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Full text of "Report of program activities : National Institute of Neurological Diseases and Stroke"

Library, .'Acquisitions Unit 
National Institutes of Health 
Building 10 
Bethesda, Maryland 20014 



ANNUAL REPORT 
OF 
PROGRMyi ACTIVITIES 
NATIONAL INSTITUTE OF NEUROLOGICAL DISEASES AND STROKE 

Fiscal Year 1974 
Volume II 



ANNUAL REPORT 
July 1, 1973 through June 30, 197^ 
Associate Director, Collahorative and Field Research 
National Institute of Neurological Diseases and Stroke 
National Institute of Health 

The goal of the Collaborative and Field Research (C&FR) program of NINDS 
is to conduct directed research of importance and relevance in the fields 
of neurological diseases and communicative disorders. This nrogram attempts 
to fill the gaps in research areas of importance to NINDS which are not 
covered by either the intramural or extramural programs of the Institute. 
The major part of this program is implemented chiefly by means of contracts 
plus some in-house effort. 

For the last year, C&FR has been heavily involved in planning all of its 
programs in an orderly fashion. Each branch and section has prepared 5 year 
programs. The program planning is a result of an internal planning aided 
and abetted by a number of outside advisory subcommittees for each major 
program area. Because of the pressure of other work it was not possible to 
start this program planning cycle on July 1, 1973- The planning cycle did 
not really start until October 1973 which caused the formulation of these 
plans to appear later in the year than anticipated. However, in spite of 
the late start, we were able to go through most of the essential steps 
which we deemed necessary for adequate planning. 

For example, we used some of the advisory subcommittees already on hand and 
established new subcommittees during the year. All of these subcommittees 
met at least once during the fiscal year. The three committees which were 
already ongoing were the Epilepsy Advisory Committee and the Science Infor- 
mation Committee (SIPAC), and the Committee on Data Analyses. The new 
subcommittees which were established and met during the year were Head Injury 
and Stroke, Infectious Diseases and Epidemiology, Communicative Disorders, 
and Biomedical Engineering and Instrumentation. 

An advisory committee for C&FR was established and met once during the year. 
This overall committee was composed of a chairman (Dr. Arthur Ward) and the 
chairmen of seven subcommittees. This committee appeared to be a valuable 
adjimct to our planning process and consequently we made a request to NIH 
and the Secretary of HEW to constitute this as a formal committee. It appears 
at this time that this committee will be approved by the Secretary of HEW. 

As a result of this planning process, a document was prepared and presented 
to the Director/NINDS on April 12, 197^- The program presented two budgets 
i. e., a core and optimum budget. A request was made to the Director for 
tentative approval of the core budget for FY 1975- This approval was 
received. 

This approval gives C&FR the ability to start planning its FY 1975 contract 
program early in the fiscal year. This should overcome the tremendous 
burden of reviewing and negotiating all contracts in the last half of the 
fiscal year. 

Is 



We are planning to start our planning for FY 1976 in July 197I+ and hope to 
"be able to be within a time frame to present the FY 1976 program and budget 
to the Director/NINDS by January 1975- 

In the past, our contract proposal reviews and negotiations have been 
carried on exclusively by our administrative officers. This has not been a 
satisfactory procedure because administrative officers have been carrying on 
a dual role and consequently neither of their jobs were receiving adequate 
treatment. In addition, since most of our administrative officers do not 
have scientific background, they were at a distinct disadvantage in 
conducting adequate technical merit reviews of proposals for contracts. 

In order to provide contracts of the highest quality possible, it was deemed 
necessary to establish a Research Contracts Office which would have both 
personnel with scientific background and those with administrative capabil- 
ities in the contract area. This office would be headed up by a scientific 
individual with management and contract experience. This individual is 
being selected and will be on board at the beginning of the next fiscal year. 
In addition, this office has two sections — one a scientific evaluation 
section and the other an administrative section. The scientific section has 
two Ph.D's with excellent scientific and management talents. They have been 
doing an excellent job. The other section is composed of contract specialists 
who work very closely with the scientific evaluation unit. The Research 
Contrads Office is also heavily involved in the program planning process with 
the various branches and sections. By the end of June 30, 197^, there will 
be a total of II6 active contracts in NINDS as compared with 87 at the end 
of June 30, 1973. These contracts will be distributed in NINDS as follows: 
C&FR - 103, IR - 9, and EP - U . 

Key personnel were added to the C&FR organization during the year. Dr. 
Lois Elliott was recruited from the Bureau of the Handicapped, Office of 
Education to head up the Communicative Disorders Section. In addition 
Dr. Christine Ludlow, a speech pathologist, was also recruited and brought 
on board for the Communicative Disorders Section. 

Dr. Mathilde Solowey and Dr. Howard Weinstein were recruited for the 
Research Contracts Office in the Scientific Evaluation Section. 

Mr. Lawrence Fitzgerald was hired as the Contracting Officer for NINDS and 
heads the Administrative Section of the Research Contracts Office. Mr. 
James VJehling and Miss Linda Waring were hired as contract specialists in the 
Research Contracts Office. 

Some key personnel also left C&FR for other positions. Dr. George Murray 
who headed up the Section on Biomedical Engineering and Instrumentation left 
the Institute during April 197^. Dr. Edgar Bering, who was head of the 
Special Programs Branchy left the Institute for private practice in 
February 197^. 

During the year, the Office of the Associate Director of C&FR was moved 
from Building 36 to Building 31. The Research Contracts Office also was 
moved to Building 31. Plans are afoot to move the Applied Neurologic 
Research Branch from Building 36 to the Federal Building on Wisconsin Avenue 
in Bethesda. 

2 s 



APPLIED NEUROLOGIC RESEARCH BRANCH 

The Applied Neurologic Research Branch is directed by Dr. J. Kiffin Penry. 
It has two sections — one on Epilepsy and one on Head Injury and Stroke. 

Section on Epilepsy 

This year there was a "breakthrough in marketing of new antiepileptic drugs. 
There have been no new antiepileptic drugs marketed in this country since 
i960. However, new drug applications were submitted for approval to the 
Food and Drug Administration by two pharmaceutical companies: One, for 
carbamazepine (Tegretol — Geigy), and the other, clonazepam (Clonopin — Roche). 
If approved by the Food and Drug Administration, these drugs will be the 
first released for epilepsy in ik years, and signal the beginning of a new 
era in therapy for seizures. NINDS sponsored clinical studies were key 
elements in the development of the new drug applications. 

NINDS has committed itself to a program of determining the feasibility of 
developing comprehensive epilepsy programs. Some 11 research contracts 
were awarded to institutions throughout the United States from urban, urban/ 
rural, and rural areas to determine if it is possible to establish compre- 
epilepsy programs providing coordination between patient services and research 
in epilepsy. If determined feasible, the Institute intends to fund at least 
one comprehensive program in the coming fiscal year to serve as a research 
resource for clinical research. Moreover, such a program could serve as a 
model for other chronic diseases which have similar problems as epilepsy in 
difficulty of diagnosis, treatment, rehabilitation and social integration. 

The section also continues to monitor and work cooperatively with others 
in development and testing of drugs of potential value in the treatment of 
epilepsy. 

In December 1973, a Workshop on the Neurosurgical Treatment of Epilepsy was 
held in Bethesda, Maryland. International leaders in the field presented 
papers on ther work which will be published by Raven Press in December 197^- 
The collation of this material will be invaluable to neurologists and others 
in the field, providing information about this mode of treatment for 
appropriate patients with seizures. 

A film. The Absence Seizure, was produced by the Section on Epilepsy to 
present clinical and electroencephalographic characteristics of absence 
seizures. Every type of absence seizure, including the electroenceph- 
alographic patterns, is illustrated by multi-camera video recordings of 
patients with absence seizures and their EEGs . 

The Pharmacology Laboratory has continued its active investigation into the 
metabolism of antiepileptic drugs, and the development of means of identifying 
and quantitating drugs and their metabolities in the blood. 

Section on Head Injury and Stroke 

Dr. G. Molinari,who heads up this section, developed a program on head injury 
and stroke with the aid of his subcommittee made up of experts from outside 
of NIH. ^3 



When Dr. Molinari came on board, he inherited the collaborative study on 
cerebral death. The final report has been submitted and is in the process 
of staff review currently. Later, an Atlas of EEG artifacts encountered 
in this difficult diagnostic category will be published by Raven Press. 
Scientific reports of specific technical aspects and center data reflecting 
specific population characteristics will be offered by individual investi- 
gators of the collaborative group, but publication will depend upon 
scientific merit and editorial policies of the professional journals. 

As a result of the cerebral death study, a new study was generated this 
year to test the validity and reliability of a portable scintillation 
angiography technique by comparison with the only universally accepted 
norm of cerebral blood flow, four- vessel aortocranial contrast angiography. 

The "Cooperative Study of Hospital Frequency and Character of Transient 
Ischemic Attack" continued to access new patients and pursue short term 
(l year) follow-up throughout the fiscal year. As of May 1, 197^, complete 
tabulations and preliminary data analyses have been completed on 878 
patients . 

Other studies initiated in FY 197^ are: (l) a test of the spatial and 
temporal resolution capabilities of computerized x-ray tomography in stroke; 
(a) a study of the feasibility of longitudinal prospective follow-up of 
stroke patients for six months; (3) a study of the prognosis after coma in 
stroke and head injury patients; and (U) a field test of a questionnaire to 
identify TIA in the general (extra institutional) elderly population. 

Remodeling for the stroke laboratory of this section was begun in October 
1973 and will be finished by the end of this fiscal year. 

SECTION OF BIOMEDICAL ENGINEERING AND INSTRUMENTATION 

This section was initiated last year under the direction of Dr. George 
Murray. Dr. Murray left this position in April 197^, for one with the 
National Heart and Lung Institute. The section was directed the remainder 
of the year from the Office of the Associate Director, C&FR. It is charged 
with overall coordination of the development of neurologically useful 
diagnostic and therapeutic technical equipment through the prototype and 
clinical evaluation stages. In-house laboratory research and development is 
combined with collaborative projects undertaken by contract in laboratories 
of other institutions. The section seeks to provide interdisciplinary 
expertise to produce or utilize technical development in the solution of 
clinical problems. 

An advisory subcommittee of experts from outside of NIH was established this 
year. This group met and reviewed the ongoing and proposed work for this 
section. 

A principal activity of the section has been initiating and monitoring 
contracts directed towards noninvasive methods of measuring regional cerebral 
blood flow. The focus in these contracts is mostly on ultrasound techniques. 

Plans are underway to hire new professional personnel for this section early 
in FY 1975. ^3 



SECTION ON COMMUNICATIVE DISORDERS 

In September 1973, Dr. Lois Elliott was recruited from the Bureau of the 
handicapped. Office of Education, to be the head of the 

Disorders Section. This section (ivelops and carries out project directed 
toward the diagnosis and treatment of disorders of hearing, speech, and 
language. Initially, the program emphasized hardware devices to assist the 
deaf (development of wearable master hearing aids and feasibility of cutaneous 
aid for profoundly deaf infants) and procedures to detect hearing impairments 
in infants and young children. In September 1973 with the arrival of a new 
program head, activities expanded into disorders of speech and language and 
problems associated with environmental noise. 

A new staff member. Dr. C. Ludlow, joined the program late in FY 197^ and will 
provide leadership for research on speech and language disorders. 

A subcommittee made up of experts from outside of NIH met once this year 
and reviewed the five year program of this section. 

The five-year program plan for the Communicative Disorders Section contains 
seven program elements as follows: 

1. Early detection of hearing disorders in infants and children. 

2. Prescriptive tailoring of hearing aids. 

3. Speech ana].yzing aid for the deaf. 

h. Speech and language disorders in children. 

5. Adult language disorders. 

6. Effects of noise people. 

7. Information Center for Hearing, Speech, and Disorders of 
Communication. 

The head of this section coordinates research on noise with the Environmental 
Protection Agency and, on an informal basis, has begiin similar coordination 
efforts with the National Institutes of Environmental Health Services and 
National Institutes of Occupational Safety and Health, with a major purpose 
being to prevent duplication of research effort. Through an interagency 
agreement, the Committee on Hearing and Bioacoustics (CHABA) of the National 
Research Counsel-National Academy of Science is assisting NINDS in reviewing 
information on the effects of noise on children. Also, the Committee is 
helping in the examination of possible research strategies for obtaining a 
better understanding of this area. 

EPIDEMIOLOGY BRANCH 

The Epidemiology Branch is headed by Dr. Jacob Brody. It has as its goals 
the identification of patterns of neurologic disease in human population and 
defining the various interactions among the affected persons, etiologic 

5 s 



agents, and environmental setting. In addition, the Branch conducts 
laboratory studies related to the epidemiology and immunology of neurologic 
diseases . 

During the past year, the Subcommittee on Infectious Diseases and Epidemiology 
reviewed that program in depth. A niimher of recommendations were made 
relative to the improvement of the work of this Branch. 

During the year, work on the following contracts were continued: A Study of 
Amyotrophic Lateral Sclerosis in Veterans; A Study of the Risk Factors in 
Stroke; and an Epidemiologic Study of Stroke in Panama. 

A new study relative to Multiple Sclerosis (MS) in the Shetland and Orkney 
Islands where the rate of MS is approximately three times higher than that 
reported anywhere else in the world will be initiated in FY 197'^- 

In September 197^+, a peer review group met for a second time on Guam and 
presented its findings and recommendations to the Directorate of NINDS in 
February 197^- Its recommendations have resulted in our renegotiating the 
Guam Agreement to continue the study for three years after October 197^- 

The rate of ALS on Guam has been declining but no genetic pattern as an 
explanation has been found. Studies on this subject are continuing. 

Early in FY 1975, Dr. Brody will be leaving this Branch to assume duties as 
the Medical Director of the Atomic Bomb Casuality Commission in Japan. 

Plans are underway to revamp the Epidemiology Branch. 

INFECTIOUS DISEASES BRANCH 

Dr. John Sever directs the Infectious Diseases Branch, whose goals are to 
carry out planned directed research programs concerned with infections which 
damage the human nervous system. 

This branch was reviewed by a subcommittee of experts from outside NIH during 
the year. The committee reviewed the program in depth and felt that the 
professional staff was highly competent and producing research of excellent 

quality. 

The program segments of this branch are: (a) perinatal, (b) acute infections, 
and (c) chronic infections. In each part of the above program, they are 
concerned with (a) etiology and diagnosis, (b) treatment, and (c) prevention. 

The contributions of outside investigators by means of contracts have doubled 
in the last two years and the trend continues in this direction. 

Some of the advances made in the three areas of the program du2'ing the last 
year are listed below: 

Perinatal : Safety of rubella vaccine . The study of possible transmission 
of rubella vaccine given during pregnancy demonstrated that only rarely was 
there transmission of the vaccine virus through the placenta. Impaired 
cellular-immunity to rubella v irus in congenital rubella . Investigations of 

6s 



cellular immunity to rubella virus showed a deficiency in the ability of 
lymphocytes to recognize rubella. Only the lymphocytes from congenitally- 
infected children showed this effect. These studies suggested that impaired 
cellular immunity may be of prime significance in the ability of children 
with congenital rubella to handle this virus infection in utero. 

Acute : Spinal Fluid Lactic Dehydrogenase (LDH for Rapid Diagnosis of 
Meningitis . A rapid 5-minutes test for spinal fluid lactic dehydrogenase 
has been developed. Preliminary testing shows that this method is quite 
satisfactory for the identification of elevated levels of lactic dehydrogenase. 
This in turn provides strong indication that the patient has acute meningitis 
infection and appropriate antibiotic therapy can be initiated. Further 
refinements of this method are in progress at the present time. 

Chronic : Multiple Sclerosis and Virus-like Particles . Reports from 
Australia have suggested virus-like particles in the brains of patients with 
multiple sclerosis. Our electron microscopy studies have also shown 
filamentous structures in brain cells in multiple sclerosis patients, however 
we are of the opinion that these are not virus particles. Our labeling 
studies with immimoperoxidase show no evidence of virus labeling with either 
measles or parainfluenza antibody systems. 

Treatment of Subacute Sclerosing Panencephalitis . A number of treatments 
have been tried for SSPE in the past and one of these have been successful. 
We have been studying the use of transfer factor in 8 patients with SSPE and 
comparing their clinical course with 8 individuals who have not received 
transfer factor. At the present time, there appears to be some possible 
benefit from the use of transfer factor. We are pursuing these studies in 
greater detail. 

OFFICE OF BIOMETRY 



The Office of Biometry is headed by Mr. William Weiss. Until this year, this 
office was primarily a central resource to NINDS for service in statistics, 
mathematics, computer technology, systems analysis and data processing. Its 
services range from brief consultations to participation as co-investigators 
on research projects. 

The Office of Biometry carries out research in mathematical statistics for 
current and anticipated use in NINDS programs. It also provides consultation 
and service to other organizations undertaking studies in the areas of NINDS 
interest. . . 

During the year, Mr. Weiss has been heavily involved with the Associate 
Director in the planning process for the C&FR five year research programs. 
He also made major contributions to the plans for the creation of an NINDS 
Office of Program Planning and Evaluation. 

This year, for the first time in its history, the Office of Biometry undertook 
the initiation and implementation of research programs. This is a program in 
disease statistics surveys. The NINDS program planning process has suffered 
over the years from a lack of precise data on the incidence, prevalence, and 
the socio-economic costs of neurological diseases and communicative disorders. 

T s 



It was decided by the Institute that this type of information was badly 
needed and that a program of this type must be initiated. The Office of 
Biometry was a natural location for such a program. 

An initial analysis was made of the surveys which would be feasible to start 
late in this fiscal year. It was decided to start the following surveys this 
year by means of contracts; intracranial neoplasms, multiple sclerosis and 
head and spinal injuries. Contracts for thes three areas have been negotiated. 

Next year, we hope to start a siirvey on stroke. 

Future plans of this office include the development of an advisory committee 
to recommend priorities for future surveys and the generation of survey 
proposals for recurrent samplings bo permit the development of trends in 
incidence and prevalence and the costs of the diseases of interest. 

The statisticians of this office continue to be heavily involved in the 
analysis of data for the ten major areas of the perinatal program. 

PERINATAL RESEARCH BRANCH 



The Perinatal Research Branch is headed by Dr. Joseph S. Drage. The basic 
objective of the Collaborative Perinatal Project is to relate complications, 
conditions and events that occur during a woman's pregnancy or during her 
labor and delivery to subsequent abnormality exhibited by the child of the 
pregnancy as the child grows and develops. 

In Fiscal Year 197^, the Collaborative Perinatal Project (CPP) will complete 
the longitudinal follow-up speech, language and hearing examinations on 
children at eight years of age. This completion will terminate CPP data 
collection. 

Last year, a Comprehensive Plan for Data Analysis and Interpretation for the 
perinatal data was developed and approved. The implementation of this plan 
which encompasses the major and ten minor areas has been underway this year. 
Good progress is being made. 

Since this program is suppose to be completed by June 30, 1976, there is a 
plan for the gradual elimination of personnel as the need for these people 
ceases. During Fiscal Year 197^ » eleven positions in the Perinatal Branch 
were declared surplus. Therefore, the personnel complement of the Perinatal 
Branch will be reduced by eleven by June 30, 197^. The remaining personnel 
in the Branch will be required for the balance of the existence of this 
program in order to accomplish the program delineated in the comprehensive 
plan. 

A s"i"udy of the relationship between intelligence at four years of age, as 
measured by the Stanford-Binet Intelligence Scale, and a variety of factors 
related to pregnancy, delivery, postnatal status, and environment, was 
completed during Fiscal Year 197^- A substantial monograph has been 
developed and has received outside review. The manuscript has undergone 
revision and publication is anticipated in six to nine months. 



The study of "Toxemia of Pregnancy" was undertaken through contract in Fiscal 
Year 197^- The objective is to establish a precisely-defined classification 
by a time-trend analysis of the data pertinent to clinical diagnosis: blood 
pressure, proteinuria, and edema. Contributory factors for each toxemia 
classification will be examined. During the first year of this contract, 
the time-trend analysis of blood pressures will be completed. 

A major contract was negotiated in Fiscal Year 197^ for an extensive analysis 
of speech, language and hearing data. 

The specific aims of the proposed contract are: l) to identify a small 
number of key speech, language and hearing variables, including indices which 
will serve to describe communicative ability or status of the subject tested; 
2) to explore the association between speech, language, and hearing variables, 
and other Collaborative Perinatal Project variables, in order to establish 
etiological and predictive relationships where they exist; and 3) to prepare 
a final manuscript in publishable form, which reports on the samples and 
methods, results, and their interpretation and potential application. 

During the year a consultant was hired to advise us on the requirement for 
microfilming the extensive files gathered during the years of this perinatal 
study. On the basis of his report, a contract is being negotiated to 
microfilm all of this material over the next two and a half years. This 
valuable material will then be in a form which will make it usefully 
available to future researchers in the perinatal field. 

RESEARCH CONTRACTS SECTION 

In last year's annual report, we indicated that the contract load of the 
Institute was increasing. At the end of June 30, 1973, we had 87 active 
contracts. At the end of June 30, 197^, we will have ll6 active contracts. 
This is an increase almost by a factor of two. Last year we indicated that 
the contract processing was not being handled uniformly nor well. We 
indicated that we needed a separate contract office with adequate personnel 
of high quality in order to handle the anticipated increased load of work and 
also for the purpose of increasing the quality of the contracts. 

Early in this fiscal year, we presented a plan for such an office with 
recommendations for an organization and personnel. We felt that in order 
to improve the quality of the contracts that good technical merit reviews 
were required and that this could only be handled by people with scientific 
training and background. We also felt that we needed a new contract officer 
and some new contract specialists. This office was approved in the fall 
of calendar year 1973 but really could not be implemented because we lacked 
people. Actually, we were not able to get our scientific people on board 
until December 1973, and a new contract officer and new contract specialists 
did not become available until January 197^- Therefore, for all practical 
purposes this office did not begin functioning until January 197^. In that 
time this office negotiated 67 contracts, 31 new contracts and 36 renewals. 
They have done an excellent job. 



9s 



We were very fortunate to get Dr. Mathilde Solowey to head the Scientific 
Evaluation Unit of this office. She obtained an assistant. Dr. Howard 
Weinstein during the latter part of Fiscal Year 197^- Dr. Solowey did a 
magnificent job and conducted 25 technical merit review committess during 
the latter part of Fiscal Year 197^- Mr. Lawrence Fitzgerald, the new 
contracting officer and his staff, did an excellent job in negotiating 
these contracts. He was ably assisted by Mr. James Wehling and Mrs. 
Teresa Mosedale, and Ms. Linda Waring. The secretarial staff also did 
yeoman service. All during the year we were attempting to recruit a high 
level scientist and manager to head up this office. While this recruiting 
has been going on. Dr. Watson Alberts who is Special Assistant to the 
Associate Director of Collaborative and Field Research assumed the role of 
Acting Head of this office. The success of this office during this period 
was in great measure due to his leadership. 

SPECIAL PROGRAMS BRANCH 

As of June 30, 197^, the Special Programs Branch was abolished. In March, 
Dr. Bering left his position as head of the branch to go into private 
medical practice. Mr. Weissberg was appointed Acting Head. 

The Branch's goals were to organize programs for the exchange of scientific 
information so that there would be a more rapid and efficient use of 
research results and, second, the development of NINDS related research in 
foreign countries through the PL i+80 program. 

A great deal of time this past year was devoted to attempting to restructure 
the Neurological Information Network. This network has been supported 
exclusively by NINDS since 196^+. The original goals for this network have 
been obscured during the years and. the network has expanded much beyond the 
original intent. The support of the network was becoming exceedingly high 
and many services not originally planned were added. 

This year with the help of the Scientific Information Program Advisory 
Committee the network was looked at very closely with the purpose of making 
the network relatively self supporting. 

A great deal of difficulty was encountered in obtaining cooperation from 
various centers relative to this goal. A very extensive campaign was 
launched by one of the centers to maintain that center in its existing 
status. Pressure was placed on Congress. As a result of this pressure, NINDS 
was ordered by Congress to maintain two of the centers at levels of funding 
close to last years funding. 

The publication " Parkinson's Disease and Related Disorders ," is a recurring 
bibliography from the NLM data base that has been distributed at no charge 
by the NINDS. In May 197^, by arrangement with the National Technical 
Information Service (NTIS) it is being distributed at a cost of twenty dollars 
($20.00) per year to subscribers. 

The Special Programs Branch will go out of business on June 30, 197^- The 
various contracts previously monitored by this branch will be distributed 
for monitoring to other branches in C&FR. 

10 s 



NEURAL PROSTHESIS PROGRAM 

Recent experimental work in animals and in humans has indicated that 
electrical stimulation of the cerebellum can terminate and possibly prevent 
certain types of seizures. However, refinement of the technique, including 
evaluation of potential side effects is needed before such an approach can 
be recommended for widespread clinical application. In addition, little is 
known about the neurophysiological mechanisms involved and the long-term 
effects of cerebellar stimulation. This year the Neural Prosthesis program 
inaugurated contract studies in non-human primates to supply information in 
these areas with the ultimate goal of improving the treatment of epilepsy 
in hiimans . 



lis 



ANNUAL REPORT 
July I, 1973 through June 30, 1974 
Communicative Disorders, Collaborative and Field Research 
National Institute of Neurological Diseases and Stroke 



Introduction: 



The program on Communicative Disorders was established in the Office of 
the Associate Director for Collaborative and Field Research on April 12, 1972 to develop 
and carry out planned directed research toward the improvement of diagnosis and treat- 
ment of disorders of hearing, speech, and language » Initially, the program emphasized 
hardware devices to assist the deaf (development of wearable master hearing aids and 
feasibility of cutaneous aid for profoundly deaf infants) and procedures to detect hearing 
impairments in infants and young children o In September 1973 with the arrival of a new 
program head, activities expanded into disorders of speech and language and problems 
associated with environmental noise o A new staff member is expected to join the program 
late in FY 74 to provide leadership for research on speech and language disorders^ 

Currently the responsibilities of the Communicative Disorders program are: 

L Planning and conduction of directed research having the goals of 

diagnosis, treatment, amelioration, and prevention of communicative 
disorders, 

2o Coordination of NINDS research on noise with the Environmental 
Protection Agency, 

3o Serving as a source of scientific expertise and consultation to other 
government agencies for topics relevant to communicative disorders. 

Directed Research: Program planning of directed research has been based on detailed 
review of research supported by the NINDS extramural program and by other Federal 
agencies. The program concentrates attention on areas of pressing research need and 
attempts to sponsor exemplary research, particularly in areas where availability of 
research expertise is limited. In this manner, the program is also attempting to exercise 
a catalytic effect on research which may be supported otherwise. 

The five-year program plan for Communicative Disorders contains seven 
program elements, which are described briefly below: 

I, Early detection of hearing disorders in infants and children, A contrac- 
tor is developing and validating screening procedures for use by parents, paraprofessionals, 
and clinical audiologists. In addition a manual for impedance testing of young children 
will be written during FY lb and then evaluated at clinical sites to be selected by 
competitive bid. Future efforts may concern development of a simple, highly portable 
device for Impedance testing which could be used by paraprofessionals. 

It 



2. Prescriptive tailoring of hearing aids. As described in the contract 
narrative section, prototype models of wearable master hearing aids have now been 
delivered. We expect to award a contract at the end of this fical year for developing 
a prescriptive protocol for fitting hearing aids to individuals who typically do not find 
aids satisfactory. Among other features, the contract will test the hypothesis that 
adaptation time is required by new aid wearers In order to select the best hearing aid 
characteristics for the Individual. If the developmental effort with one type of dlfflcult- 
to-flt hearing aid wearers proves successful, later studies will adapt the prescriptive 
procedures to other types of hearing-impaired persons. 

3. Speech analyzing aid for the deaf. As described in the contract 
narrative, a contractor is conducting feasibility studies concerning the possibility of 
augmenting visual information available on the face of the "talker" with either visual 
or tactile Information derived from electronic analysis of the acoustic speech wave 
produced by the talker. ^ -^ ^ 

4. Speech and language disorders In children. Because the richness of 
speech and language cannot be studied In nonhuman species, there are large gaps In 
understanding the neurological bases of language and, as a result, there is limited 
understanding of many disorders of language development. A contract will be awarded 
later this year under which two documents will be developed concerning the hypothesis 
of a critical age (or ages) for language learning „ Under this contract, a team of 
behavioral and biological scientists will first write a critical review, analysis and 
synthesis of existing data and then develop a position paper outlining research strategies 
needed to achieve greater understanding of disorders of language learning in terms of 
neurologic development. In addition to providing a basis for future NINDS requests for 
proposals, both documents are expected to be published in the scientific literature and, 
thereby, to encourage scientists to pursue research on this topic. 

Next year this program element will expand with planning for two new 
thrusts. There is need to develop Improved measures of children's language competence 
with sufficiently fine units of measurement for assessing small improvements among 
children with language disorders or delayed language development. The other emphasis 
will concern classification, diagnosis, and treatment of children whose language 
disorders are neurological ly based. 

5. Adult language disorders. Although speech and language therapy Is 
routinely provided to adult head injury or stroke patients, there Is little available 
Information concerning the relative effectiveness of differing therapeutic approaches 
for different types of patients. In order to evaluate therapeutic procedures and to seek 
to Improve them, It will first be necessary to develop a research tool for dfjscrlbing and 
quantifying the therapeutic process. This will be initiated this year under contract for 
which competitive bids have been soughto This research tool will be available for 
general research on aphasia treatment and will be utilized In future NINDS requests for 
proposals. 



2t 



6. Effects of noise on people. Two directed research activities will be 
supported this year concerning research on noise. Through an interagency agreement, 
we have requested the Committee on Hearing and Bioacoustics (CHABA) of the National 
Research Council -National Academy of Sciences to help us review existing information 
and plan research concerning effects of noise on children and to examine possible research 
strategies for obtaining better understanding in this area. (Although we consider these 
questions extremely important, the need to implement research without placing children 
at risk requires extra planning.) Our plan calls for completion of both these documents 
by late December 1974, after which we will advertise selected research strategies for 
competitive bid. The documents, themselves, will also be made available to the 
scientific community. 

A second directed research activity initiated this year is development of a 
new test of speech discrimination in noise. Unfortunately, pure tone tests of auditory 
sensitivity (i.e. audiograms) do not allow adequate prediction of the possible handi- 
capping" effects of hearing loss, and discrimination of speech is particularly difficult to 
predict because of the cognitive components of the task. The new test, consisting of 
sentences presented against a noise background, will be used to determine the extent of 
understanding of "everyday speech" among persons with noise-induced hearing loss and 
is expected also to be useful in the fitting of hearing aids for persons with other types of 
acquired hearing loss. The Request for Proposals for this work was advertised during the 
winter and proposals are now under review. 

Several additional noise research problems are being considered for support 
beginning in FY 75 provided that necessary resources become available. These would 
concern: effects on hearing of noise exposures of less than eight hours duration and of 
intermittent exposures; the possible interactions of metabolic, nutritional, or ototoxic 
agents with noise exposure in producing noise-induced hearing loss; and extra-auditory 
effects of noise on human health . We view each of these topics as requiring careful 
research planning prior to implementation of the full research effort. 

7. The Information Center for Hearing, Speech and Disorders of Communi- 
cation has recently been transferred to this program. Under present plans, this center 
will be phased out during FY 75 and FY 76. 

Limited funds were also provided to enable an investigator to conduct a pre- 
liminary study of athletically induced head trauma in youth and hearing loss at high 
frequencies (e.g. frequencies above 10,000Hz). Results of this pilot study indicate a 
significant positive relation between history of head trauma and high frequency hearing 
losso Since other research shows that hearing loss at these high frequencies precedes 
ototoxic drug-induced hearing loss in the audiometric frequency range typically 
tested (125-8000 Hz) by as much as 120 days, high frequency hearing testing may serve 
as a warning of Impending hearing loss of handicapping magnitude to boys and young men 
participating In sports such as boxing. 



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Several addiflonal problem areas which were not included when the five- 
year plan was developed during September and October, 1973, are now being considered 
for inclusion in revised plan. These include research on secretory otitis media In 
children and parametric investigations of the auditory capabilities (with special reference 
to speech perception) of persons with hearing loss due to aging (presbycusis) or with 
noise-induced hearing loss. 

Coordination of Research on Noise: The head of the Communicative Disorders program 
coordinates NINDS research on noise with the Environmental Protection Agency through 
participation in that group's Health Effects Panel. Coordination of NINDS noise 
research with other HEW agencies (National Institutes of Environmental Health Services 
and the National Institute of Occupational Safety and Health) has been carried out on an 
informal level and we expect to formalize these communications In order to prevent 
duplication of research efforts. 

Consultation and Other Research Activities: In addition to providing technical 
assistance to the NINDS Office of Scientific and Health Reports, the Communicative 
Disorders program has also provided consultation on a number of other activities 
supported by other agencies. These include an evaluation of services for handicapped 
children supported by the HEW Office of the Assistant Secretary for Planning and 
Evaluation; an evaluation of head start programs for handicapped children supported by 
the HEW Office of Child Development; an evaluation of preschool education programs 
supported by the Bureau of Education for the Handicapped; ana the Interagency Panel 
on Early Childhood Research and Development which is supported by the Office of Child 
Development and a number of other Federal agencies. 

FinciJy, two publications will result from activities during this reporting 
period. Galley and page proofs for the book Psychology and the Handicapped Child 
edited by John A. Swets and Lois L. Elliott have been read; publication is scheduled 
for May 21, 1974. A manuscript concerning auditory perception by persons with 
sensorineural hearing loss (described in the project narrative) was prepared and submitted 
to International Audiology . 



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ANNUAL REPORT 

July 1, 1973 through June 30, 1974 

Section on Biomedical Engineering and Instrumentation 

Collaborative and Field Research 

National Institute of Neurological Diseases and Stroke 

National Institutes of Health 



Introduction : 

The Section on Biomedical Engineering and Instrumentation carries out 
planned, directed research to develop biomedical instrumentation needed to 
advance the diagnosis and treatment of neurological and sensory disorders. 
The Section develops and directs programs to appropriately adapt and apply 
recent advances in technology to problems in identified areas of neuro- 
sciences for which biomedical instrumentation is needed. The Section 
coordinates projects with other laboratories and branches of the Institute 
and the other biomedical engineering activities of the NIH. By working 
directly with the clinicians and scientists on their instrumentation 
problems, the Section seeks to provide the necessary interdisciplinary inter- 
face between the clinical problems and the technical development to assure 
the proper management of technical and clinical input at all stages of 
development. 

The laboratory research and feasibility studies of the Section are 
designed to explore possible approaches to adapt basic techniques to 
specific clinical problems, and to investigate the feasibilities of 
alternative methods. 

The Section develops and directs prototype instrumentation engineering 
to adapt feasible approaches and techniques into practical clinical proto- 
type instruments. The Section monitors the clinical testing and evaluation 
of prototype instrumentation and provides for needed modification and 
refinement to lead to useful practical clinical tools for commercial 
development and wide clinical application. 

Work in Progress : 

During the past year the staff of BEI has assisted other branches and 
sections with their instrumentation development projects by providing 
technical assistance and advice, both in preparing and soliciting for 
proposals and serving as Project Officers on contract projects funded from 
other units. These include the following contracts: 

NOl-NS-3-2322 - Stanford Research Institute - Wearable EEG Tape 

Recorder 



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^ NOl-NS-2-232/ - Bolt Beranek and Newman - Wearable Master Hearing (') 

Aid Development 

NOl-NS-2-2316 - CBS Laboratories - Wearable Master Hearing Aid 

Development 

NOl-NS-3-2315 - Gallaudet College - Speech Analyzing Aid for the 

Deaf 

( 
In addition, the Section has initiated and supported several planned 
directed research contract projects for feasibility studies and instrumenta- 
tion R&D aimed at the development of clinical instrumentation to meet high 
priority needs for instrumentation in other fields related to the mission 
of NINDS. These include the following contracts: 

NOl-NS-2-2323 - Ohio State University Research Foundation - Semi- 
conductor Gamma Camera Development 

NOl-NS-3-2319 - Indianapolis Center for Advanced Research - Study and: 

Test Ultrasonic Techniques for Diagnosis of Cerebral 
Disorders 

NOl-NS-4-2304 - Bowman Gray School of Medicine - Study and Test 

Ultrasound Techniques for Diagnosis of Cerebral 
Disorders 

NOl-NS-2-2318 - Johns Hopkins University - Study of Cutaneous and 

Visual Patterned Stimulation for the Profoundly 
Deaf Infant 

The Section has carried out limited laboratory research aimed at 
evaluating instrumentation and techniques for potential application to 
specific research instrumentation needs of other programs of the Institute. 
Activities in the past year have included preliminary studies of the light 
scattering properties of antibodies and antibody antigen complex for the 
purposes of screening for antibody precipitate formation (NDS(CF)-73 BEI 
2047). Further development of rapid microspectrophotometer for the study 
of photo pigment kinetics (in collaboration with the Laboratory of Neuro- 
physiology, NINDS) is reported under project #NDS(l)-69 LNP/SP 1690. 



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CONTRACT NARRATIVE 
Biomedical Engineering and Instrumentation Section, C&FR, NINDS 
July 1, 1973— June 30, 1974 

JOHNS HOPKINS UNIVERSITY (NOT -NS-2-2318) 

Title : Study of Cutaneous and Visual Patterned Stimulation Communication 
Aids for the Profoundly Deaf Infant 

Contractor's Project Director : Moise H. Goldstein, Ph.D. 

Current Annual Level of Support : $26,714 

Objectives : These studies are to develop the techniques and instrumentation 
for studying and evaluating the feasibility of using cutaneous or visual 
dynamic patterned stimulation derived from deaf infant's own vocalizations 
in order to provide information feedback of those vocalizations. It is 
hoped that this will both assist in vocalization development and serve as 
a communication aid for profoundly deaf infants. Comparative studies on 
three groups of profoundly deaf infants are being carried out to determine 
the effectiveness of both the visual patterned stimulation and the tactile 
patterned stimulation on the development of, and change in, the vocaliza- 
tions by these infants during and following the training sessions. The 
basic question to be answered is whether these alternative sense modalities 
can be effectively used by severely deaf infants, and if cross-modality 
transfer can be established effectively. 

Major Findings : During the past year it was found that subjects, even in 
this age group, can be trained to attend both to the visual and to the tactile 
displays. The efficacy of these very simple pattern displays in affecting 
the vocalization has been tested on three groups of subjects: those trained 
with the visual display, those trained with the tactile display, and a 
matching control group. While the number of subjects is still very small at 
this writing, very preliminary indications are that the total number of 
utterances during and following the training sessions is not outstandingly 
different between the test groups and the control group, although they all 
increase with training. However, the type of vocalization seems to be 
affected by this training, particularly when an evaluation is made of the 
number and frequency of complex syllables being used is analyzed. While 
complete training and test sessions have been recorded on tape, only pre- 
liminary analysis of these data has been carried out to test the feasibility 
of providing, through alternative sense modalities, the feedback from one's 
own vocalization. More sophisticated analysis of these data and the testing 
of more subjects with more sophisticated displays will be required to 
establish the effectiveness of such displays for a language development aid 
for profoundly deaf infants. 



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Contract (N01-NS-2-2318) 

Significance to NINDS Programs and Biomedical Research : It has long been 
felt that normal development of language patterns and speech skills require 
both participation in speech communication using normal language patterns 
and practice in the mechanical skills of articulation and vocalization 
during normal developmental periods. The severely deaf infant is denied such 
input of acoustic speech information both from others and from his own 
vocalization efforts by virtue of his loss of hearing. It fs felt that pro- 
viding for Increased sensory input of the patterns of normal speech during 
the normal development period using cross-modality input with such prosthetic 
aids, may assist in a more normal development of language and speech 
capabilities during the early years. If such cross-modality sensory input 
proves effective, the later development of wearable communication aids based 
on such cross-modality input could help the language and speech development 
of an estimated 250,000 pre-school children who suffer from hearing 
deficiencies sufficient to impair their normal language development. 

Proposed Course of Contract : By the end of this second year, training and 
data collection will be completed on a first group of subjects which have 
been trained and tested in a minimal number of sections, in order to evaluate 
whether, in fact, the feedback loop and cross-modality transfer can be 
effected. A small group of consultants will assist NINDS in determining what 
additional data analyses of the tapes should be completed to determine effects 
of training for these children. In addition, more sophisticated training 
and testing over an extended period of time with a second small group of 
subjects may be carried out to test the effectiveness of different forms of 
display and to probe more deeply the effectiveness of these displays on 
vocalization development. 



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CONTRACT NARRATIVE 

Biomedical Engineering and Instrumentation Section, C&FR, NINDS 

July 1, 1973— June 30, 1974 

OHIO STATE UNIVERSITY RESEARCH FOUNDATION (NOl-NS-2-2323) 

Title : To Develop a Semiconductor Gamma Camera System for Nuclear Medicine 

Contractor's Project Director : Robert F. Redmond, Ph.D. 

Current Annual Level of Support : $262,128 (26 months) 

Objectives : The goal of this contract is to develop and test a new position 
sensing germanium gamma ray detector and to design, build, and evaluate a 
prototype semiconductor garrma camera based on this detector system. 

Major Findings : Studies of detector development have resulted in successful 
fabrication of position sensing solid state detectors using ultra high 
purity germanium which has been grooved in parallel strips which are othogonal 
on opposite surfaces. On each side, parallel strips have been resistively 
connected and electrically function as though the side were a linear resistive 
charge-separating continuous surface. In this configuration then, the 
detectors operate electrically just as they would had we succeeded in 
fabricating sufficiently quiet uniform resistive surfaces on both sides as 
originally proposed. Such uniform resistive surfaces, giving sufficiently 
low leakage current and hence, tolerable electric noise figures, were not 
achieved with the existing state of technology. 

A prototype camera using one such detector, a specially designed orange 
crate collimator, and all the electronic circuits necessary to read out the 
detectors, have been fabricated and evaluated. This evaluation indicates 
the spatial resolution of less than 2 millimeters intrinsic in the detector 
and energy resolutions of approximately 4kev. Both figures fit well with 
initial expectations from theoretical analysis. The use of ultrapure 
germanium detectors with ion implantation electrode surfaces as opposed to 
the initially proposed lithium drifted germanium, has resulted in a much 
more stable and rugged detector system which can withstand both temperature 
and atmospheric recycling without destroying the detector. 

Significance to NINDS Program and Biomedical Research : The gamma camera 
being developed under this contract should allow increased spatial and 
energjf resolution in diagnostic i-naging for lesions which are ■'dentifiable 
by radiopharmaceutical uptake, thus significantly improving the capabilities 
for diagnosis and serial evaluation of intracranial lesions. 



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Contract (NOl-NS-2-2323) 

Proposed Course of Contract : The current activities of design for full- 
scale clinical camera will continue into the early part of FY '75 and 
produce an engineering design drawing and preliminary evaluation and analysis 
of the anticipated capabilities and limitations of such a clinical camera. 
Should this evaluation of the clinical camera design indicate the appropri- 
ateness of construction of such a clinical camera prototype and its 
clinical evaluation, construction of the camera will be carried out under 
a new contract negotiated in FY '75. 



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CONTRACT NARRATIVE 
Biomedical Engineerinq and Instrumentation Section, C&FR, NINDS 
July 1, 1973— June 30, 1974 

INDIANAPOLIS CENTER FOR ADVANCED RESEARCH (NOl-NS-3-2319) 

Ti tl e : Study and Test Ultrasonic Techniques for Diagnosis of Cerebral 
Disorders 

Contractor's Project Director : Francis J. Fry 

Current Annual Level of Support : $154,097 (18 months) 

Objectives : This project, coordinated by and primarily performed at the 
Indianapolis Center for Advanced Research (ICFAR) with subcontract agreements 
to Bolt Beranek and Newman (BBN) and Purdue University, is studying the 
feasibility of processing ultrasound return signals to acoustically identify 
and characterize the status of intracranial structures, and is evaluating 
the potential of these techniques for identifying specific intracranial 
lesions. In addition, prototype instrumentation and on-line computer 
processing are being developed to support trials of these techniques in a 
laboratory environment. These instrumentation development efforts are 
supplemented by studies of the acoustic properties of normal and pathologic 
brain tissue to be used for interpretation of the measured acoustic character- 
istics of intracranial structures for the ultimate purpose of non- 
invasive diagnosis of lesions and serial tracking of recovery following 
treatment. 

Major Findings : The development and implementation of special transducers, 
data handling systems, and computer algorithms for signal processing have 
been carried out and preliminary evaluations begun. In addition to the 
implementation of the "impediography" algorithm from BBN, collaborative work 
between BBN and Purdue has resulted in generation of several alternative 
algorithms utilizing different interrogating wave forms. These algorithms 
are currently being implemented in the computer and comparative evaluations 
of the alternative approaches are underway to determine their resolution and 
sensitivity capabilities, and their susceptibility to multiple reflections 
and other forms of "noise". Preliminary evaluations indicate that the rf 
return signal from intracranial acoustic interrogation can be processed to 
extract measurable impedance profiles of the tissues and structures 
interrogated. 

Significance to NINDS Programs and Biomedical Research : The diagnosis and 
treatment of stroke, head injury, brain tumor, and other intracranial 
disorders is of prime interest to NINDS and the scientific fields that it 
represents. It is felt that non-invasive techniques, both for screening for 



7u 



Contract (N'Ol-NS-3-2319) 

cerebral disorders and for diagnosis of specific pathologies and their 
serial assessment following treatment, would be of great use in improving 
the clinician's capabilities in the diagnosis and treatment of these 
disorders. If successful, the techniques being investigated under this 
contract will provide an extended capability for such non-invasive assess- 
ment of brain tissue status which should be of value for such diagnosis 
and serial assessment. 

Proposed Course of Contract : In the remaining six months of the initial 
contract period, the feasibility study should be completed and an analysis 
made of the capabilities and limitations of these preliminary approaches 
to this non-invasive assessment through signal processing of ultrasound 
signals. If, as it now appears, these approaches are demonstrated feasible, 
possible extension of the workscope of the contract may be negotiated to 
explore more fully the several alternative approaches and algorithms and 
to develop and implement the necessary instrumentation to carry out clinical 
studies of the technique. 

In addition, a close cooperation and collaborative effort with Bowman Gray 
Medical School (see contract #N01-NS-4-2304) will be initiated together 
with other related ultrasound diagnostic technique projects to be funded in 
the near future. Should the techniques prove feasible and the clinical 
evaluations indicate their utility, commercial development end ir,strumenta- 
tion for wide application will be encouraged. In any event, all results of 
these studies will be published and techniques and instrumentation designs 
developed under this contract put in the public domain for wide use. 



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CONTRACT NARRATIVE 
Biomedical Engineering and Instrumentation Section, C&FR, NINDS 
July 1, 1973--June 30, 1974 

STANFORD RESEARCH INSTITUTE (SRI) (NOl-NS-3-2322) 

Title : Development of a Wearable Four-Channel EE6 Cassette Recording System 

Contractor's Project Director : Joseph H. Chadwick, Ph.D. 

Current Annual Level of Support : $135,342 

Objectives : The objective of this effort is to design, fabricate, and 
clinically test two prototypes of a wearable EEG recording system and two 
associated playback systems which will make it possible to record, from 
ambulatory patients, four channels of EEG during a twenty-four hour period 
using commercially available magnetic tape cassettes and play these back 
at high speed for computer analysis and clinical evaluation. This develop- 
ment is to include electrodes and preamps to provide for four channels of 
differential recording connected to a wearable tape recorder with appropriate 
signal processing electronics and digital encoding circuitry to provide for 
a full dynamic range and suitable bandwidth to allow for automatic processing 
of EEG data by computer. In addition to the design and development of the 
recording technique and implementation in hardware, the overall system is 
to be clinically evaluated with a selected group of patients under environ- 
mental conditions similar to those for which it will ultimately be used. 
The playback system is to transcribe the encoded data from initial tape 
cassettes and reform that data onto conventional computer compatible digital 
tape, as well as interfacing directly to a computer and providing real time 
analog outputs for strip chart recording. 

Major Findings : Electrodes and preamps have been designed and fabricated and 
suitably packaged for easy wearing on the head. Digital recording technique 
has been implemented using a modified commercially available tape recorder 
deck to provide six to twelve hours of recording on a single cassette with 
effective signal bandwidth of 40 to 80Hz and signal dynamic range of 1000 to 
1 for all four channels. Electrode resistance and calibration checkings of 
circuitry have been implemented and the playback system currently being 
developed will be completed by the end of the year. 

Significance to NINDS Programs and Biomedical Research : In evaluation of 
treatment, including anti-epileptic drug evaluations and establishment of 
appropriate medication schema, a need exists to evaluate seizure activity 
in patients while free-ranging in a natural environment subjected to normal 



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Contract (NOl-NS-3-2322) { 

everyday stresses. This capability with the high fidelity of recording 
required is not currently available through commercial equipment. Successful 
accomplishment of the goals of this project will provide for such monitoring 
capability for ANRB anti-epileptic drug evaluations. In addition, the 
system will provide a generally useful wide dynamic range, wide bandwidth, 
high bit rate, long-term lightweight portable recording system for recording 
other bioelectric potentials and other physiologic parameters which future { 
programs may wish to evaluate under such free-ranging conditions. 

Proposed Course of Contract : By the end of the first contract year, we should 
have basic techniques solved and first prototype models of the recording 
system built and evaluated. In the second year, further clinical evaluation 
and testing will be carried out to lead to improved design and development 
of a "Mark 11" wearable system. The aim of the second year effort will be 
to reduce the weight of the wearable components, possibly extend the time 
of recording, and/or increase the number of channels recorded through the 
implementation of alternate, more sophisticated coding techniques explored 
during the first year but not yet implemented. In addition, some added work 
on reduction or suppression of artifacts may be undertaken (a definite 
workscope will be negotiated on 24 April 1974). 

Collaborating Units : This project is being carried out under combined 
collaborative effort of the Applied Neurologic Research Branch and the 
Section on Biomedical Engineering and Instrumentation. The Applied Neuro- 
logic Research Branch is providing the clinical direction and the funding, 
while the Biomedical Engineering Section is providing the technical direction 
and project management. 



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CONTRACT NARRATIVE 
Biomedical Engineering and Instrumentation Section, C&FRj NINDS 
July 1, 1973--June 30, 1974 

BOWMAN GRAY SCHOOL OF MEDICINE (NOl-NS-4-2304) 

Title : Study and Test of Ultrasound Techniques for Diagnosis of Cerebral 
Disorders 

Contractor's Project Director : Ralph W. Barnes, Ph.D. 

Current Annual Level of Support : $169,274 

Objectives : These studies will measure and characterize the acoustic prop- 
erties of normal and pathologic intracranial tissues to gather a data base 
with which to study the identification of specific pathologic states based 
on analysis of signal return from ultrasound interrogation. In addition, 
the contractor will develop and clinically evaluate techniques and instru- 
mentation for non-invasive assessment of cerebrovascular dynamics through 
the use of iroving target indicator approach. These studies will be 
coordinated with those of ICFAR (see contract #N01-NS-3-2319) and other 
future contractors monitored by NINDS. 

Major Findings : This contract is a new contract negotiated in April 1974, 
and at the time of writing of this report no work has yet been undertaken. 

Significance to NINDS Programs and Biomedical Research : The diagnosis and 
treatment of stroke and other intracranial disorders is a major concern to 
NINDS and to the medical fields related to the Institute's mission. The non- 
invasive diagnostic techniques to be investigated under this project will, 
if successful, provide valuable tools for non-invasive diagnosis of major 
forms of cerebrovascular disease, as well as providing additional capabilities 
for diagnostic interpretation of ultrasound interrogation of other tissues 
and structures within the cranial cavity. 

Proposed Course of Contract : The contract recently signed, April 1974, is 
for a 24 month study beginning with the characterization of the acoustic 
properties of intracranial structures, and the development of the moving 
target indicator technique and supportive instrumentation. Evaluation of 
this procedure for its capabilities and limitations as a useful clinical tool 
in diagnosis of specific intracranial vascular disorders will be carried out. 
State-of-the-art instrumentation will also be implemented for rapid acquisi- 
tion of moving targets, such as blood vessels, and evaluated for the ability 
to find and follow blood vessels as they decrease in size. It is expected 
that during the next two years this work will be carried out in conjunction 
with related work at ICFAR and other ultrasound instrumentation projects 
both within NINDS and other Institutes of the NIH. 



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Serial No. NDS(CF)-73 BEI 2047 

1. Office of Associate Director 

2. Section on Biomedical Engineering 
and Instrumentation 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Biomedical Engineering and Instrumentation Studies 

Previous Serial Number: Same 

Principal Investigator: George C. Murray, Ph.D. 

Other Investigators: Abraham Rosner 

Cooperating Units: Section on Technical Development, NIMH/NINDS/NIH 

Man Years: 



Total : 


0.6 


Professional : 


0.1 


Others: 


0.5 



Project Description: 

Objectives : To provide the facilities and support for breadboarding 
and testing of instrumentation techniques and carry out feasibility studies 
and evaluation of possible application and adaptation of advanced technology 
to the measurement and instrumentation needs of Institute programs of neuro- 
science research and clinical neurology. 

Methods Employed : Investigation of physical and engineering data on 
potentially useful instruments and techniques and pilot studies to develop 
and evaluate needed design changes and test prototype approaches to problems 
related to the Section's instrumentation development programs. 

Laboratory facilities were developed for the Section staff to modify 
and test the capabilities of existing techniques and equipment for their 
potential development and application to specific instrumentation problems 
of the Section and other programs of the Institute. Whenever possible, 
cooperative work is carried out with the staff and facilities of the other 
labs and branches of the Institute and the other bioengineering facilities 
of the NIH. 



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Serial No. NDS(CF)-73 BEI 2047 

A simple prototype light scattering apparatus was developed using 
incandescent source monochromator and optical collimators, and incorporating 
a channeltron photomultiplier tube with specially designed and constructed 
circuitry to allow counting of individual photon events. Light scattering 
from solutions of antibodies, antigens, and mixtures of the two, were 
measured with this apparatus as a function of the angle scattered off of the 
optical axis and the time after mixture. 

Major Findings : While the measurements made during the current fiscal 
year showed some asymmetry in the light scattered both from the antibody 
antigen and the mixtures, the technique did not reliably distinguish on the 
basis of light scattering asyrrjtietry alone between stable mixture solutions 
from solutions of the components added. However, when the freshly mixed 
solutions were observed as a function of time after mixing, the light 
scattering patterns changed as a function of time with a characteristic 
asymmetry and with a time course consistent with the known kinetics of 
precipitate formation for the mixtures used. It was therefore concluded that 
more refined scattering apparatus could be used to follow the changes in 
light scattering as precipitates form, and perhaps thereby identify formation 
of precipitates. 

Proposed Course of Project : Due to lack of available time and manpower 
(the project was carried out by Mr. Rosner, a part-time student trainee), 
and a diminished interest on the part of the Infectious Diseases Branch to 
pursue this particular technique, this light scattering study under the 
general instrumentation project has been temporarily dropped. 

The overall bicengineering instrumentation studies project will be 
continued to allow initiation and carrying out of other pilot studies for 
other instrumentation as needs arise in the development of new biomedical 
programs to support clinical research of the branches and laboratories of 
the Institute. 

Honors and Awards: None 

Publications: None 



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CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Epilepsy 
July 1, 1973~June 30, 1974 



NEW CASTLE STATE HOSPITAL (NOl-NS-8-1310) 



Title : Development of a Model for Assessment of New Anticonvulsant Agents 

Contractor's Project Director : Joseph T. Brock, M.D. 

Current Annual Level : $90,000 

Objectives : To develop a model to study the efficacy, safety and bioavail- 
ability of new antiepileptic drugs in humans. During the past year, inves- 
tigations included the relative antiepileptic properties of carbamazepine, 
diphenylhydantoin, and phenobarbital administered concurrently to patients 
with generalized seizures refractory to treatment, the evaluation of possi- 
ble side effects of the drugs, and evaluation of drug serum concentrations. 
Mexiletine (Kfi 1173) will be studied during the final months of the current 
fiscal year. 

