 

6. Consider postgraduate courses designed particularly for family 
physicians, pediatricians, and other disciplines. 

7. Provide more continuation courses for otolaryngologists in 
private practice. 

NEUROLOGY: A MEDICAL DISCIPLINE TAKES STOCK 

In 1962 the Institute awarded a contract to Columbia University to evaluate 
the status of training for research and service in the neurological sciences 
and to formulate recommendations for strengthening and improving university 
and non-university activities related to such training. The study was 
carried out by Dr. Aura Severinghaus and during the period of the study 
the following three publications by Dr. Severinghaus appeared: 

Distribution of Graduates of Medical 
Schools in the United States and Canada 
According to Specialties, 1900-1964. 
J. Med. E., 40:721-736, 1965. 

A Medical Discipline Takes Stock. Arch. 
. Neurol., 17:461-470, 1967 

62w 



Neurology and Neurological Sciences 
Research and Training Study. Arch, 
Neurol., 17:471-483, 1967. 

Dr. Severinghaus' s study has been completed and the Institute has received 
his final manuscript which is being put into final form for consideration 
for possible publication. 



APPENDICES 

Following are four appendices which show how support for the Institute's 
training programs is divided among the various training areas . 

Appendix A is the anticipated number of grants and the training 
grant funds awarded for FY' 71 according to the Institute's areas of 
responsibility. This is an increase of 1 award over FY' 70. 

Appendix B is the anticipated number of Special Traineeship Awards 
and the amount for FY'71 divided among the Institute's areas of responsibility. 
This is an increase of 40 awards over FY' 70. 

Appendix C is the distribution of nine Teacher-Investigator Special 
Traineeships among the Institute's areas of responsibility. This is an 
increase of four awards over FY' 70, the initial year of this training 
activity. 

Appendix D is the anticipated number and the amount awarded for 
RCA's, RCDA's, and Postdoctoral Fellowships in FY'71. This is an increase 
of five RCDA's and 19 Postdoctoral Fellowships over FY' 70. The number of 
RCA's (12) remains unchanged from FY' 70. 



63'' 



APPENDIX A 



Distribution, by Scientific Fields, 
of Training Grants Awarded in FY 1971 



Field 



Number 



Audio logy 


6 


Cerebrovascular 


2 


Child Neurology 


15 


Communicative Disorders 


7 


Neuroanatomy 


5 


Neurobiology 


1 


Neurochemis try 


3 


Neurological Sciences 


6 


Neurology 


59 


Neuropathology 


14 


Neuropharmaco logy 


3 


Neurophys io logy 


13 


Neur or ad io logy 


10 


Neurosurgery 


24 


Neurovirology 


1 


Otolaryngology 


44 


Sensory Physiology 


3 


Speech Pathology 


3 



$ 



Amount 

368,700 

68,600 

613,900 

533 , 100 

151,200 

20,800 

102,000 

225,400 

4,197,800 

513,900 

166 , 100 

731,700 

250,300 

917,400 

47,500 

2,680,400 

128,300 

162,900 



TOTAL 



219 



$11,880,000 



April 29, 1971 



Gk 



APPENDIX B 



Distribution, by Scientific Fields, 
of Special Traineeships Awarded in FY 1971 



Field 



Number 



Audiology 


5 


Basic Neurosciences 


3 


Biochemistry 


8 


Biophysics 


1 


Cerebrovascular 


10 


Child Neurology 


46 


Communicative Disorders 


1 


Immunology 


3 


Neuroanatomy 


3 


Neurobiology 


6 


Neurochemis try 


6 


Neuroendocrinology 


5 


Neurology 


9 


Neuropathology 


16 


Neuropharmaco logy 


6 


Neurophy s io logy 


29 


Neuroradiology 


17 


Neurosurgery 


5 


Neurovirology 


4 


Oto laryngo logy 


4 


Sensory Physiology 


3 


Speech Pathology 


2 



Amount 

53,500 

48,400 

81,200 

16,700 

106,900 

500,600 

16 , 100 

35,600 

31,300 

74,700 

65,100 

49,000 

114,900 

171,700 

63,200 

293,700 

198,900 

41,600 

44,900 

47,500 

37,200 

28 , 100 



TOTAL 



192 



$2,120,800 



April 29, 1971 



65 1 



APPENDIX C 

Distribution, by Scientific Fields, 
of Teacher-Investigator Awards Granted in FY 1971 



Field 



Nvunber 



Amount 



Neuroanatomy 
Neurobiology 
Neurology 
Neurophys io logy 
Neurosurgery 
Neurovirology 
Sensory Physiology 
TOTAL 



19,500 
15,500 
35,000 
35,000 
22,200 
21,500 
15,500 
$164,200 



66 w 



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