Course of Contract ; A one year study of carbamazepine combined therapy with 
two marketed drugs, phenobarbital and/or diphenylhydantoin, was completed in 
September of 1973 . Clinical data was collected and sent to the Section on 
Epilepsy, NINDS, Bethesda, for review and preparation for computer aided 
analysis. Analysis of that data is under way. In May of 1974, patient 
trials began with mexiletine with eight patients in a preliminary study of 
the drug. A twenty week study of the efficacy and safety of mexiletine com- 
bined with other drugs is planned for the next fiscal year. 

Major Findings : Carbamazepine has been found to be as effective an anticon- 
vulsart as diphenylhydantoin or phenobarbital when used singly in generalized 
seizures resistant to treatment. Also, combinations of the drug were studied 
and found to be more effective than the drugs singly. Additional data on 
long-term usage of carbamazepine is being analyzed. Continuation of the study 
will now determine the efficacy of mexiletine as an antiepileptic drug. 

Significance to NINDS Program and Biomedical Research : The pharmaceutical 
industry has demo.istrated little interest in developing new antiepileptic 
agents. Aside from the economic factors involved, one of the industry's 
major p"oblemL 's to obtain satisfactory i-'linical studies of antiepileptic 
drugs. Through this contract and others, NINDS ha", supported clinical studies 
of antiepi]eptic drugs. Well con^.roJled studies — as conducted at New Castle 
State Hospital — will be significant indicators of therapeucic merit of new 
antiepileptic drugs and may encourage the pharmaceutical industry to develop 
promising agents for clinical trial. It is anticipated NINDS sponsored studies 
will enable drugs to reach the market more readily, and \.hus ba available to 
physicians who treat patients with seizures. 

Proposed Course of Contract : Additional evaluations of antiepileptic drugs 
and further development of the model for drug evaluation are planned. 



V 



Publications : 

Cereghino JJ, Penry JK, Brock JT: A controlled prospective study of the use 
of carbamazepine combined with other drugs in the treatment of epilepsy. 
Abstracts X International Congress of Neurology, Barcelona, September 8-15, 
1973. Excerpta Medica International Congress Series No. 296, p. 113, 1973. 

Cereghino JJ, Brock JT, Van Meter JC, Penry JK, Smith LD, White EG: Carbama- 
zepine for epileptic patients. A controlled prospective evaluation. 
Neurology (Minneap) 24^, 1974, in press. 

Cereghino JJ, Brock JT, Van Meter JC, Penry JK, Smith LD, Fisher P, Ellenberg 
J: Evaluation of albutoin as an antiepileptic drug. J Clin Pharmacol 15 , 
406-416, 1974. 



8v 



CONTRACT NARRATIVE 

Applied Neurologic Research Branch — Section on Epilepsy 

July 1, 1973— June 30, 1974 

MEDICAL COLLEGE OF GEORGIA (NOl-NS-9-2169) 

UNIVERSITY OF VIRGINIA SCHOOL OF MEDICINE (NOl-NS-9-2196) 

Title : Study of Anticonvulsant Properties of Clonazepam and Ethosuximide 

Contractor's Project Directors : Paul R. Dyken, M.D., (MCG) 

Fritz E. Driefuss, M.D. , (UV) 

Current Annual Level : $46,890 (Medical College of Georgia) 

$65,907 (University of Virginia School of Medicine) 

Objectives : To study and compare the effect of ethosuximide and clonazepam 
on the frequency and intensity of absence (petit mal) seizures in patients 
previously untreated for this disease and in patients who have failed to be 
controlled by ethosuximide; to evaluate the effect of ethosuximide and of 
clonazepam therapy on physiologic, psychometric, and other functions. 

Course of Contract : Data is collected for each patient during the initial 
hospitalization period, during an outpatient and during the final hospitali- 
zation periods^ and sent to the Section on Epilepsy, NINDS, Bethesda, for re- 
view and entry for ccanputer-aided analysis. The relocation of part of the 
study to the Medical College of Georgia has been successfully completed, and 
patient input has been good at the new location. However, due to the lack 
of patient input during the transition period, and a corresponding drop in 
new referrals at the other center, the goal of 50 patients at each center will 
not be reached this year. 

Major Findings : The procedures and protocol established during earlier studies 
were proven valuable in conducting an evaluation of an antiepileptic agent. 
EEG telemetry has been developed as a major means of evaluating absence sei- 
zure frecjuency. Preliminary findings show that the two drugs evaluated have 
about the same effectiveness in absence epilepsy. 

Proposed Course ; The current study comparing ethosuxirdde with clonazepam 
will continue until contract expiration in August 1974. 



9v 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Epilepsy 
July 1, 1973 — June 30, 1974 



UNIVERSITY OF WASHINGTON (NOl-NS-0-2281) 

Title ; Study of Carbamazepine in the Treatment of Partial Complex Seizures 

Contractor's Project Director : Allan S. Troupin, M.D. 

Current Annual Level ; $130,296 

Objectives ; To study the relative antiepileptic efficacy and safety of car- 
bamazepine and diphenylhydantoin in partial seizures; to measure drug con- 
centrations in patients' blood; and to assess patients' psychological com- 
petence and social function. 

Course of Contract : Twelve patients were studied in a preliminary evalua- 
tion of the double-blind study. However, preliminary information indicates 
that carbamazepine is as effective as diphenylhydantoin in the control of 
partial complex seizures. 

Proposed Course ; The contract will be continued with additional funds to 
complete the comparison of carbamazepine and diphenylhydantoin in patients. 
A preliminary evaluation of clorazepate dipotassium as an adjunct treatment 
in this population will be conducted concurrently. 

Publications : 

Troupin AS, Green JR, Levy RH: Carbamazepine in epilepsy—a pilot study. 
Neurology (Minneap) , 24 , 1974, in press. 

Green JR, Friel P, Amick-Corkill JA: Methods for gas-liquid chromatographic 
determinations of carbamazepine, phenytoin, phenobarbitone , and primidone. 
In; Methods of analysis of anti-epileptic drugs. International Congress 
Series No. 286. 

Green JR, Troupin AS, Halpern, LM, Friel P, Kanarek P: Sultliiame ev^iluation 
as an anticonvulsant. Epilepsia, 15^, 1974, in press. 

Reitan RIi: i?sychological testing of epileptic patients. In: Handbook of 
Clininal Neurology, 1974, in ^.ress. 



llv 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Epilepsy 
July 1, 1973--June 30, 1974 



UNIVERSITY OF WASHINGTON (N01-NS-1-22S2 ) 

Title ; Study of Experimental Antiepileptic Drugs in Animals 

Contractor's Project Director : Joan S. Lockard, Ph.D. 

Current Annual Level ; $225,000 

Objectives ; To compare the antiepileptic efficacy of carbamazepine , pheno- 
barbital, and diphenylhydantoin in spontaneous motor seizures of primates. 
Seizure frequency and behavioral toxicity are compared with drug dosage 
and drug blood ccncentration in the primate with spontaneous seizures. 

Course of Contract ; During the past year, this animal model of focal motor 
epilepsy has bean utilized to determine the propylactic efficacy of combined 
diphenylhydantoin/phenobarbital treatment on posttraumatic epilepsy; this 
study will be concluded in December 1974. Also, the basic kinetics of clona- 
zepam, dipropylacetic acid, and ethosuximide were studied in the monkey. The 
behavioral precipitance of seizures in a group of free-roaming epileptic 
monkeys instrijmented with motor seizure telemetry units was studied; this 
study will be concluded December 1974. 

Major Findings ; The half-life of clonazepam was determined to be 4 — 5 hours, 
and of dipropylacetic acid, 1/2 to 1 hour. Thus it was necessary to develop 
a system for catheterization whereby continuous administration of drugs and 
continuous monitoring of drug blood concentrations may be made. This system 
will be used in the coming year — to determine the efficacy of dipropylacetic 
acid, and clonazepam in this model. 

Proposed Course; The contract will be continued to further demonstrate and 
refine the usefulness of this animal model of epilepsy. Proph xaxis of pose- 
traumatic epilepsy, motor seizure telemetry and social behavior, deep-sleep 
EEG telemetry and spontaneous seizures in the monkey mc del will be concluded. 
The efficacy of dipropylacetic acid, clonazepam, and ethosuximide will be 
studied in the monkey model. The pharmacokinetics of dipropylacetic acid, 
clonazepam and ethosuximide will be determined in the priirate before experi- 
mentation is begun. 

Publications: 

Levy RH, Lockard JS, and Kupferberg HJ: Plasma levels of the anticonvulsant 
clonazepamin monkey. Electroenceph. Clin. Neurophysiol. 1974, in press. 

Lockard JS, Levy RH, Uhlir V, and Farguhar JA; Pharmacokinetic evaluation of 
anticonvulsants prior to efficacy testing exemplified by carbamazepine in 
epileptic monkey model. Epilepsia, _15, 1974, in press. 



13v 



Levy RH, Lockard JS and Kupferberg HJ: Pharmacokinetics of carbamazepine in 
monkey. (Abstract #67, p. 70). APha Academy of Pharmaceutical Sciences, 
1973, San Diego, California. 



14V 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Epilepsy 
July 1, 1973— June 30, 1974 

LAFAYETTE CLINIC (NOl-NS-2-2310) 



Title: Evaluation of the Anticonvulsant and Psychologic Effects of Carbama- 
zepine in Inpatients with Temporal Lobe Epilepsy and in Outpatients with 
Temporal Lobe Epilepsy and/or Other Forms of Epilepsy. 

Investigators : Ernst A. Rodin, M.D. 
Philip Rennick, Ph.D. 

Current Annual Level ; $103,580 

Objectives : To determine the anticonvulsant, electroencephalographic, and 
psychotropic effects of carbamazepine and placebo in 36 inpatients also 
treated with diphenylhydantoin and phenobarbital ; to compare the anticon- 
vulsant efficacies of carbamazepine and primidone in dOuble-blind cross-over 
fashion in outpatients with temporal lobe and/or other forms of epilepsy. 

Major Findings : Carbamazepine is markedly effective in reducing frequency 
of psychomotor seizures and grand mal seizures in inpatients with predominately 
psychomotor epilepsy. Carbama::epine did not appear to have any beneficial 
psychological effect in hospitalized patients. A mild lowering of arousal 
level was noted in 26 out of 36 patients. Carbamazepine had no consistent 
effect on the niomber of spikes noted in the EEC. It is clearly more effec- 
tive in the control of clinical seizures than in improving the EEC. 

Proposed Course ; The second phase of the study, the comparison of carbamaze- 
pine and primidone in a double-blind cross-over fashion in outpatients s some 
of whom also participated in the first part of the study, is now under way. 
Clinical seizures, psychological parameters, and the EEC are being monitored. 



15v 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section Epilepsy 
July 1, 1973~June 30, 1974 



UNIVERSITY OF KANSAS MEDICAL CENTER (NOl-NS-2-2313) 

Title : Investigation of Posttraumatic Epilepsy Prophylaxis 

Contractor's Project Director : Charles Brackett, M.D. 

Current Annual Level : $61,366 

Objectives : A pilot study to determine the effectiveness of propylac 
treatment with diphenylhydantoin and phenobarbital in persons who suffer 
head injury, and are thus liable to posttraumatic epilepsy. 

Course of Contract : Although the admission rate is less than anticipated, 
there has been excellent patient rapport and compliance in the beginning 
months of the study. Progress and quality of the study has been such that 
several of the design hypotheses may be answered at this time, before the 
study is completed. 

Proposed Course of Contract : A ■sufficient number of patients will be studied 
to answer the hypotheses proposed and determine the necessity and feasibility 
for additional studies. 



17 



V 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch--Section on Epilepsy 
July 1, 1973— June 30, 1974 

EXCERPTA MEDICA FOUNDATION (N01-N$-3-2303) 

Title : Publication of Epilepsy Abstracts , Volume 7 

Current Annual Level : $39,343 

Objectives : To scan serial publications and periodicals from approximately 
3000 of the world 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 appro- 
priate translations into English from foreign languages, classify, index, 
and store the abstracts in a computer retrievable form; and produce a 
9-tract computer tape in the NIH-PRS format, to be delivered when Volume 6 
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 cemera-ready copy, the printing, and distribution, 
the Excerpta Medica Foundation sells subscriptions to recover the cost of 
production of camera-ready copy, printing, and distribution. 

Course of Contract : Subscriptions to Epilepsy Abstracts , each at an annual 
cost of $25.00, have been acquired by Excerpta Medica at a satisfactory 
rate. Interest in the publication continues at a high level. 

Proposed Course of Contract : It is anticipated that issues will be distri- 
buted as scheduled, and that the computer tape will be delivered in 
accordance with the contract. This tape will be added to the Epilepsy 
Abstracts Retrieval System (EARS) data base. 



19v 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Epilepsy 
July 1, 1973— June 30, 1974 



ABBOTT LABORATORIES (NOl-NS-3-2314) 

Title: Synthesis and Testing of New Anticonvulsants 

Contractor's Project Director ; N. P. Plotnikoff, Ph.D. 

Current Annual Level ; $111,437 

Objectives: To synthesize a new series of organic compounds and evaluate 
their potential anticonvulsant spectra in small animals. This is to be fol- 
lowed by selecting the best candidate compounds for fxirther pharmacologic 
studies. 

Course of Contract : The contract was initiated on June 15, 1973, and work 
started immediately thereafter. Approximately 30 compounds have been syn- 
thesized and evaluated for anticonvulsant activity. Several of these com- 
pounds are now undergoing further evaluation. 

Proposed Course of Contract ; This study will continue for 1 more year at 
which time synthesis and pharmacologic evaluation of anticonvulsant activity 
will be completed. 



21 



V 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Head Injury and Stroke 

July 1, 1973— October 20, 1974 



UNIVERSITY OF NEW MEXICO (NIH-71-2316) —Coordinating Center 

VIRGINIA COMMONWEALTH UNIVERSITY (NIH-71-2315) 

OHIO STATE UNIVERSITY (NIH-71-2317) 

UNIVERSITY OF CALIFORNIA, SAN DIEGO (NIH-71-2318) 

NEW YORK UNIVERSITY (NIH-71-2319) 

NORTHWESTERN UNIVERSITY (NIH-71-2320) 

UNIVERSITY OF UTAH (NIH-71-2322) 

COLLEGE OF PHYSICIANS & SURGEONS-COLUMBIA UNIVERSITY (NIH-72-2312) 

Title : Collaborative Study of Cerebral Death 

Project Coordinator : A. Earl Walker, M.D. (UNM) 

Project Directors : Gary Suter, M.D. (VCU) 

Norman Allen, M.D. (OSU) 
Reginald G. Bickford, M.D. (UCSD) 
Julius Korein, M.D. (NYU) 
Benjamin Boshes, M.D. (NU) 
Donald R. Bennett, M.D. (UU) 
Eli Goldensohn, M.D. (CPS) 

Current Annual Level : $251,270 - completion 

Objectives : The overall objectives of the collaborative study of cerebral 
death were 1) to verify or modify the current clinical and electroencephalo- 
graphic criteria for cerebral death in relation to patient age and cause of 
coma; 2) to determine the minimal time that clinical and EEC criteria must be 
operative to indicate cerebral death in relation to age and cause of coma; 
3) to assess the neurologic deficit of patients who improve after having 
fulfilled all criteria for some length of time; and, finally, 4) to 
characterize abnormal EEC patterns seen prior to electrocerebral silence. 

Major Findings : The study is completed, and 503 cases have been analyzed. 
The report summarizing the results of the study has been received and is in 
review by NINDS staff. 

In the apneic, unresponsive patient the results show that a one-hour flat EEG 
predicts death to a high degree of accuracy. However, "respirator brain" was 
not present in all patients meecin,^ the Harvard Criteria, and, therefore^ 
cerebral death remains a clinical state without a pathological diagnosis. 

There have been two cases of drug intoxication in whi^h the LEG's have been 
flat and subsequent recovery achieved. In view of this fact, any future 
studies will have to provide an alternative means of identifying the drug- 
intoxicated cases. By further requiring fixed and dllcted pupils, these two 
cases would not have been exceptions. However, back-up tests of cerebral 
blood flow estimations need to be used. A noninvasive test of blood flow was 
proven feasible, and a separate contract-supported project has been solicited 
to establish the validity and reliability of the technique. 



The report concludes that although death under certain circumstances may be 
predicted with varying degrees of confidence, definitive criteria for brain 
death cannot be established on a statistical basis. 

Significance to NINDS Program and Biomedical Research : Considerable con- 
troversy still exists over the legal, moral, and scien' ific definitions of 
death. The criteria used for the diagnosis of brain death vary in the world 
literature. The need for reliable uniform criteria still exists in oi'der to 
spare families the emotional stress and financial burden of unnecessary 
delays in pronouncement of death, and, at the same time, to protect against 
undue haste in cessation of efforts of resuscitation. European criteria 
include assessment of blood flow by aortocranial angiography in primarily 
brain-damaged patients while the Harvard Criteria which do not require blood 
flow assessment remain the most widely accepted norms in this country. This 
study points out the need for establishing morphological criteria for 
respirator brain and irreversible anoxia as well as the need to quickly 
identify patients who have taken depressctnt drugs in the absence of clinical 
history. While the investigators conclude that some quick noninvasive blood 
flow measurement should be developed to increase the accuracy of prediction of 
death, the study also pointr; out the need for standardization and dissemination 
of drug screening procedures. 

Publication : 

Smith, A.J.K., Walker, A.E.: Cerebral blood flow and brain metabolism as 
indicators of cerebral death. Johns Hopkins Med J 133:107-119, 1973. 



24v 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Head Injury and Stroke 

July 1, 1973— June 30, 19 74 

INDIANA UNIVERSITY FOUNDATION (NIH-72-2324) 

Title : A Study of the Hospital Frequency and Character of Transient 
Ischemic Attacks — Mark L. Dyken, M.D. Principal Investigator 

Current Level : $247,406 (21 months) 

Collaborating Centers : University of Maryland 

University of Mississippi 
Institute of Medical Sciences 
University of Washington 
Massarhusetts General Hospital 
Indiana University 

Project Directors : Thomas R. Price, M.D. (UMd) 

Armin F. Haerer, M.D. (UMiss) 
Philip R. Calanchini, M.D. (IMS) 
Phillip D. Swanson, M.D. (UW) 
David C. Poskanzer, M.D. (MGH) 
Mark L. Dyken, M.D. (Ind) 

Objectives : The objectives of this study are to determine and describe 
prospectively the relative frequency of each of the etiologies of transient 
ischemic attacks (TIA's), to classify TIA symptoms, and to collate s3niiptoms 
frequency and accuracy of diagnosis as determined through short term 
follow-up. The study is limited to patients presenting for hospitalization 
or hospital outpatient clinics with. a presumptive diagnosis of transient 
ischemic attack. 

Major Findings : Total entry forms for October 1972 through December 1973 are 
590. During this period 488 new entries were made with 202 follow-ups. 
Analysis of this preliminary data indicated blacks have less TIA's of the 
carotid system. Only 28 patients have been examined during an actual TIA 
attack. Initial data also seems to indicate no significaat clinical change 
occurred using antiplatelet, anticoagulation or surgical approaches. Most 
it>:portout is the documentation that approximately 30 percent of initial TIA 
diagnoses change after critical scrutiny. 

Significance to NINDS Program and Biomedical Research : A data base for 
evaluating the character, frequency and prognostic significance of transient 
ischemic attacks will be provided by this project. The findings wil] influence 
the interpretation of data from past and future studies of the epidemiology 
and therapy of TIA. A functional or descriptive definition of TIA's may be 
developed from this data base. 

Proposed Course of Contract : With a projected data base of approximately 1200 
patients, further prognostic evaluation by either individualized or collabora- 
tive efforts over a longer follow-up period would seem desirable. However, the 
data analysis now in progress should produce specific hypotheses to be tested, 
as well a? useful classifications of TIA's for testing in fu .re studies. 

25v 



Therefore, only certain subpopulations of these cases may be involved in new 
or extended research programs, depending upon the scientific merit of the 
present program. There is a high priority need for a uniform, generally 
acceptable definition and prognostic classification of TIA's. 



26 



V 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Head Injury and Stroke 

July 1, 1973— December 29, 1973 



UNIVERSITY OF WASHINGTON (NIH-72-2325) 

Title : Fragility of Brain Tissue 

Contractor's Project Directors : James D. Chalupnik, Ph.D. 

E. C. Alvord, Jr., M.D. 

Current Annual Level : 

Objectives : The purpose of this project was to establish a fragility index 
for brain tissue in vivo, reflecting the minimum levels of stress and strain 
likely to cause histological damage. This was to be accomplished by comparing 
experimental data derived from animals to predicted data from a computerized 
mathematical model of static loading of cerebral cortex. 

Major Findings : To date there are no useful data reported from animal model 
studies. A final report has not yet been received. 

Significance to NINDS Program and Biomedical Research : As originally 
designed, this study was to use data from the Head Injury Model Project, and 
the findings would complement that study. The instrument us»d to generate the 
stress fields in brain tissue was to provide a predictable field but not 
simulate the blunt trauma caused by accidental head injury in humans. To 
establish norms of microvascular fragility remains an important goal. 

Proposed Course of Contract ; Contract was terminated in December 1973 after a 
six-month, no cost extension failed to permit successful completion of the 
project. 



27 V 



CONTRACT NARRATIVE 
Applied Neurologic Research Branch — Section on Head Injury and Stroke 

July 1, 1973— June 30, 1974 

UNIVERSITY OF WASHINGTON (NIH-72-2326) 

Title : Human Responses to Head Injury 

Contractor's Project Director : Ralph M. Reitan, Ph.D. 

Current Level : $271,274 (36 months) 

Objectives : 1. To describe prospectively the recovery pattern of psychological 
function after brain trauma. 2. To establish the time course over which 
recovery occurs. 3. To identify the sub-tests in a comprehensive neuro- 
psychological battery which have the greatest prognostic value. 

Major Findings : Based on work done to date: 1. There has been a 30 percent 
loss to follow-up among survivors of acute head injuries. 2. Patients still 
in follow-up show pr(;gress changes toward improvement and unstable EEC 
patterns at one year. 3. The loss to follow-up rate and small numbers of 
patients prohibit the prospective study from producing statistically valid 
conclusions. 4. Neuro-otological work-up of patients has not proved valuable 
in predicting deafness or permanent vestibular dysfunction in head-injured 
patients. 

Significance to NINDS Program and Biomedical Research : This study will yield 
late recovery patterns which reputedly may require up to two years of con- 
valescence for completion. The study is complementary to numerous other 
program projects addressing prognosis for survival after coma and prognosis 
for functional recovery after stroke. 

Proposed Course : Due to small numbers, statistical significance of trends 
noted in the prospective subjects alone cannot be tested. In order to achieve 
the originally stated goals and objectives, this contract will be ex*-°nded for 
one year to allow two years of prospective follow-up since psychological and 
electrophysiological parameters are not stable at one year and to allow for 
statisti;,al testing of hypotheses developed in the prospective 3tudy in the 
voluminous data bank on head-injured patients; accumulated by the project 
director. Successful completion of tliis contract is expected after one 
additional year's effort. 



29v 



Serial No. NDS (CF)-71 AN 1932 

1. Collaborative and Field Research 

2. Epilepsy Section 

3. Bethesda, Maryland 

PHS— NIH 
Individual Project Report 
July 1, 1973--June 30, 1974 

Project Title: Long-term electroencephalographic telemetry of patients 
with absence (petit mal) seizures 

Principal Investigator: J. Kiffin Pe^ry, M.D. 

Other Investigators: Bruce L. Ehrenberg, M.D. 

Fritz E. Dreifuss, M.D. 
Paul R. Dyken, M.D. 

Cooperating Units: Departments of Neurology, University of Virginia 

School of Medicine, Charlottesville, Virginia, 
and Medical College of Georgia, Augusta, Georgia 

Man Years: 

Total: 0.75 
Professional: 0.25 
Other: 0.5 

Facilities and Equipment: 

The telemetry equipment and FM tape recorder were provided by the 
Section on Epilepsy. The clinical research units were provided by the 
University of Virginia Hospital, and the Medical College of Georgia 
Hospital . 

Project Description: 

Objective s: This study evaluate''^ patterns of paroxysmal abnormal 
discharge in patients with absence (petit mal) seizures. Another 
primary objec;;ive is to determine if tnis method can be established 
as a primary mode of evaluating any absence drug. 

Methods Employed : Each patient has a 12-hour telemetered electro- 
en cephaTographic recording on the clinical research unit. The telemetry 
equipment is placed on the patient's head by an EEG technician who also 
applies the electrodes. The patient is free to move about the ward and 
the signal from the transmitters is taken to a dipole antenna and then 
to four receivers where four channels of the EEG 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 



31 V 



Serial No. NDS (CF)-71 AN 1932 

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 
physician. Patterns of discharge and effects of anti -absence drugs are 
evident when the data is processed by a 360 IBM computer and the Calcomp 
plotter. 

Major Findings : Patients with absence epilepsy have generally shown 
improvement following treatment with absence drugs. 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. 

The 12-hour telemetered EEG has been compared statistically to other 
measures of seizure frequency and found to be more reliable. A program 
has been developed on the PDP-12 computer for rapid quantification of 
the number and duration of spike-wave paroxysms in a 12-hour period. 

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 
suggested patterns of paroxysmal discharyj in patients with absence 
seizures and points a way for further investigation of the mechanisms 
of absence epilepsy. 

Honors and Awards: None 

Publication : 

Browne TR, Penry JK, Porter RJ, Dreifuss FE: A comparison of clinical 
estimates of absence seizure frequency with estimates based on prolonged 
telemetered EEGs. Neurology 24:381-382, 1974. 



32 V 



Serial No. NDS (CF)-71 AN 1933 

1. Collaborative and Field Research 

2. Epilepsy Section 

3. Bethesda, Maryland 

PHS--NIH 
Individual Project Report 
July 1, 1973-^June 30, 1974 

Project Title: Quantitation of clinical manifestations of spike-wave 
activity by a reaction time method 

Principal Investigator: J. Kiffin Penry, M.D. 

Other Investigators: Teiko Sumino, M.D. 

Susumu Sato, 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 disputed 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 
recognition 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. 0.) instanta- 
neous recognition the reaction timer is triggered and a tone is delivered 
to the subject. The subject responds by turning off the high pitch tone 

33 V 



Serial No. NDS (CF)-71 AN 1933 

with a telegraph key. Between 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 bv television, including a portion of 
the screen reserved for the reaction time from the digital clock. There 
is no age limit in selecting patients, but they must all have spike-wave 
paroxysmal discharge. A second group of patients was studied with the 
apparatus altered slightly so that the auditory stimulus was delivered 
0.5 seconds into the seizure in order to see if responsiveness becomes 
less as the seizure progresses. Osicillographic displays from magnetic 
tape recordings of spike-wave paroxysms revealed shifting asymmetries. 

Major Findings : The findings of the first group of patients suggest 
that some ability to respond early during the paroxysmal burst is main- 
tained; 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 such 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. Specif- 
ically, this study is an analysis of the relation of the patient's 
behavior to his EEG during paroxysmal electroencephalographic events. 
An understanding of this relationship is important--not only as a guide- 
post 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. The degree of generalization of 
the spike-wave paroxysms will be measured by computer. 

Honors and Awards: None 



34 V 



Serial No. NDS (CF)-72 AN 2097 

1. Collaborative and Field Research 

2. Epilepsy Section 

3. Bethesda, Maryland 



PHS— NIH 
Individual Project Report 
July 1, 1973— June 30, 1974 

Project Title: Diagnostic value of prolonged telemetered EEG in epilepsy 

Principal Investigator: J. Kiffin Penry, M.D. 

Other Investigators: Teiko Sumino, M.D. 

William L. Brannon, Jr., M.D. 

Cooperating Unit: Department of Neurology, Naval Medical Center, 

Bethesda, Maryland 

Man Years: 

Total: 0.4 
Professional: 0.2 '' ' 
Other: 0.2 

Facilities and Equipment: 

The telemetry equipment and FM tape recorder were provided by the 
Section on Epilepsy. Research was conducted at the Department of 
Neurology, Naval Medical Center, Bethesda, Maryland. 

Project Description: 

Objectives and Methods : To develop a means of detecting and 
recording interictal paroxysmal abnormalities (epileptiform) in the 
EEG of patients who have suffered a clinical convulsion. Routinely, 
a 20-30 minute electroencephalogram, which includes hyperventilation 
and photic stimulation, will detect interictal EEG abnormalities in 
30-50% of the patients who suffer convulsions; there remains th? ot^^er 
50% of the patients who are at risk for recurt-erit convulsions and may 
be suffering the onset of epilepsy. Traditionally these patients must 
await recurrence of seizures to make a diagnosis, or repeating EEG 
determinations in the hope of catching an interictal abnormality. A 
recent study by Johnson, et al . has shown that 40% of the patients who 
suffer a single generalized tonic-clonic seizure have a recurrence within 
one year. If th'sse patients could be detected they could be treated 
and seizure occurrence prevented, without treating those patients who 
will not have a recurrence. 



35 



V 



Serial No. NDS (CF)-72 AN 2097 

Therefore, a study has been designed to record a telemetered EEG for 
6 hours in order to sample over a period of time longer than the usual 
20-30 minutes and during normal activity; in some patients, the latter may 
evoke interictal paroxysmal abnormalities. The study compares the 
incidence of diagnostic paroxysmal abnormalities in the 6-hour telemetered 
EEG with those from routine conventional EEGs. 

Course of the Study : To date 30 patients have been studied. Each 
patient has had a bona fide seizure within 30 days of the recording, 
and has also had a normal electroencephalogram by ordinary diagnostic 
workup. 

Statistically, it is estimated that from 60-80 patients will be 
required to prove the hypothesis that the prolonged telemetered EEG 
during normal behavior will significantly increase the diagnostic ability 
over that of a single EEG after the first seizure. 

Major Findings : In the first 30 patients accessioned, three have 
had diagnostic abnormalities on the 6-hour telemetered EEG which were 
not recorded in the routine EEG. The accessioning of patients will 
continue over another 10 months. 

Significance : Th° ability to detect and record diagnostic epilepsy 
format abnormalities in the EEG after a single seizure will aid in the 
early treatment and long-term prognosis of patients who suffer their 
initial seizure. 



36v 



Serial No. NDS (CF)-74 AN 2098 

1. Collaborative and Field Research 

2. Section on Epilepsy 

3. Bethesda, Maryland 

PHS~NIH 
Individual Project Report 
July 1, 1973~June 30, 1974 

Project Title ; The Monaural Auditory Evoked Potential as a Measure of 
Anticonvulsant Drug Effect on Brain Function. 

Principal Investigator : Jonathan R. Wolpaw, M.D. 

Other Investigators : J. Kiffin Penry, M.D. 

Man Years: 



Total: 1.7 
Professional: 1.0 
Other : 0.7 

Facilities s Equipment : Laboratories of NINDS, NIH 

Project Description : 

Objectives; To develop the ipsilateral-contralateral peak latency dif- 
ference previously noted by others in the monaural auditory evoked response 
as a measure of acute and chronic anticonvulsant effect on central nervous 
system function. 

Major Findings ; Through an extensive study in normal control, we fur- 
ther define the origin and magnitude of the previously reported ipsilateral- 
contralateral peak latency difference. In pilot studies, moderate doses 
of ethanol and caffeine increased the ipsilateral-contralateral peak latency 
difference, often giving differences significantly higher than control values. 
At present, data is being collected from patients before and after initiation 
of anticonvulsant therapy. Results from a small number of patients allow 
no definite conclusions at this point, but it appears that phenobarbital gives 
abnormally high ipsilateral-contralateral peak latency differences. 

Proposed Course; Continued collection of data from patients pre- and 
post-initiation of anticonvulsant therapy, continued follow-up of patients 
already in study if possible, and continued ancillary control studies to fur- 
ther define the nature of the measurement and to arrive at the best possible 
method of measuring it. 



37v 



-, , Serial No. NDS (CF)-74 AN 2099 

^' ., ., J 1. Collaborative and Field Research 

... , .: 2. Epilepsy Section 

"; 3. Bethesda, Maryland 

PHS — NIH 
Individual Project Report 
July 1, 1973~June 30, 1974 

Project Title : Pharmacokinetics of Ethotoin in Epileptic Patients 

Principal Investigator : Harvey J. Kupferberg, Ph.D. 

Other I nve stigators : Wayne D. Yonekawa, M.S. 

Frederick Cantor, M.D. 

Cooperating Units : Georgetown Medical School 

Department of Neurology 

Man Years : 

Total: 0.25 

Professional: 0.20 
Other: 0.05 

Facilities : Department of Neurology, 

Veterans Administration Hospital; 
Clinical Research Unit, 
Georgetown University. 

Pro j ec t Description ; 

Objective: To devise analytical procedures for the analysis of etho- 
toin, antiepileptic drug, in patients who have seizures; to determine the 
metabolic pathway and some of the pharmacokinetic parameters of the drug. 

Major Findings : A sensitive analytical method of analysis was developed 
for ethotoin, and several other hydantoins currently being used for the con- 
trol of seizures. The method is not applicable for the analysis of barbi- 
turates. This analysis was used to cetermine steady state blood levels in 
patients receiving long term chronic ethotoin therapy. Patients were given a 
single 2 gm dose and 0.5 gm on different days The absorption and disap- 
pearance of ethotoin from planma was determined. The t/2 was not significantly 
different for the two doses; the values being 5, 8 and 10 hours in the three 
patients. One patient showed "saturation" kinetics at the 2.0 gm dose. 

Three metabolites were isolated and identified in tne urine of patients re- 
ceiving ethotoin. They are: 5- phenylhydantoin, 3 ethyl-5 parahydi oxyphenyl 
hydantoin and xts gucuronide and phenyl hydantoic acid. 

Significance ; A sensitive method for the analysis of hydantoins has 
been developed which can be used to identify their metabolites. This cannot 



39 



V 



be done by the presently used "on-column" methylation procedures. The plasma 
half-life of ethotoin has been established showing it to be a short acting 
anticonvulsant. The blood levels and pharmacokinetics are necessary in order 
to maximize dosage schedule and seizure control in epileptic patients. 

Course of Project : The study has been completed. 



40 



V 



Serial No. NDS (CF)-74 AN 2100 

1. Collaborative and Field Research 

2. Epilepsy Section 

3. Bethesda, Maryland 

PHS — NIH 
Individual Project Report 
July 1, 1973~June 30, 1974 

Project Title : Analysis of KO 1173 (Mexiletine) by electron-capture-gas- 
liquid chromatography. 

Principle Investigator : Harvey J. Kupferberg, Ph.D. 

Other Investigator : Wayne Yonekawa, M.S. 

Man Years : 

Total : 0.2 
Professional: 0.2 

Facilities S Equipment : Laboratories of NINDS, NIH. 

Project Description ; 

Objectives ; To devise a sensitive, specific method of analysis for 
K6 1173, a new anticonvulsant. The method of analysis must be sensitive 
enough to detect less than 0.2 yug of drug. 

Methods Employed ; Following extraction of drug from plasma, gas-liquid 
chromatography combined with electron-capture detection was used. The com- 
pound was then reacted with pentaf luoro-benzoyl chloride to form a stable 
benzamide derivative. 

Major Findings: The assay can detect as little as 0.01 ^g of Ktt 1173 
in a ml of plasma. A double extraction procedure of plasma prior to deriva- 
tive formation imparts a high degree of specificity to the method. 

Significance : This method can be used to measure blood concentrations 
of this drug in the clinical efficacy studies supported by the Institute. 

Course of Project: This project has been completed. 



41 V 



Serial No. NDS (CF)-74 AN 2101 

1. Collaborative and Field Research 

2. Epilepsy Section 

3. Bethesda, Maryland 

PHS — NIH 
Individual Project Report 
July 1, 1973~June 30, 1974 

Project Title ; Anticonvulsant Activity of Mephenytoin in Relationship to its 
Metabolism to Nirvanol 

Principal Investigator : Harvey J. Kupferberg, Ph.D. 

Other Invesx-igator : Wayne Yonekawa, M.S. 

Man Years : 

Total: 0.25 
Professional: 0.25 

Facilities and Equipment ; Laboratory facilities of the ANRB, C&FR, NINDS 

Project Description : 

Objective ; To study the relationship between the dealkylation of me- 
phenytoin to nirvanol and their anticonvulsant activity in mice. 

Methods Employed ; The anticonvulsant ED of mephenytoin and nirvanol 
was determined at 30 min, and 3 hr. respectively following intraperitoneal 
administration. Mephenytoin and nirvanol brain and plasma levels were deter- 
mined at various times by gas-liquid chromatography following the administra- 
tion of 40 mg/kg mephenytoin intraperitoneally. 

Major Findings ; The maximal electroshock seizure (MES) ED of mepheny- 
toin in mice following intraperitoneally administrated drug at 30 minutes and 
3 hours was 36 mg/kg and 40 mg/kg ri^spectively. The brain levels of mepheny- 
toin 30 minutes following IP administration of 40 mg/kg was .'.9.2^g/cm. The 
brain levels fell to 2.7 ^g/gm at 30 minutes to 17,7 ^g/gm at 3 hours. The 
efficacy of mephenytoin in blocking electrically induced seizures in mice at 
early tiiries after administration is due to the present drug and at later times 
due to the dealkylated metabolite nirvanol. This conclusion is further sup- 
ported by the finding that the MES ED of nirvanol 3 hours after intraperi- 
toneal administration is 30 mg/kg. 

SignificancP i; A drug's metabolism plays an important role in determining 
its duration of action. In this specific case, the metabolism of the parent 
compound to an active metabolite leads to a prolong duration of activity. The 
metabolism of nephenytoin to nirvanol occurs in man end most likely is an im- 
portant factor in this drug's usefullness as an anticonvulsant. 

Course of Project ; This project has been completed. 

43 V 



Serial No. NDS (CF)-73 AN 2051 

1. Applied Neurologic Research Branch 

2. Section on Head Injury and Stroke 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1973— June 30, 1974 



Project Title: Experimental models laboratory — head injury and stroke 

Previous Serial Number: SAME 

Principal Investigators: G. F. Molinari, M.D. 

J. Fein, M.D. (AFRRI) 

Man Years: 

Total: 1.0 
Professional: 0.5 
Other: 0.5 

Project Description: 

Objective : To establish the maximum duration of cerebral tolerance of 

ischemic anoxia and to evaluate the association of ischemia and hemorrhage in 

primate models. To determine the maximum time allowable to begin therapeutic 

intervention by a number of methods. 

Methods Employed : Having established the pathologic anatomy of models 
of ischemia and subarachnoid hemorrhage, acute experiments using hydrogen 
washout blood flow methods and EEG, the early stages of evolution of ischemia 
following subarachnoid hemorrhage have been studied. These parameters will be 
monitored simultaneously in both primary and secondary zones of disordered 
circulation. 

Major Findings : Models have been developed for segmental cerebral artery 
occlusion, Cc.rebral vasospasm, and subarachnoid hemorrhage. Hemorrhagic or 
bland infarction can be selectively produced; dysautoregulation following 
subarachnoid hemorrhage has been characterized. 

Significance to Biomedical Research and the Program of the Institute ; 
These experiments are designed to provide basic pathophysiological data 
related to secondary prevention of stroke and head injury. Whila certain 
areas of direct traumatic necrosis or total cessation of blood are inevitable, 
functional sequelae may be minimized by prevention of detrimental secondary 
vascular events in fields of collateral circulation. 

These experiments are expected to complement clinical studies of secondary 
prevention in patients supported by the Section's field research program. 

Proposed Course ; Since work on the Stroke Laboratory in Building 36, 
NIH Campus has been completed, the logistical need for the In r? ,ency 
Agreement \ ith AFFRI has abated. However, successful collabo^ -tive 

45 V 



Serial No. NDS (C&FR)-73 AN 2051 

arrangements with Dr. Fein have produced studies not within the capabilities 
of this Section's in-house laboratory, and a contract with Dr. Fein's 
institution will be negotiated to continue the ongoing research projects. 
These include use of dogs for superficial temporal middle cerebral anastomotic 
surgery studies of oxygen availability and cerebral metabolism following 
subarachnoid hemorrhage in primates. 

Publications: 

Fein, J., Molinari, G.F.: Augmentation of canine cerebral blood flow by 
microanastomosis. J. Neurosurg 1974. (In Press) 



46t 



Serial No. NDS (CF)-74 AN 2118 

1. Applied Neurologic Research Branch 

2. Section on Head Injury and Stroke 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973--June 30, 1974 

Project Title: Stroke models in the primate — an embolic technique for 
intrinsic segmental occlusion of the middle cerebral 
artery 

Previous Serial Number: None 

Principal Investigator: G. F. Molinari, M.D. 

Other Investigators: J. I. Moseley, M.D. 

J. P. Laurent, M.D. 

Man Years : 

Total: 1.5 ..,■,, ; ..•.,. 
Professional: 1.0 
Other : 0.5 

Project Description: 

Objective : 1. To develop a method of embolizing the proximal middle 
cerebral artery (MCA) in the primate without involving cranial surgery. 2. 
To apply techniques using local anesthesia which permit observation of the 
natural history of embolic infarction in conscious animals. 

Methods Employed : As developed originally in dogs, a technique 
utilizing Microfil(R) silicone latex cylinders injected into the internal 
carotid artery was adapted to the monkey. Intravenous and intraperitoneal 
pentobarbital anesthesia was used in early experiments. General anesthetic 
is not necessary once technical and surgical procedures have been standardized, 

Major Findings : A model has been developed in the primate to selectively 
occlude the proximal middle cerebra] artery (MCA) by a segmental embolism. 
Silicone rubber cylinders of 1.6 mm dianeter introduced at the carotid bulb 
cause horizontal segment occlusions in 80% of trials. Failures are due to 
faulty polymerization or injection techniques. Occlusions of the anterior 
cerebral or posterior communicating arteries or the middle cerebral branches 
which spare the horizontal segment are considered failures in this effort to 
produce a standard embolic model. 

Significance to Biomedical Research and the Program of the Institute ; 
Because the procedure requres only local anesthesia and surgery confined 
entirely to the neck, the animal's behavior and physiologic response to the 
produced infarction may be observed as the stroke evolves. Any cerebral 
recording devices may be permanently fixed to the head prior to infarction 
since the brain is not manipulated and general anesthesia does not mask 
natural, history or pathophysiology. «-, 



Serial No. NDS (CF)-74 AN 2118 



Proposed Course : Preliminary pathological results suggest that with 
slight modification, e.g., manipulation of mean ar .erial pressure, pC02s 
etc., the basic method may yield the first experimental model of intra- 
cerebral hematoma and thereby establish the relationship between ischemia 
and cerebral hemorrhage. 

Publication: 

Molinari, G.F., Moseley, J. I., Laurent, J. P.: Segmental middle cerebral 
artery occlusion in primates: an experimental method requiring minimal 
surgery and anesthesia. Stroke (In Press) 

Film: _ 

Laurent, J, P., Moseley, J.I., Molinari, G.F.: Pathogenesis of cerebral 
infarction in the awake monkey — an experimental method. The film was 
prepared for the NINDS by Drs. Laurent, Moseley and Molinari showing 
experimental procedure methodology and neurological examination in conscious 
animals. This will be used in conjunction with presentations of the 
technique . 



48v 



Serial No. NDS (CF)-74 AN 2119 

1. Applied Neurologic Research Branch 

2. Section on Head Injury and Stroke 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973— June 30, 1974 

Project Title: Natural history of experimental cerebral infarction in 
conscious monkeys 

Previous Number: None 

Principal Investigator: J. P. Laurent, M.D, 

Other Investigators: J. I. Moseley, M.D. 

G. F. Molinari, M.D. 

Man Years: 

Total: 1.5 
Professional: 1.0 
Other: 0.5 

Project Description: 

Objective : To establish clinical sequence and tempo of focal 
cerebral ischemic infarction in the conscious monkey and to correlate site 
and size of vascular occlusions with distribution and size of brain 
parenchymal lesions. 

Methods Employed : A comprehensive sensory^notor-behavioral 
neurological examination was standardized for use in primates. Using the 
cerebral infarction model of preformed silicone emboli placed via internal 
carotid catherization into middle cerebral artery, immediate rept citive 
neurological exams were conducted. Anatomical and pathological specimens 
were harvest'id for correlation with the neurological status . 

Major Findings : Proximal or distal middle cerebral artery occlusion 
resulted in distinctive clinicopathological patterns. 

Significance to Biomedical Research and the Program of the Institute : 
These experiments are designed to provide basic anatomical and patho- 
physiological data related to acute stroke. 

Proposed Course : Determination of the immediate specific 
neurological sequelae of stroke allows assessment and prediction of later 
functional recovery. Continuation of clinicopathological correlation 
combined with blood flow measurements will give more informatiou 
regarding the "safe time" during which definitive surgical or medical 
treatments might produce reversal. 

49 V 



Serial No. NDS (CF)-74 AN 2120 

1. Applied Neurologic Research Branch 

2. Section on Head Injury and Stroke 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1973— April 1, 1974 



Project Title: Evaluation of the role of barbiturate in treating acute 
cerebral infarction 

Previous Serial Number: None 

Principal Investigator: John I. Moseley, M.D. 

Other Investigators: John P. Laurent, M.D. 

G. F. Molinari, M.D. 

Man Years: 

Total: 1.0 
Professional: 0.5 
Other: 0.5 

Project Description: 

Objectives ; To determine in the primate whether a short acting 
barbiturate given for 12 hours beginning within 30 minutes after embolic 
occlusion of the middle cerebral artery reduces the clinical and/or 
pathological sequelae observed in the first five days after the infarct. 

Rationale ; Cerebral infarction might be delayed by reducing metabolic 
demand while secondary supply sources improve or are restored. 

Major Findings : Compared to a control group, animals treated with 
barbiturate regained grasp, movement ^ and some strength in affected limbs. 
Untreated animals did not improve measurably in these areas during the five- 
day observation period. Pathologically, the untreated monkeys have both 
cortical and deep infarction, while treated animals have relative cortical 
sparing. FurthermorSj the morphological changes due to ischemia develop 
at a slower rate in treated animals. 

Significance to Biomedical Research : This work partly confirms previous 
work done in the dog and rabbit. Although more evaloation is needed using 
blood gas evaluations, dosage and duration of barbiturate administration, 
this work indicates that there may be a protective effect in focal 
Infarction in the primate afforded by barbiturate treatment. Barbiturate 
drugs may prove to be a desirable anesthetic for use in cerebral re- 
vascularization and other neurosurgical procedures. 



51v 



Serial No. NDS (CF)-74 AN 2120 



Proposed Course : Additional work is planned to assess the changes in 
blood gases, circulation, and possible sites of action of the barbiturate 
during the next fiscal year. 



52 V 



ANNUAL REPORT 
JULY 1, 1973 THROUGH JUNE 30, 1974 
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 

Since the preparation of last year's report, Dwayne Reed, M.D. , 
Deputy Branch Chief, has left NIH to join the South Pacific Commission in 
New Caledonia. Mrs. Estelle Kornhauser, R.N. has retired. Dr. William Horton 
has replaced Dr. Robert Stem in the Section on Genetics in Epidemiology. 

I. Epidemiologic studies of neurologic diseases 

Multiple Sclerosis 

Our activities on the NINDS MS Task Team and the National Advisory 
Commission on Multiple Sclerosis were completed and our contributions were 
included in the Report and Recommendations of the National Advisory Commission 
on Multiple Sclerosis. 

Our in-house studies are directed toward the determination of environ- 
mental factors which would explain the geographic distribution <: 7 MS and the 
obsei-vation that the critical determinants in acquiring subsequent MS occur 
before age 20. Our studies in Kansas continue in which we are attempting to 
interview a larg^ series of patients and -controls whose mothers ere still 
available for detailed information of events occurring to the patient and 
control prior to age 20. Because of the difficulty in finding suitable 
patients we have extended our study to the Topeka area and continue working 
in the Wichita area. We continue to investigate clusters of MS in different 
areas of the country in order to determine if there are common events prior 
to age 20 which could explain the high density of MS in these areas. We have 
published our findings in a cluster in Mossyrock, Washington, where In a 
population of little more than 100 people, 6 MS patients vrere bom between 
1916 and 1920, In this area a smallpox epidemic occurred in 1924 and in the 
same year the-^e were 13 deaths ranging in age from 6 months to 47 years 
listed as being caused by measles. In Comanche County, Kansas we learned 

iw 



of 5 resident MS patients. We are writing to all people bom in this area 
who have migrated away and have learned of 4 more patients. Other areas 
in which we are collecting information from apparent clusters are Bowman, 
North Dakota, Broadus , Montana and Suffern, New York. 

Data collection for our contract on MS in Veterans with the National 
Academy of Sciences has been completed. Information on approximately 6,000 
patients and controls is now being analyzed. We are in the process of 
negotiating the contract with the Massachusetts General Hospital to conduct 
a study of MS on the Shetland and Orkney Islands where the rate of MS is 
approximately 3 times higher than that reported anywhere else in the world. 
Laboratory investigations of MS are described in Section II of this summary. 

Amyotrophic Lateral Sclerosis 

Analysis of deaths among 12,000 statesiders who worked for more than 
one year on Guam between 1945 and 1955 has been updated through 1972. During 
this phase of the investigation, the age-adjusted rates of ALS were similar 
to those of the American male population and 200-fold lower than the Guamanian 
male population. 

Death rates among Guamanians for ALS and PD in California were analyzed 
through 1965. It was found that ALS occurs as frequently as on Guam, although 
PD is apparently rare. We have negotiaced the purchase of California death 
certificates for Guamanians from 1965 to 1973 in order to update these 
findings. 

In view of reports that pancreatic insufficiency may be associated with 
ALS we have analyzed a large series of patients with peptic ulcer treated by 
various regimens in Veterans Administration Hospitals. Our findings indicate 
that while there may be a slight excess of serious neurologic disease the 
rates of ALS were no higher than expected in this population. 

We are starting an in-depth study of familial ALS in order to determine 
if different syndromes occur which are family specific. In addition, 'J'' are 
collecting neuropathologic material whenever possible for routine studies 
as well as for viral studies related to our potential new finding on Guam. 
Thus far we have received one specimen and arrangements have been made to 
secure a second. 

Familial and genetic factors have been invoked etiologically in MS but 
for idiopathic ALS there is little evidence of familial factors. We are 
studying morbidity and mortality among siblings of MS and ALS patients in 
North Carolina to determine if these are distinct patterns. At the present 
time we have data on 45 MS families and 26 ALS families. 

We have completed an extensive literature review on the epidemiology of 
familial and non-familial forms of motor neuron diseases which was published 
in the Medical Progress series of the New England Journal of Medicine. 

As part of our contract with NAS on motor neuron diseases among veterans 
(Contract PH 43-64-44, Task Order 62) we are in the process of coding the 



2W 



questionnaires on living patients and controls (patients with brain tumor). 
To date we have received 90 completed questionnaires. 

Stroke 

In collaboration with Dr. Manning Feinleib, Division of Heart and 
Vascular Disease, Epidemiology Branch, National Heart and Lung Institute, 
we have analyzed cardiovascular disease in California for the 3-year period 
around 1960 and 1970 census, comparing rates among native-born Californians 
and migrants. While we observed a general decline in cardiovascular disease 
deaths in the 10-year period, there were no significant differences between 
migrants and native-born Californians except for a small excess of stroke 
among migrants in older age groups. 

Work on our contract study of the relative role of environment versus 
heredity in black populations in Panama continues (Contract NOl-NS-3-2304) . 
The study in the Bocas del Torro area has been completed and censuses are 
being prepared for the other designated areas. Our contract with Kaiser 
Permanente in San Francisco (Contract # NINDS 72-2319) , to study a population 
which developed a fjtroke at some time after a multiphasic screening exam, 
has been extended. In this survey we are matching for known risk factors 
and attempting to determine as yet undetected risk factors among those who 
develop stroke. 

A recommendation of the Guam Peer Review was that we extend our studies 
of stroke to Guam and other Pacific Islands. We would like to implement this 
recommendation through the contract mechanism as soon as possible. 

Infections or possible infections 

Juvenile Polymyositis: 

A total of 42 patients with juvenile polymyositis and matched controls 
were studied. Patients in general tended to be at least as healthy as 
controls prior to onset of illness, and no distinj^uishing antecedent environ- 
mental factors were descernable. In nine instances the onset of polymyositis 
was preceded by a variety of acute infectious diseases. Of potential signif- 
icance was th^ fact that laboratory proved streptococcal e cposure occurred 
in 20 patients and in 13 controls. Serologic studiss are underway for series 
oi viruses and also streptococcal antigens. 

Creutzfeldt-Jakob Disease: 

We have pursued the potential lead that Creutzfeldt-Jakob disease may 
be related to the consumption of animal brains. We have been unable to 
acquire accurate information on the sale of animal brains and the type of 
animal brains sold in the United States and Great Britain. British co- 
workers were unable to determine brain consumption from their patients. In 
Israel however, a study revealed that the rate of CJD in Libyan Jews is 
approximately 30 times higher than the rest of the population. As a result 
of our inquiries it was learned that sheep brain is a common food among this 



3w 



group of people. Our protocol has been incorporated therefore in a follow-up 
study in Israel relating to CJD and animal brain consumption. 

Subacute Sclerosing Panencephalitis: 

Although it is widely accepted that measles is etiologically involved 
in some way in SSPE there have been reports of toxoplasmosis in SSPE and a 
suggestion that SV 40 virus may be involved. As a result, in collaboration 
with Dr. John Sever 's group and in our own laboratory, we are in the process 
of reviewing our SSPE and controls sera for a number of antigens. In our 
first run, we did find higher titers to toxoplasmosis in patients than in 
their siblings or in their controls or controls' siblings. This is being 
repeated with more refined techniques. The data on the SV 40 and other 
antigens are not yet available. 

Selected Studies. Outcome of Pregnancy among Nurses: 

There have been two series of studies which indicated that abnormal 
pregnancy occurred at a higher rate among certain categories of nurses. In 
one series the high rate was encountered among nurses exposed to infectious 
diseases while in another it occurred in nurses who were in operating rooms 
and particularly in contact with anesthetics. Analysis of our data on 1500 
nurses in the Washington, D.C. area revealed that nurses in general have a 
slightly elevated rate of abnormal births but no individual nursing specialty 
was at a higher risk. 

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

Lymphocyte transformation studies of Guamanians with amyotrophic lateral 
sclerosis (ALS) and parkinsonism-dementia (PD) have been completed. Using 
various CNS antigens for patients and controls, no stimulation was observed 
in lymphocytes of ALS patients. A low level of stimulation was encountered 
in lymphocytes of PD patients for all CNS antigens indicating that either an 
immune mechanism plays a role in the pathogenesis of PD or more likely that 
it reflects a secondary response to the massive destruction of CNS cells in 
the disease. 

Dr. Hilary Koprowski, Director of the Wlstar Institute, Philadelphia, 
and his associates have recently isolated parainfluenza type I from the 
brains of 2 MS paLiencs. As reported previously we were unable to demonstrate 
higher hemaggli'tinating-inhibitory (HI"; antibody titers in tha sera of MS 
patients than controls for the MS 6/94 isolate and standard pfxainfluenza 
type I. In an expansion of this study we measured hemolysis-lnhibiting (HLI) 
antibody titers of the same study group. We found HLI antibody levels to be 
essentially similar in MS patients and controls with the MS -3/94 isolate and 
standard parainfluenza type I. HLI antibody titers to measles virus however ^ 
were found to be significantly higher in sera of MS patients than control 
sera. 

In an attemp" to delineate the role of measles virus in the pathogenesis 
of MS we are currently devising hybridization techniques designed to detect 



4w 



measles virus nucleic acids in cells of patients with the disease. Radio- 
active viral nucleic acid probes are being prepared using various strains 
of purified measles virus. Cellular nucleic acids will be extracted from 
CNS tissue specimens and cell cultures of patients with MS and annealed with 
the radioaccive viral probes. If viral genetic material related to measles 
is found in cells of MS patients it will be the strongest evidence yet for 
an association of this agent with the disease. 

We have completed a collaborative study with Dr. James Ganley of the 
National Eye Institute in which lymphocytes of patients suffering from 
presumed ocular histoplasmosis were challenged with a large battery of 
antigens and mitogens. Lymphocytes of patients with the disciform lesion 
were more reactive to pokeweed mitogen than Ijrmphocytes of controls whereas 
phytohemagglutinin was found to be less stimulatory. Lymphocytes from the 
study group with the disciform lesion had higher mean counts per minute than 
those from controls with 10 of the 11 specific antigens tested. This suggests 
a hyperreactivity of the cellular immune system in patients with the disciform 
lesion of presumed ocular histoplasmosis and strengthens the hypothesis that 
the cellular immune system plays an etiologic role in the development of the 
lesion. 

During the past year our laboratory continued to emphasize the develop- 
ment and application of direct biochemical approaches to the detection of 
viral infection. Among these were the demonstration of reverse transcriptase 
(RT) enzyme activity and the detection of viral DNA or RNA by nucleic acid 
hybridization. These were applied to two model systems (minute virus of 
mice infections i^ vitro , and murine leukemia Iji vivo ) to selected neurologic 
diseases in man. The studies on MVM (minute virus of mice) involved the 
further definition of 5 tissue culture systems in which infection has per- 
sisted for 1-3 years. The investigations on murine leukemia (carried out in 
collaboration with Dr. Michael Viola) helped us to further develop techniques 
to detect and characterize oncornavirus infections. In these studies, virus- 
related nucleotide sequences were demonstrated in highest concentration in 
malignant cells (as compared with normal cells) and in normal cells from mice 
of strains having a high incidence of "spontaneous" leukemia or lymphoma (as 
compared with low incidence strains). Further, thermal stability of the 
hybrid molecules was studied to clarify the extent of relatedness between 
the "endogenous'' leukemia virus, Rauscher leukemia virus. Gross lympiioma 
virus, and the AKR leukemia virus. 

Similar methods for oncornavirus d»>.tecticn were applied (a^ain in 
collaboration with Dr. Viola) to brain tissues fjom patients with MS, ALS , 
schizophrenia, SSPE, stroke, parKineonism-dementia, and ALS of Guam. In all 
but the last two diseases, preliminary results were negative. RT enzyine 
activity associated with an oncornavirus-like particle has been demonstrated 
in several Chamorran brains, from patients with ALS, PD, and unrelated 
diseases. Preliminary characterization suggests that the enzyme is of viral 
origin but immunologically distinct from other oncornavirus RT enzymes, and 
that the viral genome has partial homology with the visra virus genome. These 
findings are consistent with high prevalence, endemic oncornavirus infection 
among Chamorros and raises the possibility of a viral etiology for Guamanian 



5w 



ALS and PD. Intensive investigation of these possibilities and a search for 
other virus-related DNA and RNA sequences in these and other neurological 
diseases is continuing. 

III. The continued activities on Guam and t'^e Trust Territories 

In September 1973, a Peer Review group met for a second time on Guam 
and then presented its findings and recommendations to the Directorate 
of NINDS in February 1974. In view of the favorable recommendations, NINDS 
is renegotiating the Guam Agreement to continue the study for 3 years after 
October 1974. 

All the data on Guam cases and the various registries has been or is 
being computerized here in Bethesda. A statistically significant decline in 
the rates of ALS and PD for both sexes has been documented during the last 
8 years. The rate of ALS still remains about 50 times higher on Guam than 
elsewhere in the world. The decline in rates has been most marked in southern 
villages in which rates previously had been highest. The decline occurred in 
all age groups and hence no cohort phenomenon was observed. A series of 
genetic registries have been updated and while familial clustering, particu- 
larly of ALS is apparent, classic genetic patterns have not emerged. Studies 
of the offspring of doubly-affected spouses while still too early to be 
definitive are suggestive that the rates will be lower than those compatible 
with a simple genetic explanation. 

Life tables for ALS have been completed and reveal that no presenting 
symptoms (even bulbar involvement) are useful in predicting duration of 
survival other than late age at onset. A similar analysis for PD is in 
Progress. 

Drug studies in collaboration with Thomas N. Chase, M.D. , Chief, Unit 
on Neurology, Laboratory of Clinical Sciences, NIMH, reveal that all patients 
with extrapyramidal features continue to benefit on L-Dopa and are much 
easier to manage on L-Dopa plus MK 486. ALS patients were treated for one 
year with L-Dopa and showed no improvement. Of interest was the observation 
that no secondary adventicious movement developed among ALS patients, ^nile 
half of the PD patients on similar doses for equivalent times demonstrated 
these secondan^ mcements. The on— off phenomenon observed in c-assic Parkin- 
son's disease patients being treated with L-Dopa apparently does not occur 
among FD patii^nts . Biochemical studies are underway to determine the cause 
cf this discrepancy. A 2-year study of inofiiplex (isoprinosine) in amyotrophic 
lateral sclerosis revealed that the drug vas of no benefit. Analysis of this 
study revealed that at least 20% of ALS patients, whether on drug or placebo 
were clinically stable for a 2-year period, emphasizing the necessity for 
careful study design in therapeutic trials of ALS. 

Neuropathologic studies continue to reveal a high percentage of 
apparently normal Guamanians with some degree of PD-like CNS degeneration. 
These studies will be expanded in the coming year. Fresh frozen material 
from the CNS and in some instances from other organs has been collected 
from approximately 50 deaths on Guam during the past year. This includes 
material from ALS and PD patients, from Guamanians who died from other 



6w 



causes and from non-Guamanians . Preliminary results from Dr. Michael Viola of 
the Institute of Cancer Research, New York, in Dr. Speigelman's group at 
Columbia, and from our own laboratories suggest the presence of an oncorna- 
like virus in the brains of ALS and PD patients, and Guamanian controls which 
is not present in non-Guamanian material, Evan if this observation proves to 
be correct, the observation of the particle in non-affected Guamanians will 
make it difficult to establish the relationship between this finding and ALS 
and PD. 

IV. Genetic studies of neurologic diseases 

The Genetics Section continues its interest in hereditary movement 
disorders, hereditary nervous system tumors, and hereditary epilepsies. 

Evaluation of various treatment programs for the torsion dystonias 
indicates neurosurgery in the hands of Irving Cooper, M. D. as the single 
most effective approach. Of the myriad drugs employed. Diazepam, is most 
helpful. Less than 5% of dystonia patients received long-term benefit from 
L-Dopa. An unusua.! number of yoimg patients reported significant sexual 
changes while on L-Dopa and this is being studied further, A clinical trial 
of Dantrolene Sodium is underway in collaboration with Dr. Stanley Fahn of 
the Neurological Institute, New York City, Columbia Presbyterian Medical 
Center and Eaton Laboratories. 

Explanation for the observation that dopamine-beta-hydroxylase, the 
enzyme which governs the conversion of dopamine to noradrenalin, is present 
in abnormal levels in individuals with autosomal dominant torsion dystonia 
has not been forthcoming. This work, carried out in collaboration with 
Dr. Fred Wooten and Dr. Julius Axelrod continues since it may lead to an 
understanding of the basic biochemical abnormalities in the dystonias and 
potentially in other movement disorders such as Parkinson's disease and 
Huntington's chorea. 

The evaluation of attitudes about Huntington's disease is complete. 
In affected families, greatest concern is related to physical and mental 
changes rather than genetic or social consequences. Most respondents regarded 
a lay organization. The Committee to Combat Huntington's Disease as their 
best source o^: information in contrast to medical specialijts or geneticists. 
Although respondents at risk were not likely to limit family size automati- 
cally, tliey were interested in predictive tests on parent or fetus. This 
study indicates that lay organizations can be a useful 5 efficient means for 
the dissemination of information and provision of psychological support for 
members of families affected with specific genetic disease. 

In the area of hereditary brain tumors we continue to explore the 
relationship between central neurofibromatosis, as characterized by hereditary 
bilateral acoustic neuroma and peripheral neurofibromatosis. Evaluation of 
melanosomes from cafe-au-lait spots of patients with bilateral acoustic 
neuroma is being carried out with the collaboration of the Department of 
Dermatology of the Massachusetts General Hospital. If these melanosomes show 
the same changes as have been described in peripheral neurofibromatosis, it 
will be strong evidence that both conditions have a common origin. It will 



7w 



also allow a means of detecting carriers of either trait in the pre symptomatic 
period. Nerve growth factor for use has been evaluated by David Siggers, 
Johns Hopkins Hospital, in one kindred with central neurofibromatosis and 
was found to be elevated in those affected and in a fraction of those at 
risk. If this change is consistent, it also could be useful in counseling 
as well as instructive in suggesting etiology and treatment. 

The first phase of the progressive myoclonic epilepsy study is completed. 
Thirty-seven affected in 19 families have been ascertained. At least five 
distinct diseases are present within this symptom complex. We continue our 
collaboration with several neurochemical laboratories in an attempt to define 
the biochemical lesion of each. Over 75 affected individuals in 15 families 
with the von Hippel-Lindau syndrome are being studied from clinical, genetics 
and chromosomal aspects. 



8V7 



CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 1974 

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: - Ended October 1973 



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 
were analyzed. 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. The updated results indicate 
a pattern of greater risk in Georgia than the Mountain States thus 
supported previous observations. However, in contrast to other studies 
the difference was entirely due to an excess of deaths in Georgia from 
hemorrhagic stroke. Ischemic stroke rates were equal in both areas. 
The data were not changed when race was controlled. 



9v? 



Contract (NIH71-2026) 

Significance to NINDS Program and Biomedical Research : The 
epidemiological patterns of stroke are poorly xmderstood, 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 : This project has been terminated. 

Publications : Nefzger, M.D. , Acheson, R.M. and Heyman, A.: Mortality 
from Stroke among U.S. Veterans in Georgia and Five 
Western States. I. Study Plan and Death Rates. J. Chron . 
Pis . 26:393-404, 1973 

Acheson, R.M. , Nefzger, M.D. and Heyman, A.: Mortality 
from Stroke among U.S. Veterans in Georgia and Five 
Western States. II. Quality of Death Certification and 
Clinical Records. J. Chron. Dis . 26:405-414, 1973. 

Acheson, R.M. , Nefzger, M.D. , and Heyman, A.: Mortality 
from Stroke among U.S. Veterans in Georgia and Five 
Western States. III. Hypertension and Demographic 
Characteristics. J. Chron. Dis . 26:417-429, 1973. 

Heyman, A., Nefzger, M.D. and Acheson, R.M. : Mortality 
from Stroke among U.S. Veterans in Georgia and Five 
Western States. IV. Clinical Observations. J. Chron . 
Dis . 26:431-446, 1973. 

Acheson, R.M. , Nefzger, M.D. and Heyman, A.: Mortality 
from Stroke among U.S. Veterans in Georgia and Five 
Western States. V. Seventh and Eighth Revisions of the 
International Classification of Diseases as They Relate 
to Stroke. Amer. J. Epid . 96:396-400, 1973. 

Nefzger, M.D. , Acheson, R.M. and Heyman, A.: Mortality 
from Stroke among U.S. Veterans in Georgia and Five 
Western States. VI. Clinical Validation of Death 
Certificates. J. Chron. Dis. In press. 



10 



w 



CONTRACT NARRATIVE 

Epideniiology Branch, C&FR, NINDS 

Fiscal Year 1974 

NATIONAL RESEARCH COUNCIL, FOLLOW-UP AGENCY (PH43-64-44 [Task Order 53] ) 

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

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

James Norman, 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, socioeconomic 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 : Data collection is proceeding. Material will be 
analyzed during the third year of this contract. Considerable effort 
is now being expended in identification and controlling of Korean War 
veterans . 

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 
socioeconomic 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 has been terminated. 



11 W 



CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 19 74 

NATIONAL ACADEMY OF SCIENCES (PH43-64-44 [Task Order 62] ) 

Title : Epidemiologic and Follow-up Study of Amyotrophic Lateral 
Sclerosis and Other Motor Neuron Diseases 

Contractor's Project Director : James Norman, M.D. 

Gilbert Beebe, M.D. 
John F. Kurtzke, M.D. 

Current Annual Level : $43,996.00 

Objectives : 1. An epidemiologic evaluation of potential risk factors 
among approximately 500 patients with various forms of motor neuron 
diseases will be conducted. All deaths among U.S. males coded 365.1 
(ALS) for 1963-1967 will be collected and those who are veterans will 
be selected for the study. Controls will be based on National Service 
Life Insurance holders in a fashion similar to the ongoing multiple 
sclerosis veterans study. Pertinent data will be collected from 
various induction and military service records and coded, and significant 
differences between patients and controls will be sought. A subsample 
of patients' records will be reviewed to confirm the death certificate 
diagnosis of ALS. 2. A study of the natural history of the various 
motor neuron diseases including ALS will be investigated from Veterans 
Hospital admissions from 1957 to 1964, who were diagnosed as having motor 
neuron disease. The total number should be approximately 500 of whom 
approximately 200 should have motor neuron diseases other than ALS. The 
epidemiologic patterns as well as the clinical course of the various 
entities now grouped under motor neuron disease will be analyzed and 
compared to see if distinctive patterns emerge. In addition, through 
this technique the first life table of ALS will be developed by using 
a stratified age of onset sampling method. 3. A detailed case-control 
study will be conducted involving 200 living ALS patients and matched 
controls. With this mechanism more specific information on life styles 
and patterns of residence, employment, etc. can be analyzed which would 
not be available through abstracting military induction of Veterans 
Administration Hospital records. The 200 patients and 200 controls 
will be selected from the same VA Hospital Service and interviewed by 
trained social workers in the VA System. 

Ma j or Fi ndi ngs : During the contract's second year all 515 cases for 
Part 1 were identical. Controls are being secured. Tapes were made 
available for the VA for Part 2. Approxim.ately 100 interviews have 
been completed for Part 3. 

Significance to NINDS Program and Biomedical Research : Regional 
differences and socioeconomic differences have been commented upon 
for amyotrophic lateral sclerosis in the United States but definitive 
data are lacking. This study will determine for a large population 



13 V7 



Contract (PH43-64-44 [Task Order 62]) 

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 of Project : This project was to run three years. 
Budget for the third year will be 57,580. Because of a disasterous 
fire in the VA Record Center this project will require a fourth year 
at $35,000. 



14 



V7 



CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 19 74 

KAISER FOUNDATION RESEARCH INSTITUTE (NINDS 72-2319 ) 

Title : Risk Factors for Stroke 

Contractor's Project Director : G. Browne Goode, M.D. 

G. Friedman, M.D. 

Current Annual Level : Project was to run from June 30, 1972 - 

December 29, 1973 at a total cost of $91,440. 
A one-year extension was granted at a cost of 
$18,150, 

Objectives : 1. Identify patients admitted to the San Francisco Kaiser 
Foundation Hospital and Outpatient Clinic, with the diagnosis of CVD, 
during the period 1965-1971, who received a multiphasic screening exam 
prior to onset of the stroke. 2. Review the patient charts for 
validation of the diagnosis and grouping of the patients into various 
categories of CVD. 3. Select two types of control patients without 
evidence of CVD; one group matched to the cases by known risk factors 
of CVD who had received the multiphasic screening exam, and also the 
entire population which received multiphasic screening stratified by 
age and sex. 4. Obtain by computer search and transfer to a separate 
computer tape the stored multiphasic examination data for the case 
group and two control groups. 5. Conduct statistical comparisons of 
the case group with the two control groups for each variable and 
measurement included in the multiphasic examination. Statistical 
tests of differences will be used for individual items, and more 
complex statistical analysis will be used to determine the interaction 
of variables which differentiate the cases from the controls. 

Major Findings : During the first year 276 cases with various types 
of stroke were identified. Because part of the study was being funded 
from other sources which were lost, the control populations have not 
been completely identified. 

Significance to NINDS Program and Biomedical Research : Stroke is the 
fourth cause of death in the United States. By studying a well-defined 
population who received prior multiphasic screening we will increase 
knowledge on the relative affects of various risk factors a^id through 
using controls subtle and previously unrecognized risk factors may 
emerge . 

Proposed Course of Project : Necessary data and analysis will be 
completed by December 31, 1974. 



15 w 



CONTRACT NARRATIVE 

Epidemiology Branch, C&FR, NINDS 

Fiscal Year 19 74 

THE UNIVERSITY OF TEXAS AT HOUSTON (NOl-NS-3-2304 ) 

Title : Epidemiological Studies of Stroke Risk Factors in Panama 

Contractor's Project Director : Stephen Bennett, M. D. 

Reuel Stallones, M.D. 
Darwin Labarthe, M.D. 

Current Annual Level : $132,701.00 (Feb. 28, 1974 - Feb. 27, 1975) 

Objectives : To design and carry out an epidemiologic survey of certain 
populations in Panama, to study the association of genetic and environ- 
mental factors with cerebrovascular risk. At the conclusion of the 
field survey, the data will be analyzed by computer and will consist 
of descriptive tabulations of study variables by appropriate subgroupings , 
and the association of genetic and environmental factors with cerebro- 
vascular risk factors, both among and within the population subgroups. 

Major Findings : Forms were developed, Panamanian medical students 
trained and procedures were tested in the field. Attempts to work in 
the Canal Zone proved frustrating and only 100 of the 500 scheduled 
patients actually cooperated. Appropriate modifications have been 
initiated. The survey in Bocas del Toro was completed with more than 
600 participants. 

Significance to NINDS Program and Biomedical Research : Rates of 
hypert£nsion and stroke are approximately twice as high among blacks 
as among whites in the United States. There is, however, little 
information related to the association of these risk factors to 
different life styles, and genetic influence. This cross-sectional 
survey would yield valuable information concerning the comparative 
prevalence of CVD precursor disease states among defined black 
populations with different origins and of different socioeconomic 
levels. Such contrast could shed light upon the relative importance 
of the genetic versus environmental factors. 

Proposed Course of Project : This project will run approximately 2 
years, but because of delays the analysis may not be completed within 
this time. 



17 



w 



Serial No. NDS (CF) - 55 E 201 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

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

Previous Serial Number: Same 

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

Dwayne Reed, M.D. 
Frank H. Anderson, M.D. 
NINDS Research Center 

Other Investigators: Jose Torres 

NINDS Research Center 
Francisco Leon Guerrero 

NINDS Research Center 
Manuel T. Cruz -. •■ 

NINDS Research Center ■' ' 

Thomas N. Chase, M.D. 
■ NIMH ^ 

Consultants: Kwang-Ming Chen, M.D. 

Guam Memorial Hospital 
Yoshiro Yase, M.D. ■ . ■ • 

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

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

The Mayo Clinic, Rochester, Minnesota 
Olivia Cruz, M.D. 

Guam Memorial Hospital • 

Cooperating Units: NINDS Research Center, Tamuning, Guam ' 

Special Chronic Disease Studies, C&FR, NINDS 
Laboratory of Slow, Latent, and Temperate Viruses, 
C&FR, NINDS 
■: ' The Mayo Clinic, Rochester, Minnesota 

. . Department of Pathology, Massachusetts General Hospital, 

Boston 
Department of Neurology, Wakayama Medical College, 

Wakayama, Japan 
Medical Research Center, Brookhaven National Laboratory, 
Upton, New York (George C. Cotzias, M.D.) 



I9w 



Serial No. NDS (CF) - 55 E 201 

Trust Territory Health Office 
Laboratory of Clinical Sciences, NIMH 
Columbia University Hospital (Dr. Spiegelman's lab - 
Michael Viola, M.D.) 



Man Years 



Total: 11/3 
Professional: 1/3 
Other: 1 

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 : Available methods for epidemiological, clinical, 
neuropathological, neurochemical, therapeutic and virologic investigations. 

Major findings : In September 1973, a peer review group met for a 
second time on Guam and then presented its findings and recommendations to 
the Directorate of NINDS in February 1974. In view of the favorable 
recommendations, NINDS is renegotiating the Guam Agreement to continue the 
study for 3 years after October 1974. 

Dr. Dwayne Reed spent calendar year 1973 on Guam completing and 
updating various registries and conducting a series of investigations. The 
results are detailed in eight reports and three papers for publication. 

As of March 1, 1974, the NINDS Research Center was following 41 ALS 
patients, 50 patients with PD and 8 with PD/ALS. In addition, we are 
following 5 ALS suspects and 19 PD suspects. There has been a definite 
decline in rates for both ALS and PD since 1965, most pronounced in the 
southern villages where the rates were highest. The rates on Rota are 
approximately the same as those on Guam, while rates among Chamorro on 
Saipan are lower than on Guam. Extensive case-control studies have been 
completed and analyzed and showed no clear pattern which could distinguish 
patients from controls. There was an impression however that patients 
tended to lead more traditional lives and be of somewhat lower socioeconomic 
levels. This would be compatible with the finding of a declining rate for 
ALS and PD since there is a marked decline in most traditional aspects of 
life on Guam. The declining rates however did not occur in a pattern which 
would be expected if the disease were a cohort phenomenon related to any 
particular past exposure. 

At present we have approximately 40 patients with PD receiving Sinemet, 
a combined medication containing MK 486 and L-Dopa. We have published our 
successful use of MK 486 and L-Dopa during the past year. The study of 
isoprinosine in ALS was terminated. We found the drug to be totally without 

20 v 



Serial No. NDS (CF) - 55 E 201 

effect but made the very interesting observation that at least 20% of ALS 
patients are stable over a period of 2 years and that even on placebo and 
encouragement 5 weight gains of up to 15 pounds were observed. L-Dopa 
treatment of ALS proved to be without benefit although we still observe 
lowered CNS dopamine metabolism in ALS patients. Further, while more than 
half of the patients with PD developed secondary movement disorders while 
more than half of the patients with PD developed secondary movement disorders 
while taking L-Dopa, this was not observed in a single instance among 11 
ALS patients who received high levels of L-Dopa for one year. In collabo- 
ration with Dr. Thomas Chase, Chief, Unit on Neurology, Laboratory of 
Clinical Sciences, NIMH, we are planning a small study on PD patients using 
melanocyte stimulating hormone release-inhibiting factor (MIF) . 

We continue to review Chamorro brains of individuals who died of 
diseases other than ALS and PD. The work is being pursued by Dr. Haruo 
Okazaki , Section of Experimental and Anatomic Pathology, Mayo Clinic. 
Recently Dr. Leung Chen, Pathologist, Guam Memorial Hospital, at routine 
autopsy, found several Guamanians whose brains were pathologically undis- 
tinguishable from those with classic PD, but in whom there was no history 
of dementia or extrapyramidal signs. Next year Dr. Frank Anderson will be 
assigned to Dr. Edward P. Richardson, Jr., Associate Professor of Neuro- 
pathology, Pathology Department, Massachusetts General Hospital to pursue 
these unusual neuropathologic findings and to attempt to establish corre- 
lations between dementia and cortical cell loss. 

Dr. Reed completed a study of the natural history of ALS on Guam and 
it was found that survival is independent of the initial symptom or the 
sequence of symptoms involved (including bulbar) but correlated negatively 
only with advance age at onset. Dr. Anderson is conducting a similar 
study of the natural history of PD. In addition to its predictive value 
and the demonstration of creating a mathematical model for analyzing a 
disease, these data will be of particular use in analyzing effect, if any, 
on the natural history of PD by L-Dopa treatment. 

Various collaborators continue their investigations of the possible 
role of manganese, calcium metabolism and parathyroid hormone in ALS and 
PD of Guam. - ■ ■. ■■ 

A series of clinical studies are underway by Dr. Anderson and Dr. 
Kwang-Ming Chen. We still have not observed the on-off phenomenon in PD 
even in patients who have been on high doses of L-Dopa for more than one 
year. Ten patients were intensively studied over an 8-hour period with 
increasing doses of L-Dopa but even in this situation, the on-off phenomenon 
was not observed. In collaboration with Dr. Robert Joynt, Professor and 
Chairman, Division of Neurology, University of Rochester School of Medicine, 
we are attempting to determine if the absence of the on-off phenomenon is 
because Guamanian patients maintain high and stable blood levels of L-Dopa 
as opposed to stateside Parkinson's disease patients. We have performed 
serial EEGs on PD patients of normalization of EEG patterns during L-Dopa 
treatment. A major, but frustrating effort is being made to better 

21 w 



Serial No. NDS (CF) - 55 E 201 

characterize the dementia in PD. We are using progression matrices, the 
porteus maze, and Benton visual retention, along with more conventional 
neuropsychological tests. 

We have completed a study of lymphocyte transformation among patients 
and controls using Guam brain material as the test antigen. There was a 
slight tendency for lymphocytes from PD patients to respond to all antigens 
a bit more than ALS patients or controls, but this was thought to be a 
result of the massive CNS destruction and not evidence of an immune mechanism 
in either of the Guam diseases. Virologic studies with Drs. Gibbs and 
Gaj dusek continue to be negative. Studies in collaboration with Dr. Michael 
Viola in Dr. Spiegelman's group at Columbia and by Dr. Lon White in our 
lab continue to suggest the possibility that a C-type particle is present 
in Guamanian brain tissue. Findings are still preliminary but considerable 
effort is going into this study and suitable materials are being collected. 

Significance to biomedical research and the program of the Institute : 
Guam has the highest incidence in the world of motor neuron disease and the 
unique disease PD. The documentation of the epidemiological, clinical, and 
neuropathological aspects of ALS and PD have added to the world's knowledge 
concerning these neurologic diseases. In fields in which there are no known 
causes and no kncnA7n 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. 

Proposed course : Major efforts will follow peer review recommendations 
and consist of carefully following our patients, further exploring the 
neuropathology and the nature of the dementia in PD, and pursue appropriate 
epidemiologic and virologic needs. Expansion into studies of other neuro- 
logic diseases such as stroke and epilepsy is contemplated when personnel 
and time become available. 

Honors and Awards: None 

Publications: Reed, D. and Brody, J.A. : Recent observations of amyotrophic 

lateral sclerosis and parkinsonism-dementia of Guam. Presented 
at the annual meeting of the Society for Epidemiologic Research 
in Houston, Texas, May 1972. 

Reed, D. , Labarthe, D. and Stallones, R. : Epidemiologic 
studies of serum uric acid levels among Micronesians. 
Arthritis Rheum . 15:381-390, 1972. 

Stanhope, J.M. , Brody, J.A. and Morris, C.E. : Epidemiologic 
features of amyotrophic lateral sclerosis and parkinsonism- 
dementia in Guam, Mariana Islands. Int. J. Epid . 1:199-210, 
1972. 



22 w 



Serial No. NDS (CF) - 55 E 201 

Reed, D. , Labarthe, D. , Stallones, R. and Brody, J. A.: 
Epidemiologic studies of serum glucose levels among 
Micronesians. Diabetes , 22:129-136, 1973. 

Chase, T.N., Holden, E.M. and Brody, J.A. : Levodopa-induced 
dyskinesias. Comparison in parkinsonism-dementia and 
amyotrophic lateral sclerosis. Arch. Neurol. 29:328-330. 
1973. 

Labarthe, D. , Reed, D. , Brody, J.A. and Stallones, R. : 
Health effects of modernization in Palau. Am. J. Epid. 98: 
161-174, 1973. — 

Holden, E.M. , Chase, T.N. and Brody, J.A. : Parkinsonism- 
dementia of Guam: Treatment with L-Dopa and methyldopa- 
hydrazine. Neurology , 24:263-265, 1974. 

Brody, J.A., Chen, K.M. , Yase, Y. , Holden, E.M. and Morris, 
C.E. : Inosiplex and amyotrophic lateral sclerosis. Thera- 
peutic trial in patients on Guam. Arch. N eurol. 30:322-323, 
1974. 

Brody, J.A. and Kurland, L.T. : Amyotrophic lateral sclerosis 
and parkinsonism-dementia in Guam. ta_ Tropical Neurology, 
Oxford University Press. In press. 

Brody, J.A. , Stanhope, J.M. and Kurland, L.T. : Patterns of 
amyotrophic lateral sclerosis and parkinsonism-dementia of 
Guam. In Contemporary Trends: Neurological Problems in 
Oceania. Editor: R.W. Homabrook, New Guinea. In press. 

Kurland, L.T. and Brody, J.A. : Amyotrophic lateral sclerosis 
with special reference to the Guam type. In Handbook of 
Clinical Neurology. Editors: P.J. Vinken and G.W. Bruyn, 
Amsterdam. In press. 

Nemo, G. , Brody, J.A. and Cruz, M. : Lymphocyte transformation 
studies of Guamanian amyotrophic lateral sclerosis and 
parkinsonism-dementia in patients. In press. 

Reed, D.M. and Brody, J.A.: Amyotrophic lateral sclerosis 
and parkinsonism-dementia on Guam - 1945-1972. I. Descrip- 
tive epidemiology. In press. 

Reed, D.M. , Torres, J.M. and Brody, J.A.: Amyotrophic lateral 
sclerosis and parkinsonism-dementia on Guam - 1945-1972. 
II. Familial and genetic studies. In press. 

Reed, D.M. , Brody, J.A. and Holden, E.M. : Predicting the 
duration of Guam ALS . In press. 



23, 



Serial No. NDS (CF) - 63 E 1103 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

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

Previous Serial Number: Same 

Principal Investigator: Frank H. Anderson, M.D. 

NINDS Research Center 

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

Guam Memorial Hospital 
Jacob A. Brody, M.D. 
Leonard T. Kurland, M.D. 

Mayo Clinic 
Dwayne Reed, M.D. 

Cooperating Units: NINDS Research Center, Tamuning, Guam 

Mayo Clinic, Rochester, Minnesota 

Man Years : 

Total: 1 1/10 
Professional: 1/10 
Other: 1 

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 and as a source for studying other neurologic diseases. 

Methods employed : Most neurologic diseases on Guam are referred to the 
NINDS Research Center, where appropriate surveys, vital statistics and 
hospital records are used for patient detection. 

Major findings : The epilepsy registry was maintained but not updated 
during the year. All stroke deaths and stroke patients seen as inpatients 
or outpatients have been documented for the years 1972-73, 

Significance to biomedical research and the program of the Institute : 
This study adds to the general body of knowledge being collected by the 



25 w 



Serial No. NDS (CF) - 63 E 1103 

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 and making these 
data available to other investigators. 

Honors and Awards : None 

Publications: None 



26w 



Serial No. NDS (CF) - 66 E 1319 

1. Collaborative & Field Research 

2. Epidemiology Branch 
■■"■'■-: ■'■■•>''"■ ■'•'•■■■■ ;■■.■•■■■ .- 3. Bethesda, Maryland 

PHS-NIH • •• ■ ''■■■■: 

Individual Project Report 
■•:■ v :■:■■■:■ i:- July 1, 1973 through June 30 , 1974 

Project Title: A search for autoimmune mechanisms in the pathogenesis of 
chronic neurological diseases by the use of peripheral 
■ " '''f ■' lymphocytes ' .:..... 

Previous Serial Number: Same --"'t:/'.;'- ■^'^-■- :...<'':■:/--■: i y- ■' ":■ ..v-.:. ■ ■■- . 

Principal Investigators: George Nemo, Ph.D. ■'■■•-■ ■■■ ^■■^'^''■■■:- .■^-•:--' >.,.''■•■..•.■••• .■ 
• . ;■ ■■^.y-'v: Jacob A. Brody, M.D. ' ' ■•■■■- ■■! :^.;.:' i -^ ••^*- ' 

Other Investigator: Harry Bartfeld, M.D. 

^;f;i.c £:;-•-. New York. University j iK:.:. .•'■.;•■■ :! ". .:;'^f'...''. ■.■.i:£i.!' 

Cooperating Unit: Department of Medicine, New York University " .,•-''■■■-".•=■ 
Man Years: :■:■/;■' ■ .;;-■ ^r-. ;■-■.'■■.;■:. \:::.-. " ' • ::■" ; ■ ■ ■■■■■■-■.■ 



Total : . 


8/15 


Professional: 


1/5 


Other: 


1/3 



Project Description: ■■.'•■.■ " ^- ■ ■ '^^ - ■•• 

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

Methods employed : Peripheral lymphocytes from patients with multiple 
sclerosis (MS) and other chronic neurological diseases 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 and is measured using liquid scintil- 
lation spectrometry. 

An alternative cellular immune procedure is currently being explored. 
Lymphocytes from MS patients and controls are challenged with brain antigens 
and tested for their ability to produce migration- inhibitory factor (MIF) . 
MIF is a pharmacologically-active substance elaborated by sensitized 
lymphocytes in response to a specific antigen. It is characterized primarily 
by its ability to prevent the migration of macrophages from the open end of 
a capillary tube. 

Maj or findings : As reported previously, lymphocyte transformation 
studies involving MS patients, stroke patients and healthy controls have 
consistently failed to yield significant differences when various brain 



27 



w 



Serial No. NDS (CF) - 66 E 1319 

antigens such as MS brain, normal brain and basic protein were tested 
in our laboratory and in most other laboratories. Positive results have 
been reported, however, for the MIF. 

It appears with the limited number of patients tested, brain degeneration 
whether it be CVA, cancer or MS exacerbation can be detected with the test 
and is most probably nonspecific in nature. 

Lymphocyte transformation studies of Guamanians with amyotrophic lateral 
sclerosis (ALS) and parkinsonism-dementia (PD) were performed using various 
CNS antigens. No stimulation was observed in lymphocytes from ALS patients. 
A low level of stimulation was encountered in lymphocytes from PD patients 
indicating that either an inimune mechanism plays a role in the pathogenesis 
of PD or more likely that it reflects a secondary response to the massive 
destruction of CNS cells in the disease. 

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 studies of MS and ALS casts serious doubt on the hypothesis that these 
disorders result from autoimmune mechanisms. 

Proposed course : Macrophage-migration inhibition tests upon MS patients 
and controls are continuing. Similar studies are planned with Guamanian 
ALS and PD patients. 

Honors and Awards : None 

i 

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 l\Tnphocytes from multiple sclerosis patients. 
New Eng. J. Med . 279:202-204, 1968. 

Nemo, G.J., Brody, J. A. and Cruz, M. : Lymphocyte transformatior; 
studies of Guamanian amyotrophic lateral sclerosis and ; 

parkinsonism-dementia patients. Neurology , In press. 



i 



28 



Serial No. NDS(CF) - 67 E 1487 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

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

Previous Serial Number: Same •"'■ ' ' " . .. 



Principal Investigators 



Dwayne M. Reed, M.D. 
Jacob A. Brody, M.D. 
William Horton, M.D. 



Other Investigators 



Jose Torres 

NINDS Research Center 
Manuel T. Cruz 

NINDS Research Center 
Francisco Leon Guerrero 

NINDS Research Center 



Cooperating Units: 



NINDS Research Center, Tamuning, Guam 
Center for Demographic and Population Genetics, 
University of Texas at Houston (Dr. William J. 



Schull) 



Man Years: n^.--~'j': ■' ■' '..-,.. 

Total- -^ >M. . 11/3 ■ ■• '■"'„ ■■ :!' '■ ' ' '^' :■;' 

Professional: 1/3 

Other: 1 

Project Description: ■ - 

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

Methods employed : As earlier pedigree studies failed to disclose a 
simple autosomal mode of inheritance for ALS and PD, a number of other 
approaches are being continued and updated. One effort involves a registry 
based upon the first 100 deaths each for ALS and PD and an equal number of 
matched control deaths. All live-born offspring from these cases and controls 
have been registered and are being followed for the development of ALS or 
PD as well as death from all causes. A second effort involves the updating 
of a family registry started in 1958. This registry consists of 136 patients 
with either ALS or PD seen at the NINDS Research Center between 1958 and 
1963, and an equal number of matched controls. All known first-degree 
relatives and spouses of these cases and controls were registered by 1963. 
Another study involves the conjugal instances of ALS and PD. To date there 



29V7 



Serial No. NDS (CF) - 67 E 1487 

have been 10 raatings of ALS x ALS , 3 niatings of PD x PD, and 14 matings of 
ALS X PD. The status of all offspring of these matings is continuously 
updated. 

In cooperation with the Center for Demographic and Population Genetics, 
University of Texas at Houston, we have begun a computer genetic analysis 
for affected families. Since 1945 as each new case of ALS or PD has been 
seen at the NINDS Research Center, a family pedigree form has been completed. 
To date we have over 560 such pedigrees. Individual index cards have been 
maintained on each relative and this file is currently being prepared for 
computer analysis. In addition, a detailed tracing of the entire village 
of Umatac back to 1835 is being completed, utilizing church and vital 
statistic records. Subsequently Merizo and other areas will be included. 

Major findings : Genetic causation has been the most consistently 
involved explanation for the apparent familial concentration of ALS and PD 
among the Chamorros of Guam and other Mariana Islands. Autosomal models 
have difficulty accounting convincingly for the sex differences in incidence 
and mortality rates of ALS and PD. Sex-linked models are also unattractive 
for a number of reasons. For example, affected women do not always have 
affected fathers and affected fathers with normal spouses do not have only 
affected daughters as a sex— linked model requires. 

For these reasons a number of new approaches were undertaken to clarify 
the genetic issue. These included a careful search for the missing women 
which would account for the uneven sex distribution. No compelling cause 
of death was found to explain these differences. Secondly, an effort is 
being made to evaluate the fertility of individuals with one or both of 
these neurologic disorders. Complete ascertainment of the status of 
offspring of the first 100 cases of ALS and PD, the original family registry, 
and of the conjugal matings together with data from previous studies should 
allow a good comparison of the frequency of illness among the offspring of 
two affected, one affected, and no affected parental matings. Slight 
differences were found but they were not significant. A major updating of 
all registries was completed and while familial grouping is apparent no 
Mendelian pattern has emerged. 

Significance to biomedical research and the program of the Institute : 
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 catalog 
of inherited neurologic diseases. 

Proposed course : The three registries (Krooth-Plato, first 100, and 
doubly-affected spouse) will be maintained and updated. The entire pedigree 
information on all cases is being coded for analysis. The pedigrees in 
Umatac, Merizo and other selected villages will be pursued. 



30 



w 



Serial No. NDS (CF) 67 E 1487 
Honors and Awards : None 

Publications: Reed, D.M. , Torres, J.M. and Brody, J.A. : Amyotrophic 

lateral sclerosis and parkinsonism-dementia on Guam - 1945- 
1972: II. Familial and genetic studies. In press. 



31w 



Serial No. NDS (CF) - 67 E 1488 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

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

Previous Serial Number: Same 

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

John L. Sever, M.D. 

Infectious Diseases Branch, C&FR, NINDS 

Other Investigator: None . ■ . . 

Cooperating Units: Infectious Diseases Branch, C&FR, NINDS 

Man Years : 

Total: 7/12 
Professional: 1/4 
Other: 1/3 

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 and carefully matched controls, families, 
twins, etc. in various studies are routinely bled and sera is stored. 

Serological analysis is conducted in the Infectious Diseases Branch, 
C&FR, NINDS using a battery of common viriis antigens by hemagglutination and 
complement fixation methods. Differences between the patient, siblings, and 
controls are 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 n£ 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. 



33w 



Serial No. NDS (CF) - 67 E 1488 



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 plan to extend our studies to the Shetland and 
Orkney Islands where MS occurs at a rate three times higher than elsewhere 
in the world and to use our current specimens for other studies as indicated. 

Honors and Awards : None 

Publications: Henson, T.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 
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 , 216:1441- 
1446, 1971. 

Brody, J.A., Sever, J.L., Edgar, A.H. and McNew J.: Measles 
antibody titers of multiple sclerosis patients and their 
siblings. Neurology , 22:492-499, 1972. 

Brody, J.A. : Epidemiologic and serologic data on multiple 
sclerosis and their possible significance. Presented to the 
Kroc Foundation Sjonposium on "Research in Mu.ltiple Sclerosis," 
in Santa Ynez, Calif., February 1972. 



34w 



Serial No. NDS (CF) - 6 7 E 1488 

Brody, J. A.: Comments on the epidemiology of multiple 
sclerosis and a possible virus etiology. Lancet , 11:173, 
1972. 

Brody, J. A.: Epidemiologic and serologic data on multiple 
sclerosis and their possible significance, ^n Multiple 
Sclerosis: Immunology, Virology, and Ultras tructure . UCLA 
Forum in Medical Sciences, #16. Eds. F. Wolf gram, G.W. 
Ellison, J.G. Stevens and J.M. Andrews. Chap. 6, pp. 127- 
141. Academic Press, New York, 19 72. 



35 



w 



Serial No. NDS (CF) - 67 E 1490 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Development of an i^ vitro serum cytotoxicity assay for 
amyotrophic lateral sclerosis 

Previous Serial Number: Same 

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

George Nemo, Ph.D. 

Other Investigator: None 

Cooperating Unit: None 

Man Years : 

Total: 1/10 
Professional: 1/10 
Other: 

Project Description: 

Objective s ; To develop a simplified diagnostic test for amyotrophic 
lateral sclerosis (ALS) . 

Methods employed : The technique recently described by Dr. Frederick 
Wolfgram involves treating explant cultures of anterior horn cells of 3— day- 
old mice with diluted ALS serum. ALS serum appears to specifically destroy 
the neurons in culture. Toxicity is determined by visually examining 
strained preparations (Holmes and Bodian staining methods) using phase- 
contrast and regular light microscopy. We are trying to duplicate this work 
and to simplify the technique. 

Major findings : None as yet. 

Significance to biomedical research and the program of the Institute : 
It is hoped that the technique when applied as an epidemiologic tool will 
serve to elucidate the pathogenic event(s) leading to the disease. 

Proposed course : This project has been terminated. 

Honors and Awards : None 

Publications: None 



37w 



Serial No. NDS (CF) - 67 E 1496 
• -■ ^ ^ •■■ 1. Collaborative & Field Research 

2. Epidemiology Branch 

'v.;l":.:.... .■'.^., ' . "'".' ' ' ,3. Bethesda, Maryland 

■ -!.'■• ^--i,.:^ PHS-NIH ' ' -h..-; ,■<..>/■■■;:•- 

Individual Project Report • '■■■•- ' ■ ■• 

July 1, 1973 through June 30, 1974 

Project Title: Sequelae of CNS diseases in childhood and perinatal period 

Previous Serial Number: Same J^i-J '< ^'^ ;-.;0 'r'-i'' v;j.;. ;;:■::.■■»■.; '■■■"■ '-.•■?u- -;.■■;.>/' .;• .; 

Principal Investigator: Milton Koch, M.D. O;; >'•'■ -"i-;- ■■ •:;•.;. ■■ -i ' ^f^ 

Other Investigator: Jacob A. Brody, M.D. -r ■ ,■•■■.■■•': .; ■,••:,.■•■•:.■;.<■■')- ''i 

Cooperating Unit: University Hospital, University of West Virginia Medical 

Center, Morgantown, West Virginia 

Man Years : 

Total: 7/20 
Professional: 1/4 
Other: 1/10 

Project Description: 

Objectives : During this year we studied possible birth abnormalities 
of the CNS in relationship to a community-wide epidemic of diarrhea caused 
by a potentially teratogenic (parvovirus) virus in Morgantown, West Virginia 
which had occurred in May 19 71. 

Methods employed : All medical records in the University Hospital from 
1967 until July 19 72 coded for any complication of pregnancy, stillbirth, or 
congenital malformation were pulled and reviewed. Baseline numbers for 
normal deliveries were also obtained for that time period. The annual 
numbers of complications were recorded. 

The numbers were compared in order to see whether there was a rise in 
any of these aforementioned complications related temporarily to the 
epidemic. This was considered possible because parvoviruses, which are 
small DNA viruses, are known to cause spontaneous abortion in cattle, and 
several other intrauterine complications, in addition to diarrheal illness 
in adult humans. 

Maj or findings : After reviewing approximately 350 charts for the 
5-year period, no increase in incidence of complications of pregnancy, 
stillbirth, or congenital malformations were noted for the 9-month period 
following May 1971. 



39w 



Serial No. NDS (CF) - 67 E 1496 

Significance to biomedical research and the program of the Institute : 
From this brief study we can say that the parvovirus associated with this 
diarrheal epidemic did not produce major abnormalities which were grossly 
detectable at birth. 

Proposed course : We will look for other situations where large groups 
of people were affected by viral illness to determine possible affects in 
utero and in early childhood, specifically Norwalk, Ohio (the site of an 
epidemic where the agent was first identified). 

Honors and Awards : None 

Publications: Pending 



40 v/ 



Serial No. NDS (CF) - 68 E 1594 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

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. 

Other Investigators: None 

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: 


3/10 


Professional : 


1/5 


Other: 


1/10 



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 dyskinetic reactions 
and 13%, Park ins on- 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 neuro- 
logical reactions. By comparing the patterns of reactions in monozygotic 
with those of fraternal twins, we can employ the usual methods of analysis to 
determine the relative importance of genetic factors in the manifestation of 
extrapyramidal signs resulting from phenothiazine induction. 



41 



w 



Serial No. NDS (CF) - 68 E 1594 

Major 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 : This project has been terminated. 

Honors and Awards : None i_ 

Publications: None 



42 



w 



Serial No. NDS (CF) - 68 E 1597 
•, • 1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report ■ . . • 

■' July 1, 1973 through June 30, 1974 V; , ; ; 

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



Previous Serial Number: Same ., . ., • ,. . . ..■.,■. , , ,,. 

Principal Investigators: Jacob A. Brody, M.D. . •...,,.; ,.., v-rr-.r-:-" 

E. Michael Holden, M.D. 
.,,.,; NINDS Research Center -,..^ , .,.,,,.■,.., ; ,. . . 

.../ . . V. ...... Dwayne Reed, M.D. 

Other Investigators: Manuel Cruz ' ••■ • ■• "■ 

...,.., .... NINDS Research Cenr.er _, ^..., .,;■, 
,,'',..' ' ", , Jose Torres ■.•■■..■-.;.■,■ 

■"'■■ ■' ,.., NINDS Research Cencer .'..";; „, ,■ ,:.: 
Francisco Leon Guerrero 

... ,. . NINDS Research Center ,;. ...,:■..,:,., 

Cooperating Unit: None ■ ' 

Man Years : 

Total: 1 1/10 
Professional: 1/10 
Other: 1 

Project Description: 

Objectives : To investigate neurologic illness occurring in the Trust 
Territories and the Pacific area. During the past few years, this involved 
the study of leprosy patients in New Caledonia, and congenital blindness 
among the people of Pingelap. Currently we are looking for ALS and PD 
among non-Chamorro groups and stroke in various populations. 

Methods employed : Through contact with the Department of Health, U.S. 
Trust Territory and other agencies involved in health in this area, we 
continue to search for unusual patterns of neurologic disease. 

Major findings : We are currently following several Carolinian patients 
on Saipan suspected of having ALS. In addition, we have hospital discharge 
diagnoses of ALS for two patients in the Truk district and one from Palau 
district. Reports from a health survey in the 19 50 's were uncovered and 
10 patients in a village of 245 in Palau were reported as having parkinsonism. 



43 



w 



Serial No. NDS (CF) - 68 E 1597 

We plan to follow up these reports. We are now defining Palauan populations 
at different levels of aculturation for stroke studies. 

Significance to biomedical research and the program of the Institute : 
By discovering population isolates with unusual patterns of neurologic 
disease, we have unusual opportunities to study the mechanisms of cause 
and control. 

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: Reed, D. , Labarthe, D. and Stallones, R. : Epidemiologic 
studies of serum uric acid levels among Micronesians . 
Arthritis Rheum . 15:381-390, 1972. 

Reed, D. , Labarthe, D. , Stallones, R. and Brody, J. A.: 
Epidemiologic studies of serum glucose levels among 
Micronesians. Diabetes , 22:129-136, 1973. 

Labarthe, D. , Reed, D. , Brody, J. A. and Stallones, R. : 
Health effects of modernization in Palau. Am. J. Epid . 98: 
161-174, 1973. 



44 w 



Serial No. NDS (CF) - 68 E 1598 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 19 73 through June 30, 1974 

Project Title: Epilepsy on Guam 

Previous Serial Number: Same 

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

Other Investigators: Manuel Cruz 

NINDS Research Center 
Jose Torres 

NINDS Research Center 
Francisco Leon Guerrero 

NINDS Research Center 
Kwang-Ming Chen, M.D. 

Guam Memorial Hospital 

Cooperating Unit: NINDS Research Center, Tamuning, Guam 

Man Years : 

Total: 11/3 
Professional: 1/3 
Other: 1 

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. 



45 



w 



Serial No. NDS (CF) - 68 E 1598 

Major findings : Epilepsy rates were slightly higher on Guam than in 
most areas where data exists. We found that clinical and hospital sources 
must be supplemented by field surveys to approach a "true" rate. While 
symptomatic epilepsy was fairly well reported through medical channels 
idiopathic grand mal epilepsy was severely underreported. Febrile convulsions 
were inadequately reported through medical channels. While field surveys were 
more complete our best method for detection was by follow-up of birth cohorts 
for given years. Rates were consistently higher in some areas but no genetic 
or environmental factors emerged to explain this. 

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 : This project has been terminated. 

Honors and Awards: None 

Publications: Stanhope, J.M. , Brody, J. A. and Brink, E.: Convulsions 
among the Chamorro people of Guam, Mariana Islands. 
I. Seizure Disorders. Amer. J. Epid . 95:292-298, 1972. 

Stanhope, J.M. , Brody, J. A., Brink, E. and Morris, C.E. : 
Convulsions among the Chamorros people of Guam, Mariana 
Islands. II. Febrile Convulsions. Amer. J. Epid . 95: 
299-304, 1972. 



46w 



Serial No. NDS (CF) - 68 E 1605 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

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

Previous Serial Number: Same ' ■ -' '" ••■''■•• 

Principal Investigators: James A. Schnur, M. D. ''''.. 'l.[ ' 

NINDS Research Center ■■■-•■ '•■:•'- 
R. Michael Scott, M.D. 

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

Joyce Cannon, R.N. 

Cooperating Units: Spring Grove State Hospital, Catonsville, Maryland 

Crownsville State Hospital, Crownsville, Maryland 



Man Years : 




Total: 


3/10 


Professional : 


1/5 


Other: 


1/10 



Project Description: 

Objectives : To determine whether increased skin pigmentation is 
associated with decreased prevalence of phenothiazine— induced Parkins on- 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 
relationshhip 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. We are still working out methods to evaluate the 
effects of duration and type of medication. 



47 



w 



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. 



48 



v 



Serial No. NHS CCF) - 69 E 1774 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bathes da, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Neurologic signs and symptoms associated with malabsorption 

Previous Serial Number: Same 



Principal Investigators: 



Milton J. Koch, M. D. 
Jacob A. Brody, M.D. 
Anne H. Edgar 



Other Investigator: 
Cooperating Units: 



Paul M. Hoffman, M.D. 

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: 


3/10 


Professional: 


1/5 


Other: 


1/10 



Project Description: 

Objectives : To determine if both clinical and subclinical malabsorption 
is associated with a high incidence of neurologic signs and symptoms, and 
deaths due to motor neuron diseases. 

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 reanastoraosis 
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 



49 



w 



Serial No. NDS (CF) - 69 E 1774 

from abstracts and hospital records where available. Patients were not 
included in the study if there was a history of carcinoma, severe diabetes 
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 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. Thorough 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 865 death certificates obtained by follow-up of 2500 cases 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. 

Proposed course : This project is completed and the data will be 
considered for use in further investigations of the relationship between 
neurologic disease and the mechanism of absorption of essential nutrients. 

Honors and Awards : None 
Publications: None 



50w 



Serial No. NDS (CF) - 69 E 1777 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 19 74 

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

Previous Serial Number: Same 

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

Other Investigators: Anne H. Edgar 

Marjorie Gillespie, R.N. 

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

Administration, DHEW 

Man Years : 

Total: 3/10 

Professional: 1/5 
Others: 1/10 

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 12,601 individuals. The Social Security Admin- 
istration searched its records to determine which of these workers had died 
and where they had died. We contacted the individual states and obtained 
the death certificates of the deceased workers to determine the cause of 
their death. 

Major findings : Of the 12,601 cards submitted to the Social Security 
Administration, 2,552 had no Social Security number or an incorrect number, 
8,091 men are still alive and 1,958 men are deceased. Death certificates 
were obtained and coded for cause of death. They were then analyzed by 
age and cause of death and compared with mortality records for U.S. white 
male population and Guam population. Analysis indicates that there was no 
excess of ALS in this group. 

51 w 



Serial No. NDS (CF) - 69 E 1777 

Significance to biomedical research and the program of the Institute : 
The initial trend of this study 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 environmental 
factors on Guam do not increase susceptibility to people whose exposure 
occurs later in life. 

Proposed course : All data have been coded and analyzed and we are 
now interpreting the results. 

Honors and Awards : None 

Publications: None - _ 



52 



w 



Serial No. NDS (CF) - 69 E 1779 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 19 73 through June 30, 1974 

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

Previous Serial Number: Same 

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

Anne H. Edgar 

Other Investigator: None 

Cooperating Unit: None 

Man Years : 

Total: 3/10 

Professional: 1/5 
Other: 1/10 

Project Description: 

Objectives : To determine death rates due to amyotrophic lateral 
sclerosis among native-bom and migrant residents of California and 
Washington. 

Methods employed : Certificates were collected for all deaths attributed 
to ALS as primary or underlying cause, occurring in California residents from 
1954 to 1964, excluding 1956, and in Washington residents from 1955 to 1964. 
Death certificates were analyzed for age at death, sex, race, place of 
residence and place of birth. The material presented was limited to American- 
born whites. Death rates for native-bom Calif omians and Washingtonians 
and for migrants to these states from the nine geographical divisions of the 
United States were calculated from the average number of deaths per year by 
region of birth and the corresponding population at risk. Because the age 
and sex composition of the migrant and native populations differed, the 
death rates were adjusted for age and sex using the population of the United 
States in 1960 as a standard. 

Major findings : During the ten-year period under study, the average 
age-sex-adjusted ALS death rates were similar for the native-born populations 
of California and Washington (0.62 and 0.57 per 100,000 respectively). Rates 
among American-bom migrants from southern and northern areas to California 
and Washington ranged from .74 to 1.03 for males and .39 to .60 for females. 



53 w 



Serial No. NDS (CF) - 69 E 1779 

There was no correlation of ALS death rates with state or area of birth as 
we had demonstrated in a parallel study of MS in the same populations. The 
results provide no suggestion of possible exogenous factors which may be 
related to the etiology of amyotrophic lateral sclerosis. 

Significance to biomedical research and the program of the Institute : 
Numerous epidemiologic studies of mortality from amyotrophic lateral sclerosis 
have attempted to uncover patterns of disease distribution which might provide 
a suggestion as to the etiology of this disease. The great proportion of 
this research has been based on national and international mortality statistics 
and, therefore, interpretation of the results must take into account geographic 
variations in medical care, diagnostic facilities, coding of deaths and death 
certification. The design of this study allows for a comparison of rates 
among people born in various areas of the country but who were subject to 
similar standards of medical care and certification at the time of death. 
The results suggest that future research into the causes of amyotrophic 
lateral sclerosis might emphasize other than those related to geographic 
location. 

Proposed course : This project has been terminated. 

Honors and Awards : None 

Publications: Edgar, A.H. , Brody, J. A. and Detels, R. : Amyotrophic lateral 
sclerosis mortality among native-born and migrant residents 
of California and Washington. Neurology , 23:48-51, 1973. 



54 w 



Serial No. NDS (CF) - 70 E 1832 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Serologic responses of multiple sclerosis patients and 

controls to a virus isolated from a multiple sclerosis case 

Previous Serial Number: Same 

Principal Investigators: George Nemo, Ph.D. 

Jacob A. Brody, M. D. 

Other Investigator: Hilary Koprowski, M.D. 

Wis tar Institute 

Cooperating Unit: The Wistar Institute, Philadelphia, Pennsylvania 

Man Years : 

Total: 1/5 
Professional: 1/5 
Other: 

Project Description: 

Objectives : To measure antibody levels in serum from MS patients and 
controls against MS virus 6/94 which was recovered from the brain of an MS 
patient and to explore the relationship between the agent and MS. 

Methods employed : A total of 48 patients, 24 females and 24 males, and 
an equal number of controls were included in the study. Each patient was 
matched with a control who was a lifelong friend of similar age and sex, who 
was born in the same community and who attended the same schools. 

Serum antibody determinations using MS virus 6/94 and a classical 
parainfluenza Type I isolated from a patient with acute respiratory disease 
were carried out employing hemagglutination- inhibition (HI) and hemolysis- 
inhibition (HLI) techniques. Serum antibody titers to measles virus were 
also measured employing the HLI test. 

Major findings : We were unable to demonstrate higher HI or HLI antibody 
titers among MS patients and controls for the 6/94 MS isolate. Similarly, no 
differences were found in antibody to standard human parainfluenza I. HLI 
antibody titers to measles virus, however, were found to be significantly higher 
in sera of MS patients than control sera. The etiologic relationship of the 
virus isolate 6/94 to MS could not be determined with these procedures. 



55v7 



Serial No. NDS (CF) - 70 E 1832 

Measles virus, however, remains as a strong candidate for possible involvement 
in the pathogenesis of MS . 

Significance to biomedical research and the program of the Institute : 
It has long been hypothesized by numerous workers that >B may be due to a 
viral etiology. The recent isolations of an infectious agent related to 
group I parainfluenza virus from co— culture of MS brain cells and indicator 
cells seemed to strengthen the hypothesis. It is the purpose of this study 
to determine the relationship of parainfluenza I virus, if any, to MS. 

Proposed course : The same serological procedures are being employed 
using a different study group. 

Honors and Awards : None 

Publications: None 



56 v7 



Serial No. NDS (CF) - 70 E 1833 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Immune mechanisms in chronic and congenital viral infections 

Previous Serial Number: Same 

Principle Investigator: Lon R. White, M.D. , NINDS (formerly with DB, NICHD) 

Other Investigators: George Nemo, Ph.D., 

Jacob A. Brody, M.D. 
Samuel Baron, M.D. , LVD, NIAID 

Cooperating Unit: Developmental Biology Branch, NICHD (HD DB - 5) 

Man Years : 

Total: 1/10 
Professional: 1/10 
Other: 

Project Description: 

Objectives : 1. To establish models of chronic, congenital, and/or 
latent viral infection in animals and in tissue culture. 2. In infected 
animals, to define the development of immunity to the infecting and to 
heterologous antigens. 3. In tissue culture, to define the effect of 
immunologic factors (antibodies, immune lymphocytes, interferon) on the 
evolution of acute and chronic infections. 

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 fluorescent and immuno- 
fluorescent 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. Interferon production is studied 
by assay of media from tissue culture preparations of cells infected with 
MVM, and by the ability of MVM infected cells to support the growth of 
other viruses. Sensitivity of MVM to interferon is tested by determining 
the yield of virus following exposure of cells in culture to known amounts 
of interferon. 

Major findings : MVM does not appear to induce interferon elaboration 
in vitro . Pretreatment of L cells with interferon produces a mild to 
moderate diminution in virus yield. 



57w 



Serial No. NDS (CF) - 70 E 1833 

Significance to biomedical research and the program of the Institut e: 
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 continued at diminished level of effort. 

Honors and Awards : None 

Publications: None 



58 w 



Serial No. NDS (CF) - 70 E 1835 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Epidemiologic and immunologic study of families of subacute 
sclerosing panencephalitis patients and families of matched 
controls 

Previous Serial Number: Same 

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

Anne H. Edgar 

Other Investigator: John Sever, M.D. 

Head, Infectious Diseases Branch, C&FR, NINDS 

Cooperating Unit: Infectious Diseases Branch, C&FR, NINDS 

Man Years : 

Total: 7/12 

Professional: 1/4 
Other: 1/3 

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. 42 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 
probands, controls and their families. A 20cc sample of blood was drawn 
from each individual in the proband and control families. 

Major findings : Analysis of the 42 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. One-quarter of 
the probands had no history of measles. Other significant associations 
were chickenpox occurring just prior to measles and a history of exposure 
to fowl and to dogs with possible distemper. No inner city patients were 



59 



V7 



Serial No. NDS (CF) - 70 E 1835 

found except for 3 in New York who were brought up in Puerto Rico and 2 in 
Chicago who had extensive contact with rural environments. Measles titers 
were elevated only among patients and not their siblings. Measles titers 
in mothers of patients with early measles did not differ from control 
mothers . 

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 SSPE 
occurs in an unusual epidemiologic pattern with rates among males far 
exceeding those of females and the absence of true inner city cases. Thus 
we have postiiLated that SSPE is caused by an unusual measles infection and 
a subsequent triggering event which is probably a zoonotic virus. 

Proposed course : Intensive search for the possible "zoonotic trigger" 
will be carried out with appropriate groups when the situation becomes 
available. We are also testing the sera for other antigens including 
toxoplasmosis, parainfluenza and SV 40. 

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 the Annual 
Meeting of the American Academy of Neurology, April 1971. 
Neurology, April 1971. Neurology , 21:439, 1971. 

Brody, J.A. , Detels, R. and Sever, J.L. : Mealses antibody 
titers in sibships of subacute sclerosing panencephalitis 
patients and controls. Lancet , 1:177-178, 19 72. 

Detels, R. , Brody, J.A. , McNew, J. and Edgar, A.H. : Further 
epidemiologic studies of subacute sclerosing panencephalitis. 
Lancet, 11:11-14, 1973. 



60 w 



Serial No. NDS (CF) - 70 E 1836 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Epidemiologic factors in Creutzf eldt-Jakob disease 

Previous Serial Number: Same 

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

A. Roger Bobowick, M. D. 
Raymond P. Roos, M.D. 
IR, NINDS 

Other Investigator: Marjorie Gillespie, R.N. 

Cooperating Unit: Laboratory of Slow, Latent and Temperate Virus 

Infections, IR, NINDS 

Man Years : 

Total: 7/12 
Professional: 1/4 
Other: 1/3 

Project Description: 

Objectives : Creutzf eldt-Jakob disease is one of the spongiform 
encephalopathies which has been transmitted to chimpanzees. There are 
3 forms of Creutzf eldt-Jakob disease which are distinct clinically. 
Nothing is known of the epidemiology of this syndrome. We wish to 
determine if there are common factors among patients who develop Creutzfeldt- 
Jakob 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 and cases were used from the series at the 
Laboratory of Slow, Latent and Temperate Virus Infections, IR, NINDS. 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 was 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 serologic studies. 

Major findings : From the series of 69 patients, we have interviewed 38 
families. The methods outlined above have proved workable. We found that 
patients with the classical or slowly progressive form of the disease 

61 w 



Serial No. NDS (CF) - 70 E 1836 

associated with amyotrophy are on an average 10 years younger than other 
Creutzfeldt -Jakob disease patients. This type of disease has not yet 
transmitted to chimpanzees. Severe upper respiratory illness merged with 
the onset of Creutzfeldt-Jakob disease in 3 patients with the ataxic form 
of Creutzfeldt-Jakob disease. No specific factors, exposures or prior 
history of disease distinguished the patients from the controls. It was 
of interest, however, that one-third of the patients and one-third of the 
controls ate animal brains. The patients tended to have eaten animal brains 
more frequently and more recently than controls. Also 10 of the 13 patients 
and only 3 of the controls stated specifically that they ate hog's brains. 

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. While there 
was no overall difference in brain consumption between patients and controls, 
the information, particularly about hog brains, may be a lead since all the 
other slov; virus transmissible spongiform encephalopathies are transmitted 
in nature through exposure to brain material. 

Proposed course : This project has been terminated. 

Honors and Awards: None 

Publications: Bobowick, A. R. , Matthews, M.A., Brody, J. A., Roos , R. and 

Gajdusek, D.C. : Creutzfeldt-Jakob Disease: A case-control 
study. Am. J. Epid . 98:381-394, 1973. 



62, 



Serial No. NDS (CF) - 70 E 1837 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Multiple sclerosis distribution and patterns 

Previous Serial Number: Same 

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

Dwayne Reed, M.D. 
Milton Koch, M.D. 

Other Investigator: Marjorie Gillespie, R.N. 

Cooperating Units: None 

Man Years : 



Total: 


7/12 


Professional: 


1/4 


Other: 


1/3 



Project Description: 

Objectives : To conduct epidemiologic investigations of unusual clusters 
of cases of multiple sclerosis. To- study possible risk factors before age 
20 which distinguish cases from controls. 

Methods employed : This project involves morbidity studies of affected 
persons and controls in stable communities, and routine epidemiologic 
investigations of clusters of multiple sclerosis cases. 

Major findings : We now know of five apparent foci of multiple sclerosis, 
one in Comanche County, Kansas, a second in Mossyrock, Washington, a third 
in Suffern, New York, a fourth in Broadus , Montana, and a fifth in Bowman, 
North Dakota. We have worked up the first 2 and intend to continue with 
descriptive studies of the other areas, in hopes of finding common environ- 
mental factors. In Comanche County we know of 7 cases in a population of 
3,000 and have written to 1,253 graduates of the three high schools. In this 
manner, we have picked up 5 more patients which increases the potential 
significance of this cluster. In Suffern, New York we learned '.hat 4 members 
of the Class of 1949 of Suffern High School have MS. In Broadus, Montana we 
have been informed of 10 cases of MS in the past 10 years in a population of 
628. In Bowman, North Dakota we learned of 8 women in the graduating Class 
of 1952 (total of 26 members) who have MS. At present we are in the process 
of verifying the Suffern, Broadus and Bowman clusters. In the areas around 
Wichita and Topeka, Kansas we are in the process of studying a large series 
of patients and controls who were born in Kansas. We are concentrating on 

63v; 



Serial No. NDS (CF) - 70 E 1837 

the sequence of infections and exposures which occurred before age 20, 
through questioning the mothers about the patients and controls, in a 
detailed manner. As would be expected, we are having difficulty finding 
enough patients and controls whose mothers are available for interview. 
We expect, however, to have enough cases for analysis within the year. 

Significance to biomedical research and the program of the Institute : 
Some aspect of geographic latitude has consistently been associated with the 
risk of multiple sclerosis both in the United States and throughout the 
world. To date these are the most important epidemiologic clues to the 
etiology of multiple sclerosis. The continuing search and investigation of 
communities may provide further clues to the etiology of multiple sclerosis, 
and clues to undertanding the factors contained in the broad concept of 
geographic latitude. 

Proposed course : These studies will be continued and expanded through 
prevalence and case-control studies. 

Honors and Awards : None 

Publications: Koch, M.J., Reed, D. , Stem, R. and Brody, J.A. : A multiple 
sclerosis cluster in a small northwestern U.S. community. 
JAMA. In press. 



64. 



Serial No. NDS (CF) - 70 E 1838 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 19 73 through June 30, 1974 

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: 3/10 

Professional: 1/5 
Other: 1/10 

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. 

65w 



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 : This project has been terminated. 

Honors and Awards: None 

Publications: None 



66w 



Serial No. NDS (CF) - 70 E 1839 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 19 73 through June 30, 1974 

Project Title: 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 Investigator: Lon R. White, M. D. , NINDS (formerly with DB , NICHD) 

Other Investigators: George Nemo, Ph.D. 

Jacob A. Brody, M. D. 

Cooperating Unit: Developmental Biology Branch, NICHD 

Man Years : 

Total: 1/5 
Professional: 1/5 
Other: 

Project Description: 

Objectives : a. To localize within dividing mouse cells in vitro , 
■particularly in relation to chromosomes, the site of residence of the 
infecting viral genome (of the minute virus of mice - MVM) . 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, 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 hydroxyapatite column chromato- 
graphic demonstration of DNA - DNA hybridization using chromosomal DNA and 
radioisotope "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 immunofluorescent and immunohisto- 
chemical methods. 

Major findings : a. A "fate" study in mouse spleen cells using tritium- 
labeled virus preparations showed no difference in autoradiographic grain 



67w 



Serial No. NDS (CF) - 70 E 1839 

localization between control and virus infected cultures. These were carried 
out with "tagged" viruses of relatively low infectivity and specific activity. 
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 sperraiogenesis . c. Several animals 
initally infected in the perinatal period have produced normal litters. 

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 addition experiments. It was found 
that tritium labeling of viral DNA did not provide a sufficiently high 
specific activity to allow for autoradiographic localization of viral DNA 
in infected cells. An effort is being made to obtain high specific activity 
by labeling the DNA with I^^^ rn vitro . 

The DNA hybridization technique has been used to detect viral nucleic 
acid in whole cell DNA preparation from animals and from cells in culture. 
Current work is focused on the use of chromosomal DNA in order to determine 
if the viral genome may be integrated into or associated with the host DNA. 

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. Tlie interaction 
of noncy toly tic , non-onrogenic 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 : This project has been terminated. 

Honors and Awards : None 

Publications: None 



68 v; 



Serial No. NDS (CF) - 70 E 1843 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Causes of death among siblings of MS and ALS patients 

Previous Serial Number: Same 

Principal Investigator: Milton Koch, M.D. 

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

Cooperating Unit: Department of Neurology, Duke Univer'sity Medical Center 

Department of Neurology, University of North Carolina 
Medical Center 



Years: 




Total: 


7/20 


Professional: 


1/4 


Other: 


1/10 



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 
imusual illnesses possibly related to immunologic defect. The diseases 
we are interested in include collagen diseases, thyroid diseases, and 
rheumatoid arthritis. 

Methods employed : We will control the siblings of MS patients with 
the siblings of ALS patients. We have contacted siblings of at least 44 
ALS and 84 MS cases recently diagnosed in North Carolina. In addition, we 
have information on approximately 100 cases of ALS from North Carolina, who 
had this diagnosis on death certificates. 

Major findings : Earlier attempts to locate siblings of 100 MS and 100 
ALS patients from the autopsy file of Montifiore Hospital, New York City, 
have been unsuccessful. To date, we have received completed questionnaires 
from 25 ALS and 45 MS patients. There is no indication yet that there is an 
increase in collagen-vascular disease in the siblings of MS patients, as 
compared to ALS. 

Significance to biomedical research and the program of the Institute : 
There is no firm evidence that an autoiimnune mechanism or a viriis is the 
cause of MS. There is, however, accumulating suggestive evidence that this 

69 w 



Serial No, NDS (CF) - 70 E 1843 

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 
have illnesses related to immunological mechanisms we would have valuable 
evidence that these mechanisms are involved in the etiology of MS. 

Proposed course : Attempts using this new patient source are being made 
in hopes of getting better access to follow-up data on siblings. 

Honors and Awards : None 

Publications: None 



70 



w 



Serial No. NDS (CF) - 70 E 1844 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Amyotrophic lateral sclerosis in veterans 

Previous Serial Number: Same 

Principal Investigators: John Kurt zke, 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 
James Norman, M.D. 

National Research Council of the National 

Academy of Sciences 
The ran J. Fry 

Social Work Service, Veterans Administration 

Central Office, Washington, D.C. 



Other Investigators: 



Jacob A. Brody, M.D. 
Milton Koch, M.D. 



Cooperating Units: Veterans Administration 

National Research Council of the National Academy of 
Sciences 



Years : 




Total: 


1/5 


Professional: 


1/5 


Other: 






Project Description: 

Objectives : We will determine if there are distinctive epidemiologic 
features related to amyotrophic lateral sclerosis using the veterc.n 
population. We will also determine if service in Guam and the Mariana 
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 



71v/ 



Serial No. NDS (CF) - 70 E 1844 

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. We directly participate 
in the "interview sample" phase of this contract (see Contract #PH 43-64-44, 
Task Order 62) . 

Major findings : A feasibility study of the extensive questionnaire 
was successful and we have now received approximately 91 questionnaires 
from patients and controls. 

Significance to biomedical research and the program of the Institute : 
Regional differences and socioeconomic 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. The 
contract commenced on July 1, 1972 and will run for at least 3 years. 

Honors and Awards: None 

Publications: None 



72 w 



Serial No. NDS (CF) - 70 E 1845 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

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: Marjorie Gillespie, R.N. 

Cooperating Units: National Parkinson Foundation, Inc., Miami, Florida 

Medical Care Insurance Corporation, Saskatchex^ran, Canada 



Man Years : 




Total: 


3/10 


Professional: 


1/5 


Other: 


1/10 



Project Description: 

Objectives : To determine the long-term effects of L-Dopa on the 
natural history of Parkinson's disease and possible harmful effects of 
this medication over time. 

Methods employed : While a case-control study would be the optimum way 
of determining the long-term effects of L-Dopa it would not be ethical at 
the present time to withhold the drug from a large number of cases. We 
have therefore been trying to identify populations in which there was 
sufficient information on a pre-L-Dopa cohort to determine a life table which 
could be compared with a cohort on L-Dopa. We have worked with the National 
Parkinson Foundation, Inc. in Miami and with the New York Hospital, but in 
each case, either follow-up has proven impossible or self-selection of 
patients in the pre-L-Dopa cohort made comparison with the post-L-Dopa cohort 
invalid. During the past year we have been attempting to determine if a 
valid pre- and post-L-Dopa cohort could be identified in Saskatchewan, The 
Province of Saskatchewan began a socialized medicine program in 1962, with 
the unique feature of having a central recording system. L-Dopa was not 
used until 19 70. The overall population in one million with a relatively 
high proportion in older age groups. Thus, it might be possible to utilize 
this population for a suitable study. 

Major findings : Previous efforts in the U.S. have not been successful 
(see above). We reviewed data available in Saskatchewan and have proposed 
a feasibility study. 

73w 



Serial No. NDS (CF) - 70 E 1845 

Significance to biomedical research and the program of the Institute : 
It is not known whether L-Dopa provides only symptomatic relief of Parkinson's 
disease or whether it actually affects the pathologic process which 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 : This project has been terminated. 

Honors and Awards : None 

Publications: None 



74W 



Serial No. NDS (CF) - 71 E 1917 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

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 Hospital, Washington, D.C. 
Zdenek Hrubec, Sc.D. 

Follow-up Agency, NAS-NRC 
Marjorie Gillespie, 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: 


7/12 


Professional: 


IM 


Other: 


1/3 



Project Description: 

Objectives : The abundant epidemiological literature of multiple sclerosis 
has indicated beyond reasonable doubt that critical environmental factors 
influence the rate of disease. Prevalence 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 bom during the years 1917 to 1927. 
Information was collected concerning: 1) pertinent medical history and 
neurological exam, 2) an indepth epidemiologic Interview concentrating on 
events prior to age 20, and 3) blood samples. 



75 v/ 



Serial No. NDS (CF) - 71 E 1917 

Major findings : Initial contact has been made with all of the twin 
pairs where possible. Nine twin pairs have been visited. Others could 
not be located, diagnosis could not be confirmed, or the patient refused. 

Significance to biomedical research and the program of the Institute : 
Although exhaustive studies have been conducted to invoke specific etiologic 
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 unrelated 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 : Termination pending completion of laboratory data 
and analysis. 

Honors and Awards : None 

Publications: None 



76 V' 



Serial No. NDS (CF) - 71 E 1920 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: A study of pregnancies among nurses; high risk pregnancies 
with possible viruses exposure 

Previous Serial Number: Same 

Principle Investigators: Anne H. Edgar 

Marjorie Gillespie, R.N. 

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

Cooperating Unit: None 

Man Years: 

Total: 3/10 

Professional: 1/5 
Other: 1/10 

Project Description: 

Objectives : This project was developed to determine if nurses with 
frequent virus exposure have increased risk of abnormal pregnancy. 

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 1125 nurses registered in the Maryland and 
District of Columbia area. 

Major findings : Of the 1125 questionnaires sent we have received 
863 replies. From these replies there were 1652 pregnancies of which 221 
ended in miscarriage or stillbirth. 

Significance to biomedical research and the program of the Institute ; 
This study will determine if pregnant women working with people with infec- 
tious disease have an elevated risk of fetal damage. 

Proposed course : The coding and preliminary analysis have been completed, 
More extensive tables and statistical tests are now underway. 

Honors and Awards : None 

Publications: None 

77w 



Serial No. NDS (CF) - 71 E 1921 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: A search for measles virus nucleic acid in cells of patients 
with multiple sclerosis (MS) 

Previous Serial Number: Same 

Principle Investigators: George J. Nemo, Ph.D. 

Jacob A. Brody, M.D. 

Other Investigators: Lon R. White, M.D. , NINDS (formerly with DB, NICHD) 

Randolph Taylor 

Cooperating Unit: None 

Man Years : 

Total: 8/15 
Professional: 1/5 
Other: 1/3 

Project Description: 

Objectives : To devise hybridization techniques to detect measles virus 
nucleic acid in cells of patients with MS. 

Methods employed : Radioactive viral nucleic acid probes will be prepared 
using various strains of purified measles virus. Cellular nucleic acids will 
be extracted from CNS tissue specimens and cell cultures of patients with MS 
and annealed with the radioactive viral probes. 

Major findings : None 

Significance to biomedical research and the program of the Institute : 
Serologic evidence obtained in our laboratory and others consistently show 
that MS patients possess elevated measles antibody titers when compared to 
appropriate controls. If viral genetic material related to measles is found 
in cells of MS patients it will be the strongest evidence yet for an associ- 
ation of this agent with the disease. 

Proposed course : Measles virus preparations are currently being purified 
for use as viral probes. Preliminary experiments designed to measure the 
effectiveness of these probes will be performed. 

Honors and Awards: None 

Publications: None 

79W 



Serial No. NDS (CF) - 71 E 1922 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 19 7A 

Project Title: Search for a negative DNA strand in cells infected with the 
minute virus of mice (MVM) 

Previous Serial Number: Same 

Principle Investigator: Lon R. White, M.D. , NINDS (formerly with DB, NICHD) 

Other Investigators: George Nemo, Ph.D. 

Jacob A. Brody, M.D. 

Cooperating Unit: Developmental Biology Branch, NICHD (HD DB - 15) 

Man Years : 

Total: 1/5 
Professional: 1/5 
Other: 

Project Description: 

Objectives : a. To demonstrate a "template" DNA (negative strand) 
complementar}'^ to the single-stranded DNA (positive strand) of the MVM virion, 
b. To determine if the negative strand is associated with the cell's chromo- 
somal DNA. c. To investigate the kinetics of negative strand synthesis 
relative to cellular DNA replication, cellular division, virus infection, and 
virus production. d. To utilize this method to detect a latent or cryptic 
viral infection in the cells of experimental animals and in tissue culture, 
e. To characterize the virion DNA, the "negative strand," and replicative 
forms of MVM DNA. 

Methods employed : These studies utilize radioisotope "tagged" viral 
DNA. Annealing with "cold" DNA from virus infected cells is demonstrated by 
liquid scintillation detection of double- and single-stranded DNA in fractions 
eluted from a hydroxy apatite column. 

Major findings : Contrary to expectations, DNA extracted from purified, 
concentrated virus preparations is inhomogeneous and partially double- 
stranded. A significant amount of additional duplex formation results when 
the initially single-stranded DNA is allowed to "self-anneal." That these 
sequences are truly viral and not from the host cell is shown by the failure 
of added mouse or rat DNA to influence the rate or extent of annealing. 
Denaturation and reannealing kinetics of the initially double-stranded 



81 v 



Serial No. NDS (CF) - 71 E 1922 

suggests the presence of one or more loop or hairpin molecular configurations. 
Nucleic acid hybridization reactions using total cell DNA and a "stripped" 
(all double-stranded parts removed) viral DNA "probe," has now been used to 
detect complementary' sequences in DNA from both acutely infected L cells 
(tissue culture) and newborn mice. Negative results were found with normal 
mouse (3 strains), rat, and guinea pig DNA, as well as with DNA from 2 
tissue culture lines. Unexpectedly, positive results were found with DNA 
from a leukemic rat. Current work is focused on the utilization of this 
method to detect a latent or persistent parvovirus infection in animals and 
tissue culture. 

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 
infections. Congenital, chronic, or latent virus infections may be involved 
with the etiologies of certain reproductive, developmental, and chronic 
diseases whose causes are now unknown. This experimental approach may prove 
to be directly applicable to a search for a parvovirus DNA in the cells in 
animals and persons with such diseases. 

Propose d course: To be continued at a decreased level of effort. 

Honors and Awards: None 

Publications: None 



82 



v 



Serial No. NDS (CF) - 71 E 1923 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Development of in vitro cellular immune tests for viral 
antigens 

Previous Serial Number: Same 

Principal Investigators: George Nemo, Ph.D. 

Jacob A. Brody, M. D. 

Other Investigator: Randolph Taylor 

Cooperating Unit: None 

Man Years : 



Total: 


8/15 


Professional: 


1/5 


Other: 


1/3 



Project Description: 

Objectives : To develop lymphocyte transformation assays using human 
lymphocytes and highly purified measles virus and vaccinia virus as test 
antigens . 

Methods employed : We have developed purification procedures for 
various strains of measles virus (e.g. SSPE, low and high passage Edmonston) 
and vaccinia virus which includes differential centrif ugation, ammonium 
sulfate precipitation and density gradient techniques. Lymphocytes are 
cultured employing standard procedures. Purified viral preparations either 
live or UV-irradiated are added and serve as viral antigens. Tritiated 
thymidine is added 5 hours prior to harvest and uptake of the isotope is 
measured by liquid scintillation spectrometry. 

Major findings : We have been successful in our attempts to develop an 
in vitro lymphocyte transformation assay in humans using highly purified 
preparations of measles and vaccinia virus. Our experiments have: shown that 
the peak of lymphocyte stimulating activity for both measles and vaccinia 
occurs on day 7. This agrees well with other specific antigens such as PPD 
and Candida which also peak on day 7. Density gradient fractions of measles 
virus were capable of stimulating lymphocytes with the peak of stimulatory 
activity coinciding with the peak of virus infectivity. UV-irradiated 
preparations of measles and vaccinia are also capable of stimulating human 
lymphocytes . 



83W 



Serial No. NDS (CF) - 71 E 1923 

Significance to biomedical research and the program of the Institute : 
MS patients have been shown to possess elevated measles antibody titers 
when compared to appropriate controls. Furthermore, the recent finding 
that spinal fluid of MS patients contains antibody to vaccinia virus has 
prompted the inclusion of purified vaccinia virus as a test antigen in our 
cellular assay. We are particularly interested in measuring the cellular 
immune reactivity of MS patients and controls to purified measles and 
vaccinia antigens in hopes that the relationship of these agents to MS may 
be determined. 

Proposed course : We have begun to measure lymphocyte responsiveness to 
these particular antigens in large groups of MS patients and controls. 

Honors and Awards : None 

Publications: None 



84 w 



Serial No. NDS (CF) - 71 E 1924 

1 . Collaborative and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Biochemical and pharmacologic studies in torsion dystonia 

Previous Serial Number: Same 

Principal Investigator: Rosweli Eldridge, M.D. 

Other Investigators: G. Frederick Wooten, M.D. 

Thelma Koerber 
William Horton, M.D. 
William Kanter 

Cooperating Units: Laboratory of Clinical Science, NIMH 

Man Years: 

Total : 2/5 

Professional: 1/5 
Other: 1/5 

Project Description: 

Objectives: Torsion dystonia consists 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 hereditary form of torsion dystonia. Through appropriate analysis of response 
to drugs, such as those influencing neurotransmission the most effective treatment for 
each form may be ciafified. 

Methods employed: Plasma has been collected from affected patients, quick 
frozen, and assayed for various catecholamine levels in the search for a specific 
enzyme abnormality. Double blind, crossover drug trials using promising agents 
such as Dantrolene Sodium (Eaton Laboratories) will be undertaken. 

Major findings: Observations in 40 patients with torsion dystonia indicated that 
the enzyme dopamine beta hy Hr>-»x,'iase (DBH), which governs the conversion of 
dopamine to ncradrenalin, is present in abnormally high levels in individuals with the 
dominant form of torsion dystonia^ 



85w 



Serial No. NDS (CF) - 71 E 1924 

Theurapeutic measures which have been most successful in the opinion of patients 
include brain surgery as performed by Irving Cooper and use of the muscle relaxant. 
Diazepam . 

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 achieve- 
ment in the quest for inborn errors of metabolism. In addition, important information 
may develop regarding biochemistry and physiology of movement control . Specific 
treatment and pre-clinical detection useful in the genetic counseling of these families 
may be more Immediate rewards. In addition, because of the recognized association 
of increased Intelligence and the recessive form of dystonia, we stand to gain infor- 
mation regarding the development of Intelligence. Pin-pointing the most effective 
treatment will afford immediate relief to those affected and may suggest the biochemi- 
cal basis . 

Proposed course: Continued biochemical and pharmacologic investigation of patients 



from the genetics registry. Addition of more sophisticated biochemical techniques to 
the analysis of various tissues and evaluation of promising treatment regimens as they 
become available. 

Honors and Awards: None 

Publications: Eldridge, R.: The torsion dystonias : Literature review: Genetic and 

clinical studies. In the torsion dystonias (Dystonia musculorum defor- 
mans). Editor, Roswell Eldridge. Neurology, suppl. 20:11, part 2, pp. 
1-78, 1970. 

Chase, T.N.: Biochemical and pharmacologic studies of dystonia. In 
the torsion dystonias (Dystonia musculorum deformans). Editor, Roswell 
Eldridge. Neurology , suppl. 20:11, part 2, 1970. 

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

Wooten, F., Eldridge, R., Axelrod, J., and Stern, R.: Elevated plasma 
dopamine beta hydroxylase activity in autosomal dominant torsion dystonia. 
N. Eng. J. Med. 288:6, pp. 284-287, 1973. 

Wooten, F., Eldridge, R., Gordon, EoK., and Axelrod, J.: Studies 
of catecholamine metabolism in two heieditary forms cf dystonia. Trans. 
Amer. Neurol. Ass. vol. 98, 1973. 



86 V' 



Serial No. NDS (CF) - 71 E 1924 

Eldridge, R., Kanter, W., and Koerber, T,: Levodopa in dystonia, 
The Lancet , 11:7836, pp. 1027-1028, 1973. 



87W 



Serial No. NDS (CF) - 71 E 1925 

1„ Collaborative and Field Research 
2o Epidemiology Branch 
3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Intelligence in Israeli patients with torsion dystonia 

Previous Serial Number: Same 

Principal Investigator: Roswell Eldridge, MoDo 

Other Investigators: Morris Gross, M.D. 

Richard Goodman, M.Do 
Thelma Koerber 



Cooperating Units: 



Department of Education 
Herbert H. Lehman College 
Bronx, New York 



Man Years: 

Total: 3/10 

Professional: l/lO 
Other: 1/5 

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 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 drama- 
tic 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 <, 



89w 



Serial No. NDS (CF) - 71 E 1925 
Major findings: Although we have known for several years of the presence of over 



20 cases of torsion dystonia in Israel we have not been able to arrange for visitation. 
Neurologists, psychologists, and educators in Israel have been alerted and indicated 
their interest in participating in this study. 

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. Elucida- 
tion of this basic chemical abnormality would suggest a method for enhancing intellec- 
tual 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 but travel 
funds have not been available for over the past 3 years. 

Honors and Awards: None 

Publications: Eldridge, R., Harlan, A.,, Cooper, I.S., and Ricklan, M.: Superior 
intelligence In recessively inherited torsion dystonioo The Lancet, 
1:7637, pp. 65-67, 1970. 



90W 



Serial No. NDS (CF) - 71 E 1926 

1 . Collaboral-ive and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The difficulty of diagnosing acoustic neuroma in young patients 

Previous Serial Number; Same 

Principal Investigator: Roswell Eldridge, M.D. 

Other Investigators: Thelma Koerber 

Cooperating Units: Department of Neurosurgery 

Columbia Presbyterian Hospital 
New York 

Man Years: 

Total : 2/5 

Professional: 1/5 

Other: 1/5 

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 indi- 
viduals we have been impressed with the unusual length of time and unusual number 
of special ists consulted before diagnosis of acoustic neuroma became a consideration. 
This period was often one of unusual unnecessary 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 neuro'no were 
questioned in detail regarding physicians consulted and diagnostic procedures 
employed . 

Major findings: In general, the younger the individual when first symptomatic, 
the longer the period before diagnosis, the greater the number of physicians consulted 
and the more expense to the family. 



% 



Serial No. NDS (CF) - 71 E 1926 

Significan ce to biomedtcal reserach and the program of the Institute: Such do- 
cumentation should impress the medical community about the resistance of this con- 
dition in young individuals thereby reducing time, expense and turmoil to patient, 
family and physician when a new case develops. Through such publicity it is pos- 
sible that our acoustic neuroma registry will be expanded. 

Proposed course: This project has been terminated. 

Honors and Awards: Presentation at Middle Atlantic Neurosurgery Society 

Publications: Allen, J., and Eldridge, R.: Genetic study of early onset acoustic 

neuroma. Medical Genetics Today. The Johns Hopkins Press, Summer 
1974 



92 



w 



Serial No. NDS (CF) - 71 E 1927 

1» CollaboraHve and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report- 
July 1, 1973 through June 30, 1974 

Project Title: Clinical, genetic, biochemical and pathologic studies in hereditary 
early onset acoustic neuroma 

Previous Serial Number: Same 



Principal Investigator: Roswell Eldridge, M.D. 

Other Investigators: Robert Stern, M.D. 

Martin Mihm, M.D. 



Cooperating Units: 



David Siggers, M,D„ 

Department of Dermatology 
Harvard Medical School 

Department of Medicine 
Johns Hopkins Hospital 



Man Years: 



Total : 

Professional 

Other: 



2/5 
1/5 
1/5 



Project Description: 

Objectives: To clarify the relationship of classical "peripheral" neurofibromatosis 
to early onset hereditary bilateral acoustic neuroma be selected clinical and laboratory 
techniques o To evaluate these findings as possible diagnostic marker for the former 
disease. 

Methods employed: Obtain skin punch biopsies from individuals with, or at risk for, 
early onset bilateral hereditary acoustic neuroma. Examine the histopathology of these 
tissue specimens comparing them to be found in "normal" individuals and those found 
in neurocutaneous syndromes., 

Obtain blood for determination of Nerve Growth Factor levels. 



93 



w 



Serial No. NDS (CF) - 71 E 1927 

Major findings: Abnormal melanosome staining reaction to L-DOPA was found In 
some but not all of those with hereditary acoustic neuroma. 

Significance to biomedical research and the program of the Institute: Through the 
study of the histopathology of cafe-au-lait spots, it may be possible to define the re- 
lationship between hereditary bilateral acoustic neuroma and neurofibromatosis and also 
provide a genetic marker for hereditary acoustic neuroma. This latter information 
would be of great aid for purposes of genetic counseling and indicating those relatives 
that require maximum surveillance. Similar implications hold for studies of Nerve 
Growth Factor. Nerve Growth Factor has been studied in one kindred and found ele- 
vated in those affected and about half of those at risk. 

Proposed course: Proceed with investigations as outlined. 



Honors and Awards: None 

Publications: Allen, J., Eldrldge, R., and Koerber, T.: Acoustic neuroma in the 
last months of pregnancy. Amer. J. Obst. Gyn . (In press). 



94 



v/ 



Serial No. NDS (CF) - 71 E 1928 

1. CollaboraHve and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Biochemical and chromosome studies in retinoblastoma 

Previous Serial Number: Same 

Principal Investigator: Roswell Eldridge, M.Do 

Other Investigators: Frederick Wooten, M.D. 

David Kitchen, M.Do 
Robert Stern, MoD. 

Cooperating Units: Eye Institute 

Columbia Presbyterian Hospital 
Nev/ York City 
Laboratory of Clinical Science, NIMH 

Man Years: 

Total : 2/5 

Professional: 1/5 
Other: 1/5 

Project Description: 

Objectives: Retinoblastoma, a hereditary tumor of the eye, affects children in the 
first year or two of life. Early diagnosis is critical and can lead to life and sight sav- 
ing therapy. Diagnosis now depends on complete retinal examination of at risk indivi- 
duals as does evaluation of therapy. A screening test involving a specific tumor meta- 
bolite in blood or urine is needed to determine newborns who carry the retinoblastoma 
gene and in treated youngsters to evaluate if therapy has been adequate. Neuroblas- 
toma, a nervous system tumor sharing same histologic similarities with retinoblastoma 
produces elevation in dopamine beta hydroxylase activity in plasma of many instances. 
We are studying dopamine beta hydroxylase to determine if it is elevated in retinoblas- 
toma as well . 

Methods employed: Treated and untreated patients and their sibs attending the Eye 
Institute comprise the sample population. Infants and children with other eye disorders 
comprise the control population. Blood samples are obtained in the morning, red cells 

95V? 



Serial No. NDS (CF) - 71 E 1928 



are spun down and the plasma Fs frozen. DBH is defermined In the laboratory of Dr. 
Julius Axelrod . 

Major findings: Approximately 15 patients have been studied to date; although 
several seem to have high levels, thorough analysis is not yet completed. 

Significance to biomedical research and the program of the Institute: If patients 
with active retinoblastoma have significantly higher DBH a valuable tool for early 
detection and monitoring therapy will be available. Also, the confusing genetics 
of retinoblastoma may be clarified. 

Proposed course: This project has been terminated. 



Honors and Awards: None 
Publications: None 



96 



Vv 



Serial No. NDS (CF) - 71 E 1930 

1. Collaborative and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project' Report- 
July 1, 1973 through June 30, 1974 

Project Title: Von HIppel-Lindau syndrome: Clinical, genetic and 
biochemical aspects 

Previous Serial Number: Same 

Principal Investigator: William Horton, M.D. 

Other Investigators: Roswell Eldridge, M.D, 



Cooperating Units: 



Armed Forces Institute of Pathology 
Johns Hopkins Hospital, Baltimore, Maryland 
Columbia Presbyterian Medical Center, New York 
University of Kansas Medical Center, Kansas 



Man Years: 

Total: 3/10 

Professional: 1/5 
Other: l/lO 

Project Description: 

Objectives: The von Hippel-Lindau syndrome is an autosomal dominant disorder 
characterized by multiple CNS and visceral lesions of which the primary lesions in- 
clude retinal angiomata, cerebellar hemangioma, spinal hemangioma, renal cell car- 
cinoma, pheochromocytoma, and pancreatic and renal cysts. All current data concern- 
ing the spectrum of clinical manifestations, therapy, course and prognosis are based 
on single family studies, since no multi-family study has been done. We intend to 
evaluate these parameters from a large number of families throughout the country to 
determine if distinctive syndromes exist under this several classification . 

Since treatment is available for nearly all the manifestations of the syndrome, 
early diagnosis is essential. Indirect ophthalmoscopy and chromosome analysis are 
two methods which have been reported to detect affected individuals prior to symptoms. 
We intend to evaluate these methods in terms of usefulness as screening techniques. 

Methods employed: Families will be ascertained through neurologic and ophthal- 



97 



w 



Serial No. NDS (CF) - 71 E 1930 

mologic deparlrnents of selected medical centers and the Armed Forces Institute of 
Pathology. Contact with these families will be mode through their personal physician 
Data will alternately come frompersonal interviews, physical examinations, hospital 
records, autopsy reports and other family records. Indirect ophthalmoscopy will be 
performed on all at risk as well as affected individuals and of chromosomal analysis 
will be performed on a limited number of affected individuals. 

Major findings: None 

Significance to biomedical research and the program of the Institute: The results 
of this study should clarify the present understanding of the clinical spectrum and 
natural history of this disease as well as the effect of various kinds of treatment. 

Proposed course: See "Methods employed". 

Honors and Awards: None 

Publications: None 



98w 



Serial No. NDS (CF) - 72 E 1977 

1. Collaborative and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

•-• ■• PHS-NIH 

Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Selected genetic and clinical aspects of Huntington's Disease 

Previous Serial Number: Same 

Principal Investigator: Roswell Eldridge, M.D. 

Other Investigators: William Norton, M.D. 

Marjorie Guthrie 
Thelma Koerber 

Cooperating Units: Committee to Combat Huntington's Disease 

Man Years: 

Total: 2/5 

Professional: 1/5 
Other: 1/5 

Project Description: 

Objectives : To explore past and present attitudes of individuals with Huntington's 
disease and their relatives, and to determine the sources of information about the disease 
and family planning. 

Methods employed : Historical research concerning viev^s prevailing when this trait 
was first described; questionnaire survey of members of the Committee to Combat Hun- 
tington's Disease (CCHD).. 

Major findings: The chief concern is at present with the physical and mental aspects 
of this disease rather than the social, or genetic. For information, family members rely 
on CCHD more than on medical sources o 

Significance to biomedical research and the program of the Institute: Lay groups 
such as CCHD can provide an efficient means for disseminating information and hope 
to patients and families confronted with distressing hereditary disease, in addition 
they can provide an atmosphere permitting those affected and at risk to come forward 
and discuss their attitudes and concerns. 

99w 



Serial No. NDS (CF) - 72 E 1977 

Proposed course: Trace views toward Huntington's disease since the initial aware- 
ness in the United States. Add indepth analysis of patients, spouses, and at risk indi- 
viduals regarding genetic counseling and screening procedures. 

Honors and Awards: None 

Publications: Eldridge, Ro, and Stern, R.: Huntington's Disease - Attitudes Toward 
Genetic Counseling. The American Society of Human Genetics, 
October 1973, Atlanta, GA. 



lOOw 



Serial No. NDS (CF) - 72 E 1980 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 19 73 through June 30, 1974 

Project Title: Development and application of methods for the study of the 
minute virus of mice (MVM) 

Previous Serial Number: Same 

Principal Investigator: Lon R. White, M.D. , NINDS (formerly with DB, NICHD) 

Other Investigator: George Nemo, Ph.D. 

Cooperating Unit: Developmental Biology Branch, NICHD (HD DB - 16) 

Man Years : 

Total: 1/5 
Professional: 1/5 
Other: 

Project Description: 

Objectives : To develop improved methods for the propagation, purifi- 
cation, and quantitation of the virus and its nuclei acid. To investigate 
basic biological, biochemical, and structural properties of the virus and 
its DNA in order to develop techniques to investigate its persistence in 
congeni tally and chronically infected experimental animals. 

Methods employed : The agent is propagated in tissue culture and detected 
by cytopathic effect or antigen production. Its DNA is radioisotope "tagged" 
during replication. It is purified by differential centrifugation and 
by chemical means. The virus, its proteins, and its DNA are characterized 
by velocity sedimentation in sucrose gradients, electron micrographic 
appearance, salt and thermal elution characteristics from hydroxyapatite 
columns, and polyacrylamide gel electrophoresis. 

Major findings : Previous reports have described the growth of MVM in 
only two cell lines, both primary, with CPE in only one of these. We have 
extended this to 4 continuous cell lines, 2 of which show CPE. We have 
shown that whereas maximum virus production demands that the host cell divide, 
infection can occur in non-dividing cells and the virus can remain "latent" 
without apparent injury to the cell until it divides. In addition, we have 
established chronic infection in four cell lines, two of which have been 
producing virus in high concentration continuously for 2 years. We have 
accomplished a 100-1000 x improvement in yield of virus and hemagglutinating 
antigen and have shown that purification by isopycnic ultracentrif ugation 
is associated with moderate virus disruption and loss of infectivity. 

10 Iw 



Serial No. NDS (CF) - 72 E 1980 

Additional studies have revealed no significant loss of hemagglutination 
or infectivity titers with 3 cycles of f reeze-thawing or with heating at 
56 °C for up to 90 minutes. UV inactivation progresses in approximately one 
logj^g decrements per second under standard conditions. The in vitro 
infectivity titration method has been improved from a rather unreliable 
one-month procedure (which works poorly with "wild" strains of MVM) to a 
sharply reproducible, reliable one-week procedure usable with all strains. 
The nuclei acid, extracted by NaOH treatment and by a pronase-phenol- 
chloroform method, has been found to be extremely labile and "sticky," 
adsorbing to plastics, millipore filters, and dialysis tubing. Results 
of hydroxy apatite chromatography suggest that it may have a "snap back" 
area where it is double-stranded, that some virions may contain some 
"negative" strand DNA, and that the DNA may be packaged in segments. A 
method has been developed for the demonstration of viral DNA in infected 
cells and are being used to study the course of MVM infection in vitro and 
in vivo . The virus has at least 3 proteins, whose molecular weights are 
between 50,000 and 80,000, and whose relative concentrations most closely 
resemble the pattern observed with a bovine parvovirus. Differences in 
relative concentration have been observed with cultivation in different 
cell lines; these are currently under study. 

Significance to biomedical research and the program of the Institute : 
MVM is representative of a group of small, single-stranded DNA viruses (the 
parvoviruses) which are characterized by ubiquity, generally low virulence 
and cy topathogenicity , an apparent preference for dividing cells, and a high 
frequency of transplacental infections. The study of this agent is expected 
to lead to a better understanding of mechanisms and the possible role of 
viruses in the causation of certain genetic, reproductive, and developmental 
abnormalities. MVM has been relatively unstudied, presumable because of 
technical problems. The improved methods which we have developed, and the 
basic information obtained by the application of these methods now make such 
studies feasible. 

Proposed course : This project has been terminated. 

Honors and Awards : None 

Publications: None 



102w 



Serial No. NDS (CF) - 73 E 2045 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Epidemiology of polymyositis in children 

Previous Serial Number: None 

Principal Investigators: Milton Koch, M.D. 

Marjorie Gillespie, R.N. 
Jacob A. Brody, M.D. 

Other Investigator: None 

Cooperating Units: Various neurologic and pediatric centers 

Man Years : 

Total: 7/12 
Professional: 1/4 
Other: 1/3 

Project Description: 

Objectives : Polymyositis in children under 16 years of age will comprise 
the first phase of this investigation. Recently, several reports have 
indicated the presence of myxovirus— like particles in muscle biopsy of 
affected patients. We wish to determine, by administration of a questionnaire 
to a case-control population, if we can detect distinctive antecedent factors 
which may be predisposing or causative in this disease. 

Methods employed : We have contacted several medical centers requesting 
the names of patients with documented diagnosis of polymyositis, under age 
16, diagnosed within the last 5 years. The parents of these patients were 
contacted by us to obtain permission to administer a questionnaire, and to 
find a suitable control. Then, this questionnaire was administered both to 
the parents of the patients and the controls. Blood was drawn from both 
patients and controls to obtain serum for serologic studies of viral antibody 
levels, and other tests that may be indicated as data from the questionnaires 
are obtained. 

Major findings : Forty-two patients and controls have been interviewed. 
Parents of the 42 cases of childhood polymositis were interviewed along with 
parents of the controls matched for sex and age. Extensive review of past 
medical history, animal exposure history, residential and family history, and 
immunization history failed to reveal any significant differences between the 



103 w 



Serial No. NDS (CF) - 73 E 2045 

2 groups. The only suggestive difference was exposure to bacteriologically 
confirmed streptococcal diseases in 20 cases as compared to 13 controls. 

Significance to biomedical research and the program of the Institute : 
This disease has been considered possibly infectious or immunologic. The 
observation of virus-like particles in patients lends support to this 
assertion. If the disease is an acquired infection, our case-control study 
may have developed leads as to causation or predisposing factors. 

Proposed course : This study was completed in early 1974, and publica- 
tion is pending. Viral serologic data is currently being collected. 

Honors and Awards : None 

Publications: Pending 



104W 



Serial No. NDS (CF) - 73 E 2046 

1 . CollaboraHve and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project- Report- 
July 1, 1973 through June 30 , 1974 

Project Title: Clinical and genetic study of the hereditary epilepsies 

Previous Serial Number: Same 

Principal Investigator: Roswell Eldridge, M.D. 

Other Investigators: William Norton, M.Do 

Salvador DeMoura, M.D. 
Anatole Dekaban, M.D. 

Cooperating Units: Department of Neurology 

University of Pennsylvania School of Medicine 
Collaborative and Field Research 

National institute of Neurological Diseases and Stroke 

Man Years: 

Total: 3/10 

Professional: 1/5 
Other: 1/10 

Project Description: 

Objectives: To study the clinical and genetic aspects of the hereditary epilepsies, 
such as the progressive myoclonic epilepsies, permitting their biochemical definition 
and specific prevention or treatment. 

Methods employed: It has been possible to ascertain over 30 families with a 
diagnosis of progressive myoclonic epilepsy with dementia (PME D) in the United 
States. Examination of affected individuals and their relatives, documentation of 
family history and review of medical records provided the basis for an initial classifi- 
cation of these disorders. Laboratory studies including analysis of polyglucosan and 
mucopolysaccharide in various tissue are in progress. 

Major findings: On the basis of the examination of thirty-seven affected indivi- 
duals in nineteen families the following classification of PME D is suggested: 



105 w 



Serial No. NDS (CF) - 73 2046 

Type I (Lafora) Childhood onset, rapid progression, autosomal recessive inheri- 
tance, Lafora bodies often present. 

Type II Childhood onset, slow progression, autosomal recessive inheritance, 
no Lafora bodies. 

Type III (Unverrlcht-Lundborg) Late adolescent onset, autosomal recessive inheri- 
tance, moderate or slov/ progression, occasional Lafora bodies. 

Type IV (Hartung) Late onset, variable progression, autosomal dominant inheritance, 

Type V Late adolescent onset, moderate to slow progression, autosomal dominant 
inheritance, neural hearing loss. 

Significance to biomedical research and the program of the institute: Distinction 
between the various diseases producing progressive myoclonic epilepsy and dementia 
Is a pre-requlsite for determining the specific metabolic defect In each disease. In a 
practical sense this classification should be of interest to clinicians faced with diagno- 
sis of disorders characterized by myoclonic epilepsy and dementia. Documentation of 
inheritance pattern in these disorders permits Improved genetic counseling for members 
of these families. In addition, such a study provides a model for the genetic study of 
seizure disorders . 

Proposed course: Continued ascertainment of affected families and facilitate study 
of abnormal tissue by appropriate laboratories. 

Honors and Awards: None 

Publications: Stern, R., and Eldrldge, R.: Clinical and genetic study of the pro- 
gressive myoclonic epilepsies. J. Amer. Soc . Hum. Genet, pp. 76A, 
1973. 



106 



w 



Serial No. NDS (CF) - 74 E 2113 

1. Collaborative and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Side effects of L-DOPA on pre-pubital patients with dystonia 

Previous Serial Number: None 

Principal Investigator: William Kanter 

Other Investigators: Roswell Eldridge, M„D. 

William Horton, M.D. 
Thelma Koerber 

Cooperating Units: None 

Man Years: 

Total: 3/10 

Professional: l/lO 
Other: 1/5 

Project Description: 

Objectives: It has been established that dopamine is in high concentration in the 
median eminence of the hypothalamus and probably is the principal mediator in the 
release of many hypothalamic releasing factors. An increase in grov^th hormone, FSH, 
LH, and a decrease in proactin have been demonstrated following the oral administra- 
tion of L-DOPA. Theoretically, the administration of L-DOPA to pre-pubital patients 
might therefore interfere with normal growth and sexual maturation o 

Children with dystonia represent probably the only group with pre-pubital patients 
in which L-DOPA has been used. We therefore intend to evaluate these patients in 
terms of the long term effects of L-DOPA on growth and sexual maturation. 

Methods employed: Patients ascertained through previous studies of dystonia will be 
evaluated through a questionnaire regarding the effects of L-DOPA on growth through 
sexual development. Each matched dystonia patient never having been treated by L-DOPA 
and age matched patients will serve as controls. If the questionnaire suggests any signi- 
ficant differences in these groups, confirmatory biochemical studies will be performed o 



107w 



Serial No. NDS (CF) - 74 2113 

Major findings: Results to date show 5 percent of dystonia patients surveyed noticed 
changes In sexual Interest or development concurrent with levodopa treatment. We are 
initiating further communication with these individuals to assess the relationship between 
levodopa and changes noted. 

Significance t o biomedical research and the program of the Institute : Since the 
beneficial effects of L-DOPA in the treatment of dystonia are somewhat controversial , 
the finding of Interference of normal development would support the argument that this 
drug should not be used at least not In this age group. 

Proposed course: See "Methods employed". 



Honors and Awards: None 
Publications: None 



108w 



Serial No. NDS (CF) - 74 E 2114 

1 . Collaborative and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The study of familial Parkinson's disease 

Previous Serial Number: None 

Principal Investigator: William Norton, M.D. 

Other Investigators: Roswell Eldridge, M„D. 
Jacob Brody, M.D. 

Cooperating Units: None 

Man Years : 

Total : 2/5 

Professional : 1/5 
Other: 1/5 

Project Description: 

Objectives: Parkinson's disease is characterized clinically by a variety of extra- 
pyramidal signs including muscular rigidity, tremor, and pausity of movement; biochemi- 
cally by diminished dopamine in the substantia nigra and corpus striatum and pathologi- 
cally by degenerative changes in the basal ganglia. Parkinson's disease has traditional- 
ly been classified by etiology into etiopathic postencephalitic and arterosc I erotic types. 
Recently several investigators have emphasized that a positive family history is often 
obtained in as many as 16 percent of the idiopathic type. Autosomal dominant and more 
recently multifactorial inheritance have been postulated to explain the familial nature 
of this disorder. We intend to examine this group in which a positive family history is 
obtained to see if there are any clinical and pathologic features which distinguish it 
from the more common sporadic disorder. In addition, genetic analysis of a large number 
of families may clarify the current understanding of the genetics. We also will compare 
this disorder to the Parkinsonism-Dementia syndrome endemic on Guam. 

Methods employed: Families in which two or more members are affected by Parkin- 
sons imwilTbeascertai ned from selected medical centers. These families will be com- 
pletely evaluated in terms of presence and extent of this disorder within family members. 
Further studies will be done when indicated . 

109w 



Serial No. NDS (CF) - 74 E 2114 

Major findings: None 

Significance to biomedical research and the program of the Institute: Delineation 
of the complete clinical spectrum of Parkinsonism may reveal several discrete disorders 
showing features of Parkinsonsim but genetically and biochemically distinct. This would 
allow more specificity in terms of treatment as well as more accuracy regarding diagno- 
sis and prognosis . 

Proposed course: See "Methods employed" . 

Honors and Awards: None 

Publications: None 



llOw 



Serial No. NDS (CF) - 74 E 21 15 

1. Collaborative and Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Familial amyotrophic lateral sclerosis: Clinical, pathologic 
and genetic study 

Previous Serial Number: None 

Principal Invesitgator: William Horton, M.D. 

Other Investigators: Roswell Eldrldge, M.D. 

Jacob Brody, M.Do 

Cooperating Units: None 

Man Years : 



Total : 


2/5 


Professional : 


1/5 


Other: 


1/5 



Project Description: 

Objectives: The term amyotrophic lateral sclerosis currently refers to a neurologic 
disease characterized by widespread degeneration of motor neuron and anterior horn 
cells of the spinal cord, motor nuclei of the brain stem, Betz cells of the cerebellar 
cortex as well as demylination of descending pyramidal tracts. The manifestations of 
weakness, wasting, fasciculations, and long tract signs are variable and dependent 
upon the predominant sites of involvement. In the last 20 years, over 60 families 
have been described with the familial form of this disorder which is inherited as 
autosomal dominant. Is clinically indistinguishable from the sporadic form, however 
pathologically many patients show distinctive changes. The clinical and pathologic 
features have varied significantly and have reported families and there have been 
no consistent pathologic correlations. The object of this study Is to review the 
clinical, pathologic and genetic features in a large number of families including 
those currently ascertained as well as those previously reported to determine if 
distinctive patterns emerge such that specific sub-types might be identified. Because 
of the Involvement of slow virus infections in familial Creutzfeldt-Jakob disease, we 
intend to initiate appropriate laboratory studies to evaluate this possibility and to con- 



lllw 



Serial No. NDS (CF) - 74 E 2115 

siderways In which a viral infecHon could possibly present an aul-osomal dominant 
trait . 

Methods employed: Families with ALS will be ascertained from Epidemiology 
Branch files as well as selected medical centers throughout the country. Families 
will be visited to verify the diagnosis and note the range of clinical manifestations, 
to screen at risk individuals for early manifestations and to derive a complete 
family history and to secure blood, brain and other tissue for appropriate laboratory 
studies, A detailed genetic analysis will be made of all previously reported and 
current families with this disorder. 

Major findings: One frozen brain specimen from a familial ALS case has been 
securred for viral studies. 

Significance to biomedical reserach and the program of the Institute: To deter- 
mine if more than one type of familial ALS exists and to evaluate possible role of 
virus in this disorder. 

Proposed course: See "Methods employed". 

Honors and Awards: None 

Publications: None 



112W 



Serial No. NDS (CF) - 74 E 2116 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30 1974 

Project Title: Nucleic acid studies on selected oncornaviruses 

Previous Seri.^'- Number: None 

Principal Investigator: Lon R. White, M.D. , IR, NINDS 

Other Investigators: Michael Viola, M.D. , Institute for Cancer Research 

Columbia University School of Medicine 

Marvin Frazier, Ph.D., Institute for Cancer Research 
Columbia University School of Medicine 

Gurmid Aulakh, Ph.D., NCI 

Clarence J. Gibbs , Jr., Ph.D., IR, NINDS 

Jacob A. Brody, M.D. 

Cooperating Units: Institute for Cancer Research, Columbia University 

School of Medicine, New York, New York 
Laboratory of Tumor Cell Biology, NCI 
Laboratory of Central Nervous System Studies, IR, NINDS 

Man Years : 



Total: 

Professional : 
Other: 



1/5 
1/5 




Project Description: 

Objectives : To investigate selected oncornaviruses: their relatedness 
and their occurrence in vivo , particularly in certain strains of mice. 

Methods employed : Viruses studied include Rauscher leukemic virus (RLV) , 
the AKR murine leukemia virus, the SJL murine lymphoma virus, the 1504 virus 
(a wild neurotropic leukemogenic agent), and visna virus. Most studies 
employ nucleic acid hybridization techniques, 

Maj or findings : In a study of mouse leukemia virus-specific DNA 
sequences utilizing ~T1— DNA probes made from the AKR virus and RLV, the AKR 
probe showed more extensive hybridization with all mouse cell DNA preparations 
tested. The percentage of the AKR probe hybridized was greatest with lymphoma 
DNA and successively less with normal AKR, Balb/c and NIH Swiss mouse DNA. 
Thermal stability characteristics of the various duplexes formed suggested 
a close relationship between the AKR probe and the endogenouse virus-related 
sequences of Gross lymphoma and normal AKR cells. 



113w 



Serial No. NDS (CF) - 74 E 2116 

Significance to biomedical research and the program of the Institute : 
These studies illustrate the utility of using molecular techniques and 
reagents produced from known viruses to investigate the pathogenesis of 
chronic diseases associated with related agents. With the emergence of 
information suggesting a pathogenic role for oncornaviruses in certain 
chronic neurological diseases in man, and because of the apparent ability 
of a "wild" oncornavirus (the 1504 virus) to produce an amyotrophic lateral 
sclerosis-like disease and lymphoma in mice, it has become important to 
develop methods for the detection of such infections. The studies described 
represent development and application of such methods. 

Proposed course : To be continued. 

Honors and Awards : None 

Publications: Viola, M.V. and White, L.R. : Differences in murine leukemia 
virus— specif ic DNA sequences in normal and malignant cells. 
Nature, 246:485-487, 1973. 



114W 



Serial No. NDS (CF) - 74 E 2117 

1. Collaborative & Field Research 

2. Epidemiology Branch 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Search for biochemical evidence of virus involvement in 
chronic neurological diseases 

Previous Serial Number: None 

Principal Investigator: Lon R. White, M.D. , IR, NINDS 

Other Investigators: Michael Viola, M.D. , Institute for Cancer Research 

Columbia University School of Medicine 

Marvin Frazier, Ph.D., Institute for Cancer Research 
Columbia University School of Medicine 

Clarence J. Gibbs , Jr., Ph.D., IR, NINDS 

Jacob A. Brody, M.D. 

George Nemo, Ph.D. 

Cooperating Units: Institute for Cancer Research, Colimibia University 

School of Medicine, New York, New York 
Laboratory of Central Nervous System Disease, IR, NINDS 

Man Ye ars : 

Total: 1/5 

Professional: 1/5 
Other: 

Project Description: 

Objectives : To search for virus-related enzymes, nucleic acids, and 
antigens in tissues of patients with chronic neurologic diseases. 

Methods employed : In most cases, brain and spinal cord tissue are 
studied. These are obtained at autopsy from patients with parkinsonism- 
dementia (PD) , Guamanian amyotrophic lateral sclerosis (G-ALS) , stateside 
ALS , subacute sclerosing panencephalitis (SSPE) , multiple sclerosis (MS), 
schizophrenia, and other diseases. These are examined for particle- 
associated reverse transcriptase (RT) enzyme activity, and virus-related 
nucleotide sequences in cell DNA and/or RNA. Viruses of particular interest 
include herpes I, herpes II, visna virus, Rauscher leukemia virus, and the 
1504 wild mouse neuropathogenic agent. 

Major findings : RT activity has been demonstrated in brain homogenates 
from both normal and diseased (PD and G-ALS) Chamorran (Guamanian) patients. 
The enzyme is associated with an oncornavirus-like particle, has a molecular 



115 



w 



Serial No. NDS (CF) - 74 E 2117 

weight similar to that of other oncornavirus enzymes, but appears to be immuno- 
logically distinct from other RT enzymes. RNA homologous to the DNA generated 
by the RT has been extracted from Chamorran brain cells. 

Significance to biomedical research and the program of the Institute : 
This approach complements more standard approaches to the detection of viruses 
and offers an alternative way of demonstrating an association between a virus 
and neurologic disease. The results described are consistent with an associ- 
ation between an oncornavirus- like agent and high risk for development of PD 
or G-ALS in the Chamorran population. The demonstration of such an association 
is prerequisite to future definition, control, treatment, or eradication of 
these diseases if they do have a viral etiology. 

Proposed course : To be continued. 

Honors and Awards : None ^ 

Publications: None 



116 w 



ANNUAL REPORT 
July 1, 1973 through June 30, 1974 
Infectious Diseases Branch, C&FR 
National Institute of Neurological Diseases 
and Stroke 
John L. Sever, M.D. , Ph.D. 



Introduction 



The goal of the Infectious Diseases Branch is to carry out planned 
directed research programs concerned with infections which damage the human 
nervous system. The Branch is divided into three sections: 
1) Immunochemistry and Clinical Investigations, 2) Virology and Bacteriology, 
and 3) Experimental Pathology. These sections utilize the techniques of 
immunology, clinical investigations including human volunteers and clinical 
trials; virology, bacteriology, mycoplasmaology, neurovirology and electron 
microscopy; as well as experimental pathology with non-human primates. 

The program segments are: a) Perinatal, b) Acute, and c) Chronic. In 
each segment we are concerned with 1) etiology and diagnosis, 2) treatment, 
and 3) prevention. 

The research activities in the program segments include: 

1 . Perinatal 

Develop and utilize large scale methods to study the relation 
between viral, bacterial, mycoplasma and protozoa infections in the perinatal 
period and birth defects, related abnormalities and pediatric neurological 
diseases. Investigate approaches to early diagnosis, treatment and 
prevention using combined laboratory and clinical studies. 

2. Acute 

Investigate agents which may be responsible for acute neurological 
diseases such as meningitis, encephalitis. Bells' Palsy, and tic douloureux 
as well as possible methods for rapid diagnosis, treatment and prevention. 

3. Chronic 

Study chronic neurological diseases such as multiple sclerosis, 
amyotrophic lateral sclerosis and subacute sclerosing panencephalitis using 
combined tissue culture, immunological, serological, electron microscopic 
and clinical approaches for possible infectious etiologies. Whenever 
possible explore methods for early diagnosis, treatment and prevention. 



A. Findings 

1. Perinatal 



a. Safety of Rubella Vaccine - The study of the possible 
transmission of rubella vaccines given during pregnancy demonstrated that 
only rarely was there transmission of the vaccine virus to the placenta. 
Transmission to the fetus occurred at a very low rate. These studies were 
conducted with the cooperation of women who were susceptible to rubella and 
undergoing therapeutic abortions for other purposes. The women received 
rubella vaccine and then two weeks later, at the time of therapeutic abortion, 
the products of conception were tested for presence of rubella virus. 

b. Perinatal Infections and Epidemiology of Anencephaly 

and Spina Bifida - Anencephaly and spina bifida could not be demonstrated to 
be associated with any of the more than dozen perinatal infections studied. 
A difference in epidemiology of the diseases among whites and blacks was 
noted and was studied in relation to the hypothesis proposed by British 
investigators that anencephaly and spina bifida might be related to the 
ingestion of blighted potatoes. Because of the differences in the frequency 
of anencephaly and spina bifida among white and black patients, it was clear 
that factors other than blighted potatoes alone must be considered in the 
study of the cause of ASB. 

c . Impaired Cellular Inmunity to Rubella Virus in Congenital 
Rubella - Investigations of cellular immiinity to rubella virus showed a 
deficiency in the ability of lymphocytes to recognize rubella-infected 
cultures. Only the lymphocytes from congeni tally-infected children showed 
this defect. Later in life, however, when the children were subsequently 
exposed to either rubella vaccines or natural rubella, their lymphocytes 
acquired the ability to detect rul)ella. These studies suggested that 
impaired cellular immunity may be of prime significance in the inability 

of children with congenital rubella to handle this virus infection in utero . 

d. Immune Responses in Fetal Monkeys - Studies of the immune 
response in fetal monkeys demonstrated that early in gestation, these ,_ 
animals are linable to produce significant amounts of interferon. Later 

in gestation, however, their ability to produce interferon is similar to 
that of adult animals. It is apparent then that the interferon mechanism 
of protection from viral infections is unavailable to the early fetus and 
thus renders it at risk beyond that of older fetuses and adults. 

e. Experimental production of hydrocephalus with Influenza-A 
and Venezuelan Equine Encephalitis Viruses in Monkeys - The inoculation of 
fetuses at 100 days gestation, with two different viruses, was studied. 
Influenza A resulted in the production of hydrocephalus in 50 percent of the 
fetuses. This is of particular importance in view of live vaccines which 

are now under stut^ in man. The use of the attenuated Venezuelan EncephaJ-itis 
vaccine resulted in 100 percent of the animals having the hydrocephalus 
and bilateral cataracts. These findings are consistent with clinical reports 
from Central and South America relating maternal infection with VEE Virus 
to the production of children with porencephalic cysts and cataracts. 

2 X 



f • Cytomep;alovirus Infections in Males - Recent reports 
from Florida have indicated high rates of infections of cytomegalovirus 
in males. This would be of considerable importance in relation to the 
possible etiology of cancer of the prostrate and transmission of 
cytomegalovirus in semen to the fetus. Our studies of specimens obtained 
at vasectomy failed to show any evidence of cytomegalovirus infections 
in over 100 males studied. 

g- Cervical Herpes Infections - High rates of cervical 
herpes infections have been reported in studies conducted in Birmingham, 
Alabama. These rates are particularly increased at the time of delivery. 
Our investigations of women at the Naval Medical Center and at the 
Kaiser Hospital in Los Angeles showed the presence of cervical CM\'' virus 
at a low levelj, but we had no evidence of cervical herpes infections. It 
appears, then, that there is considerable Influence of race and/or 
socioeconomic group in relation to the frequency of infection with both 
cytomegalovirus and herpesvirus in the area of the cervix during pregnancy. 

2. Acute 

A. Spinal Fluid Lactic Dehydrogenase (LDH) for Rapid 
Diagnosis of Meningitis - We have developed a rapid 5-iiiinute test for 
spinal fluid lactic dehydrogenase . Preliminary testing shows that this 
method is quite satisfactory for the identification of elevated levels of 
lactic dehydrogenase. This in turn provides strong indication that the 
patient has acute meningitis infection and appropriate antibiotic therapy 
can be initiated. Further refinements of this method are in progress at 
the present time. : .';;..:■::.•..■ .- ■ -■ ■ ■ " . 

B. Neonatal Deaths - Congenital Malformations - Studies of 
children dying in the first days of life with associated congenital 
malformations have revealed a number of previously unrecognized congenital 
infections. These infections appeared to be most frequent with organisms 
of the herpesvinas group. ■ ■■ -. .' . . 

3. Chroni c -• 

A. Multiple Sclerosis and Virus-like Particles - Reports 

from Australia have suggested virus-like particles in the brains of patients 
with multiple sclerosis. Our electron microscopy studies have also shown 
filamentous structures in brain cells in multiple sclerosis patients, 
ho\iever we are of the opinion that these are not virus particles. Our 
labeling studies with immunoperoxidase show no evidence of vir.'us labeling 
with either measles or parainfluenza antibody systems. 

B. Subacute Sclerosing Panencephalitis and Measles Vaccines - 
The use of measles vaccines in the last ten years has suggested the 
possibility these vaccines are producing higher rates of subacute 
sclerosing panencephalitis or bringing on clinical disease earlier in 
children who receive the vaccine. Our study of over 250 children with 
subacute sclerosing panencephalitis showed no shortening of incubation 
period or earlier age of onset of SSPE among children who received the 
various measles vaccines. Interestingly, the incubation period between 

3x 



natural measles and onset of SSPE was quite consistently found to be 
six years plus or minus 2-5 years. This was for all age groups of 
onset of measles between 1 year and 6 years of age. 

C. Treatment of Subacute Sclerosing Panencephalitis - 

A number of treatments have been tried for SSPE in the past and none of 
these have been successful. We have been studying the use of transf er 
factor in 8 patients with SSPE and comparing their clinical course with 
8 individuals who have not received transfer factor. At the present 
time, there appears to be some possible benefit from the use of transfer 
factor. We are pursuing these studies in greater detail. 

D. Cellular Immunity and Chronic Infections - The 
studies of Cellular Immumty and Subacute sclerosing panencephalitis 
indicate that there is a serum factor which blocks the immune lymphocytes 
from attacking the infected cells. This may be of key importance in the 
progression of SSPE in children. Further identification of the blocking 
factors is in progress. 

E. Studies and Progress 

1. Perinatal 

a. Collaborative study - phase 2 studies 

Clinically recognized perinatal infections - We 
have identified the more 1h an 8000 women with clinical infection during 
pregnancy who are part of the 58,000 study pregnancies in the collaborative 
perinatal .investigation. We are in the process of correlating the clinical 
infections with the data for the 8 year follow-up for these pregnancies. A 
work and progress report was given and further studies will be completed 
during this year. 

2. Serological Studies ^lOOO Abnormals, ^000 Controls - 
We are now completing the serological studies using 11 different antigens 
for infectious agents in the stuc^ of infections in UOOO pregnancies that 
resulted in abnormal children and 4000 matched controls . With the 
completion of the serological data, we will analyze with the use of the 
computer, the clinical findings for the 8 year follow-up for the abnormals 
compared to the controls. We will be looking for higher frequencies of 
antibody or higher titers among the abnormals than the controls. 

3- High IgM and Abnormal Babies - We have identified 
the approximately 2000 babies in the Collaborative Study who have high 
I^ in the cord blood. We have also completed the specific testing for 
cytomegalovirus and toxoplasmosis IgM antibody in these cord sera. We 
are now in the process of testing the same sera for antibody to syphilis. 
With this data, we will conflate the recognized congenital infections 
with the clinical findings for these pregnancies. Furthermore, we will 
extend the testing to include rubella and herpes type II as soon as we 
have perfected these methodologies. 



X 



b. High Risk Babies 

We are studying the high risk babies at the 
University of Tennessee to identify the infectious causes of disease in 
this high risk population. Serial blood samples are obtained for IgM 
levels as well as virus isolation tests. This data is being correlated 
with the clinical course of the child to determine the affect of these 
infections. 

c. Infertility and Abortions 

In studies at the Kaiser Hospital in Los Angeles, 
we are analyzing infections f otind in the cervical area and antito dy studies 
of women who havB infertility and/ or abortions. These data are being 
utilized to determine the extent to which infection is responsible for 
these two abnormal outcomes. 

d. Mycoplasma Infections-Abortions and Prematvirity 

Several reports have suggested that mycoplasma 
infection during pregnancy may result in abortion and prematurity. Our 
preliminary studies have indicated that there are high rates of infection 
particularly in lower socio-economic groups dviring pregnancy. We 
are now studying 250 women for specific mycoplasma infections with general 
mycoplasma and T-strains. This data is being correlated with the frequencies 
of abortions and prematurity in the population. 

e. Experimental Animals - Studies of Virus Infections 
in Monkeys . 

We are now extending our studies of Influenza A and 
Venezuelan encephalitis virus both of which produce central re rvous 
system malformations in the rhesus fetus. Current investigations will 
include mumps, reovirus, and cyclopromine . 

f . Alphafetoprotein and Anercephaly 

Elevated levels of alphafetoprotein appear to 
be frequently found in the serum of mothers carrying fetuses with anencephaly 
or spina bifida. We are studying the reliability of this assay as a method 
of diagnosing these diseases in utero by the 20th week. We hope this may 
result in the method for recognizing the damaged fetus before gestation 
proceeds past the point which abortion can be considered. 

g. Malnutrition and Immune Response 

Malnourished individuals appear to have a deficient 
immime response. With the availability of a number of severely malnourished 
monkeys, we are studying the cellular and antibody immune response in 
these animals and comparing the data with that obtained with fully nourished 
animals. This will provide a definitive study of the affect of malnutrition 
on immune responses. 

5x 



2 . Acute 

a. Spinal Fluid IDH and Limolis Tests for Meningitis 

These tests appear to be able to detect 
bacterial meningitis within minutes after the spinal fluid is obtained. 
¥e are studying both of the methods at tte Childrens Hospital in 
Washington. The limolis test is particularly useful for identifying 
gram negative meningitis. 

b. Radioimmune Assay for Antibody to Eeip es I and II 
and Measles Antibody . 

The radioimmune method is being perfected for 
use with human sera for the detection of antibody to Herpes 1 and 2 and 
Measles. This method is extremely sensitive and should permit us to 
make the rapid identification of developing antibody in patients with 
various acute neurological diseases. 

c. Meningitis and Encephalitis Leading to Mental- 
Motor Retardation . 

We are conducting studies with Collaborative 
Perinatal Research material to determine the role of acquired meningitis 
and encephalitis in the production of mental and motor retardation in 
children. It is already apparent that infections of the central nervous 
system are a leading cause of cerebral palsy - mental and motor retardation. 
The studies will be pursued and ccmpleted during this year. 

d. Simian Hemorrhagic Fever and Disease 

We have been investigating Simian Hemorrhagic 
Fever in monkeys. This disease produces a severe hemorrhagic disease which 
is almost always fatal to the monkey. The disease is somewhat similar to 
lymphocytic choriomeningitis in that immune complexes are formed and 
disseminated coagulation occurs. Further information on the disease should 
provide a better understanding of the mechanism pathogenesis of these 
types of Ariral infections. 

e. Toxoplasmosis in Squirrel Monkeys 

We have been able to demonstrate that squirrel 
monkeys become infected with toxoplasmosis and that this disease is readily 
reproducible. This now permits us an opportunity to study the possible 
means of transmission of toxoplasmosis as well as therapy fo":* the disease. 

f . Immunosuppression and Infection (ETO) 

The drug ENU results in rapid imrn'onosuppression. 
It also produces congenital malformations in several species and cancer in 
other species. We are currently studying the combined effects of this 
and infection in the production of central nervous system defects and 
malignancies . 

6x 



3. Chroni c 

A. Subacute Sclerosing Panencephalitis Eegistry - 
We continue to support the SSPE Registry to identify new patients with 
this disease. This provides us a source of patients for studies of 
treatment of SSPE as well as a monitor for the epidemiology of the disease. 
It now appears that vaccines for measles do not significantly increase 

the rate of SSPE among children who have had previous exposure to measles. 
We will continue these studies of epidemiology and therapy. 

B. Immune Electronmicroscopy and Multiple Sclerosis • 
In joint investigations with Dr. Kapikian who will use the immune 
electronmicroscopy approach to study tissues from patients with multiple 
sclerosis. This should permit us an opportunity to use this new tool 

to identify the possible presence of virus particles in multiple sclerosis 
brain. 

C. Treatment of Subacute Sclerosing Panencephalitis - 
In joint studies with Dr. Michael Blase of the National Cancer Institute 
and Dr. Fenichel at Vanderbilt University^ we are studying both the use 

of transfer factor and the use of the drug Tilorone in the treatment of 
patients with Subacute Sclerosing Panencephalitis. These studies will 
be extended. 

D. Virus Isolation Studies - Multiple Sclerosis and 
Amyotrophic Lateral Sclerosis. 

Using combined tissue culture and electron 
microscopy approaches, we are looking for the presence of viruses in 
chronic progressive neurological diseases. These studies involve the 
direct assay for virus in the original tissue as well as the cultivation 
of the tissue and a variety of methods, looking for the presence of 
virus. These studies will be extended by a contract on the use of 
homology studies to detect virus genomes during the next fiscal year. 



E. Japanese Subacute Sclerosing Panencephalitis and 



Cellular Immunity. 



A unique chronic SSPE cell line from Japan is 
being investigated in relation to the cellular immune responses in patients 
with this disease. This particular cell line carries the SSPE measles 
virus and produces plaques in tissue culture, but does not release virus. 
This affords us an opportunity to study both homoral and cellular immunity, 
since we can expose the virois containing cells to antibody and immune 
lymphocytes to determine the affect of these on the plaque i eduction of 
the cultures . 

F. Multiple Sclerosis and Cellular Immunity 

Recent investigations at the Rockefeller 
Institute have demonstrated inability of lymphocytes from multiple sclerosis 
patients to recognize measles antigen. We are pursuing these studies 



with other antigens to determine if this is a unique activity in relation 
to measles or is a more general depression of ability to fully detect 
antigen. 

G. Varicella, Chronic Infection of Nerves 

Recurrent shingles in patients demonstrates a 
persistence of varicella virus in certain individuals, probably in the 
dorsal roots of the peripheral nerves which are affected. With studies 
at the St. Jude's Hospital in Memphis, Tennessee, we are examining the 
dorsal roots from a niomber of children ^d-th known shingles. These 
studies are done at the time of autopsy and the first specimens are flown 
to the NIH for the investigations . 

H. Congenital SSPE 

Study of unique patients in which the mother 
has SSPE at the time of pregnancy and delivery are being pursued. We are 
interested in determining if there is congenital transmission of the 
infection to the fetus and then the development of disease in the child. 
Patients of this type are shown to us and we are pursuing these 
investigations . 

I . Separation of CSF Proteins in Multiple Sclerosis 
New techniques on highly refined protein separation are being pursued 
using spinal fluid specimens from patients with multiple sclerosis. We 
hope these tests will make it possible to diagnose the disease and follow 
the course of the disease. 

J. SSPE Nucleocapsids 



Electron microscopy studies demonstrate 
differences in the nucleocapsids and measles virus in the brains of 
patients with subacute sclerosing panencephalitis. Studies are being 
pursued on the nature of the differences and the reason for the inhibited 
development of the fuzzy nucleocapsids in relation to the progression 
of the disease. 

K. Tilorone for Herpes Viinas Hominis 

Studies in hamsters have demonstrated the possible 
value of tilorone for the treatment of herpesvirus hcminis. We are 
extending these investigations in other animal species to determine if this 
might be a useful dmg for the treatment of hejrpes encephalitis in m.an. 

L. Freeze Etching Techniques for Subacute Sclerosing 
Panencephalitis, Visna, Scrapie, and Multiple Sclerosis . 

The new method of freeze etching for electron 
microscopy have provided us the opportunity to investigate the presence 
of virus-like particles in a variety of diseases. We are concentrating 



these new techniques on stdies of subacute sclerosing panencephalitis, 
vlsna, scrapie, and multiple sclerosis. 

M. Immunoperoxidase , Electron Microscopy and 
Multiple Sclerosis, Visna, Amytrophic Lateral Sclerosis, and Parkinson's 
Disease . 

New techniques have been perfected by Dr. Monique 
Dubois in our laboratory for the further utilization of immunoperoxidase 
in the study of the possible presence of viruses in multiple sclerosis, 
visna, amytrophic lateral sclerosis, and Parkinson's disease. These 
studies will be extended with these more refined techniques. 



9 X 



CONTRACT NARRATIVE 

Infectious Diseases Branch, C&FR, NINDS 

Fiscal Year 1974 

THE JOHNS HOPKINS UNIVERSITY (PH43-68-710) 

Title : Longitudinal Surveillance of Children with Congenital Infection 

Contractor's Project Director : Dr. Janet Hardy, M.D. 

Current Annual Level : Terminated 

Objectives : The contractor maintained longitudinal surveillance of 
children born during the 1964 epidemic of rubella who were identified as 
having congenital rubella. These children were originally studied in 
conjunction with the Infectious Diseases Branch of the National Institutes 
of Health in 1964-1966. Serial specimens were obtained from the children 
for virus isolation and antibody determinations. Clinical evaluation is 
being obtained repeatedly so as to determine the total clinical findings 
for these children throughout a twelve-year period of observation. 
Present surveillance is being maintained by the investigator without 
contract support. 

Major Findings : During the first four years of observation, it became 
obvious that many children who were initially thought to be normal showed 
damage after the passage of time. In addition, high antibody levels 
found in the first year of life frequently decreased to low or 
undetectable levels, thus making the retrospective diagnosis of the disease 
impossible. The present findings indicate much higher rates of damage 
than had been anticipated. Most of this damage is related to the more 
subtle rr.cinifestations of mental motor retardation and mild to moderate 
hearing deficits. 

Significance to the NINDS Program and Biomedical Research : Congenital 
rubella infection is a preventable disease. The wide spread use of 
rubella vaccines should prevent the occurrence of epidemics and thus 
greatly reduce the frequency of the disease. Damage due to the infection 
however, has only been documented in detail in recent years. Severe 
congenital infection has been noted since 1941 to be associated with 
severe manifestations of the disease. More subtle disease entities 
however, are now being recognized and it is evident that many of the 
children heretofore classified as "etiology unknown" are the result of 
unrecognized intrauterine infection with rubella. 

Proposed Course of the Project : The project will continue through the 
twelve years of observation with a repeated clinical and laboratory 
findings for final presentation in 1976. 



10 



CONTRACT NARRATIVE 
Infectious Diseases Branch, C&FR, NINDS 
Fiscal Year 1974 

THE UNIVERSITY OF CALIFORNIA, LOS ANGELES (NIH-NINDS-69-4) 

Title : Infections in Pregnancy and Infertility 

Contractor's Project Director : Dr. Margaret Jones 

Current Annual Level of Support : $33,405 

Objectives : Four studies are included in this contract. 

1 . Effect of Viral Infections During Early Pregnancy 

This is a study of 4000 pregnancies. The study is unique in that 
pre-pregnant sera as well as sera obtained throughout pregnancy are 
available. Tests have been completed for rubella. Flu A, herpes, mumps, 
and cytomegaloviruses. Cord sera are now being tested for IgM levels 
(NINDS) - analysis is in progress. 

2. Toxoplasmosis 

A special study of 1500 women registered early in pregnancy serial 
blood specimens have been tested for toxo antibody using the IHA method. 
A total of 200 abortuses have been tested for presence of the organism. 
Cord sera are being tested for toxo specific IgM (NINDS). Children are 
being examined longitudinally. 

3. Infertility Study 

Eighty patients and eighty controls are being studied serologically 
for evidence of Herpes 1, II and cytomegalovirus infections. Analysis 
is in progress. 

4. Cytomegalovirus Special Studies 

Two thousand women registered in the first trimester of their 
pregnancies (1967-1971) have been studied. Serological tests have been 
performed. Urine, cervical swabs and throat swabs have been tested for 
CMV. Cord sera are being tested for IgM. Analysis is in progress. 
Children are being followed longitudinally. 

Major Findings : Data has been tabulated for patients with sero- 
conversions and four fold increases in antibody to influenza, cytomegalo- 
virus, rubella, mumps, and herpes. This is being analyzed in relation 
to time in pregnancy and pregnancy outcome. Matched controls are being 
used in this analysis. Multiply infected women are being analyzed 
separately. In addition, the prevalence of antibody is being deter- 
mined for each season and the persistence of antibody is being evaluated. 

Children from the toxoplasmosis and cytomegalovirus studies are being 
observed longitudinally and tested in detail. Analysis of these studies 
are in progress. -'■•^ x 



Specimens frcin women who are being seen because of infertility are 
under test. A total of 80 women have been studied to date. Blood specimens, 
throat swabs, cervical swabs and endometrial biopsies are being used. 

Significance to NINDS Program and Biomedical Research: Congenital infec- 
tion is a preventable cause of severe central nervous system and other 
diseases in children. Elimination of this risk factor is an exceedingly 
worthwhile approach to the prevention of damage to children and the related 
social and economic problems resulting from this type of damage. Infertility 
is often related to viral and chronic intrauterine infection and is directly 
associated with the production of damaged children. These studies, therefore, 
will provide us opportunities for determining which agents are the most 
frequent causes of infertility and fetal damage. 

Proposed Course of Project: The project will run for two more years. The 
emphasis will continue on analysis of the clinical data for the former 
studies. We will complete cord IgM data during the next year. Children 
born to mothers in the toxoplasmosis and CMV studies are being followed 
longitudinal ly. 



12 X 



CONTRACT NARRATIVE 
Infectious Diseases Branch, C&FR, NINDS 
Fiscal Year 1974 

DEPARTMENT OF PEDIATRICS, UNIVERSITY OF TENNESSEE (PH43-68-17) 

Title: Identification of Specific Antibodies in the Serum IgM 
Fraction of High Risk Infants 

Contractor's Project Director : Dr. Sheldon Korones, M.D. 

Current Annual Level: $25,720 

Objective : The study is designed to assess the validity and clinical 
usefulness of specific tests for serum IgM for the identification of 
antibodies to rubella, cytomegalovirus, toxoplasmosis, herpesvirus and 
syphilis. When these diseases are identified as being present in the 
children, appropriate therapeutic approaches are used. 

Major Findings : Approximately 2,000 infants have been tested to date. 
Serial blood specimens are collected twice each week from each study 
infant for the duration of the nursery stay. Venous blood is obtained 
from infants with IgM concentration is in excess of 20 mgm percent in 
the first two weeks of life and 30 mgm percent between the third and 
fourth weeks. Virus cultures are obtained from all infants with high 
IgM levels. Clinical evaluation data is being maintained for correlation 
with the serologic findings. 

Significance to the NINDS Program and Biomedical Research : The detection 
of high risk infants with congenital infections is hampered by the lack 
of specific tests to identify these children. Recent studies have 
indicated that elevated IgM levels are present in at least 80 percent of 
children with congenital infection. The present studies will provide 
direct information as to the usefulness of specific IgM antibody 
determinations for five major infectious agents as a means of detecting 
not only the possible presence of congenital infection but the specific 
infection which requires treatment. 

Proposed Course of Project : The project is completing its second year. 
We anticipate at least two additional years of study during which a 
total of 5000 infants will be examined. 



13 X 



Serial No. NDS (CF)-57 ID 402 

1. Infectious Diseases Branch 

2. Section on Immunochemistry and Clinical Investigations 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Immunology of Perinatal Infections 

Previous Serial Number: Same 



Principal Investigators: 



Dr. John L. Sever, IDB, NINDS 

Dr. David A. Fuccillo, IDB, NINDS 

Mrs. Anita Ley, IDB, NINDS 

Mrs. Renee Traub, IDB, NINDS 

Mrs. Mary Ruth Gilkeson, IDB, NINDS 

Mrs. Flora Moder, IDB, NINDS 

Dr. Jonas Ellenberg, OB, NINDS 



Other Investigators; 



Cooperating Units; 



Dr. Gabriel Castellano, MBA 

Dr. Janet Hardy, Baltimore, Md. 

Dr. Margaret Jones, Los Angeles, California 

Dr. Paul Terasaki, UCLA, Los Angeles, California 

Collaborating Institutions in the Perinatal Research Study 

Laboratory of Infectious Diseases, NIAID 

NICHD 



UCLA Medical School 

Johns Hopkins University Medical School 

Kaiser Hospital, Los Angeles 



Man Years : 



Total: 13.5 
Professional: 5.5 
Other: 8 

Project Description : 

Objectives : The purposes of this study is to determine insofar as possible the 
role of infections and immunity in the production of abnormal pregnancy out- 
comes. To accomplish this, 12 collaborating institutions in the Perinatal 
Research Study plus two special cooperating groups in separate studies have 
been obtaining specimens of blood and tissue throughout pregnancy, at delivery, 
post partum, and at set intervals thereafter. These sera are tested to 
determine the antibody responses of the patients during pregnancy and post 
partum and then to relate this serological information to the clinical data 
for the pregnancy and child. In addition, serum specimens from the children 
were obtained at one year of age from 10,000 study pregnancies. Sera, throat 
swabs, and urine specimens were also obtained from approximately 5,000 



15 X 



Serial No. NDS (CF)-57 ID 402 

pregnancies. Placental specimens were obtained from 2,500 pregnancies. In 
special cases when congenital infection is suspected on the basis of clinical 
or laboratory findings, throat swabs and blood specimens are obtained from the 
children. Immunoglobulin determinations are performed with the cord blood 
specimens from the children and specific antibody determinations are also made 
with these specimens. 

Methods Employed : To accomplish this program, blood specimens were obtained 
from pregnant women at set intervals throughout pregnancy and post partum. 
Completeness of the sets of sera is determined at the Serum Center of the 
Infectious Diseases Branch. Data for the 58,000 patients in the Collaborative 
Perinatal Research Study show that specimens are available from 94.2% of the 
patients. An average of 5 blood specimens is available for each patients. 
Each specimen consists of 4 vials with 3 ml of serum in each. For this study 
then, there are approximately 300,000 serum specimens and almost a million and 
a half vials of sera. There are an additional 5,000 patients studied to date 
at the Kaiser Hospital in Los Angeles and approximately 3,000 under study at 
the Johns Hopkins Medical School in Baltimore, Maryland. All specimens are 
stored at -20°C until tested and complete filing record concerning basic 
patient information and the status of the serum available is maintained 
through a computer system by the Serum Center of the Branch. 

In addition to the serum specimens, serial urine and throat specimens were also 
obtained on a large majority of the patients in the two special studies. These 
are being studied for direct virus isolation along with swabs obtained from the 
children at the time of birth. 

To date, approximately 55 publications have resulted from the analysis of the 
data from these studies. The serological method most frequently employed is 
the complement fixation test with the use of viral antigens. The test is very 
versatile and can be performed rapidly and provides broad coverage for a great 
many of the more than 130 viruses which are known to be of importance to man. 
Antigens were prepared for most of these viruses and tests of specificity were 
conducted with animal sera. In addition to the complement fixation method, 
hemagglutination inhibition tests are used for many viral serological determina- 
tions when greater specificity is needed, neurtalization methods are employed. 
Indirect fluorescence is also used for some of the studies. Virus isolation is 
conducted with tissue culture or inoculation of experimental animals. 

All tests are reproduced completely and a minimum of 95% agreement within two- 
fold variation as required. All sera showing significant changes in antibody, 
together with any sera which did not reproduce are tested the third time. 

At present, we are engaged in a large scale two-year study designed to inves- 
tigate infection and immunity in relation to 4,000 normal children in the study 
and 4,000 matched controls. The crude print-out of abnormal patients has been 
obtained from the Collaborative Perinatal Research Study (PRB) and this is 
being reviewed in detail by nurses and physicians from the IDB for more complete 
information. 



16 



Serial No. NDS (CF)-57 ID 402 

From study records, the specific abnormalities under study include abortions, 
stillbirths, cataracts, congenital heart disease, neonatal deaths, low birth- 
weight (1,000-1,500, 1,500-2,000 grams), IQ below 50, IQ 50-69, enlarged liver, 
malformations, retarded gross motor development, retarded fine motor develop- 
ment, hearing deficit in both ears, visual impairment, cranial or peripheral 
nerve damage, cerebral palsy, delayed motor development, hypotonia with poor 
deep tendon reflexes, non-febrile seizures, dyskinesia and ataxia, hearing 
deficit in one ear, and elevated bilirubin. The specimens from the mothers 
of these children and from the children themselves along with carefully matched 
controls are being studied for antibody to 11 antigens. These antigens include 
influenza A, rubeola, rubella, mumps, Coxsackie B3, Coxsackie B^ varicella, 
toxoplasmosis, cytomegalovirus, herpes Type I, and herpes Type II. All of 
these agents are known or suspected to be responsible for damage in the 
perinatal period. All laboratory work is being performed under code. The 
date is being analyzed by Mrs. Gilkeson and Dr. Ellenberg. 

A second approach is the study of reported viral, bacterial, and protozoal in- 
fections in pregnant women in the study. Serological tests are used to document 
these reports. The data is then correlated with the pediatric findings. 
Approximately 2,500 cases of reported viral infections, 3,000 bacterial infec- 
tions and several hundred protozoal infections, are under investigation. 
Clinical data is being abstracted, serological tests are being performed in 
order to document these infections. There are also approximately 1,200 patients 
identified with a postive serology for S5rphilis. These are being studied in 
detail. 

The third approach is to identify the children with elevated IgM levels in the 
newborn period and then to correlate these findings with pregnancy outcome, 
clinical performance of the child, and specific serological tests for IgM anti- 
body. Almost 32,000 cord sera have been tested for IgM antibody and approxi- 
mately 2,000 show elevated levels. These are now being studied in detail. 

Major Findings ; These reports of work "in progress" were given at the Inter- 
national Meeting on Congenital Malformations, Vienna, 1973: 

1. Maternal Clinical Infections and Pregnancy Outcomes - The frequency of 

clinical viral, bacterial, fungal, and parasitic infections was determined 
for the 58,828 pregnancies in the Perinatal Research Study. There were 
8,180 clinically infected pregnancies (13.9%). The infections included: 
viral 3,401p bacterial 4,539; fungal 102, and parasitic 138. Influenza 
"flu"-like illness occurred at the rate of 270 cases/10,000. Specific 
infections such as mumps, rubella, chickenpox/zoster and measles occurred 
at rates of 2 to 19 cases per 10,000 pregnancies. Bacterial infections 
of the kidneys - ureteral - bladder were very frequent (336/10,000 
pregnancies). Gonorrhea occurred at a rate of 32/10,000 pregnancies. 
Pinworm infections were the most frequent of the parasitic infections 
(12/10,000). 



17 X 



Serial No, NDS (CF)-57 ID 402 

2. Serological Studies of Perinatal Infections and Pregnancy Outcomes . 
Nine abnormal pregnancy outcomes were studied: abortions, stillbirths, 
cataracts, microcephaly, congenital hearts, neonatal deaths, low birth- 
weight (1001-1500 gms), low birthweight (1501-2000 gms) and low IQ 

( ^50). Eleven antigens were used. More than 100,000 antibody 
titrations were performed. Several leads have already appeared: 
1) Herpesvirus II in relation to microcephaly and abortions; 2) 
toxoplasma gondii in relation to stillbirths, neonatal deaths, and 
low birthweight; 3) Coxsackie B„, CMV, and mumps in relation to 
certain forms of congenital heart disease and mental retardation. 

3. Cord IgM Levels and Specific IgM for the Detection of Congenital 
Infections and Fetal Damage . Neonatal serum IgM levels are usually 
elevated in congenital infections. Cord blood samples 31,926 chil- 
dren had high IgM (^20 mgm %) . Specific tests for rubella, 
cytomegaloviruses and syphilis are now in progress. There were 8 
children with definite congenital syphilis. One was a stillbirth. 
There are 14 cases of suspected congenital syphilis. 

Cytotoxic (HLA) antibodies present in the serum of women with various abnormal 
pregnancy outcomes and matched controls were studied with Dr. Paul Terasaki, 
UCLA. A total of 3.5% of women who were pregnant for the first time had 
antibodies, however, this increased to 61 to 21 percent when the gravidities 
were three or more. These studies indicate that antifetal tissue antibodies 
are present in these pregnancies. 

Collaborating Studies : A primary deficit in the data from the Collaborative 
Perinatal Research Study has long been recognized as the late registration of 
the study'd patients. Since only 20% of the patients registered during the 
first trimester of pregnancy, it is impossible to document adequately the 
infectious disease experience of the patient's during the first trimester. 
To provide data on the first trimester of pregnancy and high-risk pregnancies, 
an additional cooperative study was initiated with the Infectious Diseases 
Branch. This is a study of viral infections in pregnancy of Dr. Margaret Jones 
UCLA, at the Kaiser Hospital, Los Angeles, California. 

A second study was initiated at the Kaiser Hospital, Los Angeles, California, 
to study infection in relation to infertility. Approximately 50 women are 
being studied each year. In each case sera, throat swabs, cervical swabs 
and endometrial biopsy specimens are obtained and sent to the IDE laboratories 
for detailed study. 

A second deficit of the Perinatal Research Study has been the lack of serial 
blood and tissue specimens from children with known congenital infection. For 
this reason, a special investigation was initiated with Dr. Janet Hardy at the 
Johns Hopkins Medical School to provide longitudinal surveillance of children 
with congenital infections to determine the clinical course, as well as the 
antibody levels, immunologic responses, and shedding of infectious agents in 
these children. A new study of HLA antibody in pregnant women and its effect 
on the baby was initiated with Dr. Paul Terasaki, UCLA. 



X 



Serial No. NDS (CF)-57 ID 402 

Significance of the Program to the Institute : The use of micro-serological 
techniques for a large group of new viruses provides an opportunity to inves- 
tigate the disease cause by viruses which are either difficult to isolate or 
resistant to evaluation because the clinical effects are delayed until a long 
time after the infection has subsided. In addition, the availability of new 
immunologic techniques provides the unique opportunity to detect immunologic 
deficits and to determine the presence of intrauterine infections on the basis 
of immunologic response. This data can then be correlated and analyzed as in 
relation to the possible causes of birth defects. The application of this 
type of analysis has provided valuable information on the epidemiology of 
virus infections in relation to abnormal pregnancy outcomes and is constantly 
giving us new insights into the causes of damage to the central nervous system 
and possible means of prevention of this and other damage to the developing 
fetus and newborn. 

Proposed Course of the Project ; The combined immunologic virologic program 
will continue at a very intense rate during the next year. During that time 
we plan to perform the great majority of tests which are needed in relation 
to the Cc ' " iborative Perinatal Research Study. Subsequent testing will then 
be limited : o special studies in depth and the intensive investigations in 
relation to the cooperative study at the Kaiser Hospital in Los Angeles and 
at the Johns Hopkins Medical School. 

The three approaches which are being emphasized now include: 

1. A special two-year commitment to perform serological tests on 
4,000 abnormal pregnancies and 4,000 matched controls using 11 
antigens. The abnormal children have been identified and the 
laboratory is now approximately 75% of the way through the testing. 

2. Special testing of IgM levels from 32,000 cord sera from children 
in the Collaborative Study and in the cooperative studies. This work 
provides an index for identifying children with possible congenital 
infections so that more specific testing can then proceed. These in- 
vestigations are now well under way and will be continued during the 
next year. Approximately 2000 cord sera have high IgM levels. These 
are being tested for specific antibody to rubella, toxoplasma, cyto- 
megalovirus, herpes II and syphilis. 

3. A correlation of clinically reported infections in pregnancy with 
serological findings for the pregnancy, immunologic determinations, and 
pregnancy outcome. These studies are now in progress and should be com- 
pleted for the most part in the next fiscal year. 

New rapid methods for determining IgM levels and specific IgM antibody will be 
investigated further. 



19 X 



Serial No. NDS (CF)- 7 ID 402 

Publications : 

Sever, John L,, Horta-Barbosa, L. Fuccillo, David A.: Viral and 
Protozoan Infections of the Newborn . Resuscitation of the Newborn 
Infant. Third edition, pp. 336-345, 1973. 

Sever, John L.: Viral Teratogens . Pathobiologv of Development, 1973, 
pp. 97-104. 

Sever, John L. : Efectos de infecciones sobre el riesgo en la gestacion . 
De la Rama de Enfermedades Infecciosas del Instituto de Enfermedades 
Neurologicas y Apoplejia, Institutos Nacionales de Salud, Bethesda, Md. 
Page 225-233, March 1973. 



20 



Serial No. NDS (CF)-61 ID 835 

1. Infectious Diseases Branch 

2. Section on Immunochemistry and Clinical Investigations 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Clinical Investigations with Human Volunteers and Other 
Populations Using Viruses, Vaccines, and Chemotherapeutic 
Agents. 

Previous Serial Number: Same 

Principal Investigators: Dr. John L. Sever, IDB, NINDS 

Dr. Jerome Kurent, IDB, NINDS 

Other Investigators: Dr. David A. Fuccillo, IDB, NINDS 

Dr. Michael Blaese, NCI 
Dr. Paul McCallin 

Kaiser Hospital, Hawaii 

Cooperating Units: Bureau of Prisons, Dept. of Justice 

Petersburg Federal Reformatory 

Man Years: 

Total: 1 
Professional: .5 
Other: .5 

Project Description : , ,■ 

Objectives : To study new prophylactic and therapeutic materials for the 
prevention and control of infectious diseases. To study the safety, 
antigenicity, communicability, and immunogenicity of candidate vaccines. 

Methods Employed : Human volunteer studies are conducted in collaboration 
with the Federal Bureau of Prisons. These studies are reviewed and approved 
by the Clinical Research Committee and the Medical Board of the National 
Institutes of Health and the Medical Review Board of the Federal Bureau of 
Prisons. 

Vaccines and chemotherapeutic agents which appear to warrant further 
investigations are then utilized in clinical studies of other special patient 
groups. In addition, clinical studies of epidemic are investigated as a 
means of identifying the role of infectious agents in the production of 
perinatal and pediatric disease states. 



21 



X 



Serial No. NDS (CF)-61 ID 835 

Major Findings : The immune response of volunteers with rubeola and rubella 
vaccines have been compared with the immune responses of patients with SSPE 
and MS. For SSPE there appears to be a serum inhibitor which blocks cellular 
immunity to measles. These investigations will be expanded. 

Transfer factor has been given to 8 patients with SSPE. These patients have 
been followed for 2 years. It appears that the patients have not had the 
expected rapid progression of disease. These studies will be extended. 

Significance of the Program to the Institute : Volunteers studied provide the 
basic data necessary to evaluate potential chemotherapeutic agents and 
vaccines. They also provide invaluable information on the course of 
infection in man. Utilization of this information then in clinical studies 
provides the necessary bridge to the implementation of therapeutic approaches 
which originated in the laboratory, are brought through the volunteer studies 
and then finally are taken to patient population. 

Proposed Course of the Project : The additional studies will be performed 
utilizing rubeola vaccines to determine antibody response. Rubeola antibody 
in SSPE patients will be studied in detail. Clinical studies will continue 
on utilization of various drugs for the prevention of treatment of perinatal, 
acute and chronic infections in man. The value of transfer factor for the 
treatment of SSPE will be studied with additional patients. In addition, 
the use of new methods for determining spinal fluid lactic dehydrogenase and 
endotoxin levels in CSF will be studied in relation to the early diagnosis of 
meningitis. 

Honors and Awards : None 

Publications : 

Sever, John L.; Krebs, Helen; Ley, Anita; Barbosa, L. H.; 

Rubinstein, David: Serological Tests for Measles for the Diagnosis 
of Subacute Sclerosing Panencephalitis. The Journal of the 
American Medical Association. (In Press) 



22 



Serial No. NDS (CF)-72 ID 972 

1. Infectious Diseases Branch 

2. Section on Experimental Pathology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Experimental Animal, Tissue Culture, Histopathological 
and Serological Investigation of the Role of Viruses 
and other micro-organisms in the Perinatal Period 

Previous Serial Number: Same 

Principal Investigators: Dr. William T. London, IDE, NINDS 

Dr. Amos E. Palmer, IDE, NINDS 

Dr. John L. Sever, IDE, NINDS 

Dr. David A. Fuccillo, IDE, NINDS 

Other Investigators: Mrs. Anita C. Ley, IDE, NINDS 

Mrs. Blanche Cur f man, IDE, NINDS 

Cooperating Units: Dr. Stephen Kent 

Department of Pathology 
George Washington University 
Washington, D.C. 

Dr. Neal H. Levitt 

Virology Division 

U.S. Army Medical Research Institute of 

Infectious Diseases 
Fort Detrick, Frederick, Maryland 21701 

Man Years : 

Total: 1.8 

Professional: o.8 

Other: 1.0 

Project Description : 

Objectives : To study the role of viruses and other microorganisms in the 
perinatal period, the infection of gravid and nongravid. animals of several 
different species by parenteral routes with various viruses and other micro- 
organisms to determine the effects of these agents on the animals and their 
fetal tissues. 

Attempt to recover inoculated agents from the various animals and fetal 
tissues and the correlation of these reisolation with time (in gestation) 
of inoculation and dosage given. Relate these findings with gross and 



23 



Serial No. NDS (CF)-72 ID 972 

histopathological findings. Correlate all of this information with 
serological findings. 

Methods Employed : An investigation of the role of viruses and other micro- 
organisms in the perinatal period by the continual use of experimental 
animals; tissue culture techniquea; histopathological studies; and serological 
testing. 

Special quarantine facilities are maintained to prevent feral animals going 
through quarantine from becoming infected with endemic diseases such as 
rubeola and cytomegalovirus. A serological defined breeding colony of 
Rhesus monkeys is maintained to provide susceptible pregnant females for 
inoculation. 

Pregnant animals are being inoculated by various routes with viruses and 
other microorganisms, and held in isolation chambers throughout the experi- 
ment. These animals are being observed and serum samples, liver biopsy, 
spinal fluid and throat swabs are checked for evidence of disease and/or 
effects on fetal tissue. 

Virus isolation investigations utilizing tissue culture to recover viruses 
from tissues and fluorescent antibody technique to study the location of 
virus infection produced in experimental animals are being employed. 

Major Findings : Fifteen Rhesus monkey fetuses were inoculated intracerebrally 
through the uterine wall during maternal laparotomy at 103-107 days of ges- 
tation. Nine were given mumps virus and 6 received control material. 
Persisting infection was demonstrated by isolation of mumps virus from 
the liver of one fetus 55 days postinoculation. Virus was not isolated from 
the maternal throat swabs, placenta, or amniotic fluid. Antibody to mumps 
was consistently present in the virus inoculated mothers and babies, but 
was higher in the babies. Three of the 9 infected animals and none of the 
controls showed hydrocephalus at necropsy. The hydrocephalus was bilateral 
and asymmetrical in two cases, and bilateral and symmetrical in one case. 

Significance to the Program of the Institute : A program using experimental 
animals, tissue culture techniques, and histopathological investigations 
compliments the strict serological approach being used on human sera obtained 
from the Collaborative Study and thus balances the investigations of the role 
of viruses and other microorganisms in the perinatal period. It presents the 
direct means of investigation of these agents which may contribute to human 
perinatal pathology. 

Proposed Course of the Project : Further studies using Reo and Visna Viruses 
inoculated into pregnant monkeys are in progress. In addition, we are using 
known teratogens. Influenza A Virus and Venezuelan Equine Encephalitis Virus 
(VEE) inoculated into pregnant animals and then delivering the fetus by 
cesarean section within a few days postinoculation to determine the acute 
pathogenic effects of these agents on the fetus. 



24 X 



Serial No. NDS (CF)-72 ID 972 
Honors and Awards : None 

Publications : None 



25 



X 



Serial No. NDS (CF)-65 ID 1270 

1. Infectious Diseases Branch 

2. Section on Immunochemistry and Clinical Investigations 

3 . Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1, 1973 through June 30, 197^1- 

Project Title: Toxoplasmosis: Serological and Clinical Studies 

Previous Serial Number: Same 

Principal Investigator: Dr. John L. Sever, IDB, NIMDS 

Other Investigators: Dr. Joseph S. Drage, PRB, NIEDS 

Cooperating Units: Section on Pediatric Neurology, PRE, NINDS 

Man Years: 

Total: .h 
Professional: .2 
Others : .2 

Project Description : 

Objectives : This study relates to antibody titer in pregnant women to 
abnormal pregnancy outcomes. A total of 23,000 women have now been studied. 

Methods Employed : The computer analysis of multiple variables relating to 
toxoplasmosis in pregnancy outcome has been used. The serologic data is 
beijog correlated with pregnancy outcomes. 

Major Findings : Within the group of hj patients with high titers or signi- 
ficant increases in antibody titers, 5 were found to have definite 
toxoplasmosis and 10 were suspected of having possible toxoplasmosis. 
Theire were increased rates of stillbirths and neonatal deaths. The 
acquisition of antibody to toxoplasmosis was strongly correlated with 
age and race . 

Proposed Course of the Project : Publication of complete analysis will be 
pursued this year. Data for 7-8 year IQ levels will be analyzed. 

Honors and Awards : None 

Publications: None 



27 X 



Serial No. NDS (CF)-67 ID 1503 

1. Infectious Diseases Branch 

2. Section on Immunochemistry and Clinical Investigations 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Epidemiologic Studies of Perinatal Infections 

Previous Serial Number: Same 

Principal Investigators: Dr. John L. Sever, IDE, NINDS 

Dr. Jerome E. Kurent, IDB, NINDS 

Mrs. Dorothy Edmonds, R.N., IDB, NINDS 

Other Investigators: Dr. Jonas Ellenberg, OB, NINDS 

Cooperating Units: Collaborative Perinatal Research Study 

Kaiser Hospital, Los Angeles, California 
Kaiser Hospital, Hawaii 
Kaiser Hospital, Oakland, California 
Dr. Thomas Waldmann, NCI 

Man Years: 

Total: 1 
Professional : 1 ■ • 
Other: . t •; : 

Project Description : . ; ■ • ' ' 

Objectives : Utilize information from collaborative and cooperative studies 
to identify pregnancies complicated by maternal infections or infections in 
childhood. To further delineate these cases by serological testing and to 
determine the outcome of these pregnancies in relation to the infections 
which have been defined. 

Methods Employed : Primary material utilized for these studies are the 
clinical records, serum and tissue specimens obtained during pregnancy and 
the pediatric period of observation. Serologic data developed by the 
immunology laboratories is correlated with the clinical information from the 
collaborating groups. Data is analyzed in cooperation with the Office of 
Biometry, NINDS. 

Major Findings: At least a dozen viruses have been identified as being of 
importance in the perinatal period. The specific rates of the occurrence of 
these infections appear to be excess of 5 percent of the total pregnancies. 
As many as 10 percent of the children with microcephaly and mental 
retardation may be associated with these perinatal infections. 



29 



X 



Serial No. NDS (CF)-67 ID 1503 

Preliminary studies of maternal alpha-fetoprotein levels indicate that this 
is elevated in high risk pregnancies. 

Significance of the Program to the Institute : The identification of 
clinically reported cases of infection provides an immediate tool for 
analyzing the effects of perinatal and pediatric infections on the develop- 
ment of the child. The epidemiologic studies which are possible because of 
the large numbers of patients in the collaborative and cooperating studies 
provide the necessary data base for these types of investigations. 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 analysis 
of data for neonatal sepsis. There are approximately 50 cases of each with 
5-8 year longitudinal followup observations which will be analyzed. 
Alpha-fetoprotein levels will be determined for pregnant women with various 
abnormal outcomes. This method appears to be of value in the detection of 
pregnancies of high risk. 

Honors and Awards : None 

Publications: 



Sever, John L.: Effects of Infections on Pregnancy Risk. 
Clinical Obstetrics and Gynecology, Vol. 16, No. 1, 
March, 1973. 

Sever, John L.: Potatoes and Birth Defects: Summary. 
Teratology, Vol. 8, No. 3, pp. 319-320, December, 1973. 

Kurent, Jerome E., Sever, John L.: Perinatal Infections 
and Epidemiology of Anencephaly and Spina Bifida. 
Teratology, Vol. 8, No. 3, pp. 359-361, December, 1973. 



30 X 



Serial No. NDS (CF)-69 ID 1731 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Isolation of Infective Agents From Chronic Diseases 

Previous Serial Number: Same 



Principal Investigators: 



Dr. Luiz H.-Barbosa, IDE, NINDS 

Dr. David A. Fuccillo, IDE, NINDS 

Dr. William T. London, IDE, NINDS 

Dr. John L. Sever, IDE, NINDS 

Dr. Monique DuBois, IDE, NINDS 

Dr. Jerome Kurent, IDE, NINDS 



Other Investigators: 



Cooperating Units: 



Mrs. Rebecca Hamilton, IDB, NINDS 
Mr. Otto Gutenson, IDB, NINDS 
Mrs. Anita Ley, IDB, NINDS 
Mr. Leonard Moore, IDB, NINDS 

University of Tennessee (Dr. J. T. Jabbour) 
University of Vermont (Dr. George Schumacher) 
Indiana University Medical Center (Dr. Wolfgang Zeman) 
Wilmington General Hospital (Dr. George Boines) 
Cornell University, N.Y. (Dr. Fred Plum) 
UCLA, Los Angeles (Dr. Gary Gitnick) 



Man Years: 

Total: k,Q 
Professional: 1.5 
Other: 2.5 

Project Description : 

Objectives : To establish whether persistent or tolerant viral infections are 
associated with chronic diseases such as Bell's palsy, amyotrophic lateral 
sclerosis, Creutzfeldt-Jakob disease, progressive multifocal leukoencephal- 
opathy, and multiple sclerosis. 

To purify and concentrate PML and SSPE viruses and examine these agents 
electron microscopically, biochemically, and antigenically in parallel with 
the conventional forms of these viruses. 

To attempt the transmission of MS to SPE mice and chimpanzees using intra- 
cerebral inoculation of 10% homogenate prepared with "plaque" areas from 
brain autopsy material. 



31 X 



Serial No. NDS (CF)-69 ID 1731 

131 
To trace the possible anti-brain activity of MS immunoglobulin, using I 

labeled MS serum to inoculate Rhesus monkeys for brain autoradiography. To 

develop an animal model system to study the immunopathogenesis of SSPE as well 

as possible therapeutic approaches. 

To conduct epidemiological surveys concerning the occurrence of SSPE during the 
1965-1975 decade. 

To evaluate the role of cellular immunity, humoral immunity, and interferon in 
protecting experimental animals against neurotropic measles virus. 

Methods Employed : Brain specimens from well-documented cases of MS, PML, ALS, 
SSPE, etc., were homogenized to a 10% suspension and inoculated intracranially 
in groups of 5 Rhesus monkey fetuses during the first one-third of gestation 
when the animals are expected to be immunologically immature and hence more 
susceptible to infectious agents. These fetuses were carried to term and, 
immediately after birth, one animal was killed and examined histopathologically 
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 from patients with encephalopathies, muscle from patients with 
polymyositis, and lymph nodes from individuals with SSPE and MS were examined 
by electron microscopy, fluorescent microscopy and, whenever possible, 
cultured in vitro. These tissue cultures were submitted to virologic and 
serologic assays, inoculated into experimental animals, and cocultivated with 
human diploid and heteroploid cell lines in efforts to isolate and identify 
intracellular agents. 

Brain autopsies from patients with PML and known to contain intranuclear 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 
papovavirus were used to immunize guinea pigs in order to produce monospecific 
antisera intended to identify the virus. The peroxidase-coupled antibody 
technique was perfected for measles virus and it is expected to become a 
valuable tool when utilized with these guinea pig antisera and PML brain 
tissue. Similar approach, but using zonal centrifugation to purify the virus, 
was planned for SSPE and is awaiting approval of the necessary contract. The 
same technique is expected to be utilized to study particulate fractions of 
MS brain. 

Comparative studies between SSPE measles and prototype measles virus were 
conducted in efforts to establish whether or not the SSPE isol^.tes have 
markers to distinguish them from laboratory strains of measles. 

A questionnaire concerning the occurrence of SSPE since 1965 was sent to 
neurologists, pediatricians and pediatric neurologists at 104 medical centers 
throughout the USA. Data was analyzed in our laboratory and at the University 
of Tennessee. 



32 



Serial No. NDS (CF)-69 ID 1731 

In order to investigate a possible impairment of cellular immunity in SSPE 
patients, an in vitro lymphocyte assay was conceived and efforts initiated 
to obtain a measles chronically-infected cell line necessary to evaluate 
lymphocyte measles-specific delayed hypersensitivity. 

Major Findings : Primary and secondary MS brain cultures were extensively 
tested for the possible presence of viruses. No evidence of an infectious 
agent was found in three MS brain cultures studied by immunofluorescence, 
electron microscopy, hemadsorption, cocultivation, and biochemical "shocking." 

An immunoper oxidase electron microscopic test for measles virus was developed 
and used to study SSPE strains of the virus. This assay is expected to be 
useful on the search for antigens in MS, PML, and other neurological diseases. 
A papovavirus was purified from a PML brain autopsy and subsequently used to 
inoculate laboratory animals. The purification was achieved by banding the 
virus in CsCl (1.6 grs/ml) . Spectrophotometry and electron microscopy 
monitoring resulted in a rather pure virus preparation of approximately 10 
intact particles/5 grams of tissue. 

Both PML tissue suspensions and the purified virus were utilized to inoculate 
a variety of tissue culture systems which, after months of careful follow-up, 
were considered free of PML virus as monitored by cellular changes and/or 
presence of intranuclear particles. 

The search for biological markers to distinguish SSPE isolates from conventional 
measles virus proved unsuccessful. Marked differences between SSPE strains 
were noted by various parameters, indicating strain dissimilarities as great 
or even greater than those previously observed between laboratory strains of 
measles virus. 

Epidemiological studies involving over 250 SSPE patients and dozens of medical 
institutions produced the following major findings: 1) the frequency of SSPE 
was approximately 1:1,000,000 during 1960-1970; 2) concentration of patients 
in the southeastern part of the USA; 3) the age of onset of the disease was 
7.2 years; 4) a 3.3:1 male/female ratio; and 5) early measles infection 
<.3 years of age in the majority of patients. 

Significance to the Program of the Institute: The development of mixed cell 
cultures to unmask latent infections provides an excellent methodology for 
the study of chronic diseases of possible viral etiologies. 

The understanding of slow virus infections of the CNS will depend upon 
purification of the suppressed form of these agents followed by careful 
biochemical and biophysical analysis. To accomplish this goal, the use of 
zonal centrifugation for cellular fractionation should be emphasized. 

Proposed Course of the Project : Special emphasis will be placed upon a brain 
culture research program. Investigation of the mechanism of pathogenesis 
and possible immune deficiencies in patients with neurological diseases will 
be conducted. Our selection of patients with multiple sclerosis, Parkinson's 



33 



Serial No. NDS (CF)-69 ID 1731 

Disease, progressive multifocal leukoencephalopathy, and amyotrophic lateral 
sclerosis is supported by the existing data which suggest possible viral 
etiologies for each of these diseases. Tissue specimens and blood from 
patients will be provided through collaborative-contract arrangements with 
investigators throughout the country. 

The possible viral etiology for MS will be exhaustively investigated. Since 
the pathology of Visna and MS are similar, and reverse transcriptase was 
found in tissues infected with Visna virus, we intend to determine whether 
this viral enzyme is present in spinal fluid and brain tissue of MS patients. 
In addition, molecular hybridization studies with MS tissue and tissue culture 
will be initiated to search for virus "fingerprints" such as messenger RNA, 
virus specific DNA or RNA. These homology assays would be conducted with MS 
purified DNA and RNA in presence of labelled measles RNA and possibly 
labelled varicella-zoster DNA. The latter viruses are at present the best 
candidates since serologic surveys indicated that MS patients have elevated 
measles and/or zoster antibody levels. 

In a second approach, MS gamma globulins will be coupled to tracers 
(fluorescein and peroxidase) and used in a direct antigen-antibody test with 
MS brain tissue culture. Using the electron microscope and UV microscope, we 
can determine and identify antigenic sites in the MS brain tissue culture. 

Third, in a more direct fashion, we will utilize experimental animals known 
to be surgically acceptable to slow neurotropic agents (scrapie, visna) to 
inoculate MS brain preparations. These animals will include SPF mice and 
primates. 

Utilizing the mixed culture technique, we hope to determine whether 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 CNS. Antibody levels and competence of lymphocytes from patients will 
be examined, using the standard techniques. The SSPE strain of measles virus 
will be studied in laboratory animals and efforts will be directed at the 
development of an animal model system for this disease. 

Honors and Awards : None 

Publications : 

Dietzman, D. , and Horta-Barbosa, L. : Slow Virus Infections. In 
Brenneman's Practice of Pediatrics. Harper & Row Publishers, Inc., 
Vol. II, Chapt. 43, September 1973. 

Farmer, G. , Vincent, M.M. , Fuccillo, D.A., Horta-Barbosa, L., 
Ritman, S., Sever, J.L., and Gitnick, G.L. : Viral Investiga- 
tions in Ulcerative Colitis and Regional Enteritis, Gastro- 
enterol. 65: 8-18, July 1973. 



34 X 



Serial No. NDS (CF)-69 ID 1731 

Horta-Barbosa, L., Hamilton, R.: Virological Studies with 
Multiple Sclerosis Brain Tissue, Lancet, June 23, 1973, pp. 1415- 
1417. 

Vernon, M.L. , Krebs, H.M. , Horta-Barbosa, L., Fuccillo, D.A. , 
and Sever, J.L.: Birbeck Granules (Langerhans Cell Granules) 
in L3miph Nodes from "Normal" and Diseased Patients. Am. J. 
Path,, 60: 771-779, December 1973. 

Sever, J.L., Krebs, H.M. , and Horta-Barbosa, L. : Availability 
of Serological Tests for Measles for Diagnosis of SSPE,(jAMA 
in press) . 

DuBois-Dalcq, and Horta-Barbosa, L. : Immunoperoxidase Stain 
of Measles Antigen in Tissue Culture. J. Virol. 12: 909-914, 
October 1973. 



35 X 



Serial No. MDS (CF)-69 ID 1732 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 
3- Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^1 

Project Title: Investigation of the Role of Mycoplasma spp. in 
Perinatal and Neurological Diseases 

Previous Serial Number; Same 

Principal Investigators: Dr. David L. Madden 

Dr. David A. Fuccillo 
Dr. John L. Sever 

Other Investigators: Mrs. Aurella Krezlewicz 

Cooperating Units: National Naval Medical Hospital 

Man Years : 

Total: 1.00 
Professional: .25 
Other: .75 

Project Description : 

Objectives : To study the role of mycoplasma in diseases of man, particixLarly 
those associated with perinatal disease, infertility, or neurological disease. 
To develop animal models and to determine the pathogenesis of these diseases. 
To develop new tests for identification of mycoplasma. 

Methods Employed ; Urine samples and small pieces of vas deferens were 
collected from men undergoing vasectomy. These samples were cultured for 
presence of mycoplasma and positive cultures were identified by standard 
techniques . 

Small pieces of biopsy material from a variety of neurological diseases and 
secondary tissue cultures are also being studied for presence of mycoplasma. 
These include such diseases as Subacute Sclerosing Panencephalitis, Alateral 
Sclerosis, Multiple Sclerosis, and Creutzfeldt -Jakob disease. These tissues 
are being cultured in standard mycoplasma media, and the positive isolated 
cultures are being identified by standard techniques. 

Major Findings ; A mycoplasma associated with a nonproductive virus culture 
has been serologically identified as related to Mycoplasma orale type I. 
This isolate, however, has biological characteristics not associated 
with this strain. Inoculation of both cultures into brains of newborn 
hamsters did not produce clinical signs of the disease. Organisms have 
been recovered from apparently normal inoculated hamsters up to six months 

37 X 



Serial No. NDS (CF)-69 ID 1732 

post inoculation. Growth of infected tissue culture cells in high levels 
of specific mycoplasma antibody and antibiotics have not been successful 
in elimination of the organism from the tissue culture. 

The study on occurrence of mycoplasma in ma2es has been continued. In 
matched samples^ classical strain mycoplasma and T-strain mycoplasma have 
been isolated from both semen and urine but not from the vas tissue. 
This further indicates that mycoplasma in normal males are not present 
in the vas deferens . 

Significance of the Program to the Institute ; A program devoted to studying 
the effects of mycoplasma in various diseases complements the virological 
studies currently being done. This study and its support given to other 
investigators may help to more accurately define the role of mycoplasma and 
other agents in disease. 

Proposed Course of the Project ; Efforts are underway to initiate a study 
to determine the role of mycoplasma in infertility and repeat abortion. 
Attempts to define the role of mycoplasma in neurological diseases will be 
continued. Continued efforts will be made to apply new techniques in the 
identification of mycoplasma and mycoplasmal diseases. 

Honors and A'.^ra.rds ; None 

Publications ; 

Traub, R. G., Madden, D. L., Fuccillo, D. A., and McLean, T. W.; The 

Male as a Reservoir of Infection with Cytomegalovirus, Herpes and Mycoplasma. 

IT Engl. J. Med. 289; 697, 1973- 



38 



Serial No. MDS (CF)-71 ID 1903 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 
3' Be the s da, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 197^ 

Project Title: Investigation of the Etiology and Effect of Serum and 
Infectious Hepatitis in the Perinatal Period 

Previous Serial Number: Same 

Principal Investigators: Dr. David L. Madden, IDB^ NINDS 

Dr. John L. Sever, IDB, NINDS 

Other Investigators: Dr. Benedict Nagler 

Mrs . Betty Dunlop ... 

Dr. Robert Pur cell 
Mrs. Mary Krasny 

Cooperating Units: Lynchburg Training School and Hospital 

Montgomery County Public Health SerAn.ce 
' ■ Abbot Laboratories 
Electro-nucleonics 

Many Years: 

Total: 0.50 
Professional: O.25 
Other: ■ G.25 

Project Description : 

Objectives : To determine the etiology of Australia antigen associated 
(serum) and infectious hepatitis. To determine the relationship of hepa- 
titis/congenital jaundice and postnatal jaundice. To develop animal models 
and new diagnostic tests for these diseases. 

Methods Employed : A large epidemic of infectious hepatitis occurred in the 
Lynchburg Training School and Hospital during the summer of 1970. Serial 
samples of feces and serum were obtained from many patients prior to 
development of disease at the time of acute disease and post infection. 
Serum samples have been obtained from these patients at 6-, 12- and I8 months, 
Hepatitis B antigen and antibody detenninations have been performed on all 
patients with Down's Syndrome on eleven wards and matched controls. In 
cooperation with Electro-nucleonics, Co., three serial samples of serum and 
many fecal samples have been examined by electron microscopy and immuno 
electron microscopy techniques for presence of Hepatitis A antigen. 



39 X 



Serial No. MDS (CF)-71 ID I903 

Major Findings : A major epidemic of hepatitis A in an institution for 
mentally retarded was studied. The effect of hepatitis B in the institu- 
tion was studied. Serological typing of hepatitis B antigen in the patients 
indicated that the predominant type in the Lynchburg Training School and 
Hospital was ad. The serotypes did not seem to be random among the wards 
nor the patients. The ad serotype occurred more often in patients housing 
wards which house older patients and the ay more prevalent in younger patients . 
The ad serotype was more prevalent in the males than the females. 

Virus particles similar in size to those observed in experimental hepatitis 
A infections have been observed in the feces of patients from the I^ynchburg 
Training School and Hospital epidemic. A seroconversion against the hepa- 
titis A agent has been found. Data from the Lynchburg Study and the Perinatal 
Study indicates that there is no association between the formation of genetic 
Down's syndrome and occurrence of hepatitis B prior to or during pregnancy. 

Significance of the Program to the Institute ; Hepatitis in pregnancy has 
been associated with neonatal jaundice, Down's syndrome and other forms of 
mental retardation, among the institutionalized mentally retarded patients 
\^D.th trisomy 21 karyotypes have a high rate of chronic hepatitis B 
antigenemia. This study provides additional depth to determining the 
role of infectious agents in perinatal disease and mental retardation. 
Data obtained from this study will clarify the association of Down's syndrome 
to hepatitis A infections. 

Proposed Course of the Project : Studies on the effect of hepatitis B in a 
mentally retarded population are being completed. The relationship between 
exposure to hepatitis B antigen and neonatal jaundice, hepatitis and mental 
retardation will be detejrmlned. Efforts are being made to initiate studies 
vrLll well determine the relationship of Down's syndrome and hepatitis A. 

Awajds and Honors : None 

Publications : 

Dletzman, D. E., Madden, D. L., Sever, J. L., Dunlop, B.: Family Contacts 
of HAA Positive Persons. N. Engl. J. Med. 288: lij-09-lUlO, 1973- 

Matthew, E. B., Dletzman, D. E., Madden, D. L., Newman, S. J., Sever, J. L., 
Nagler, B., Bouton, S. M., and Bpstaflnski, M.: A Major Epidemic of 
Infectious Hepatitis in an Institution for the Mentally Retarded. Am. J. 
of Epid. 98: 199-215, 1973. 



40 X 



Serial No. NDS (CF)-72 ID 1981 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Immunoglobulin M and Congenital Infections 

Previous Serial Number: Same 

Principal Investigators: Dr. Luiz H. Barbosa, IDB, NINDS 

Dr. John L. Sever, IDB, NINDS 

Other Investigators: Miss Helen Krebs 

Mrs. Rebecca Hamilton 
Mr. Otto Gutenson 

Cooperating Units: University of Tennessee (Dr. Sheldon Korones) 

Electro-Nucleonics, Inc. (Dr. Edward Bond) 

Man Years: 

Total: 0.8 
Professional: 0.2 
Other: 0.6 

Project Description : - • 

Objectives: To study the antibody specificity of cord and newborn serum IgM 
as an investigational screening procedure to identify intrauterine or 
neonatal infections. High risk pregnancies will be included in this study, 
using the elevated IgM level as index for possible perinatal infections. 

Methods Employed: Part of this investigation is being studied in 
collaboration with Dr. Korones at the University of Tennessee. In these 
studies, serum specimens are obtained bi-weekly from newborn infants in the 
high risk nursery. Elevated IgM levels (>20 mg. %) will be used as 
indication of possible congenital or neonatal infection. Sera with high IgM 
content are then passed through prepared sepharose columns in order to 
separate the 7S from the 195 gamma globulins. Purified IgM samples are 
subsequently tested by the indirect immunofluorescent assay against CMV, 
HVH, Rubella, Toxoplasma, and Treponema Pallidum which are known to be the 
main causative agents of perinatal infections. Infected tissue cultures are 
used as the subs rate assay for the three viruses and toxoplasma and 
treponema smears are utilized for determining IgM antibody elicited by 
toxoplasmosis and syphilis, respectively. A fluorescein conjugated anti-IgM 
serum is employed to stain antigen-antibody complexes. A careful 
longitudinal follow-up of the children under study will be conducted by 

41 X 



Serial No. NDS (CF)-72 ID 1981 

Dr. Korones to correlate serological data with clinical observations, and 
when indicated, clinical specimens will be sent to the laboratories of the 
Infectious Diseases Branch. 

A second part of this project involves the separation and antibody determina- 
tion of the IgM in approximately 12,000 cord serum samples from babies with 
elevated IgM. These infants' samples are from the Collaborative Perinatal 
Study and are products of conception from mothers with serological findings 
indicating infections during pregnancy. Antibody specificity of the purified 
cord serum IgM samples are tested in the same manner described above. 

Major Findings : The novel technique of antibody-coupled sepharose particles 
was perfected in collaboration with Dr. Bond (Electro-Nucleonics) and found 
to be useful to separate immunoglobulins from body fluids. This technique 
provides an unexpensive and rapid method to purify IgM in contrast with the 
elaborate, time-consuming preparative ul tracentrifuga.tion in sucrose 
gradients. 

Measles specific Igf1 was found to be present in the serum of SSPE patients. 
This observation correlates with an early report by Connoly, et al., 
describing rubeola antibody in the IgM fraction of SSPE patient's spinal 
fluid. 

Significance to the Program of the Institute : Several infectious agents are 
known to produce intrauterine infection and fetal damage. A great number of 
mentally retarded children are a consequence of congenital infection 
affecting the CNS. Cord and newborn serum IgM levels are usually elevated 
with congenital infections. Therefore, the quantitation and qualitation of 
cord and newborn serum IgM can be applied as an investigational screening 
procedure to identify high risk infants, or in studies of newborn in 
distress. More specific procedures can then be used to determine the cause 
for the elevation of this immunoglobulin. 

Proposed Course of the Project : Comparison studies between specific 
immunofluorescence and latex agglutination techniques will be conducted to 
evaluate degree of sensitivity and specificity of cord IgM antibodies and 
candidate antigens (Toxoplasma, syphilis, CMV, rubella, and Herpes). Once 
the assay is perfected to a desirable reproducibility, approximately 1,800 
high IgM and serum samples from the Collaborative Perinatal Research Study 
will be tested for specific IgM antibodies. The results will be correlated 
with the mother's serum antibody findings as well as mother's clinical 
history and the infant's findings. 

Honors and Awards : None 

Publications: None 



42 



X 



Serial No. NDS (CF)-72 ID 1982 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Delayed Hypersensitivity in Chronic Viral Diseases 

Previous Serial Number: Same 

Principal Investigator: Dr. David A. Fuccillo, IDE, NINDS 

Other Investigators: Mrs. Renee Traub, IDB, NINDS 

Cooperating Units: Dr. Bellanti and Dr. Russell Steel 

Georgetown University School of Medicine 
Washington, D. C, 
Mr. Monty Vincent 
Microbiological Associates, Bethesda, Md. 

Man Years: 

Total: 1 
Professional: 0.5 
Other: 0.5 

Project Description : 

Objectives : To determine the role of delayed hypersensitivity (cellular 
immunity) in chronic viral infections. 

Methods Employed : A microassay technique for cell mediated immunity was 
developed which will increase the number of specimens that can be done with 
the added advantage of requiring less blood. 

A lymphotoxin assay to be used in this study for rubella has been developed. 
Other tests for CMV and Herpes are beinq v/orked on at the present time. 

A rubella chronically infected cell line was found to be very useful for 
the lymphotoxin assay. This assay was developed and tested against 
imnunized rubella children and normal results indicate a workable system is 
now available. 

A rubeola chronically infected cell line was developed. This system is now 
being used in the study of SSPE patients to determine their cellular immune 
capabilities against this virus. 



43 



Serial No. KDS (CF)-T2 ID I982 

51 

Major Findings : A Cr microassay procedure for the in vitro measurement 

of cell mediated immunity to rubella was described and published. Specific 
cell-mediated immunity resources to rubella virus were studied in 12 children 
with documented congenital rubella employing this microassay cell mediated 
immunity was only demonstrated in 9 of "the children. These results suggest 
that congenital rubella infection produces an impairment of cellular immunity 
and could account for the continued isolation of virus in the presence of 
antibody. 

Significance of the Program to the Institute ; Prior to the development of 
the above test^ no reliable tests for measuring cellular immunity in humans 
to viral antigens existed. Antibody tests to viral antigens ha-ve been 
developed and used in this and other laboratories for many years. This 
represents a test for only part of the body's immune response. The 
development of lymphotoxin assay into a clinically useable test for virus 
infection now allows both forms of the body's protective immune mechanisms, 
humoral and cellular, to be tested simultaneously. 

Proposed Course of the Project ; To complete the above mentioned specific 
projects and to develop additional tests to elucidate how the cellular 
immune system is involved with other viral diseases especially those that 
cause congenital infection. Congenitally infected infants excrete virus in 
the presence of high antibody titers, possibly indicating that their cellular 
immune system is impaired. The effect of complement as well as the function 
of cells other than lymphocytes can be studied with this system. 

This test will also be used to determine the cellular immune capabilities of 
patients with MS and other neurological diseases. 

Honors and Awards ; None 

Publications ; 

Steel, R. W., Hensen, S. A., Vincent, M. J., Puccillo, D. A., 
and Bellanti, J. A.; A 51cr Microassay Technique for Cell- 
Mediated Immunity to Viruses. J. Immunology, vol. 110, 1502-1509^ 
1973. 

Fuccillo, D. A., Steel, R. W., Hensen, S. A., Vincent, M. M., 
Hardy, J. B., and Bellanti, J. A.: Impaired Cellular Immunity 
to Rube3J.a Virus in Congenital Rubella. Infection and Immunity; 
Vol. 9, 81-8 ij-, ±9lh. 

Steel, R. W. , Hensen, S. A., Vincent, M. M., Fuccillo, D. A., and 
Bellanti, J. A.; The Development of Specific Cellular and Humoral 
Immune Responses in Children Immunized with Live Rubella Virus 
Vaccine. J. Infectious. Diseases (in press). 



44 



Serial No. NDS (CF)-72 ID 1983 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through Jane 30, 1974 



Project Title: Chronic Viral Infections 

Previous Serial Number: Same 

Principal Investigator: Dr. David A, Fucc111o 



Other Investigators: 



Cooperating Units: 



Dr. John L. Sever, IDE, NINDS 
Dr. William T. London, IDB, NINDS 
Dr. David L. Madden, IDB. NINDS 
Dr. Luiz H. Barbosa, IDB, NINDS 
Mrs. Flora Moder, IDB, NINDS 
Mrs. Renee Traub, IDB, NINDS 

Dr. L. Johnson 

Harvard Medical School, Boston, Massachusetts 
Dr. Mark May 

McMllllan Hospital, St. Louis, Mo. 
Dr. Gary L. Gitnick 

Dept. of Medicine, UCLA Center for Health Sciences, 

Los Angeles, California 
Mr. Monty Vincent, Microbiological Association 
Dr. Bllllngsley, Naval Medical Center, Bethesda, Md. 
Dr. McLean 



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 herpes simplex 
virus (Type I, "oral" and Type II, "genital") cytomegalovirus, varicella, in 
the causation of human disease including carcinoma, ulcerative colitis. Bells' 
palsy, facial paralysis, herpes zoster and chronic neurological diseases. 

Methods Employed : The principal methods employed are: (1) the indirect 
hemagglutination test, by which type specific herpesvirus antibody is 
identified; (2) mass serological surveys of materials from selected patients 
with special disease entities; (3) virus isolation, titration, and 



45 



Serial No. NDS (CF)-72 ID 1983 

characterization procedures; (4) monolayer cultivation of tissue with 
cocultlvation; (5) fluorescent antibody studies of monolayer cultures for 
the presence of latent infection. 

Major Findings : In collaboration with Dr. Gitnick, a higher frequency of 
cytomegalovirus antibody and higher CMV titers were found in ulcerative 
colitis patients. Three of 6 ulcerative colitis patients and 1 of regional 
enteritis had CMV isolated from biopsy tissues. The tissue culture of one 
patients underwent spontaneous cellular transformation. The etiology of 
ulcerative colitis still remains unknown, but this work may help determine 
the relationship of this virus to the pathogenesis of this disease. 

In collaboration with Dr. McLean a study was done to determine if the 
male was a reservoir of infections for cytomegalovirus, herpes and mycoplasma. 
Data from this study indicates that the vas deferens was free of demonstrable 
viral and mycoplasma but the latter were found to be present in a number of 
urine specimens. Results were reported. 

Significance of Biomedical Research and the Program of the Institute ; These 
investigations attempt to elucidate the pathogenesis of viral infections of 
the adult and fetus using immunological and virological techniques. Herpes 
simplex virus, CMV, and varicella have been agents which have received 
particular attention in these studies, since they have significant 
neurotropic capabilities in terms of newborn and adult encephali tides. There 
is considerable speculation that these viruses 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 per- 
mitted more definitive establishment of such capabilities. Furthermore, the 
therapeutic potential of experimental drugs in animals with these infections 
may prove useful for future treatment of humans. 

Proposed Course of Project : Expanded study of antibody and isolation of 
virus from women with carcinoma in situ and carcinoma of cervix in collabora- 
tion with Dr. Johnson. Serological tests utilizing neo or early herpes 
antigens will be attempted to determine if any relationship exists between 
these antigens and cancer. In collaboration with Dr. May we will serologi- 
cally screen acute and convalescence serum from Bells' palsy patients for 
IgM antibodies to CMV, herpes zoster and herpes simplex. Attempts to cul- 
ture agents from tissue and screen monolayer cultures by fluorescent anti- 
body technique for latent virus. 

In collaboration with Dr. Billingsley we will culture hysterectomy specimens 
for the presence of herpes, CMV and mycoplasma. This will help determine the 
focus of chronic infection and clinical importance in controlling infection 
to the fetus or newborn. A larger number of vasectomy specimens will also 
be cultured for the presence of virus inorder to determine if the male may 
be involved In the transmission of these viruses. 

Honors and Awards: None 



46 



Serial No. NDS {CF)-72 ID 1983 
Publications ; 

Farwr. G., Vincent. M. M.. FuccIHo. D. A.. Horta-Barbosa. L.. 

Ritman. S.. Sever, J. L., and Gitnlck. G. L.; Viral Investigations 

Ic T^^f^.J.^ Colitis and Regional Enteritis. Gastroenterology: 
65:8-18, 1973. 

Traub, R. G., Madden, D. L.. Fucclllo. D. A., and McLean. T. W.: 
The Male as a Reservoir of Infection with Cytomegalovirus, Herpes 
and Mycoplasma. New England Journal Medicine: 289: 697-698. 1973. 

Fucclllo. D. A.. Kurent, J. E., and Sever. J. L.: "Slow Virus 
Diseases", Annual Review of Microbiology (In press), vol. 28, 
1974, 



47 



Serial No. NDS (CF)-72 ID 1984 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1» 1973 through June 30, 1974 

Project Title: Maternal Infection and Pregnancy Outcome 

Previous Serial Number: Same 

Principal Investigators: Dr. David A. Fuccillo, IDB, NINDS 

Dr. John L. Sever, IDB, NINDS 

Other Investigators: Dr. William T. London, IDB, NINDS 

Mrs. Renee Traub, IDB, NINDS 
Mrs. Mary Ruth Gilkeson, IDB, NINDS 
Mrs. Flora Moder, IDB, NINDS 

Cooperating Units: Pennsylvania Hospital (Dr. Corson and Dr. Bolognese) 

Naval Medical Center, Bethesda, Md. (Dr. Billingsley 

Man Years: 

Total: 4.5 
Professional: 1.0 
Other: 3.5 

Project Description ; 

Objectives : Of the 3.5 million children born each year in the U. S. 
approximately 3% are mentally retarded. It appears that as many as 10% of 
these cases may be attributed to infectious disease. Our objective Is to 
utilize various virological techniques in an intensive study of viruses to 
determine their role in the production of birth defects and related 
abnormalities. To develop virological techniques necessary for the 
investigation of the natural course of the disease as caused by the infectious 
agents. 

Methods Employed : New virus Isolation techniques and serum neutralization 
tests are used for large scale testing in the study of pregnant women and 
their children. The development of new techniques, such as the IHA test for 
Herpes hominis Types I and II and a new test for cytomegalovirus will now 
permit the determination of the frequency of virus experience among study 
populations along with the presence of antibody and change in antibody titer 
to the virus. These tests are being used to establish the reliability of 
other tests and to deternine their sensitivity and specifdcity. A seroloqic 
study utilizing these tests on sera collected on the Collaborative Study 
population v/as conducted to deternine the frequency of antibody changes 



49 



Serial No. NDS (CF)-72 ID 1984 

during pregnancy and the effect of these infections on the developing embryo. 

The use of a fluorescent, specific anti-IgM test is proving to be a 
valuable method for identifying congenital infections. The use of affinity 
chromatography is also studied for its value in separation of IgM. 

Rubella vaccine is being given to pregnant women negative for rubella (at the 
Pennsylvania Hospital) who are subsequently being aborted. This will help 
determine the effect of vaccine on the fetus. 

Pregnant women will be studied for genital infection with herpesvirus and 
cytomegalovirus. The effects of virus on pregnancy during gestational 
period will be studied. Amniotic fluid will be taken during pregnancy to 
determine presence of virus and IgM determinations performed on the fluids 
to ascertain the presence of this globulin and its value in the early 
detection of in utero infection. Follow up on patients with herpes will be 
done for abnormal Pap smears and cancer of cervix. 

Major Findings : Previously three new tests were developed for specific 
virus serology. These were hemagglutination tests for Herpes I, Herpes II, 
and Cytomegalovirus. Recently another test for varicella was developed. 
The tests now provide highly specific, reliable, rapid, micromethods for 
virus serology with these agents. This is of particular value because 
previous methods were particularly laborious and expensive or not specific. 
Also, since these are hemagglutination tests, the anticomplementary effects 
found in many Perinatal Research Study sera can be avoided and the sera are 
now useable. In order to conserve sera for future studies a new microtest 
using .01 ml volumes were developed and recently published. 

Virus isolation studies of women for herpesvirus show transient as well as 

persistent infections. Isolation rates have been rather low in the 

population studied so a new socio-economic group is now being studied along 

with a local population to eliminate possible transportation ^problems. 

The fetal abortion studies, after rubella vaccinations, have been extended 
and results have been reported. Virus was isolated from products of con- 
ception in three patients. In collaboration with Dr. Smith, Tulane University, 
a persistent rubella virus infection of chondrocyte cell cultures was reported. 
A comprehensive review on viral teratology was published. 



Si gnii-lcance to the Prograi 
help determine what effect 



am of the Institute: The results from these studies 



virus infection has on abnormal pregnancy outcomes 
and also provide valuable information on the epidemiological aspects of virus 
infections. The relationship of these in utero infections to various degrees 
of neurological dysfunction in later years of life is still unknown. 

Proposed Course of the Project : Studies are still In progress on cytomegalo- 
vlrus infections during pregnancy and in several population groups in Maryland. 
Additional studies are being conducted on Herpes Type I and II infections in 
women. Drug evaluation of Cytosine Arabinoside for CMV and 5-IOU for Herpes 
hominis is being conducted in monkeys. Experimental drugs are also being 
tested. Approximately 16,000 sera from abnormal pregnancy outcomes are being 
tested for Herpes I and II and CMV. 50 



Serial No. NDS (CF)-72 ID 1984 



Honors and Awards ; None 
Publications: 



FucclUo, D. A., and Sever, J. L.: Viral Teratology. 
Bacteriological Reviews, vol. 37, No. 1, 19W31, 1973. 

Bolognese, R. J., Corson, S. L., Fucclllo, D. A., Sever, J. L., 
Traub, R.: Evaluation of Possible Transplacental Infection 
with Rubella Vaccination During Pregnancy. Am, J. Obst. S 
Gynecol, vol. 117: 939-941, 1973. 

Fitzgerald, S. C, Fucclllo, D. A., Moder, F., Sever, J. L.: 
Utilization of a Further Miniaturized Serological Micro- 
technique. Applied Micro. Vol. 27, 1974. 

Smith, J. L., Farley, E. M., London. W. T., Fucclllo, D. A., 

and Sever, J. L.: Persistent Rubella Virus Production In 

Embryonic Rabbit Chondrocyte Cell Cultures. Proc. Soc. Exp. 
Biol. & Med. Vol: 143. 1037-1041, 1973. 



Six 



Serial No. NDS (CF)-72 ID 1985 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1. 1973 through June 30, 1974 

Project Title: Isolation and Identification of Viral Antigens and 
Antibody for Rapid Identification of Virus Strains 
and Diagnosis of Diseases 

Previous Serial Number: Same 

Principal Investigators: Dr. David L. Madden, IDB, NINDS 

Other Investigators: Dr. David A. Fuccillo 

Dr. Luiz H. Barbosa 
Dr. Gabriel Castellano 
Mrs. Mary Krasny 
Mrs. Aurella Krezlewicz 

Cooperating Units: Microbiological Associates, Inc. 

Man Years: 



Total : 


1.50 


Professional : 


i50 


Other: 


1.00 



Project Description 

Objectives : To isolate and identify several viral antigens from each of 
several viruses. To utilize these antigens for more specific, rapid, 
sensitive identification of antibody, and/or a more accurate Identification 
of the viral agent. 

Methods Employed : High-titered virus preparations have been prepared 
using tissue culture techniques. The viral proteins have been concentrated 
by use of ammonium sulphate precipitation, protamine sulphate and strepto- 
mycin B precipitation, alcohol precipitation, polyethylene glycol and 
gradient techniques. The various classes of antigens will be separated 
and identified by use of gel filtration, polyacrylamide gel, electrophore= 
sis, and iso-electric focusing. These antigens will be inoculated into 
animals to produce specific antisera. Hiqhly nurified antinens and specific 
antibodies will be labelled with iodine 125, tridium, and chromium 51. 
Radioimmunoassay and inmune cellular release studies have been undertaken 
on measles, herpes I and sinian hemorrhaqic fever virus. 



53 



Serial Mo. MDS (CF)-72 ID 1985 

Major Finding s: Application of precipitation, gel filtration and polyacry- 
lamide gel electrophoresis indicates that the HA activity of measles can 
be concentrated. Herpes I virus has been precipitated from tissue culture 
fluid by alcohol precipitation. Simian hemorrhagic fever viruses have been 
purified and concentrated by polyethylene glycol and sucrose gradients 
Application of a sandwich-type radioimmunoassay for detection of measles 
antibody has not been successful. 

Significance of the Program to the Institute ; The development of more 
specific antigens or antibodies which measure more accurately the immuno- 
logical status of an individual is needed (e.g. in predicting the outcome 
of pregnancy following exposure to known teratogenic agents). Highly 
specific antigens or antibody may help identify the biological differences 
between nonpathogenic and pathogenic strains of these organ<sms. 

Proposed Course of the Project : Further studies will be done to identify 
the antigens associated with the measles and rubella. Herpes I, Herpes II, 
and CMV, and SHF viruses. Efforts will be made to utilize the immune 
electron microscopic technique in detection of agents associated with 
multiple sclerosis and other neurological diseases of unknown etiology. 

Honors and Awards : None 

Publications: None 



54 



Serial No. NDS (CI>-72 ID 1986 
1. Infectious Diseases Branch 
■" '■ '■'■'"' 2. Section on Experimental Pathology 

""' ' '3. Bethesda, Maryland 

' -.-'^.; PHS-NIH 

,;;. ._.....,. Individual Project Report 
■ "' July 1, 1973 through June 30, 1974 

Project Title: Intrauterine Inoculation of Fetal Monkeys With Tissues 
From Patients With Chronic Diseases and Infections 

Previous Serial Number: Same .,,^ ,■.,. ..!-.;;■. 

Principal Investigators: Dr. William T. London, IDB, NINDS 

Dr. Amos E. Palmer 
Dr. Luiz H. Barbosa, IDB, I^INDS 

Other Investigators: Dr. David A. Fuccillo, IDB, NINDS 

Dr. John L. Sever, IDB, NINDS 
Mrs. Blanche Curfman, IDB, NINDS 

Cooperating Units: None 

Man Years : 

Total 0.7 
Professional: 0.2 
Othei : 0.5 

Project Description : 

Objectives : The study of chronic diseases, such as Subacute Sclerosing 
Panencephalitis, Creutzfeldt-Jakob disease. Amyotrophic Lateral Sclerosis, 
Progressive Multifocal Leukoencephalitis, and Multiple Sclerosis following 
the inoculation intracerebrally of fetal rhesus monkeys. 

Methods Employed ; In many diseases, the fetus and young are more susceptible 
than the adult. The possibility of exploiting this greater susceptibility of 
the neonate was the basis of the design to inoculate the fetuses of pregnant 
monkeys at 100 days of gestation with CNS material taken from patients show- 
ing clinical signs of the above diseases. Animals are subsequently delivered 
by cesarean section and held in isolation chambers for 3 to 5 years. Animals 
are observed daily for abnormal signs. Sera and spinal fluid are collected 
for study every three months. 

Major Findings: The inoculated animals are now 26-38 months of age, to date, 
no abnormal signs have been observed in any of the monkeys. 

Surveys of sera and cerebrospinal fluid for elevated antibody titers to a 
battery of antigens revealed no differences between inoculated animals and 
controls. 



55: 



Serial No. NDS (CF)-72 ID 1986 

Significance to the Program of the Institute : A short time ago, the hypothesis 
that chronic or subacute degenerative diseases of the nervous system might 
be transmissible would have been an unbelievable conception to most medically 
trained pathologists or neurologists. To date, using animal models, at least 
two chronic degenerative conditions seen in humans have been transmitted. As 
these diseases become more fully understood, we may have new ideas for the 
study of other degenerative diseases of the brain and central nervous system. 

Proposed Course of the Project : Will be continued along the parameters out- 
lined above. 

Two animals from each disease group will be immunosuppressed with cyclophospha- 
mide when they are 24-30 months of age in an attempt to activate any latent 
infection. 

Honors and Awards : None 

Publications: None ' '•' ' •-'-■■'''■ 



56 



X 



Serial No. NDS (CF)-72 ID 1987 

1. Infectious Diseases Branch 

2. Section on Experimental Pathology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Caloric and Protein Restriction in Pregnancy and Their Effect 
on Measurements and Biochemistry of Newborn Rhesus Monkeys 

Previous Serial Number: Same 

Principal Investigators: Dr. William T. London, IDE, NINDS 

Dr. Amos E. Palmer, IDB, NINDS 
Dr. Donald B. Cheek, Dept. of Pediatrics, 
Royal Children's Hospital 
Victoria, Australia 3052 

Other Investigators: Dr. John L. Sever, IDB, NINDS 

Dr. Jonas Ellenberg, OB, NINDS 
Dr. John G. Bieri, NIAID 

Cooperating Units: Dr. Dominick Purpura 

Albert Einstein Medical Center 
Bronx, New York 10461 

Dr. William Nyhan 
■■. University of California - San Diego Branch 
P.O. Box 109 ' 
LaJolla, California 92037 

Dr. Ernest Bueding ' 

School of Hygiene 
. .: . Johns Hopkins Hospital 

Baltimore, Maryland 21205 

Man Years ; ■ 

Total: 1.3 
Professional: 0.3 
Other: 1.0 

Project Description : 

Objectives : Initiate a nutritional study of non-human primates using pregnant 
rhesus monkeys to test the hypothesis that there is no causal relation between 
maternal nutrition during pregnancy and certain sensory, pathological, immuno- 
logical and biochemical characteristics of the infant. It is considered that 
a monkey trial in which the concept of randomness is permitted is a necessary 
preliminary to a human trial. 



57 



X 



Serial No. NDS (CF)-72 ID 1987 

Methods Employed : In the nonhuman primate nutritional study, pregnant rhesus 
monkeys are maintained throughout pregnancy on one of three purified diets. 
One diet was restricted in protein, one deficient in both protein and calories, 
and the third was the control diet. The pregnant animals are delivered at 158 
days of gestation by cesarean section and the infant's tissues are processed 
for biochemical analysis. The nutritionally deprived female monkeys are 
continued on their respective diets and studies for immunological responses 
to various antigens such as tuberculin, rubella-mump virus vaccine, vaccinia 
virus, and standard diphtheria-pertussis-tetanus vaccine will be carried out. 

Ma j or Find ings : Twenty-four viable fetuses were delivered from monkeys main- 
tained on one of 3 diets. 

Group 1 - Control diet (8 fetuses) 

Group 2 - Protein deficient-Calorie (8 fetuses) 

sufficient diet 
Group 3 - Calorie deficient-Protein (8 fetuses) 

deficient diet 

All organs were weighed at sacrifice and anthropometric measurements taken. 
When the fetal measurements (body weight, crown- rump, crown-heel and head 
circumference) were compared, no significant differences were found between 
the fetuses from monkeys maintained on the three diets. 

Significance to the Program of the Institute : General obstetric practice in 
the United States is to restrict weight gain during pregnancy except where 
specifically contraindicated. Investigators analyzing the maternal weight 
gain/birthweight data from the Perinatal Research Program have concluded that 
this practice may be actively increasing the low birthweight rate, and possibly 
the perinatal mortality rate. Indeed, this practice may produce babies who 
are smaller than expected for gestation and who may be poor performers post- 
natally and of lower intelligence. 

Proposed Course of the Project : Tissue collected from the 24 fetuses will be 
processed by Dr. Cheek's group using various biochemical assays. Dr. Purpura 
will examine histological sections of fetal brains to determine if any patho- 
logical lesions are present. All findings will then be reported. The 
nutritionally deprived female monkeys will be tested for immunological 
responses to various antigens outlined above. 

Honors and Awards : None 

Publications: None 



58 X 



Serial No. NDS (CF)-72 ID 1988 

1. Infectious Diseases Branch 

2. Section on Experimental Pathology 

3. Bethesda, Maryland 

: .-i^: PHS-NIH 

Individual Project Report 
July 1, 1973 through June 30, 197A 

Project Title: Herpesvirus Induction of Cervical Cancer in Cebus Monkeys 

Previous Serial Number: 

Principal Investigations: Dr. Amos Palmer, IDE, NINDS 

Dr. William T. London, IDE, NINDS 
Dr. Andre J. Nahmias 

Dept. of Pediatrics, Emory University 
•■;..:. r%.;.- ,;, School of Medicine, Atlanta, Georgia 
;.;..: ■ Dr. John L. Sever, IDE, NINDS 

Other Investigators: Dr. David A. Fuccillo, IDE, NINDS 

Mrs. Renee Traub, IDE, NINDS 

Cooperating Units: Dr. John Vema 

Meloy Laboratories 
Springfield, Virginia 

Man Years: 

Total: 1.0 

Professional: 0.5 

Other: 0.5 

Project Description : ■■. , ■•.•..•...; ■.•.■. 1-L 

Objectives : Several studies reported in the literature have indicated that 
herpes simplex virus (HSV) type II may be etiologically related to hviman 
cervical cancer. One important way to establish causal relationship between 
genital herpes infection and cervical cancer would be to determine whether 
cervical cancer can be produced in monkeys that are infected by genital inocula- 
tion with herpes simplex type II virus. 

Methods Employed : We have shown that of several monkey species in which genital 
infection with HSV type II was attempted, only the cebus monkey was susceptible. 
The infection in the cebus monkey closely resembled the infection in the human. 
Since single infections may not cause cellular transformation and progressive 
neoplasia, we are in the process of repeatedly inoculating 225 female cebus 
monkeys genitally with HSV type II and 75 female animals with noninfected tissue 
culture material. 

Major Findings : A primary genital infection can be established in female 
cebus monkeys with the development of genital lesions in approximately half 
of the inoculated animals. The lesions are typically herpetic and develop 



59 



Serial No. NDS (CF)-72 ID 1988 

on the vulva and the cervix, without neurological or visceral complications, 
permitting follow-up studies for the development of cervical neoplasia. As 
in humans, spontaneous recurrence with virus shedding and genital lesions 
occur. Reinfection of previously infected animals is more difficult than with 
the primary infection. Venereal transmission from infected female monkeys to 
males has been observed. Several animals have shown cytological abnormalities 
consistent with early developmental carcinoma. However, these findings must 
persist and progress to be of significance. 

Significance to the Program of the Institute : The possible role of genital 
herpesvirus type II infection in fetal and neonatal diseases, cervical cancer, 
and chronic neurological disease stimulated this research in nonhuman primates. 

Proposed Course of the Project : Persistent reinfection of female monkeys at 
6-month intervals will continue with virological, cytological, serological and 
clinical evaluation of all animals immediately following each reinfection 
attempt. Colposcopic evaluation of the cytologically abnormal animals will 
be periodically performed to follow the progression or remission of the condi- 
tion with the colposcope. 

Honors and Awards : None 

Publications : 

Nahmias, A.J. , London, W.T., Catalano, L.W. , Fuccillo, D. A., and 
Sever, J.L. Genital Herpesvirus Hominis type 2 Infection: An 
Experimental Model in Cebus Monkeys^ Science, Vol. 171, 1970 , pp. 
297-298. 

London, W.T., Catalano, L.W. , Jr, Nahmias, A.J., Fuccillo, D.A. , 
and Sever, J.L. Genital Herpesvirus Hominis type 2 Infection in 
Monkeys . Obst. & Gyn. Vol. 37, No. 4, 1971, 501-509. 

London, W.T. , Nahmias, A.J., Niab, Z.M., Fuccillo, D.A., et al. : A 
Non-human Primate Model for the Study of the Cervical Oncogenic 
Potential of Herpes Simplex Virus type 2 . To be published in Cancer 
Research, May 1974. 



60 



Serial No. NDS(CF)-72 ID 1989 

1. Infectious Diseases Branch 

2. Section on Experimental Pathology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Transmission of Hepatitis B Virus to Sub-Human Primates 

Previous Serial Number: New 

Principal Investigators: Dr. William T. London, IDB, NINDS 

Dr. Robert H. Purcell, IDB, NIAID 

Other Investigators: Mrs. Blanche Curfman, IDB, NINDS 

Cooperating Units: None 

Man Years: 

Total: 1.2 

Professional: 0.2 

Other: 1.0 

Project Description : 

Objectives : The study and characterization of hepatitis B Virus in Rhesus 
monkeys , 

Methods Employed : An infectious pool of rhesus monkey-adapted, hepatitis B 
virus (HBV) is inoculated into rhesus monkeys in order to do biological 
characterizations of the agent. 

Major Findings : Hepatitis B viruses were not successfully transmitted to 
laboratory animals until 1972, when we successfully infected rhesus monkeys, 
and Maynard, et al, and Barker, et al, successfully infected chimpanzees. The 
success of these transmission experiments was due in large part to the develop- 
ment of very sensitive tests for detection of hepatitis B antigen (HB Ag) and 
hepatitis B antibody (HB Ab). Hepatitis B infection of Rhesus monkeys is 
entirely inapparent: infection is detected only by the development of a tran- 
sient HB antigenemia followed, in some cases, by the development of HB Ab. 
Rhesus monkeys are quite resistant to HBV infection unless a strain of virus 
adapted to growth in rhesus monkeys is used. Although less susceptible to HBV 
infection, rhesus monkeys provide a useful alternative to chimpanzees for 
experiments designed to characterize the virus. Thus, a combination of studies 
carried out in chimpanzees and rhesus monkeys provides a useful (and at present 
the only) means of characterizing the hepatitis B virus and the illness it 
causes. 



61 



Serial No, NDS(CF)-72 ID 1989 

Recently, our efforts have concentrated on the biological, biophysical, and 
immunological characterization of HBV, using rhesus monkeys as an indicator 
system for detecting the presence of the virus. 

In addition, we have continued to search for a species of non-human primate 
that will combine the advantages of chimpanzees and rhesus monkeys (i.e., high 
degree of susceptibility, development of hepatitis following inoculation, 
small size and ready availability). To date, only woolly monkeys have been 
found to be suceptible to HBV infection. These animals fulfill all criteria 
except ease of maintenance and ready availability. Because of the time and 
expense involved, we will not continue our search for new species of susceptible 
animals, but will concentrate our efforts on studies in rhesus monkeys. 

Significance to the Program of the Institute : Infectious Hepatitis (Hepatitis 
A) and Serum Hepatitis (Hepatitis B) remain two of the major human viral 
diseases for which adequate control measures have not been developed. This 
is so, because, despite heroic efforts, neither virus has been isolated in 
tissue culture and, until recently, neither virus could be transmitted to 
laboratory animals in which it could be studied. Although some important 
information was obtained through the use of human volunteer studies during the 
1940 's and 1950' s, an appreciation of the potential danger of such studies, 
coupled with changing concepts of medical ethics, has greatly curtailed the 
experimental transmission of viral hepatitis in volunteers. This study devel- 
oped in part as the results of serological surveys in the perinatal research 
program. 

Proposed Course of the Project : The ultimate goal of our hepatitis studies is 
the control of hepatitis A and hepatitis B through passive or, preferably, 
active iiumunization. Passive immunization for the prevention of hepatitis A 
is an accepted medical fact. Passive immunization for the prevention of 
hepatitis B is more controversial, but studies to determine its efficacy can 
and are being carried out in human populations. However, attempts to develop 
vaccines against hepatitis A and hepatitis B must be carried out, to the 
greatest extent possible, in animals, and it is to this purpose that much of 
our effort in animal studies will be put. Rhesus monkeys will be used to 
characterize hepatitis B virus and to evaluate various means of inactivating 
the virus. These findings will be applied to an attempt to prepare a hepatitis 
B vaccine from hepatitis B antigen purified from human blood. Chimpanzees will 
be used to evaluate the efficacy and safety of such vaccines. 

Honors and Awards : None 

Publications: None 



62 X 



Serial No. NDS (CF) 72 ID 1990 

1. Infectious Diseases Branch 

2. Section on Experimental Pathology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Chronic Lead Poisoning in Rhesus Monkeys 

Previous Serial Number: Same 

Principal Investigators: Dr. William T. London, IDE, NINDS 

Dr. Bernard C. Zook, D.V.M. 
Assistant Prof, of Pathology 
Department of Pathology 

George Washington Univ. School of Medicine 
Washington, D.C. 

Other Investigators: Mrs. Blanche Curfman, IDE, NINDS 

Cooperating Units: None 

Man Years: 

Total 0.7 

Professional: 0.2 

Other: 0.5 

Project Description : .■■■-.-■ 

Objectives : Establish the Rhesus monkey as an animal model for study of 
lead poisoning as it occurs in children. 

Methods Employed ; Adult and juvenile monkeys are given finely ground leaded 
paint orally via a stomach tube three times a week. Weekly doses range from 
50-600 mg. lead/kg. of body weight. All animals have had base line hematology, 
urinalyses, urine delta-aminolevulinic acid, coproporphyrine III, serum vitamin 
Ei2 and blood lead tests. All tests are run at monthly intervals to monitor 
changes in these values. Radiographs of the right wrist have been taken each 
month of all immature monkeys. 

Major Findings : Lead-containing paints were administered orally to 8 adult, 
5 juvenile and A infant rhesus monkeys for periods of 177 to 667 days. The 
monkeys developed Burtonian lines, elevated blood lead levels, and usually 
marked anemia terminating in a moribund state. One adult and three juveniles 
survived 557 days of paint feeding wherein daily doses exceeded several hun- 
dred times the estimated daily toxic dose for children. Adult primates were 
generally the most susceptible as they had higher blood lead values and 
developed anemia and became moribund more rapidly than infants or juveniles. 



63 



Serial No. NDS (CF)-72 ID 1990 

The relatively high doses of lead required to induce signs of toxicity, the 
resistance of young rhesus, the long course and the lack of neurologic dis- 
orders were unexpected findings. The rhesus monkey does not mimic the human 
condition and therefore is not an ideal model. 

Significance to the Program of the Institute : Lead intoxication continues to 
be a serious neurological hazard among children in the urban areas. An animal 
model is needed to study the dose-s3Tnptom relationships, since sequelae often 
develop even in the absence of overt symptoms. New therapeutic measures to 
remove excessive lead from the tissues from the onset of irreversible damage 
to the central nervous system can be more easily studied in the animal model 
than in children. 

Proposed Course of the Project : This project was closed as of December 31, 
1973. 

Honors and Awards : None 

Publications : 

Experimental Lead Paint Poisoning in Rhesus monkeys (Macaca mulatta) : 

Paper I. Clinical Findings and Excretion. B. C. Zook, W. T. London, 
J. L. Sever, and R. M. Sauer. 

Paper II. Clinical Pathologic Findings. W. T. London, B. C. Zook, 
J. L. Sever, and R. M. Sauer. 

Paper III. Pathologic Findings. B. C. Zook, W. T. London, J. L. Sever, 
and R. M. Sauer. 

All of the above papers are in Press, Journal Pediatric Research. 



64 X 



Serial No. NDS (CF)-72 ID 1991 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Immunologic Studies of Congenital Infections and Chronic 
Infections 

Previous Serial Number: Same 

Principal Investigators: Dr. Luiz H. Barbosa, IDB, NINDS 

Dr. David A. Fuccillo, IDB, NINDS 

Dr. William T. London, IDB, NINDS 

. Dr. John L. Sever, IDB, NINDS 

Dr. Jerome Kurent, IDB, NINDS 

Dr. Monique Dubois-Dalcq, IDB, NINDS 

Other Investigators: Mrs. Rebecca Hamilton, IDB, NINDS 

Mr. Otto Gutenson, IDB, NINDS 
.. : Miss Helen Krebs, IDB, NINDS 
Mr. Leonard Moore, IDB, NINDS 
Mrs. Anita Ley, IDB, NINDS 

Cooperating Units: Wilmington General Hospital (Dr. George Boines) 

Georgetown University (Dr. Joe Bellanti) 
University of Vermont (Dr. George Schumacher) 

Man Years: 

Total: 3.3 - 

Professional: 1.0 
Others: . ,, 2.3 

Project Description : 

Objectives: To conduct comparative study of the role of delayed hypersensi- 
tivity, humor immunity and interferon in neurotropic infections. To study 
the development of the fetal immune system. To develop immunologic 
methodology for evaluating intrauterine infections and CNS infeccions. To 
investigate the immunopathology of Multiple Sclerosis and other neurologic 
diseases. To conduct serologic survey with spinal fluid and sera from 
patients with chronic diseases of the CNS. 

Methods Employed : Rhesus monkeys were inoculated in utero with various 
viruses at the first, second, and third periods of gestation. At appropriate 
intervals, the fetuses were removed and blood collected for immunological 
evaluation. Antibody levels were determined by CF tests; lymphocyte trans- 
formation was measured by uptake of tritiated thymidine in the presence of 



65 



X 



Serial No. NDS (CF)-72 ID 1991 

virus antigens; interferon production was established by Sindbis HA 
inhibition assays. Concomitantly, the pathogenesis of mumps virus was 
studied in some fetuses to determine the fetal susceptibility at various 
stages of gestation and to correlate these observations with the 
immunological data. 

Sera from patients with various chronic infections were selected for immuno- 
globulin analysis. The sera were precipitated by the ammonium sulfate method 
and concentrated ten times in saline. Then preparations were ultra- 
centrifuged in linear gradients of sucrose and 7S gamma globulins separated 
from 19S molecules. The purified immunoglobulin fractions were serologically 
evaluated against different antigens to determine antibody specificity of 
early and late immune responses which might provide clues as to the nature of 
the causative agent. In addition, studies employing Sepharose columns were 
initiated to purify IgM from the sera prior to indirect immunofluorescent 
assays with specific anti-19S fluorescein-conjugated serum in tissue cultures 
infected with different agents. Since a constant residual amount of specific 
IgM seems to be associated with persistent viral infections and since the FA 
assay is the most sensitive serological procedure, it is conceivable that 
such approach could give us leads as to whether a given agent is associated 
with chronic infectious diseases. 

Sera and spinal fluid from patients with MS were compared with specimens from 
patients with other neurological diseases using the immunodiffusion 
technique and neutralization tests. Highly concentrated measles virus 
antigens were utilized for the gel diffusion tests and the Edmonston strain 
of measles virus was employed for neutralization assays. 

Lewis' rats were immunized with UV killed SSPE isolates and prototype 
measles virus combined with Freund's adjuvant and after antibodies developed 
and delayed hypersensitivity become manifest, these animals served as serum 
and lymphocyte donors to groups of histocompatible, non-immunized animals 
which were subsequently challenged with IOLDcq neurotropic measles virus. 

Major Findings : Animal experiments indicated that great antigenic 
differences exist between the 5 SSPE isolates obtained in our laboratory. 
Some of the isolates were highly antigenic, eliciting extraordinary levels 
of HI and CF antibodies, whereas other strains proved to be poor antigens. 
Great variability was also found in their ability to induce delayed 
hypersensitivity. 

Chronological evaluation of the interferon response of the fetal rhesus 
monkey following Chikungunya virus inoculation showed that in the first 
period of gestation the fetuses are uncapable of synthesizing interferon. 
In the second and third periods good levels of interferon were obtained. 

Measles-specific IgM was detected in the sera of SSPE patients. The macro- 
globulin was separated in Sepharose column and its specificity determined by 
HI test. This finding suggested that the suppressed measles encountered in 
brains and lymph nodes of patients produces a continuous antigenic stimulus. 



66 X 



Serial No. NDS (CF)-72 ID 1991 

Neutralization test was found to be at least 16-fold more sensitive than 
both HI and CF in determining measles antibody values in CF and MS patients. 
This finding warrants the re-evaluation of rubeola titers in the CSF of MS 
patients, which, according to previous studies by HI and CF, have very low 
or trace values not encountered in the normal population. 

Significance of the Program of the Institute : The successful development of 
a simple diagnostic test for SSPE now warrants further immunological studies 
with cerebrospinal fluid from patients with other neurological diseases in 
an effort to define antibody patterns of diagnostic value. Of utmost 
importance is the use of more sensitive (qualitative and quantitative) assays 
to study the increased gamma globulin levels encountered in the spinal fluid 
of MS patients. A careful serological investigation of the type and 
specificity of the immunoglobulins present in the brain of MS patients and 
appropriately matched controls could provide important clues to the possible 
infectious nature of the disease. On the other hand,, an antibody mediated 
or autoimmune theory on the etiology of MS could also be investigated by 
virtue of purifying CSF gamma globulins followed by l'-^' labelling and 
inoculation into monkey brains. The animals would be killed at 3-hour 
intervals and autoradiography of the brain sections could conceivably show 
antibody-antigen reaction sites. Another approach would be a direct immuno- 
fluorescent assay using fluorescein-coupled CSF gamma globulin with MS brain 
tissue cultures. 

Proposed Course of the Project : Special emphasis will be given to the study 
of the fetal immune system in an effort to understand the relationship 
between the maturation of the fetus immunological defenses and its resist- 
ance to transplacental infection. We will focus in the rhesus monkey model 
system as this non-human primate is known to be susceptible to intrauterine 
viral infections followed by severe malformations of the fetus. 

Continued attempts to concentrate and identify specific gamma globulins 
from sera and spinal fluid of MS patients may prove of value in establishing 
whether or not an infectious agent is associated with MS. 

Honors and Awards: None 



Publications: 



Barbosa, L. H. , London, W. T., Hamilton, R. , and 
Buckler, C: Interferon Response of the Fetal Rhesus 
Monkey Following Viral Infection. Proc. Soc. Exp. 
Biol . Med. (in press) . 

Hamilton, R. , Horta-Barbosa, L., and Dubois-Dalcq, M. : 
SSPE Measles Virus: Study of Biological Markers. 
J. Virol. 12:632-642, Sept. 1973 



67x 



Serial No. NDS (CF)-72 ID 1991 
Publications : Con' t 

Dubois-Dalcq, M. , Barbosa, L. H. , Hamilton, R. , and 
Sever, J. L. : Comparison Between Productive and Latent 
Subacute Sclerosing Panencephalitis Infection In Vitro . 
Lab. Invest. (In press) 

Raine, C. S., Feldman, L. A., Sheppard, R. D., Barbosa, 
L. H., and Bernstein, M. B. : Subacute Sclerosing 
Panencephalitis Virus: Observations on a Neuroadapted and 
Non-Neuroadapted Strain in Organotypic CNS Cultures. 
Lab. Invest, (in press) 



68 



X 



Serial No. MDS (CF)-72 ID 1992 

1. Infectious Diseases Branch 

2. Section on Immuno chemistry and Clinical Investigations 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30^ 1974 

Project Title: High Risk Pregnancies and Perinatal Infections 

Previous Serial Number: Same '.•. . ; ■,;. 

Principal Investigators: Dr, John L. Sever, IDE, NINDS 

Dr. David A. Fuccillo, IDE, NINDS 

Dr. Luiz H. Earbosa, IDE, NINDS 

Dr. Sheldon Ko3?ones, University of Tennessee 

Other Investigators: Dr. Gabriel Castellano, MBA ... 

Cooperating Units: University of Tennessee , 

Man Years: '■, . -^ 

Total : 1 

Professional: .5 
Other .5 

Project Description ; 

Objectives : The purpose of this investigation is to study high risk 
pregnancies in relation to perinatal infections. To accomplish this, a 
special study was developed at the newto m nursery at the University of 
Tennessee. This is a high risk nursery where infants from the Memphis 
Metropolitan Area are brought for special newborn care and procedures . 
Serial blood specimens are obtained and tested for immunoglobulin levels 
as a means of identifying those children with possible intrauterine 
infection. In addition, other laboratory tests and specimens are 
obtained to further define the infections which may be present. This study 
focuses on a group of infants which is most likely to have difficulties 
in the newborn period and is most frequently involved with infections. 

Methods Employed : Serial heel prick blood specimens are obtained during 
the first weeks of life. These specimens aie tested for IgM levels using 
radial immunodiffusion techniques. Infants showing elevated IgM levels 
are then further tested using venous blood specimens, throat swabs, anal 
swabs and other tissue specimens. These latter specimens are forwarded 
to the laboratories of the Infectious Diseases Branch for study. Virus 
isolation procedures and specific antibody tests are used. The data is 
then analyzed in relation to the clinical findings in the child and 
treatment is instituted wherever appropriate. 



69 



X 



Serial Wo. KDS (CF)-T2 ID 1992 

Major Findings ; Approximately 1000 children have been studied this 
year. We find that an unusually high rate of IgM elevation in this 
population exists. Investigations are in progress to define the 
specific organisms which may be responsible for the congenital infections. 

The Significance of the Program to the Institute : In our effort to 
identify the causes of neurological and other damage to the developing 
child; this population provides the unique opportunity of studying high 
risk pregnancies and thus developing a great deal of information with a 
minimal amount of study and testing. We are focusing in on a population 
which is at high risk for congenital infections. 

Proposed Course of the Project ; The study will continue during which a 
total of 2500 patients will be studied. Combined serological and 
virus isolation techniques will be used. 

Honors and Awards; None 



Publications ; Smith, J. L., Early, Elizabeth M., London, William T., 
Fuccillo, David A., and Sever, John L.: Persistent Rubella Virus 
Production in Embryonic Rabbit Chondrocyte Cell Cultures (37^65). Proc. 
Soc. Exp. Biol, and Med., Vol. 1^3, No. h, Sept. 1973, 1037-10^^1. 



70 



X 



Serial No. NDS (CF)-72 ID 1993 

1. Infectious Diseases Branch 

2. Section on Immunochemistry and Clinical Investigations 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Amniotic Fluid, Fetal Infection, Antibody and Infection 

Previous Serial: Same 

Principal Investigators: Dr. John L. Sever, IDE, NINDS 

Other Investigators: Dr. David Madden, IDS, NINDS 

Dr. David Fuccillo, IDB, NINDS 

Cooperating Units: D. C. General Hospital, Washington, D. C. 

Man Years: 

Total: 1 

Professional: .5 
Other: .5 

Project Description : 

Objectives : This is an investigation of amniotic fluid obtained at 
therapeutic abortion or amniocentesis to determine immunoglobulin levels and 
to attempt to isolate infectious agents. We are also investigating placental 
mycoplasma infections and their effect on birthweight. 

Methods Employed : Amniotic fluid specimens were obtained at amniocentesis 
for therapeutic abortions or serial determinations of bilirubin levels. 
Specimens are immediately brought to the laboratory for IgM, IgG and IgA 
determinations using the radial diffusion method and counter current 
electrophoresis. Aliquots of the specimens were also given to our other 
laboratories for virus isolation and mycoplasma isolation. Serum specimens 
are available from the mothers for subsequent antibody determinations. 
Placental specimens will be obtained for mycoplasma, isolation and 
histological studies. 

Major Findings : A number of fetal tissue specimens have shown mycoplasma. 
Infection of the fetus with mycoplasma was found in 70% of the births. We 
will now investigate placental infection with mycoplasma to determine if this 
infection is responsible for low birthweights. 

Significance of the Program to the Institute : This is the first study of 
intrauterine immunoglobulin levels and the first attempt at isolation of 
infectious agents from the uterus. Previous studies have dealt with 
specimens obtained at term. This data is confusing both in terms of 



71 X 



Serial No. NDS (CF)-72 ID 1993 

antibody levels and the presence of microorganisms since vaginal 
contamination can and does occur. 

Proposed Course of the Project : We dre in the process of completing this 
data for reporting during this next fiscal year. 

Honors and Awards : None 

Publications: None 



72 X 



Serial No. NDS (CF)-72 ID 1994 

1. Infectious Diseases Branch 

2. Section on Immunochemistry and Clinical Investigations 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Clinical Studies of Chronic Infections of the Central 
Nervous System 

Previous Serial Number: Same 

Principal Investigators: Dr. John L. Sever, IDE, NINDS 

Dr. David A. Fuccillo, IDB, NINDS 

Dr. Jonas Ellenberg, Office of Biometry, NINDS 

Other Investigators: Dr. J. T. Jabbour, Memphis, Tennessee 

Dr. George Schumacher, Vermont 

Dr. Jacob Brody, EB, NINDS 

Dr. Milton Alter, Minneapolis, Minn. 

Dr. John Kurtzke, VA Hospital, Washington, D.C. 

.. Dr. Luiz H. Barbosa, IDB, NINDS 
Miss Helen Krebs, R.N., IDB, NINDS 



Cooperating Units: 



University of Tennessee 

Veterans Administration Hospital, Washington, D.C, 

University of Vermont 

University of Minnesota 



Man Years: . :;; ^■■-.^^■■- ■ ■-■ ' 

Total: 2 

Professional: 1 - > 

Other: . . ,., .1 , ^/ 

Project Description : 

Objectives : Clinical studies are being conducted on chronic infections of 
the central nervous system. During this year, the investigations have 
primarily centered on subacute sclerosing panencephalitis, multiple sclerosis 
and amyotrophic lateral sclerosis. These studies have epidemiological and 
therapeutic components. They involve collaboration of a number of groups 
through the United States. , ■ , 

Methods Employed: A registry for subacute sclerosing panencephalitis was 
initiated as a joint effort with Dr. J. T. Jabbour at the University of 
Tennessee. This registry now has data for more than 350 cases of SSPE. The 
reporting of patients is being continued to determine if there will be a 
change in the number of cases per year related to the widespread use of 
measles vaccine. This investigation has provided us with opportunities to 



73 X 



Serial No. NDS (CF)-72 ID 1994 

study unusual patients such as one individual with hypogammaglobulinemia, as 
well as SSPE in one of two identical twins. Additiona studies have been 
conducted on the epidemiology of multiple sclerosis Serum and spina fluid 
specimens are being tested for antibody to a variety of viruses. Similar 
studies of multiple sclerosis and amyotrophic lateral sclerosis are in 
progress. 

Major Findings: Analysis of the data for 250 patients with SSPE has brought 
out a number o f new findings: D The "incubation" period between measles 
and onset of disease is 6.5 years + 2.5 years. 2) This interval is 
suprising-constant for all ages of measles ( 1 year through 6 f ^»:s)- 
3) The administration of measles vaccine after measles did not sign fcantly 
change this interval. 4) Subacute sclerosing panencephalitis (SSPE) is 
rarely seen in Blacks in this study population although most patients come 
from rural South United States. 

Patients with multiple sclerosis are being tested for cellular immunity in 
relation to measles and other viruses. 

Significan ce of the Program to the Institute : Clinical studies of SSPE, MS 
ALS and other chronic infections of the central nervous system permit direct 
investigation of the possible causes of these diseases, and provide us with 
an opportunity to study unique "experiments of nature' which often provide 
very valuable insight into the disease process. These studies are 
designed to take advantage of both the epidemiology as well as the direct 
laboratory approaches to the problems of chronic infections of the LNS. 

Proposed Cou rse of the Project : We plan to continue this SSPE registry to 
determine the effect of the widespread measles immunization program, which 
has been in effect in the United States for more than 10 years. If the 
vaccine influences the rate of SSPE, this change should occur in the next 
few years. 

Our special studies of multiple sclerosis, amyotrophic lateral sclerosis 
and other neurological diseases will utilize a combined laboratory approach 
with tissues as well as serum specimens. • 

Honors and Awards: None 



Publications: 



Vernon, Mina Lee, Fountain, Lisabeth, Krebs, Helen M. , 
Horta-Barbosa, L., Fuccillo, David A., and Sever, John L.: 
Birbeck Granules (Langerhans' Cell Granules) in Human 
Lymph Nodes. Amer. Jour. Clin. Pathology, Vol. 60, 
No. 6, pp. 771-779, December, 1973. 

Sever, John L.: "The Biology of Paramyxoviruses," 
Chapter 7, In Slow Virus Diseases (Editors- Wolfgang Zeman, 
Edwin H. Lennette, and Joel G. Brunson), The Williams & 
Wilkins Co., Baltimore, Md., 1974. 

74 X 



Serial No. NDS (CF)-72 ID 1994 

Sever, John L., Krebs, Helen, Ley, Anita, Barbosa, L. H., 
and Rubinstein, David: Serological Tests for Measles for the 
Diagnosis of Subacute Sclerosing Panencephalitis. 
JAMA (In Press). 



75 



Serial No. WDS (CF)-73 ID 203U 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda;, Maryland 2001^ 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, I974 

Project Title: Electron Microscope Immunoperoxidase Studies of Measles 
and SSPE Viruses 

Previous Serial Number: Same , . 

Principal Investigators: Dr. M. Dubois -Dale q_^ IDB^ NINDS 

Other Investigators: Dr. Luiz Horta-Barbosa^ IDE,, NINDS 

K. Worthington^, IDB^ NINDS 
0. Gutenson, IDB^ NINDS 

Man Years : ■ : 

Total 1 . 
Professional:© .5 • ■ 

Other: 0-5 . "' 

Project Description : 

Objectives : To develop a specific stain for viral antigen at the 
ultrastructural level in vivo. 

To explore the differences between productive and non-productive SSPE 
infection in brain by this technique. 

Methods Employed : Newborn hamsters inoculated intracerebrally with SSPE 
strains were studied. The direct IP technique using labeled SSPE gamma 
globulin (Dubois, et al, J. Virol 12, 9^9} 1973) was applied to 
formaldehyde-fixed thick sections. These were either chopped after 
perfusion of the animal or cryocut after quick freezing of the tissue 
and then fixed. The latter procedure allowed good correlation with 
light microscopic IP or fluorescent antibody staining on paired thin cryo- 
stat sections, whereas, the former procedure resulted in easier handling 
of the sections and better ultrastioictural preservation. 

Major Findings : Under EM, the plasma membrane of the nerve cells was 
often labeled on its inner side and in areas where nucleocarjsids 
accumulate to form buds . The membrane antigen appeared to be more 
focal than in productive SSPE infection in vitro. Within the cytoplasm 
most of the tubular inclusions were stained with the label located on the 
external granular coat of the nucleocapsid. Nonstructural antigen was 
also detected in the rough ER membrane, often on the side of a tubular 



77 X 



Serial No. NDS (cT)-Y;^ ID 'ro ]h 

inclur,ion, in the cytoplasmic matrix -rnd, moro raro.ly in tho nucl'juc oT 
giant cells. It thus seoms that SGPE p;.-unma F;lobulin '^an react wi t}i 
viral protein close to their site of synthesis. The detection of 
intracellulai' viral antigen was highly speciTlc and reproduc i.ble 
in the present system. 

Gignificance of the Prograjn to the Institute: The immunopoj-oxidase 



method allows good resolution of viral antigenic sites at high 
magnifications under EIM and may be of value in studicv. of -fh<^ 
immunopathogenesis of GSPE and other chronic vi.ral infections of 
the CNS, like Multiple Sclerosis. Indeed, intracellular viral 
components can be specifically labeled and detected even when 
conventional EM studies have not revealed the presence of virus. As 
the isolation of virus from chronic brain infections in humans 
encounters so many pitfalls, this technique mighl- help to identU'y 
viral antigen in those tissues . 

Proposed Course of the Project: The- acute hamster e n co ph;iJ. i t is 
induced by GSPE measles virus wllJ. b(} compared to the chronic SSl'E 
like disease occurring sometimes in weanling hamsl.ers using Uje 
immunoper oxidase technique. When properly fixed, SSPE human material, 
from biopsy or early autoj^sy will also be studj.ed wi tli tliis melhod. 

Honors and Awards : None 

Publication s: 

M. Dubois-Dalcq, L. TI . Barbosa, D. Fuccillo, and M. Shelanski. Immuno- 
peroxidase stain of measles antigen in tissue culture. Abstract 73rd 
annual meeting of the American Society of Microbiology. Mi;uni , May 
1973, p. PhP^' 

M. Dubois-Dalcq, and L. H. Barbosa. Lmmunoperoxidase stain of measles 
antigen in tissue culture. J. of Virology, 12, 909-918, 1973. 

M. Dubois-Dalcq, L. Horta-Barbosa, and J. J,. Sever. F>1 and Emmuno- 
peroxidase studies of productive and latent SSPE vims Infection in 
vitro. Abstract, H9th annuaj. meeting of the American Associate of 
Neuropathologists, Behamas, June, 1973, P- 3^- 

M. Dubois-Dalcq, L. H. Barbosa, R. Hnmilton, and J. L. Sever. Compari- 
son between productive .-md lai.ent suba'^ul.o sclercising panencephalitis 
viral infection in vitro . In press in Jiab. Invest. 197't-. 

M. Dubois-Dalct}, J. H. Coblentr., A. B. Pi.ont. S.uiiacutc S*- I.efo:-, i n^ P-i.n- 
encephalitis . A case study and evaln.-i.ti oti of p.-itliogf'nes Is at the ultra- 
structural levf'l. Submitted to Archives of Neurology, l'-)7H . 



78 



X 



Serial No. NDS (CF)-73 ID 203^ 

M. Dubois-Dalcq., K. Worthington, 0. Gutenson^ and L. H. Barbosa: 
Immiinoperoxidase labelling of subacute sclerosing panencephalitis virus 
antigen in hamster acute encephalitis. Abstract. Submitted for the 50th 
annual meeting of the American Association of Neuropathology. Boston, 
June, 19 7U. 



79 



Serial No. NDS (CR)-73 ID 2035 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 200lU 

PHS-mH 
Individual Project Report 
July 1, 1973 through June 30, 19TU 

Project Title: Electron Microscopic Studies of Multiple Sclerosis 

Previous Serial Number: Same 

Principal Investigators: Dr. M. Dubois -Dalcq, IDE;, NIWDS 

Other Investigators: Dr. G. Schumacher, Vermont University, Burlington, Vt. 

Dr. M. Morariu, Neurology Service, VA Hospital, 

Minneapolis, Minnesota 55^17 • 
K. Worthington, IDE, NINDS 



Man years; 



Total: .8 
Professional: .h 
Other : ,h 



Project Description : 

Objectives : To search for viral material under EM in chronic cases of 
Multiple Sclerosis. 

Methods Employed : Two hours after the patient's death, slices of brain 
from plaque areas were fixed in gluteraldehyde and processed for EM. 

Significance of the Program to the Institute : Multiple Sclerosis is one 
of the most widespread neurological diseases among young people in this 
country and its etiology is still unknown. Recent reports have suggested 
that MS may be associated with a viral infection. This may induce an 
autoimmune demyelination process. 

Major Findings: The study of these chronic cases was focused on active 
lesions with cellular infiltrates and on shadow plaques. In the first, 
nuclear strands were found in some mononuclear cells as previously described 
in Acute M.S. (Lancet II, 1^08, 1973); in the second, different abnormal 
structures were found: Hirano Body, cytoplasmic tubulo-reticular structures. 
So far, none of these structures has been found to be specific for 
measles virus antigen using the EM immunoperoxidase technique, but labeling 
of gamma globulin with the same method have revealed the fine stmctural 
distribution of these antibodies present in the active lesions. 

Proposed Course of the Project : Further brain specimens will be studied 

if other patients die of the disease in Burlington or Indianapolis Neurology 



81 



X 



Serial No. EDS (CR)-T3 ID 2035 



Departments. This material is rare and precious. Other EM studies are 
thus necessary to confirm or disprove the presence of virus-like material 
and to better understand the immunopathogenesis of the demyelinating 
areas (plaques). Also^ the immunoperoxidase technique would help in the 
identification of a possible infectious agent by the use of different 
antisera against different viruses. 

Honors and Awards : None 

Publications : M. Dubois-Dalcq, C.S. Raine, G. A. Schumacher, and J. L. Sever. 
Ulstrastucture of Central Nervous System Plaques from a case of acute 
multiple sclerosis. Abstract, i|-9th Annual Meeting of the American 
Association of Neuropathologists. Bahamas, June 1973' P* 59- 

M. Dubois-Dalcq, G. Schumacher, and J. L. Sever. Acute Multiple Sclerosis. 
Electron Microscopic Evidence for and Against a Viral Agent in the Plaques. 
Lancet 11^ itoS, 1973- 



82 X 



Serial Wo. NDS (CF)-73 ID 2036 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 
3- Bethesda, Maryland 

;■•'■';■ ■ PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^^- 

Project Title: Iinmunopathology of Chronic Neurologic Infections 

Previous Serial Number: None 

Principal Investigators: Dr. Luiz H. Barbosa 
"."""' Dr. Jerome E. Kurent 

'■'"'"■■■ ■'■ '■"".."■■■ Dr. Monique Dubois 

Other Investigators: Mr. Otto Gutenson ■'.' ' ,''^-'' ' ' ; ' 

■ ;. Mr. Leonard Moore •■/'■_■ ''•;"" ' ' ' ' ' 

Cooperating Units: None 

Man Years: ■ '^ >'''■:■■'■" ■- ■''"'■/'■■■■ ■■'•■' '\ '.'"'''.-. ".'"'' 

: •"' ■ '■'■ Total: -"■•-■■■■.■,: 0.9' ['■ ■■ y^;^;'-- ^■\';^"^- ■'""'• '•_; ' ■■■■■■•. 

Professional: O.U ' ■■■ ■■'^' ',_'''''•■'■■/' ''"."' ' ' '. 

Other: 0.5; ■■'■■•' '^ •■■■'■;; ''' '\:''''' '"' ''•''■-'":_" 

Project Description : ' '--'•- • ■ - .-..■'.! ■.■^. ■■..■■■'■; ■.,..!■ ^. .:.^ ■..■•,.. . •, 

Objective : To study the immunopathology of SSPE-like illness in hamsters. 
To establish the role of antibody and/ or cellular immunity in the 
development of chronic neurotropic measles infection in the newborn 
hamsters. To evaluate the use of different chemotherapic agents on the 
treatment of SSPE-like syndrome of the hamster. 

Methods Employed ; Recent studies indicate that newborn hamsters with 
matemal antibody survive a lethal dose of HNT (Hamster Neurotropic 
measles virus) and, eventually a certain percentage of the animals 
develop a latent brain infection which resembles SSPE in man. The 
same phenomenon occurs with weaning hamsters, and it has been suggested 
that the presence of antibody associated with an immature cellular immunity 
is crucial for the production of the SSPE-like syndrome. This study 
proposes to manipulate these two components of the immune system in order 
to establish the roles of antibody and delayed hypersensitivity in the 
hamster model system. 

Hyperimmunized animals served as donors of serum and splenocytes. Litters 
were divided into k groups of 5 litters each: 

a) Virus alone 

b) Virus and antibody 

c) Virus and splenocytes 
d) Virus and antibody and splenocytes 



83 



X 



Serial Wo. NDS (CF)-T3 ID 2036 

Conclusions will be drawn after a careful follow-up of all animals during 
wbich will be critical to distinguish between survivors with no evidence 
of virus and survivors undergoing suppressed measles brain infection 
(SSPE-like illness). This distinction must be done through immunofluores- 
cence, HSiE staining, EM and cocultivation. 

The hamster model system is intended to be used to evaluate the effect of 
anti-viral drugs such as Ara-A, Ara-C, lUDR, and others. 

Major Findings: Passive immunization protected 3 out of 10 animals and 
delayed the death of 7 out of 10 hamsters . Splenocytes alone protected 
2/10 and prolonged the course of the disease in 8/IO. The combination 
antibody + splenocytes appeared to be more effective protecting 5/IO 
hamsters and delaying death of 5/lO- Survivors are presently under 
observation for clinical signs and antibody pattern. Based on results 
obtained with this pilot experiment, we will design further protocols 
to conduct a thorough analysis of the hamster SSPE-like model system. 

Significance of the Program to the Institute : The understanding of the 
InvolArement of the immune system in the development of the newborn hamster 
SSPE-like illness is of critical importance. The application of this type 
of analysis will provide valuable information on the immunopathogenesis of 
chronic viral infections of the CWB and possible means of prevention and/or 
control of the disease in animals. These new insights into i±ie causes of 
damage to the CWS will provide guidelines for new experimental therapeutic 
approaches for the human counterpart of the disease. 

Proposed Course of Project : The combined immunologic and virologic study 
will continue for at least one more year during which we plan to evaluate 
our methodology and specific testing. Once they prove satisfactory a larger 
number of animals will be utilized in the experiments in order to give us 
solid statistical interpretation of results. 

Honors and Awards : None ....... 

Publications: None 



84 X 



Serial No. NDS (CF)-73 ID 2037 

1. Infectious Diseases Branch 

2. Section on Experimental Pathology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Perinatal Carcinogenesis in Erythrocebus Patas Monkeys 
Previous Serial Number: None ■ , . 

Principal Investigators: Dr. William T. London, IDB, NINDS 

Dr. Amos E. Palmer, IDB, NINDS 

■■">■■•;■< . • i • ■ •., Dr. Jerry Rice, EPB, NCI 

Other Investigators: Mrs. Blanche Curfman, IDB, NINDS 

Cooperating Units: Mr. William Rickman, Corbel Laboratories 

Rockville, Maryland 

Man Years: 

Total: 0.8 '.['...':: „ "\." ','■".■- ,' . . 
Professional: 0.3 ,,,,.... 

Other: 0.5 

Project Description : ■. 

Objectives : To establish a non-human primate model for chemical transplacental- 
perinatal Carcinogenesis. Then use this model to study the greater sensitivity 
of fetal and neonatal animal tissues to chemical carcinogens, as well as the 
roles of factors other than the carcinogens themselves in enhancing or suppres- 
sing chemical carcinogenesis in experimental animals during intrauterine life 
and infancy. 

Methods Employed : Pregnant patas monkeys will receive single or repeated 
injections of 0.1 mmoles/kg of 1-ethyl-l-nitrosourea (ENU) , either intravenously 
in ph3 isotonic citrate buffer or intraperitoneally in trioctanoin. The 
compound will be administered after the 50th day of gestation to reduce chance 
of abortion. 

Major Findings : Although importation of breeding monkeys has been slower than 
originally anticipated, the nucleus colony has been sufficient for preliminary 
studies of dosage. 

Transplacental carcinogenic studies have shown that pregnant females tolerate, 
without interruption of pregnancy, up to 6 successive intravenous injections 
of ENU, each of 0.1 mg ENU/kg body weight, for a total weight-adjusted dose 
which is at least 10 times that necessary for a 100 percent incidence of nerve 
tumors in rats. However, no tumors of any kind have yet been observed in any 



85; 



Serial No. NDS (CR)-73 ID 2037 

of 28 monkeys exposed to ENU in utero and observed up to 18 months after 
birth, or necropsied 1 to 14 months after birth. 

Significance to the Program of the Institute : Cancer is the third most 
frequent cause of death among infants and children in the United States 
and most industrialized countries, taking a lesser toll than either accidents 
or infectious diseases, but claiming nearly as many lives as the latter. Many 
of the specific varieties of cancer encountered in children are found uniquely 
or predominately in this age group and not infrequently develop so early in 
life that a prenatal origin is either certain or highly probable. With the 
recognition of the association between exposure to diethylstilbestrol in utero 
and the development of vaginal adenocarcinoma during the second decade of 
life, the possible significance to human health of exposure to transplacental 
chemical carcinogens has become increasingly a matter of concern. Experimental 
studies on transplacental chemical carcinogenesis have been limited to rodent 
species which differ significantly from man in many ways likely to be of 
importance for understanding the different consequences of exposure to chemical 
carcinogens during fetal versus adult life. Among these are the very much more 
rapid rates of fetal and neonatal development and maturation in rodents, a 
factor which may be responsible for the failure so far to observe any tumors 
strikingly different from those inducible in adults in rodents exposed in 
utero to potent chemical carcinogens. This project will provide empirical 
data indicating whether adult or pediatric types of tumors are more readily 
induced transplacentally in primates. 

Proposed Course of the Project : Will continue as outlined above. This project 
is an exploratory program to demonstrate transplacental chemical carcinogenesis 
in a subhuman primate species. Particular emphasis will be placed on defining 
the periods of maximal sensitivity of different organ systems and demonstrating 
parallels between both the morphological and clinical behavior of corresponding 
human tumors, and the results obtained in rodents with the same carcinogen. 
Comparative pharmacologic studies of the rate and extent of transplacental 
passage and organ/tissue localization of ENU in rodent species and in patas 
monkeys will also be made. 

Honors and Awards : None 

Publications: None 



86 



X 



Serial No. NDS (CF)-73 ID 2038 

1. Infectious Diseases Branch 

2. Section on Immuno chemistry and 

Clinical Investigations 

3. Bethesda, Maryland 



PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 197^ 



Project Title: 



Combined Clinical Viral and Immunological Investigations 
of Chronic and Subacute Disorders of the Central 
Nervous System. 



Previous Serial Number: Same 



Principal Investigators: 



Dr. Jerome Kurent 
Dr. Luiz H. Barbosa 
Dr. John L. Sever 
Dr, David A. Fuccillo 



Other Investigators: 



Mrs . Anita Ley 
Mrs . Flora Moder 



Cooperating Units: 



Medical Neurology Branch;, NINDS, NIH , '■ .r. 

Dr. King Engel 
University of Tennessee, Pediatric Neurology 

Dr. J. T. Jabbour 
University of Vermont, Dept . of Neurology 

Dr. George Schumacher ■ „; ;; .. 



Man Years: .-..■■ ,•■•■. 

Total: 1.0 ■ • .• • 

Professional 0.5 ■ 

Other 0.5 '.:, ■ 

Project Description : 

Objectives : To establish the possible viral etiology of multiple 
sclerosis (MS), amyotrophic lateral sclerosis (AI^ , and Guillain-Barre 
syndrome and to determine the pathogenesis of chronic brain infection 
in the hamster as a model for subacute sclerosing panencephalitis (uSSPE) . 

Methods Employed : ■ ■ 

1) Seroepidemiology : Semm and CSF specimens from patients with 
MS, ALS, and Guillain-Barre syndrome and controls matched for age, sex, 
and race shall be studied. The presence of serum and CSF viral antibodies 
shall be determined by one of three methods depending upon the specific 
virus. These shall include indirect hemagglutination, complement 
fixation, and sertun neutralization. Antigens shall include polio virus 



87 X 



Serial No. WDS (C3i')-T3 ID 2038 

Types I, II, and IIIj varicella-zoster; herpes Type I and II; 
cytomegalovirus; mumps; rubeola; rubella; influenza A, coxsackie 
B3 and B^l-. 

2) Immunofluorescence : The i.mmuno fluorescent technique shall be 
utilized in a search for viral anti^ n in cryostat-cut brain sections 
from MS and AIB . In addition, AI£ serum shall be studied for the 
presence of activity against viral antigen in CHS tissue sections 
frcrn mice with motor neuron disease associated with C-type virus 
particles. Similarly, mouse serum activity against AI£ tissue shall 
be studied with the Immunofluore scent technique in an attempt to 
demonstrate immunological cross reaction between these two types 

of motor neuron disorders. 

3) Tissue Culture: Muscle biopsy specimens from five ALS patients 
shall be grown as explant cult\rres and observed carefully for possible 
cytopathic effects. Immunofluore scent assays shall be conducted with 
patient's serum and if indicated with pooled ALS sera and normal serum. 
Other tests to detect the presence of an unconventional virus will 
include interferon assays with supernatant fluids of the cultures, 
hemadsorption tests with human Type 0, monkey, guinea pigs, and sheep 
erythrocytes, and a test for the presence of intrinsic interference 

by superinfection of cultures with Coxsackie A9. Attempts to rescue 
defective virus will be canried out according to three basic techniques; 
1) cocultivation of the muscle cells with WI38, HeLa, and human 
spongioblast; 2) "shocking" of the muscle cultures with 5-iododeoxyuridine; 
and 3) forced fusion of monolayers vri. th lysolecithin. 

h) Experimental SSFE in Hamsters : Evaluation of the importance of 
a deficient or immature immune system in the pathogenesis of chronic viral 
encephalitis shall be undertaken utilizing a hamster model for SSPE. 
Suckling hamsters will be treated with either immune serum, immune 
splenocytes, or both, while receiving intracerebral measles virus. It is 
expected that this treatment shall render some degree of protection from 
acute fatal encephalitis, and that a proportion of protected animals 
shall develop chronic encephalitis. Depending upon which group(s) 
survives, conclusions will be drawn regarding relative importance of the 
immune system in the production of chronic viral encephalitis. 

Major Findings: These studies are currently in progress. 

Significance of the Program to the Institute; The search for -"/Iral 
etiologies of chronic and subacute human CKS disease by means of immuno- 
logic and virologic techniques is in keeping with the major objectives 
of the Infectious Diseases Branch. Implication of a virus in the etiology 
of these disorders would help provide the rationale for their prevention 
and treatment, as well as to set the stage for an understanding of the 
pathogenesis of chronic ne\n"ological disease. 



Serial No. WDS (CF)-T3 ID 2038 

PropcBed Course of the Project ; If serological investigation of MS, 
AL£>, and Guillain-Barre serum and CSF specimens reveals as association 
between a particular -virus and either of these disorders, more 
sophisticated efforts could be made to establish an etiological 
relationship. Tissue culture approaches utilizing conventional techniques 
as well as explant cultures of diseased neural tissues would be 
employed. Special procedures to allow defective viral agents to 
express their infectivity would also be utilized as described with ALS 
muscle biopsies. If results of chronic CWS infection by measles vims 
in the hamster suggest a specific role for a compromised immunologic 
response in the pathogenesis of chronic viral encephalitis, attempts to 
adapt this model to a primate system would be made. Antiviral 
chemotherapy could be instituted in chronically infected animals, and 
favorable results possibly applied to the treatment of SSPE in man. 

Honors and Awards : None 

Publications: None 



89 



X 



Serial No. HDS (CF)-73 ID 2039 

1. Infectious Diseases Branch 

2. Section on Immuno chemistry and 

Clinical Investigations 
3- Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Blighted Potatoes and Birth Defects 

Previous Serial Numher: Same 

Principal Investigators: Dr. Jerome Kurent, IDE, NINDS 

Dr. John L. Sever, IDE, NINDS 
Dr. William T. London, IDE, NINDS 

Other Investigators: Dr. Ken Deahl 

Cooperating Units: Dr. Steve Sinden • • . ■"'■'', 

Dr. Ramon Webb 

Plant Genetics and Germ Plasm Institute 
Vegetable Laboratoiy 

Range 1 Agricultural Research Center West 
Beltsville, Md. 20705 



Man Years : 

Total: 0.2 
Professional: 0.1 
Other: 0.1 

Project Description : 

Objectives: To determine if any relationship exists between maternal 
contact with blighted potatoes and the production of anencephaly and 
spina bifida. 

Methods Employed: The approach to this problem is two-fold: 

(1) Animal feeding studies utilizing blighted potatoes (potatoes infected 
with the fungus of late blight, Phytophthora infestans) and swine, and 

(2) Analysis of prospectively collected case reports of anencephaly and 
spina bifida from the C&FR data. 

Major Findings : Analysis of the C&FR data for anencephaly and spina 
bifida cases suggested that the existence of anencephaly and spina bifida 
could not be attributed to a single environmental agent. More specifically, 
blighted potatoes could not be responsible for the production of 
anencephaly and spina bifida, i.e., other environmental determinants 
must be contributory if potatoes are involved at all. 



91 



Serial No. EDS (CF)-T3 ID 2039 

Animal feeding trials are still in progress. However, preliminary- 
results with both hog feeding experiments and hamster experiments 
suggest that blighted potatoes are not able to cause neural tube 
defects in the animal study. 

Significance of the Program to the Institute ; Results of this study 
shall hopefully support the hypothesized etiologic relationship between 
maternal contact with blighted potatoes and the production of severe 
birth defects. This would help to establish guidelines for potato 
avoidance in order to reduce the incidence of these defects, and would 
also establish the teratogenic potential of blighted potatoes for the 
central nervous system. 

Proposed Course of the Project : At the completion of the hog feeding 
experiment, this study will be terminated. 

Honors and Awards : None 

Publications ; Perinatal Infections and Epidemiology of Anencephaly and 
Spina Bifida. Jerome E. Kurent and John L. Sever: Teratology 8, 
p. 359, 1973. 



92 



X 



Serial No. NDS (CF)-74 ID 2067 

1. Infectious Diseases Branch 

2. Section on Immunochemistry and Clinical Investigations 

3. Bethesda, Maryland 



'■'--- - PHS-NIH 

Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The Antenatal Diagnosis of Anencephaly and Spina Bifida by 
Maternal Serum Alpha Fetoprotein 

Previous Serial Number: None .-,,.,..■ - 

Principal Investigators: Dr. Jerome E. Kurent, IDB, NINDS 

Dr. John L. Sever, IDB, NINDS 

Other Investigators: Mrs. Dorothy Edmonds, IDB, NINDS 

Mrs. Mary Ruth Gilkeson, IDB, NINDS 

Cooperating Units: Dr. Thomas Waldmann, NCI 

Man Years: 



Total: 


0.3 


Professional : 


0.2 


Other: 


0.1 


Project Description: 





Objectives : To establish the value of maternal serum alpha fetoprotein (AFP) 
determinations in the antenatal diagnosis of anencephaly and spina bifida. 

Methods Employed : Maternal serum AFP determinations of pregnancies resulting 
in anencephaly and spina bifida fetuses will be studied over the gestational 
period 12-20 weeks. Serum levels will be compared with pregnant women 
matched for gestational age and race. AFP determinations will be performed 
by the double antibody radioimmunoassay. 

Major Findings : Results of a pilot study indicate that maternal serum 
obtained from anencephalic and spina bifida pregnancies have significantly 
elevated AFP concentrations in the majority of cases studied. 

Significance of the Program to the Institute : If the clinical usefulness of 
maternal serum AFP determinations is established as a practical screening 
measure for the presence of neural tube defects, it will represent a valuable 
adjunct in the antenatal diagnosis of these severe congenital serological 
defects. 



93 X 



Serial No. NDS (CF)-74 ID 2067 

Proposed Course of the Project : In view of the strong association between 
elevated maternal serum AFP concentrations and neural tube defects, 
additional case studies will be performed. In addition, other abnormal 
fetal conditions shall also be evaluated, since there is evidence that AFP 
may be elevated in other unrelated fetal outcomes. The purpose of extending 
this study to include other abnormal fetal conditions in addition to neural 
tube defects would be to fully define the clinical usefulness of maternal 
serum AFP determinations during pregnancy. 

Honors and Awards : None 

Publications: None 



94 X 



Serial No. NDS (CF)-7A ID 2068 

1. Infectious Diseases Branch 

2. Section on Experimental Pathology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Simian Hemorrhagic Fever in Patas and Rhesus Monkeys 

Previous Serial Number: None 

Principal Investigators: Dr. William T. London, IDE, NINDS 

Dr. Amos E. Palmer, IDB, NINDS 
Dr. John L. Sever, IDB, NINDS 

Other Investigators: Dr. David A. Fuccillo, IDB, NINDS 

Dr. David Madden, IDB, NINDS 
Dr. Jerry Rice, EXP, NCI 

' Mrs. Blanche Cur f man, IDB, NINDS 
Man Years : 

Total: 1.5 
Professional: 0.5 
Other: 1.0 

Project Description : 

Objectives : In 1972, an epizootic of simian hemorrhagic fever (SHF) at a 
facility under contract to the Section on Experimental Pathology was responsible 
for the deaths of 212 rhesus monkeys. The SHF virus was found in the blood 
serum of clinically normal patas monkeys held at the same facility. Approx- 
imately 10 months later, an epizootic of a chronic disease began to effect 
patas monkeys in the colony. The disease in patas monkeys evolved as a chronic 
disease with a prolonged course over several weeks to several months. Some 
animals recover, others seem to recover, then relapse and die. 

The disease In the patas monkey is characterized by fever, depression, sloughing 
footpads, anorexia, dehydration, diarrhea, and persistent proteinuria. In late 
stages there are often pathological bleeding tendencies which result in the 
animal's death. 

Experimental infection of patas monkeys results in a transient disease which 
lasts 5 to 10 days, with the animal undergoing complete recovery, but maintain- 
ing a persistent viremia. 

There are at least 2 strains of SHF virus. One strain (LVR) was isolated in 
cell culture from epizootics in rhesus monkeys at the NIH and at Sukhumi, USSR, 
in 1964. Since that time there have been at least 3 epizootics from which the 
virus has not been cultivated outside the host animals (this includes the recent 
epizootics in rhesus and patas monkeys described above) . 



95 



Serial No. NDS (CF)-74 ID 2068 

This disease presents an infectious disease model in which resembles 1) immune 
complex diseases (such as lymphocytic choriomenengitis (LCM) , 2) disseminated 
intravascular coagulopathy (DIG), and 3) the hemorrhagic fevers of man. We 
intend to study SHF in patas and rhesus monkeys in an attempt to better char- 
acterize its pathogenesis, prevention and treatment. 

Methods Employed : There is presently no procedure available to detect the 
carrier state in clinically normal patas monkeys other than the inoculation 
of rhesus monkeys. We are making progress toward the development of labora- 
tory diagnostic techniques for this purpose. We have attempted immunodiffusion, 
complement fixation, and radioisotope assays. 

We are also exploring the following aspects of the disease: 

1. The site (organs or tissues) responsible for the replication 
of the virus in the chronically infected patas monkey. 

2. Determine whether there is a virus-antibody complex in the 
serum of infected patas monkeys and, if so, whether this 
complex is responsible for the disease. 

3. Continue attempts to cultivate the causative agent on cell 
or tissue culture. 

A. To evaluate immunosuppressive agents, corticosteroids, and 
anticoagulants as therapeutic agents to prevent or reverse 
the course of the disease. 

5. Attempt to develop and test inactivated antigens as immunizing 
agents to protect animals against a challenge of virulent virus. 

Major Findings : SHF is readily transmitted to rhesus monkeys from infected 
animals by direct contact, fomites, aerosol or parenteral inoculation. The 
disease in the rhesus monkey is invariably fatal. In patas monkeys, the 
disease can be transmitted by direct contact and parenteral administration, 
however, aerosol transmission is suspected but has not yet been conclusively 
demonstrated. Experimentally infected patas monkeys have not died from the 
disease. 

Naturally infected patas monkeys develop persistent proteinuria, abnormal 
hematological values and elevation of several serum chemistry values. The 
animals may undergo recovery or may appear to recover and relapse with progres- 
sively more severe clinical signs which progress to death. There is often 
severe pathological bleeding which results in the affected animal's death. 

Occasional epileptiform convulsions occur which may disappear spontaneously or 
may persist and cause the death of the animal. 

Significance of the Program to the Institute : SHF offers a potential animal 
model which may permit research on several diseases of significance to humans. 



96 



Serial No. NDS (CF)-74 ID 2068 

Proposed Cause of the Project : The development in vitro tests for detecting 
the carrier state of SHF in patas monkeys will be pursued vigorously. This 
test is necessary to identify all carriers and to permit progress in the related 
studies. Studies in patas monkeys will include virus cultivation attempts in 
cell cultures, attenpts to isolate and identify the virus-antibody complex and 
to demonstrate its presence in affected organs of infected animals, evaluation 
of immunosuppressants, corticosteroids and anticoagulants as therapeutic agents 
and evaluation of the convulsions as to their cause and treatment. 

Honors and Awards : None 

Publications : 

London, William T. An Outbreak of Simian Hemorrhagic Fever , 
Primate Zoonoses Surveillance Report No. 10, April-June 1972, 
issued February 1973. 



97 



Serial No. NDS (CF)-74 ID 2069 

1. Infectious Diseases Branch 

2. Section on Virology and Bacteriology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Cell Mediated Immunity and SSPE 

Previous Serial Number: None 

Principal Investigator: Dr. Luiz H. Barbosa, IDB, NINDS 

Other Investigators: Mrs. Rebecca Hamilton, IDB, NINDS ' ,, V 

Cooperating Units: Dr. Michael Blaese, NCI 

Man Years: 

Total: 1 
Professional: 0.5 
Other: 0.5 

Project Description: 

Objectives : To develop an in vitro test to evaluate measles delayed hyper- 
sensitivity in SSPE patients. 

Methods Employed : The Vr/60 SSPE chronically infected cell line will be 
used. This line has a characteristic CPE consisting of uniformly distributed 
small plaques. Cell-free virus has not been obtained under varying 
conditions, however, the nature of the cytopathic agent has been clearly 
defined as that of measles virus. 

Lymphocytes from SSPE patients and normal individuals will be obtained by 
the Fycoll gradient method. 

The target cells Vr/60 will be split and planted in petri dishes at a 
concentration of 2 x lO^-'^. As soon as cells attach the media will be poured 
off and cultures overlayed with lymphocyte suspensions using a lymphocyte/ 
target cell ratio of 10:1. These cultures will be placed onto a rocker 
platform and lymphocytes will be allowed to interact with Vr/60 cells for 
24 hours. After this period of time the cultures will be refed with fresh 
medium; when control Vr/60 show typical plaques all petris will be fixed and 
stained and the procedure evaluated based on plaque counts. 

Major Findings : This investigation is now in its preliminary stages. 



99 



Serial No. NDS (CF)-74 ID 2069 

Significance to the Program of the Institute : To date no reliable test for 
measuring measles delayed hypersensitivity exists. In addition, recent 
findings suggest a possible measles-specific defficiency of the cellular 
immunity in SSPE patients. The development of a plaque-reduction assay to 
evaluate measles cellular immunity will be of great value to give us a 
greater understanding of the immunopathogenesis of SSPE and thus permit new 
approaches to the prevention and treatment of the disease. 

Proposed Course of the Project : At least 10-20 SSPE cases will be studied 
in parallel with normal measles-positive and measles-negative individuals. 
This test will also be used to evaluate the cellular immune capabilities of 
MS patients since this disease is suspected to be associated with an 
aberrant measles virus infection. Finally, the assay will be utilized to 
measure the degree of measles specific delayed hypersensitivity conferred 
following experimental administration of Transfer Factor. 

Honors and Awards : None 

Publications: None 



100 



X 



:'■■■■. Serial No, KDS (CF)- 7^ ID 2070 

1. Infectious Diseases Branch 
' ■ ■ --'2. Section on Immuno chemistry and 
■"■ ' ■ Clinical Investigations 
3. Bethesda^ Maryland 

PHS-WIH 
■ ■ Individual Project Report 

. ■ ;■' " July 1, 1973 through June 30, 197^ 

Project Title: Affinity Chromatography^' .-■_-■•.;■■•■■: ■■, 

Previous Serial Number: None 

Principal Investigators: Dr. Richard S. Johnannes, IDB, NINDS 

Other Investigators: None 

Cooperating Units: None 

Man Years : 

Total: 0.25 
Professional: 0.25 
Other: 0.00 

Project Description ; 

Objectives : To obtain separation techniques using a solid matrix of 
sepharose to which a virus or its antibody could be attached. Owing 
to the fact that one member of -the antigen-antibody reaction is fixed 
the other could be selectively eluted from the column in pure form. 
This would represent a rapid and specific way of producing in pure form 
vimas or antivirus antibody. We will apply these techniques first to 
SSPE and MS. 

Methods Employed ; Separose beads will be activated with cyanogen bomide 
and then linked to the ligand (virus or antibody) via 4 to 6 Angstrome 
side arms. Varied techniques of elution will be tried in hopes that virus 
and antibody will be eluted and still retain biologic activity when removed 
from the column. 

Major Findings ; To date columns have been hampered by the relative purity 
of virus and antibody. Even so, first runs using a column associated with 
antibody did elute a protein containing faction distinct from the void 
voliime which had viral activity as measured by HI. Attempts are in progress 
to culture those same factions looking for live virus . 

Significance of the Prograin to the Institute : As many slow viral infections 
involve the central nervous system and as many viral diseases exhibit signs 
and symptoms in the central nervous system the study of these agents is of 



101 



Serial No. KDS {CF)-lk ID 2070 

great importance. This method of affinity chromatography offers real 
hope as a technique for obtaining the antigens and the globulins directed 
against them in pure form. 

Proposed Course of the Project : To continue to further investigations to 
develop a method of purifying virus via affinity chromatography. This 
would allow a method of rapid preparation of relatively pure -viral sus- 
pensions from solutions of mixed components . If this can be done with 
retention of biologic activity, efforts can be made to look at globulins ■ 
against relative pure forms of measles virus. 

Honors and Awards : Hone 

Publications: None 



102x 



Serial Wo. NDS (CF)-7^ ID 2071 

1. Infectious Diseases Branch 

2. Section on Immunochemistry 

and Clinical Investigations 

3. Bethesda, Maryland 

PHS-KIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Cerebrospinal Fluid Electrophoresis- por Diagnosis and 
Prognosis of MS, SSPE and Stroke 

Previous Serial Number: None 

Princiapl Investigators: Dr. Richard S. Johannes, IDB, NINDS 

Other Investigators: None '. ^ ■.■''.-.. . 

Cooperating Units: None ■ ^ - . ':■■.- ; • '...,• ; 

Man Years : 

Total: .75 

Professional: .75 ' ■ ,.-. . 

Other: 

Project Description ; 

Objectives: To examine isoenzymes of creati-ve phosphokinase and lactate 
dehydrogenase in cerebrospinal fluid. These offer significant potential 
for indices of central nervous system cell damage in diseases such as MS, 
SSPE, and strokes. Fresh cerebrospinal fluids will be examined as made 
available. The globulin fraction will be examined with particular attention 
to a group of possibly homogeneous globiilins accounting for the olgoclonal 
aspect. It is hoped that these proteins can be isolated and obtained in pure 
foim. 

Methods Employed : Free zone electrojhoresis will be used to examine the 
isoenzymes of creatine phosphokinase and lactate dehydrogenase. These will 
be pre concentrated by ultrafiltration. DISC Acrylamide gel electrophoresis 
will be performed using the methods of Jovin and Chrambach in order to 
systematically alter the mobility of each of the various species of ion 
involved. Radioimmunodifusion will be used in order to determine IgG, IgA, 
and I^ levels in cerebrospinal fluid. 

Major Findings : To date, only control systems have been used to examine 
LDH in CPK. The main thrust has been to obtain the proper methodology 
for preparation of the gamma globulin In acylamide gel. The problem is one 
of being able to get all of the globulins to form a thin stack in acrylamide 
prior to entering the separation gel. 



103 



Serial Wo. MDS (CF)-72 ID 2071 

Significance of the Program to the Institute : The importance of demon- 
strating the diagnostic and/or prognostic significance to enzyme levels in 
cerebrospinal fluid lies in the fact that these test are minor modifications 
of assays already present in many hospitals. Should they be found to have 
clinical meaning their application could be readily effected. The value 
of obtaining pure forms of the globulin fraction of CSF is that a family 
of proteins in this region has a close correlation with the clinical states 
of multiple sclerosis, and subacute sclerosing panencephalitis. Whether 
they represent primary or secondary pathological effects can not be known 
until they are obtained in pure form. 

Proposed Course of the Project: To continue to examine varied techniques 
of purifying and subseparating proteins which migrate in the gamma globulin 
region. It is hoped that such efforts will further understanding of the 
socalled oligoclonal aspect. Concurrently we will examine isoenzymes of 
CPK and LDH in cerebrospinal fluid in hope that these biochemical tests 
in conjunction with close inspection of the gamma globulins will result 
in useful diagnostic tests for miiltiple sclerosis and subacute sclerosing 
panencephalitis . 

Honors and A^^^fards : Wone 

Publications: None 



104 



ANNUAL REPORT 
For Period July 1, 1973 through June 30, 1974 
Office of Biometry 
Collaborative and Field Research Program, National Institute of Neurological 

Diseases and Stroke 

The Office of Biometry is a Branch within the Collaborative and Field Research 
Program, NINDS, and operates under the direction of the Associate Director, 
C&FR. 

Its program responsibilities include surveys of neurological diseases and 
communicative disorders. It is responsible for the design, pretesting, and 
accomplishment of such surveys - whether under contract, in-house, or in 
cooperation with other government agencies - to obtain disease statistics 
for NINDS program planning, and in the establishment of research priorities. 
It is establishing a current, centralized, disease statistics data repository 
in NINDS for providing disease statistics reports. 

The Office of Biometry is a central resource to NINDS for service in statistics, 
mathematics, computer technology, systems analysis and data processing. Its 
services range from brief consultations to participation as co-investigators 
on major projects. 

The Office of Biometry carries out research in mathematical statistics for 
current and anticipated use in NINDS programs. It also provides consultation 
and service to other organizations undertaking studies in the areas of NINDS 
interest. 

The Office of Biometry consists of the Office of the Chief, the Section on 
Mathematical Statistics, the Section on Experimental Statistics, the Section 
on Applied Statistics, and the Section on Systems Design and Data Processing. 

The report on the projects and services performed by the Office of Biometry 
is listed by major Program area, and followed by brief descriptions of 
Research, Miscellaneous Activities, Publications, and Future Plans. 

OFFICE OF THE DiRilCTOR, NINDS 

1. Automatic Data Processing (ADP) Evaluation 

A staff member of the Office of Biometry continues to serve as Chairman 
of the ADP Evaluation Task Force of the Institute, which includes other 
members of the Office. This is the coordinating and advisory group to the 
Branches and Laboratories which use ADP, for updating their five-year estimate 
of financial needs, •''or current inventory of equipment, and for other program 
information concerning data processing, for the HEW annual ADP report. In 
addition, the Branch provided advice to the Director's staff on matters re- 
lated to the operation, continuation or acquisition of computers, and on the 
development of an Energy Crisis reporting form, and the best meanb of process- 
ing the data 



COLLABORATIVE AND FIELD RESEARCH PROGRAM 
Office of the Associate Director, C&FR 

1. NINDS Reorganization Plan 

The Office of Biometry has continued to participate in the development 
of a comprehensive reorganization plan of the NINDS. During this reporting 
period particular emphasis was placed on: 

A. The planning process for the NINDS five-year research program. ' 
Included were the identification of the steps required in the 
development of detailed project plans and in the NINDS five-year 
research program plan, in the implementation of the research program, 
and finally, in the evaluation and review process for the research 
program. 

B. Plans for the creation of an NINDS Office of Program Planning and 
Evaluation. Included were details for the proposed structure of this 
Office, the responsibilities of the Office and its various Branches, 
and their personnel requirements. 

2. New Program in Disease Statistic s Surveys ! 

I 

The Office of Biometry, in response to recommendations by the Deputy 
Director and the Associate Director, C&FR, that the NINDS program planning 
process suffered from a lack of precise data on the incidence, prevalence, 
and the medical and economic costs of neurological diseases and communicative 
disorders, introduced a proposal for an NINDS program in disease statistics. < 

Since budgetary limitations-precluded iimediate monetary support of the 
program, OB prepared a proposal to use 1974 HEW Evaluation Plan set-aside 
funds to develop a Brain Tumor Survey and a Stroke Survey in order to plan 
and evaluate the methodologies required to obtain precise data in these i 
problem areas. The proposed budget was $275,000 in FY 74, and $iaO-f^OO in 
FY 75. 

The proposal was approved by the Director, NINDS. and was submitted to 
DHEI-' OS part of the recamendations for the NINDS Evaluation Plan. A new 
DHEH review of its total Evaluation Plan delayed any approvals for use of 
these funds to the latter part of FY 74. With a release of additional 
funds to NINDS from OMB, the Institute approved the expenditure of $250,000 
to initiate the disease statistics program. Shortly thereafter DREW approved 
the NINDS Evaluation Plan and released the reouested $275,000 for the surveys. i| 

With funds now available in the amount of $525,000 for FY 74, and with 
the lik:?lihood of additional support in FY 75 from DHEW set-aside funds and 
from NINDS, the disease statistics program was reviewed to consider new 
possibilities for expansion. After consultation with scientists w'thin and 
outside NINDS, and with recommendations of the Director and members of the 
Executive Staff, the decision was made to dpvelop proposals for FY 74 contract'! 
for surveys of brain tumors, for head and spinal cord injuries, and for ! 
multiple sclerosis. Since it is planned to involve the Nationa"* Center for i 



Health Statistics and the Bureau of the Census in the stroke surveys, and it 
was not possible to obtain the necessary exchange of funds agreements with 
these agencies in time to initiate a stroke survey in FY 74, it is now planned 
to initiate a stroke survey in FY 75. 

Since the specific methodologies to develop data in each disease area will 
be different, multi -disciplinary advisory committees will be created (as part 
of the contractual agreements) to develop the survey designs, to determine 
the appropriate definitions, to create the questionnaires, to determine the 
examinations to be performed, and to develop the plans for the pilot studies. 

An advisory committee for the stroke survey will develop the required 
protocols in advance of any contractual agreements, since the time frame will 
permit this first step. 

Additional activities in the disease statistics program are underway, and 
planned. A disease statistics repository for NINDS is being established, with 
an annotated bibliographic reference file being created to support these data- 
Activities include the collection, organization, summarization and evaluation 
(by scientists in relevant disciplines) of research, current or published, 
relevant to these disease statistics. Special reports or monographs will be 
prepared for the diseases within NINDS responsibilities for use in Institute 
program planning, and for dissemination to appropriate legislative and 
scientific bodies, and to the pubMc. 

Plans are being fonmilated to utilize foreign data sources where social- 
ized medical systems include the routine collection of medical data, in order 
to develop very precise estimates of the Incidence and prevalence of the 
diseases of interest. For example, Britain, Sweden, and Finland collect data 
that would be of considerable interest to NINDS, not only for incidence and 
prevalence, but with regard to studies of etiology as well. 

U.S. data from prepaid medical plans, such as Kaiser Permanente, the 
Health Insurance Plan (HIP) of New York, and others, can provide useful 
information, especially concerning the costs of diseases and the utilization 
of medical personnel and facilities. 

Initial inquiries indicate that the puMic health exp^irts responsible for 
ths collection and yse of the compiled social medicine data which are avail- 
atle outside the U. S., those in the U. S. who are similarly responsible for 
the prepaid health plan data, and research organizations who could carry 
oi;t contract proposals to utilize these data, are rr<ost willing to cooperate 
in these research efforts. 

Future plans also include: 

1. the development of an advisory committee to recommend priorities 
for future sur.'eys, and to plan tSieir design. 

2. the generation of survey proposals for recurrent samplings to 
permit the development of trends ir, incidence and prevalence, 
and the costs of the diseases of interest. 



3. Comprehensive Control and Information System for NINDS Contracts 

At the request of the Associate Director, C&FR, the Office of Biometry 
prepared a description of a comprehensive system for providing information 
about the NINDS research activities which use the contract method. 



c 



The major function of the system would be to provide information for 
the purposes of NINDS program planning and evaluation, and for broader informa- 
tion dissemination to advisory bodies, higher executive echelons, the Congress, 
and the public. The system would also provide data required for monitoring . 
the contracts for administrative control. * 

The computerized system would be operated by the personn«»l of the Research 
Contracts Office. Technical support for the system would be provided by the 
Office of Biometry. 

As part of this evaluation, other systems, such as CRISP, and IMPAC, were 
analyzed to determine the possibilities of their use as part of an NINDS 

system. 

Office of Biometry 

1. Fetal Growth Patterns in the Human (N.J. Obstetrical Study) 

Principal Investigators : Drs. P. Shaughnessy, DiGiacomo and McCann 

The investigators are working with a New Jersey obstetrical group to 
construct standard profiles (under several different sets of circumstances) 
for fetal growth patterns throughout pregnancy. The sampling design has 
been constructed, data collection instruments have been developed, pretesting 
is completed and data collection is now ongoing. It is expected that pre- 
liminary results based on data analysis will be available in about six months. 

2. Multivariate System 

Principal Investigators : Dr. P. Shaughnessyjir. Richter and Mr. Talbert 

In order to provide stronger support to Institute investigators in the 
area of statistical analysis, the Office cf Biometry has undertaken to develop 
a closely integrated system of multivariate statistical analysis programs. 
The development ar.d planning stages of the system have now been completed, 
as has been a feasibility study on ^-he general orifentation of the system. 
Several programs are now completed on the in-house NIH computer system as 
well as on a coroercial computer system. Although the system is still in 
its developmental stages. Office of Biometry statisticians have used it to 
good advantage on several occasions. 

3. Study of Primate Prematurity. Neonatal and Perinatal Mortality 

Principal Investigators : Drs. P. Shaughnessy and R. DiGiacomo 

This project is a continuation of the work begun while Dr. DiGiacomo was 
associated with NINDS. The study complements the Collaborative Perinatal 
Project it that one of its strengths is the accuracy of data collected on 



gestation. Extensive statistical collaboration has been provided to the 
investigator. Report of the study will be submitted for publication. 

4. The Guatemala Perinatal Study 

Principal Investigators : Drs.P. Shaughnessy and J. Mata (University 

of Washington) 

The purpose of this study is to examine fetal growth, mortality, duration 
of pregnancy, several maternal variables, infant variables, and environmental 
variables. The study yery likely will confirm the findings of the Collabora- 
tive Perinatal Project. Multiple regression analysis is the primary method- 
ological tool being applied. There are approximately 40 variables which are 
being investigated, they include various infant measurements at birth, family 
composition, maternal characteristics and pregnancy history, complications 
during the pregnanc.v, gestation, birthweight, socio-economic index, capital 
index, housing index, hygiene index, education index, and various nutritional 
measurements during pregnancy. 

Perinatal Research Branch 



1 . Minimal Brain Dy s function (MBD) Study 

Principal Investiiators : Drs. P. Nichols and T.C.Chen 

The purpose of tUs study is to describe an MBD syndrome (Phase I) 
and to identify the precursors of MBD (Phase II) in the Collaborative 
Perinatal Project. The study team has achieved the following during the 
past year: (1) Selection of MBD study cohort: an extensive list of pediatric 
conditions to be excl tiled from the 7-year file has been developed to generate 
a study cohort suitabls for MBD investigation. (2) Development of new MBD 
outcome variables: cogiltive and perceptual -motor, and learning disability 
variables in the 7-yeir data were examined for their distributions and re- 
lationships with IQ. Standardized deviation scores of these variables were 
derived to better des'ribe MBD symptoms. (3) Selection of MBD variables - 
26 variables involvincj learning disorders, behavior deviations, cognitive 
and perceptual -motor defects and neurological soft signs were selected for 
use in describir.g MBD t/pes. (4) Developing an MBD data file: 
system for the ^electel variables plus exclusions and missing 
has been derived. A d.ca file was created for Phase I study, 
methodology for variou; tasks involved in this study has also 
gated. 

2. The Mental Retardation Study 

Principal Investigatirs : Drs. S. Broman, P. Shaughnessy and P. Mi< 

The purpose of this .tudy is to identify precursors of mental retardation 
at age seven in a large : ample of children who have been followed longitudi- 
nally since the prenatal period. A time-phased research strategy has been 
laid out and the study ttam is adhering reasonably closely to the original 



A coding 
values, etc., 

(5) Statistical 
been Investi- 



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plan. Activities during the past year included: submission of a work file 
request for a pilot study, specifications and programming necessary to create 
the work file, writing of a comprehensive descriptive analysis protocol, pro- 
grarriTiing appropriate to generating the computer output for the descriptive 
analysis, the descriptive analysis (first screen^ itself, statistical method- 
ology development including multivariate computer program specifications, a 
detailed time-phasing of all planned statistical analyses, and additional 
refinement procedures with respect to the activities outlined by the general 
plan. Preliminary results will be available shortly and the study team hopes, 
by virtue of the pilot study, to further refine testable hypotheses regarding 
mental retardation. Several statistical investigations have been required 
in order to mitigate problems in data analysis methodology. In particular, 
problems in the area of missing values, control group selection, variance 
heteroscedasticity, and multivariate testing are all under study at the 
present time. 

3 . Learning Disorders Study 

Principal Investi:iators : Drs. S. Broman, P. Shaughnessy and P. Nichols 

The purpose of the Learning Disorders Study is to identify and describe 
children with learning disorders at age seven in the Collaborative Perinatal 
sample and to isolate antecedents of poor school performance among children 
of average intelligence. Progress oi. this project parallels that on the 
Mental Retardation Study since the same Office of Biometry statistician and 
research team are collaborating on the two studies. The time-phased plan 
for the Learning Disorder Study is the same, in terms of its major components, 
as the Mental Retardation Study. Thus, progress and accomplishments during 
the past year are similar to those in the area of the Mental Retardation 
Project. 

4. Cerebral Palsy Study 

Principal Investigators : Drs. K. Nelson and J. H. Ellenberg 

The Office of Biometry has made significant contribution to the analysis 
of ths cerebral palsy study. In collaboration with PRB staff, a great deal 
v^as accc:,-.piishsc: during the current fiscal year. First, all cerebral palsy 
ca^as have b-^en identi.ied, based on a uniform and currently acceptable 
medical dG-"i nitron, and a special tape is being created for possible use by 
other project areas. Second, a thorough examination of available pediatric 
variables yielded inconsistencies and errors v^hich have been accounted tor, 
and a well -documented pediatric variable list is now available for use in 
other project areas. Finally, the first phase of the data analysis of the 
cerebral palsy study is now in the form of a data processing request, and 
is being reviewed before submission for approval . 

5 . Convulsive Disorders Study 

Princ-Jpal Investigators : Drs. K. Nelson and J.H. Ellenberg 

In the initial data analysis phase, it was efficient to use similar 
approaches in the two studies. After -,he first phase data processing request 
is completed the two studies will diverge ir their goals and r> |Ui ed 

analyses. 6y 



A major portion of the convulsive disorders study and the cerebral palsy 
study activities overlapped, since the same Office of Biometry and Perinatal 
Research Branch staff were involved in both studies. 

6. Cormiunicative Disorders Study 

Principal Investigators : Dr. P. LaBenz, J. Oliver and D. Rubinstein 

The Office of Biometry participated in all aspects of the Communicative 
Disorders workgroup which culminated in the preparation of a comprehensive 
RFP. The Office of Biometry representative was the chief contributor to the 
analysis plan, a co-author of the RFP, and a support person for the merit 
review. Other Speech, Language and Hearing activities include an improvement 
of the analysis and layout of quality control data for SLH quality control 
tests, a sharpening of the definitions of three proposed SLH indices with 
respect to neurological aspects, auditory .processing, and communicative 
effectiveness, and review of specifications and computations of extensive 
summaries of 8-Year SLH data. 

7. Visual Abnormality Study 

Principal Investigators : Dr. R. Feinberg and E. Jackson 

A member of the Office of Biimetry has participated in designing the study 
plan and the detailed tabulation specifications. 

Preliminary investigations are under way to determine the extent of various 
visual problems reported, the patterns of use of the special seven-year examina- 
tions (that is, the visual screening and ophthalmology consultant's examina- 
tions), and the proper source of various types of information. The ophthal- 
mology consultant's reports are being reviewed and coded for use in the data 
analysis. 

The next phase will be an analysis of the relationship between a large 
number of demographic, obstetric, and pediatric variables and various visual 
abnormalities. The details of this study are currently being dt ;igned. 
Further studies in additional depth will be made of those variables which 
appea'^ important. 

3, 5ir thweiqht-Ges*ation S^udy 

Principe 1 Investigators : Drs. J. Hard^ and D. Mel 1 its 

The Office of Biometry has participated in the development of a proposal 
"or a litudy of prematurity based on the CPP data. 

An orderly progression of analyses has been developed to provide insight 
into the characteristics, causes, and consequences cf prematurity. 

The first phase is underway, to study the relationship between birthweight 
and gestation, and to evaluate the effect of other measures, such as placental 
weight, to determine whether their addition to the combination will more 
precisely define prematurity. 

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New indices of risk, based on combinations of these 'prematurity' variables 
win be created and tested to determine their utility in assessing the etiology 
of prematurity. 

9. "The First Year of Life", a second volume continuing the presentation 
of the basic data of the CPP . 

Principal Investigators : Brs. J, Hardy and S. Drage, E. Jackson 

A member of the Office of Biometry is participating in the preparation of 
the book. The effort includes data analysis, the procurement of computerized 
graphics, and the preparation and review of the text. 

10. Four-Year IQ Study 

Principal Investigators : Drs. S. Broman, P. Shaughnessy and P. Nichols 

The Office of Biometry analyzed some of the discriminant function results 
which were obtained for the Four-Year IQ Study and which are currently in 
manuscript form. There were some questions of interpretation attributable 
to misclassification rates with varying group proportions. Consultation was 
provided in the area of theoretical statistical considerations and computer 
applications. 

11 . Birthweight, Infant Development, and Pre-school IQ Study 

Principal Investigator : Dr. S. Broman 

The Office of Biometry provided technical support in both statistical 
theory and computer program development. The basic statistical techniques 
used have involved one-way analysis of variance methods with multiple com- 
parison procedures to test for pair-wise mean differences within birthweight 
groups on IQ and other infant test measures. 

12. Study of Familial Resemblance between Sibs and Twins on Bayley Scores 

Principal Inv es tigator : Dr. P. Nichols 

Methodological assistance was provided in contrasting the relation 
between sibs and twins in terms of 8-mcnth Bayley mental and motor scales. 
Statistical techniques involved the use of intra-class correlation methods, 
contingency table methods, exact conditional tests for comparing proportions, 
and approximate tests for proportions. The study results will be published 
in May, 1974. j 

1 3 . Congenital Malformations Studies 

Principal Investigetors : Drs. N.C. Myrianthopoulos, A. Naylor 



The Office of Biometry reviewed a study proposed by Dr. M.L. Liibs on 
the relationsh-'p of ABO and Rh blood groups of mothers and children on the 
outcome of malformations. As a result of this review additional and more 



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penetrating analyses were proposed and accepted. A member of the Office of 
Biometry reviewed the manuscript "Genetics of Obstetrical Variables" by 
Drs. Naylor and Warburton and suggested some constructive changes. Statistical 
reviews of other congenital malformations studies are planned for the remain- 
der of this fiscal year and the following. 

14. Microfilm Report 

The Office of Biometry updated its previous report and recoiisnendations 
for microfilming the Collaborative Project data. It has provided liaison 
with the Bureau of Standards to implement the microfilm storage of PRE files. 

Infectious Diseases Branch 

1 . Maternal Clinical Infections and Pregnancy Outcomes Study 

Principal Investigators : Dr. J.L. Sever, M. Gilkeson, D. Edmonds and AoL©y 

In the Collaborative Perinatal Research Study, clinical infections were 
reported in approximately 5,000 of 58,282 cases studied, with several blood 
specimens taken during pregnancy. The children born of these pregnancies 
have been followed for 5-8 years. Data for all of the infections are being 
studied in terms of their relation to pregnancy outcome, with the Office of 
Biometry collaborating on the design and analysis. The infected mothers 
will be matched with noninfected controls and comparisons made of rates of 
specified outcomes. Other relevant variables not accounted for in the 
matching procedure, will be evaluated. A preliminary report was made at 
the 4th International Conference on Birth Defects and will be published 
subsequently in a monograph, with a member of the Office of Biometry as a 
co-author. 

2. Analysis of Cord IgM Levels from Births in the Collaborative Perinatal 
Study 

Principal Investigators : Drs. J.L. Sever and L. Barbosa 

The general goals of this study are the examination of the association 
of cord IgM levels with specific mother (and/or child) abnormalities, and 
with etiologic characteristics of the cohort. The Office of Biometry has 
collaborated on the design and analysis of the study. It includes the 
development of a normal IgM distribution, stratified for key relevant variables 
such as sex, race and maternal age. The next step is a comparison of the 
cord IgM distributions for abnormal groups with that of the normal IgM distri- 
bution, accounting for as many related factors as sample size will allow in 
a multidimensional fashion. A preliminary report was presented at the 4th 
International Conference on Birth Defects and will be published subsequently 
in a monograph, with a member of the Office of Biometry as a co-author. 

3. Serological Study of Perinatal Infections and Pregnancy Outcomes 

Princ ipal Investigator ; Dr. J. L. Sever 

■'"his study involves the determination of antibody responsss to 1 J infec- 
tioui agents (mumps, rubella, herpesvirus, etc.) usi.ig serial sets of maternal 



sera which have been obtained prospectively from 4,000 pregnant women with 
abnormal pregnancy outcomes and a similar number of matched controls. The I 
general purpose of the study is to obtain information on the effect of 
maternal, serologically confirmed subclinical infections, o.i the fetus and 
child. The Office of Biometry is participating in the planning of the 
analysis of this data and preparation of data processing requests. Prelimin- 
ary analyses have been presented at the 4th International Congress on Mal- 
formations with a member of the Office of Biometry as co-author. The Office 
of Biometry also critically reviewed the paper "A Seroepidemiological Study 
of Epstein-Barr Virus" by Byrne, et al . This paper contained analyses 
relevant to the Serological Study. j 

4. Primate Maternal Flu Infection and Hydrocephaly in Neonates 

I 

Principal Investigators : Dr. W. T. London, Kent, Fuccillo and Palmer j 

I 
Office of Biometry staff is currently involved in the planning stage I 
of a series of animal experiments to examine the possible relationship of 
maternal flu infection with hydrocephaly in the newborn. The studies will 
be designed to demonstrate not only a strong association if it exists, but 
to generate information dealing with the rate and site of infection, as 
determinants of the degree of hydrocephaly. The primary responsibility of j 
the Office of Biometry to date, has been the design of the experiment and | 
determination of appropriate sample -jizes. I 

5 . Investigation of Incubation Period for Subacute Sclerosing Panencephalitis if 

Principal Investigator : Dr. J. Sever 1 

Subacute sclerosing panencephalitis (SSPE) is a slowly progressive brain i 
disease which is uniformly fatal. This study involves an epidemiologic 
investigation of approximately 200 SSPE patients, with information on date 
of onset of SSPE, date of measles infection, measles antibody titers and 
administration of measles vaccine. The analysis will focus on the interval 
between onset of measles and onset of SSPE and the effect on this interval 
of administration of measles vaccine after measles infection. 

6 . P eri^atal Infections and Epidemiology of Anericc-phaly anc' Spina Bifida 

f .-ir.c i :ia l Investigators : D'^s. J.E.Kurent and J. L. Sever 

The cases of anencephaly and spina bifida occiirring in the Collaborative 
Perinatal Study were analyzed for maternal and pediatric indices of intra- 
uterine infection as well as by demographic characteristics {race» sex, etc.), | 
and other available clinical data (e.g. maternal diabetes). The Office of i 
Biometry provided support in the analysis of these dcita dni reviewed the ' 
manuscript submitted for pi'blication. 



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1 ' study of Alphafetoprotein (AFP) in Maternal Serum of Anencephalic 
and Spina Bifida Pregnancies 

Principal Investigators : Drs. J.L. Sever and J. Kurent 

Office of Biometry staff are currently involved in the planning of an 
experiment to study maternal AFP levels, as a possible predictive factor of 
bad outcomes in newborns from the Collaborative Perinatal Study. The study 
is designed to demonstrate not only a strong association, if it exists, but 
also to generate (presently unavailable) normal values of AFP in pregnant 
women. Office of Biometry primary responsibility to date has been the 
design of the experiment and determination of appropriate sample sizes. 

8. Identification of Microcephalic and Hydrocephalic Children in the 
Collaborative Perinatal Project 

Principal Investigators : Drs. K. Nelson and J. L. Sever 

The basic otgectives of this study are to identify children with large 
head sizes (hydrocephaly) and those with small head sizes (microcephaly) 
based on an age adjusted definition of normal head size. The Office of 
Biometry has designed a procedure for comparing each child's head size at 
the several time points of measurement during its first seven years of life, 
to age, sex, race, institution and gestational age adjusted standards. The 
resultant identified children will be used as the cohort of microcephaly and 
hydrocephaly for the Collaborative Study Program and Dr. Sever 's work on 
clinical infection. 

9. Normal limits of Hematological Measurement of Cebus Monkeys 

Principal Investigators : Drs. W. London and A. Palmer ^^ 

The Office of Biometry is collaborating with the Infectious Diseases 
Branch in a project to determine the normal limits of various hematological 
measurements of Cebus monkeys. Office of Biometry participation involves 

(1) deternii nation of the sample size (number of monkeys) for this research, 

(2) investigation of the statistical nature of the frequency distributions 
of hem^itolt-gicai measurements, (3) isolation of the technical and biological 
variations from experimental data and determination ef the size of their 
components, (4) setting of confidence limits for the means of measured 
variables and (5) establishment of control limits for individual measurements, 
The detailed plan of the extensive statistical analysis has been developed. 

A computer program for some parts of the analysis has been prepared. The 
statistical work is expected to be completed in several months. 

10. Birthweight and IgM Level 

Principal Investigator : Dr. J. L. Spver 

This study is a part of the overall objectives of 1) assessing the 
strength of the association, if any, between maternal infection and prematur- 
ity (as defined by birthweight-gestation), 2) the determination of normal 
levels of cord IgM in term, premature and abnormal babies, 'id 3) the prog- 

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nosis of premature babies with low IgM or high IgM (e.g. lack of IgM results 
in greater susceptibility for the premature). The Office of Biometry assisted! 
both in the preparation of the data processing request for this preliminary 
investigation and in the analysis of the results. The basic data generated 
from this request will be used to design further studies to reach the primary 
objectives. 

11. Primate Maternal Nutrition Study 

Principal Investigator : Dr. W. T. London 

This project involves the study of caloric and/or protein restriction 
in pregnancy and its effect on biochemistry, and neurological deficits of 
newborn Rhesus monkeys. The second pilot study is nearing completion, with 
22 out of 24 monkeys successfully impregnated and maintained on their speci- 
fied diets. Approximately 600 measurements relating to the newborn and the 
mother will be analyzed. The Office of Biometry provided systems operation, 
data processing support, and computer graphics output. A manuscript is in 
preparation for the Journal of Nutrition, which will deal primarily with the 
many previously uninvestigated areas of research in methodology of primate 
nutrition with a member of the Office of Biometry as co-author. 

12. Herpesvirus Induction of Cervical Cancer in Cebus Monkeys 

Principal Investigator : Dr. W. T. London 

Several studies have indicated that herpesvirus (HSV) type 2 may be 
etiological ly related to human cervical cancer. This study is investigating 
the possible causal relationship between genital herpes and cervical cancer 
by attempting to induce cervical cancer in monkeys inoculated genital ly with 
the virus. The statistical design of the study was produced by the Office of 
Biometry in FY 72. Continual monitoring of the statistical aspects of the 
study design have been necessary due to the development of heretofore unknown 
conditions relating to the interaction of the HSV-2 virus in the Cebus model. 
A manuscript has been submitted for publication to the Journal of the National I 
Cancer Institute, regarding the methodological approach to the problem.with 
a member of the Office of Biometry as co-author. 

13. Primate Information Retrieval System 

The Primate Information Retrieval System (PIRS) is a fully computerized 
system for storing data on a large monkey colony, ""or updating the data file, 
and for rapidly obtaining any one cf 42 standard output reports. 

The PIRS permits the colony to be monitored closely, and the primate 
experiments to be carefully controlled, with savings in expenditure of time 
and clerical support. 

The Office of Biometry has completed the systems analysis and programming 
for the entire system. It has developed manuals and operating procedures for 
the system and has trained Infectious Disease Branch personnel in its opera- 
tion. It aideu the implementation of all phases of the operation. 

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Epidemiology Branch 

1. Epidemiological Study of Multiple Sclerosis in the Shetland and Orkney 
Islands 

Principal Investigators : Drs. D. Poskanzer and J. Brody 

David Poskanzer, M.D., Department of Neurology, Massachusetts General 
Hospital, in collaboration with the Epidemiology Branch, C&FR, developed a 
proposal for an epidemiological study of MS in the Shetland and Orkney Islands, 

At their request, members of the Office of Biometry reviewed the literature 
on previous MS epidemiological studies, and developed an extensive set of 
questionnaires for possible use in the Study. 

More than 17 forms, each related to a specific etiologic series of quest- 
ions, were created. Forms included a demographic control form, a current 
environment form, forms pertinent to pet and animal exposures, current and 
past occupations, diet and foods, personal information, previous residences, 
etc. 

The questionnaires were intended to be all-inclusive to be pared and con- 
densed according to the data priorities developed by the principal investiga- 
tor. 

Dr. Poskanzer may use OB for portions of the data processing and data 
analysis. 

2. Computer Applications 

The Office of Biometry provided consultation on computer applications and 
carried out data processing for a variety of studies conducted by members of 
the Epidemiology Branch, including the: 

1. V.A, Ulcer Study 

2. Polymyositis, Derma toryostatis Study 

3. PHS Guam Construction Workers Study 

4. Guaii; ALS-PD Study, Control Pairs 

5. Guam ALS Clinical Study 

6. Sub Acute Sclerosising, Pan Encephalitis Study 

7. Gt'am Control, file creation and lists 

Office of the Associate Director for Extramural Programs 

1 . Otolaryngology Survey 

A member of the Office of Biometry served as a member of the Manpower 
Committee of the American Council of Otolaryngology, which has a contract 
with the Extramural Program, The Office of Biometry provided statistical 
and data processing guidance to the group, and a staff me:nber also served 
as systems consultant to the Committee 



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The OB programming staff completed all the data processing and tables 
required for the surveys of Otolaryngologists in practice and in training. 
The staff also processed the results of a special study of training facilities 
and otolaryngologist-researchers. 

2. Computer Applications for NANDS Council 

The Office of Biometry provided special graphics for use at presentation 
before the NANDS Council. It trained an Extramural programmer in the use of 
the graphics equipment. 

INTRAMURAL PROGRAM 
Surgical Neurology Branch 

1 . Immunochemotherapy in an Intracerebral Murine Glioma Model 

Principal Investigator : Dr. L.A. Albright 

Statistical consultation and assistance was provided in a study of the 
therapeutical effect of various combined treatments of preimmunization, 
chemotherapy and immunotherapy on glioma induced in mice. The experiment was 
designed with a 2x2x3 factorial design. Data analysis involved evaluation of 
the median survival time (MST) of each experimental group and the use of a 
median test to compare the MST of each treatment group with that of the control 
group. Significance levels were computed for all comparisons. The analytical 
results and an experimental plan for further investigation were discussed with 
the investigator. An analysis of variance of the factorial experiment was 
completed. It involved arcsin transformation of the percentage data of 
survivors of 60 days or longer for each experimental group, with the transform- 
ed data analyzed in a factorial analysis of variance. Tests of srignificance 
were performed for the main effects, interactions and individual comparisons. 
Dose-response relationship of the chemotherapy under various combinations of 
preimmunization and immunotherapy was examined on a probit scale. The effect 
of preimmunication was estimated as an equivalent to a certain dose of chemical 
treatment. 

2. Study of the Effect of Lobectomy and Craniotomy on Epileptic Patients - 
Elect rode Imp ) ants 

Principa. Investigators : Dr. J.M. Van Buren, D. SadowsKy, Drs.N. Mutsuga 

and C,A. i^lariian 

The data on approximately 100 variables (etiological, neurological, medi- 
cation, EEG, socioeconomic, family history, etc.) obtained from 18 electrode- 
implanted and 18 non-implanted temporal lobe epileptic patients are involved 
in analyses. Comparisons for both groups inv-lude summaries i counts and per- 
cents) of about 85 pertinent \'ariables; graphing (histograms); and computa- 
tions (means, standard devietions, etc.), where applicable. Non-parametric 
statistical techniques permitted comparisons of (1) preoperative and post- 
operative variables to measure the extent of their association and (2) cases 
and controls to determine the effects of the electrode implants. 



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3. Study of the Effect of Lobectomy and Craniotomy on Epileptic Patients 

Principal Investigators : Dr. J.M. Van Buren, D. Sadowsky, Drs.N.Mutsuga 

and C. A. Marsan 

The data on the same variables (see electrode implants) obtained from 
150 patients (including the 36 patients in electrode implants) are being 
used in analyses and comparisons. These include: (1) summaries, graphs, 
computations; (2) non-parametric techniques to compare average and maximum 
fit frequencies with approximately 50 other variables (psychic change, motor 
type ictus, age at onset of seizures, seizure clusters, head injury severity, 
etc.) and preoperative and postoperative data from approximately 30 variables 
to determine their extent of association; (3) development of a method of 
determining "incidence rates" of seizure abolishment and/or improvement of 
70% or better; (4) the reporting of results of the analyses and comparisons. 
This project is still under way. 

4. Study of Various Therapies on Malignant Gliomas 

Principal Investigators : Dr. A.K.Ommaya, D.Sadowsky and Dr. J.M, Van Buren 

An Office of Biometry statistician was proposed as the chairman of an NIH- 
Georgetown University Brain Tumor Research Group for purposes of coordinating 
and analyzing all the data reco'^ded for both phases of a program of diagnosis 
and management of malignant gliomas. 

Phase I includes NINDS patients having surgery and conventional X-Ray 
therapy (not more than 3 months before trial entry). These patients will 
receive chemotherapy alone or combined immunochemotherapy at NIH. 

! Phase II includes Georgetown University patients who have no radiation 
therapy. They will receive fast neutron radiotherapy, after which they will 
receive, at Georgetown University Hospital, either chemotherapy alone, 
inmunochemotherapy, or no further therapy. 

For both phases, random selection for therepy and sequentia"" analycis 
plans were instituted. The analyses will include (1) "in-vivo" response 
indices for decision? on "clinical quality of survival"- {?.) ACTA Scanner 
responses tor quantitative (changes in volume of tumor) decisions; (3) sur- 
^nv?.] times from first tissue diagnosis (or entry into trial) for determin- 
ing best survival rates; and (4) further analyses of drugs, therapies, etc. 

5 . Study of Alteration 1n the Dentate Nucleus jec ondary to Lesions of the 
Diencephalon 

Pri ncipal Investigators : M.Levine, D. Sadowsky and Dr. J.M. Van Buren 

This is a continuing project. Additional data have been collected for 
comparison of the results observed from microscopic examination of brain 
tissue slides on five accidentally or surgically induced dienc3phalic- 
lerioiied and seven non-brain-affected patients. Two papers with the proposed 



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titles (1) Quantitative Study of the Dentate Nucleus in the Human (Levine,M.G. 
Sadowsky, D.A., and Van Buren, J.M.) and (2) Alterations in the Human Dentate 
Nucleus Secondary to Lesions of the Diencephalon (Van Buren, J.M., Levine.M.G., 
and Sadowsky, D.A.) will be prepared for publication. 

6. A Study of the Behaviors of Epileptic Patients after Temporal Lobectomy 

Principal Investigators : Dr. A.K. Ommaya, D. Sadowsky and Dr. D.B. Ahlberg 

The Office of Biometry provided statistical assistance for a study of the '| 
surgical effect on the behavioral and psychological variables, intelligence, 
ability to carry on daily living, and socioeconomic changes of the epileptic 
patients after operation. This will round out the projects on these data, 

7. An Assessment of Parkinson Patients after Thalamotomy 

Principal Investigators : D. Sadowsky, Drs. W: Henderson, J.M. Van Buren 

and D. Shapiro 

The final part (methodology) of this work has been completed, and a report 
(in which an OB statistician is a co-author) has been published - Henderson ,W.G., 
Sadowsky, D.A., Shapiro, D.Y., and Van Buren, J.M.: Quantitative Testing and 
Its Usefulness in the Evaluation of Therapy in Neurological Diseases, Confinia I 
Neurologica, 1973. 

Electroencephalography Branch u 

1 . A Study of Drug-Induced Seizures in Animals ' 

Principal Investigator : Dr. D. Lewis 

Statistical assistance has been provided in analyzing the EEG response 
during seizure, and oxydation of the test drug in the brain under local 
application, at various dose levels. 

Development and Metabolic Neurology Branch 

1 . A Study of the Drug Serum Concentration of Anticonvulsant Drugs 

Principal Investigator : Dr. A. Dekaban 

Nine children and 16 adult epileptic patients are involved in a study of 
the drug serum and dose levels of phenobarbital (PB), Dilantin (DPH), and | 
Primidon (PRM). The data were analyzed statistically to measure the drug 
serum level - dose level (DSL-DL) relationship. 

In a second study, DSL-DL relationships were obtained and compared for 
(1) (a) 11 adults on l'B+DPH(NINDS), (b) 13 adults + 9 children on PB+DPH+PRM 
(NINDS), and (c) 19 adults on PB alone (data recalculated for appropriate units 
from P. Lous, 1954); and (2) (a) 16 adults on PB+PRM+DPH(NINDS), (b) 14 adults 



I 



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on PB+DPH(NINDS), and (c) 26 adults on DPH alone (data read from Fig. 3, 
Svensmark, Schiller, and Buchthal , 1963). Comparisons were between different 
drug combinations, adults and children, and NINDS data and literature data. 

Laboratory of Neuropathology and Neuroanatomical Sciences 

1. Study of Neurosecretory Cells in the Brain Ganglia 

Principal Investigators : Drs. M. Brightman and Prescott 

A procedure was established for examining the number of fusion sites 
(locations on the cell membrane which are thought to be related to the expul- 
sion of a certain hormone) in two kinds of cells, namely those in a resting 
basal state and those in a stimulated state, A matched pair sequential 
sampling scheme has been suggested tentatively in this investigation in order 
to minimize the laboratory time and yet to maximize certain statistical 
considerations. 

Laboratory of Neural Control 

1 . Study of 'Sided Localization' in the Cerebellum 

Defining random localization to be the phenomenon whereby a vertical 
electrode probe is no more likely to result in the generation of a "left side 
impulse" rather than a "right side impulse" for a given brain cell, the null 
hypothesis is that random localization exists in those vertical areas where 
a probe results in more than one cell being activited. Assistance and consult- 
ation are being provided in the area of statistical techniques to test the 
hypothesis of sided localization. The statistical techniques, however, will 
be general izable to the anterior and posterior portions as well as quadrants. 

Laboratory of Neurochemistry 

1. A Study of the Effect of Electrical Stimulation on Inactivated Muscles 
in Cats 

Principal In vestigator : Dr. D. Riley 

Statistical analysis of two more sets of experimental data on this subject 
was completed this year. The analytical results of the current experiment 
agree quits well with the results of the previous experiments. Collaboration 
in the project involved developing an appropriate statistical method for testa 
of significance, preparing computer programs, carrying-out statistical 
calculations, and consulting in the preparation of a study report. 

Medical Neurology Branch 

1 . Radio-Irrmiunoassays of Prostaglandins 

A statistical analysis of the reliability of the method of radio- 
immunoassay of prostaglandins developed by Hazel ton Laboratories has been 
completed. A variance analysis technique was employed to examine the sources 



"(7^ 



of variation in the data. The analytical results revealed an unsatisfactory 
low reliability of this method. A decision was made on the basis of the 
statistical analysis to cancel the use of the Hazelton procedure. 

OTHER ORGANIZATIONS 

Laboratory of Nutritional Chemistry, NIAMDD 

1 . Bioassay of Vitamin E 

Principal Investigator : Dr. J. 6. Bieri 

Statistical consultation was provided in a study which involved two types 
of vitamin E to determine the anti -oxygenic effect of one type on the other. 
Significant recommendations were made for the design of the experiment for 
future assay work. 

The Washington Clinic 

1 . Developing a Screening Method of Internal Carotid Occlusive Diseases 

Principal Investigators : Drs. W. Howell and T.C. Chen 

The Office of Biometry has been involved ir. this continuing study screen- 
ing data obtained from the photoelectric plethysmography recordings. Analysis 
of data from an additional group of COD patients demonstrated a high consist- 
ency of the test efficiency to that previously observed. Further study will 
investigate the efficiency of applying this test to bilateral COD patients. 
Preparation of a publication is underway. 

Tho National Advisory Cottinission on Multiple Sclerosis 

1 . Multiple Sclerosis Survey 

The National Advisory Commission on Multiple Sclerosis contracted with 
Tracer, Inc., to conduct a survey, by interview, of all known MS p?.ti<^nts in 
Monroe County, N, Y. The survey was considered to be preliminary to a nationa". 
survey of MS patients; an RFP has been adverti""ed for the latter.' 

D- . \^dirry Weaver, Executive Director of the Corrmission, asked the Office 
of Biometry to perform several analyses which he believed would provide a most 
importanr addition to the Commission's final reoort to the Secretary, HEW. 
These included the development of multivariate regreision analyses to identify 
those characteristics and conditions of the surveyed MS patients which might 
be associatad with the medical and economic costs of MS, so that modified life 
tables could be constructed to provide estimates of the individual's lifetime 
costs of MS. 

Tracor provided the punched card data to OB, ani the latter, in the limited 
time of three weeks, using both complex cross-;:abulations and regre?sion 
techniques, identified the significant conditions and characteristics of the 
disease which were associated with the total :osts of MS, prepared the appro- 
priate modified life tables, and wrote the comprehensive report of this researcl 

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The Report of the National Advisory Commission on MS quoted extensively 
from the NINDS report, with relevant tables and charts. 

Mathematical Research Branch, NIAMDD 

1 . Lumping in Linear Compartments 

Principal Investigators : Dr.J.B.Hearon and R. B. Marimont 

A study of lumping in linear compartmental systems was begun in collabora- 
tion with Dr. John Z. Hearon, Chief, Mathematical Research Branch, NIAMDD. 
Lumping is the name given to an apparent reduction in the number of compart- 
ments in a system; i.e., the reactions in one or more compartments are 
appropriate to a system of fewer compartments than are actually present. 
Presence or absence of lumping depends on values of certain constants as well 
as connectivity of the system in ways not wholly understood. The sensitivity 
of the lumping behavior to small departures from the theoretical values of 
the constants is also not known. Theoretical and numerical studies of these 
questions will be carried out. The PDPIO computer has been tentatively 
selected for the numerical studies. 

RESEARCH 

1. Detection of Outliers •• ■ 

Dr. J. H. Ellenberg of the Office of Biometry is currently doing research 
on the problem of detection of outliers. The problem of detection of outliers 
in general linear regression was presented in the December 1973 issue of the 
Journal of the American Statistical Association. The focus of the current 
work is in two areas, first to examine the power function of the test for 
outliers in general linear regression and second to extend the results to 
the detection of multivariate ou-tliers. Linear regression is a frequently 
used statistical analytical method in neurologic research. 

2. Unequal Cell Counts in ^he Analysis of Variance 

Dr. Ellenberg is also currently doing research on the methodtiogy of 
analysis of variance for the case of unequal cell counts. This type of 
problem arises frequently in the analysis of biomedical date. The current 
methodology is confusing, due to the nonorthogonal nature of the data. It 
is hoped that an analysis can be developed that is fr&e from the confusing 
attributes of the standard techniques. 

3. Homogeneity of Variance 

Dr. Ellenberg is continuing work on the problem of homoge»ieity of variance 
in a two-way layout, that is, whether 0*- not the elements of a row or column 
are sampled from a population with the same variance as the rest of the rows 
or colur.r.s. To date, the literature dealinq with, this problem gives tests 
which are based on large sample approximation theory. An exact test is being 
developed which can be used for small samples. 



19y 



i 



<i 



4. Number Theory 

R. B. Marimont has solved an enumeration problem in elementary number 
theory, involving Pythagorean triangles. It was found that T(n) the number 
of primitive triangles of which the integer n can be a leg depends on r the 
number of distinct prime factors of the integer. 

r-1 
T(n) =2 if n ^ 2 (mod 4) 

T(n) =0 if n = 2 (mod 4) 

A paper was written and submitted to the American Mathematical Monthly. 

5. The Effect of Missing Values on Computed Correlation Coefficients 

D. Rubinstein has compared five methods of computing correlation co- 
efficients. Missing observations were generated by random numbers (not 
necessarily independent) on some artificially generated data and data collected:! 
in an experiment on 2277 monkeys. The results are not clear-cut since the 
random mechanism responsible for missing data has an influence on the outcome, 
as does the bivariate distribution of the random variables. On the whole the 
least distortion is introduced when the correlation coefficient is computed , 
for pairwise complete data. An algebraic approach utilizing Taylor's expan- i 
sion is under study which may provide more clarification than the numerical i 
simulations. This investigation has direct implications for the Collaborative ' 
Perinatal Study. 

i 

6. Evaluation of Screening Tests When the Criterion is in Error « 

D. Rubinstein has also considered the problem of test selection and test 
evaluation when the criterion is in error. The proposed methods utilize a 
criterion developed by earlier authors and the notion of conditional indepen- 
dence. It is shown that the criterion is especially effective in comparing 
competing screens. A paper on this subject is to be rewritten in light of 
reviews of an earlier manuscript. This research was motivated for evaluation 
of hearing test screens for infants with hearing performance at childhood 
taken as the criterion. 

7. Generation of Random Normal Deviates 

D. Rubinstein has studied the problem of finding the most efficient 
transformations in statistical simulation studies which are carried out in 
many medical and scientific areas. Some of the latter require large numbers 
of random normal deviates generated by the computer. One conventional method » 
of generating a single random normal deviate consists of summing a considerable 
number of uniform random variables which the computer generates with relative 
economy. The required number of uniform variables can be drastically re- 
duced by easily implementable transformations of the uniform random variables. 
There are several variations on this scheme. 



P 



20 y 



MISCELLANEOUS ACTIVITIES 

A. Computer Progranming and Related Activities 

1 . CPU Time Optimization Study 

Most FORTRAN programs in use in the Office of Biometry, both those 
written in-house and those appearing in analysis packages, use formatted 
I/O in reading and writing disk files. This is an extremely inefficient 
practice and is very costly when large files are being processed. In this 
study several more efficient reading techniques are being coded into test 
programs and run on the NIH 370 system in order to measure relative CPU 
times. So far, methods have been developed which are up to 20 times as 
efficient as formatted file I/O and are easy to use. 

The aim of the study is to make available to the user good estimates 
of the efficiency of various I/O methods and to provide dear instructions 
as to their application. Future plans include time comparison runs for 
optimization of FORTRAN code other than I/O. Optimization tips are scattered 
through the manuals, but the amount of payoff is never divulged and the method 
of application is not always clear. Thus, the user is not motivated to try 
these techniques. 

2. A Special Purpose Data Processing Program 

An assembler language program has been written whose purpose is to 
prepare data for input to the correlation programs of the Interactive Multi- 
variate Package. This program reads a raw data file of integers in EBCDIC 
storage mode, computes a double precision mean for each variable, writes the 
means to a disk file, and writes a new data file in binary storage mode on 
disk or tape. The program is designed to operate extremely efficiently in 
order to process very large data files at moderate cost. Coding for a pre- 
liminary version is complete and debugging is underway- 

3. Chi -Square Goodness of Fit Program 

A FORTRAN program was written to test normality of frequency data 
by means of chi -square goodness-of-fit tests. A special feature of the pro- 
gram is partial control over intervals by the user. The program determines 
the observed frequencies for the ncTiirial (%) intervals specified by the user. 

4. Correlation Program Fam ily 

As part of the Integrated Multivariate package a group of several 
correlation programs have been written. These programs compute means, 
standard deviations, and correlation coefficients, and offer variable selec- 
tion, field selection, and missing data handling in various degrees of complex- 
ity. Three of these programs are coded, debugged and working. The most 
comprehensive of the three is being reorganized to make future revisions 
easier and cheaper. 

Since this program will process \fery large data files, a fast version 
using centered integer data is now being planned. This version will have 

21y 



assembler language subroutines to do some of the repetitive, time consuming 
tasks, in order to speed up execution. If sufficient efficiency is attained 
it will be possible to retain only the most comprehensive of these programs. 

5 . Development or Implementation of Various Computer Software Packages 

a) Maintained the IMSL (International Mathematical and Statistical 
Library) on the NIH computer. 

b) Developed a special update and retrieval system to simplify use 
of IMSL. 

c) Obtained and installed at the NIH facility the BMD-P series 
software package for statistical processing. 

d) Developed an automated Job Control Language generator to permit 
non-programmers to enter jobs into the NIH computer job stream. 

B. Outside Activities 

1. A staff member of the Office of Biometry participated in site visits 
to review grant proposals involving computer applications. 

2. A staff member of the Office of Biometry presented the design of the I 
Primate Information Retrieval System at the Annual Session of the American ' 
Association of Laboratory Animal Science in Miami, Florida. 

3. He presented a report on Automated Information Systems for Laboratorisj 
at a conference at the National Bureau of Standards. 

4. He also gave a number of lectures on Systems Design at the U. S. Naval 
Academy. 

PUBLICATIONS | 

Marimont, R.B.: How can the deaf learn to speak? Some fundamento questions. 
VoUa Review 76:223-230. 1974 

G-nniayii. A^K., Albright, L.A., and Sadowsky, D.A.: Combination chemotherapy 
for man'^nant gliomas of the brain. Journal of Neurosu*-gery , in press 

Sever, J.L., Krebs, H.L., Ley, A., Barbosa, "..H., and Rubinstein, D.: 
Serological tests for measles for the diagnosis o"' subacute sclerosing 
panencephalitis. Accepted by the Journal of thp American Medical Association . 

FUTURE PLANS 

1. The Office of Biometry is planning further activities in the area of 
surveys of neurological diseases and communicdti/e disorders. For example, 
the Office of Biometry is now meeting with officials of the Natirnal Center 
for Health Statistics and the Bureau of the Census to determine possible 
cooperative arrangements among the three Ci'gamzations in support of a major 
survey of the incidence, prevalence, =ind medical and economic costs of stroke. 



t 



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2. The Office of Biometry plans to participate more fully in the design, 
analysis, and interpretation of epidemiological studies of neurological 
diseases and communicative disorders. 

3. The Office of Biometry plans to provide additional support, as required, 
in the planning and development of the program planning process in NINDS. 



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ANNUAL REPORT 

For Period July 1, 1973 through June 30, 1974 

Perinatal Research Branch 

ColIaboraHve and Field Research 

National Institute of Neurological 

Diseases and Stroke 

National Institutes of Health 



TABLE OF CONTENTS 



Page No, 



General Summary . 1 

Contract Narratives 

University of Michigan (NOl-NS-1-2390) 

Genetic Typing in Children in NIH Perinatal Project .... 9 

University of Wisconsin (NOl-NS-2-2302) 

Genetic-odontometric Study of Pre- and Neonatal Grov/th . . 11 

University of Michigan (NOl-NS-2-2320) 

Bivariant Physical Growth 13 

Johns Hopkins University (NOl-NS-2-2321) 

Ancillary Studies 15 

Boston University (NOl-NS-2-2322) 

Maternal Drug ingestion and Fetal Abnormalities ..... 17 

Pennsylvania State University (NOl-NS-3-231 1) 

Analysis of General Pathology and Placental Pathology of 

the Collaborative Perinatal Data 19 

Children's Hospital Medical Center, Boston, Massachusetts 

(NOl-NS-3-2312) 

Combined Neuropathologic and Epidemiologic Study .... 21 

Beth Israel Hospital, Boston, Massachusetts (NOl-NS-3-2320) 

A Dynamic Time Trend Analysis of Collaborative Perinatal 

Project Data and Patterns of Blood Pressure, Edema and Proteinuria 

(Toxemia) as Related to Pregnancy Outcomes 23 



TABLE OF CONTENTS (confd) 



Page No. 



III. Individual Project- Reports 25 

IV. A Comprehensive Plan for Analysis and Interpretation of Collaborative 
Perinatal Project Data 

Primary Data Analysis Areas 

Cerebral Palsy 89 

Mental Retardation 99 

Communicative Disorders 71 

Visual Abnormality 103 

Convulsive Disorders 85 

Learning Disorders 105 

Minimal Brain Dysfunction 97 

Congenital Malformations 109 

Birthweight-gestational Age 93 

Pathology 

General 19 

Neuropathology 21 

Secondary Data Analysis Areas 

Toxemia 23 

Maternal Infection During Pregnancy* 

Labor and Delivery 55 

Neonatal Hyperbilirubinemia 27 

Maternal Anesthesia-analgesia During Labor 
and Delivery - (to be developed) 

Four Year IQ 67 

Physical Growth and Development 13 

Twins - (to be developed) 

Genetic and Socioeconomic Factors 41,53,63 

Drugs Taken During Pregnancy 17 



^Reported by Infectious Diseases Branch, C&FR, NINDS 



ANNUAL REPORT 

For Period July 1, 1973 through June 30, 1974 

Perinatal Research Branch 

Collaborative and Field Research 

National Institute of Neurological 

Diseases and Stroke 

National Institutes of Health 



GENERAL SUMMARY 



INTRODUCTION: 



In Fiscal Year 1974, the Collaborative Perinatal Project (CPP) will complete the 
longitudinal follow-up speech, language and hearing examinations on children at 
eight years of age. This completion will terminate CPP data collection. The basic 
objective of the CPP is to relate complications, conditions and events that occur 
during a woman's pregnancy or during her labor and delivery to subsequent abnor- 
mality exhibited by the child of the pregnancy as the child grows and develops. As 
data collection has approached completion, the major emphasis of the project has 
shifted to data analysis and Interpretation for publication of reports on the CPP research 
findings. In order to accomplish this, a coordinated effort has been implemented to 
meet the basic data analysis objectives of the CPP. To provide a framework for this 
effort, a Comprehensive Plan for Analysis and Interpretation of Collaborative Peri- 
natal Project Data has been approved and implemented. 



II. DATA COLLECTION: 



The Collaborative Perinatal Project was initiated In 1959 when the women began 
registering at each of the collaborating institutions during their pregnancies. Women 
continued to enroll In the study through December of 1965, The babies were delivered 
between 1959 and 1966 and received several examinations during their first year of 
life, at 3 years of age, and at 4 years of age. The 7-year extensive battery of exami- 
nations Included a pediatric-neurologlcal examination; a battery of psychological 
tests, including an IQ determination; and a visual screening test. By June of 1974, 
an assessment of speech, language and hearing development will be completed 
on children eight years of age at five of the collaborating institutions. Completion of 
the eight-year examination will terminate follow-up of CPP children. 



III. A COMPREHENSIVE PLAN FOR ANALYSIS AND INTERPRETATION OF COLLABO- 



RATIVE PERINATAL PROJECT DATA: 



Broadly stated, the Collaborative Perinatal Project is concerned with the identifica- 
tion of prenatal factors that have a sufficiently high association with adverse pregnancy 
outcome and subsequent neurological and mental development of the child to provide 

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leads to the etiologies of the abnormalities and thus to the development of strategies 
for prevention and intervention. After a careful review/ of the objectives of the Col- 
laborative Perinatal Project, the data available for analysis and the work in progress, 
it was recommended that major efforts in analysis and interpretation are needed in ten 
primary areas in order to meet the basic objectives of the project. Monograph reports 
in book form are planned in each of the following primary areas: 

Cerebral Palsy (See Individual Project Report - 

Serial No. NDS (CF)-73 PR/PSC 2059) 

Mental Retardation (See Individual Project Report - 

Serial No. NDS (CF)-74 PR/PSC 2106) 

Communicative Disorders (See Individual Project Report - 
Serial No. NDS (CF)-72 PR/PSC 1971) 

Visual Abnormality (See Individual Project Report - 

Serial No. NDS (CF)-74 PR/PSC 2107) 

Convulsive Disorders (See Individual Project Report - 
Serial No. NDS (CF)-73 PR/PSC 2058) 

Learning Disorders (See Individual Project Report - 

Serial No. NDS (CF)-74 PR/PSC 2108) 

Minimal Brain Dysfunction (See Individual Project Report - 
Serial No. NDS (CF)-73 PR/PSC 2062) 

Congenital Malformations (See Individual Project Report - 
Serial No. NDS (CF)-74 PR/PSC 2109) 

Birthweight-Gestational Age Relationships (Prematurity) (See Individual 
Project Report - Serial No. NDS (CF)-73 PR/PSC 2060) 

Neuropathology, General Pathology and Placentology (See Contract 

Narratives, Contracts NOl-NS-3-231 1 & NOl-NS-3-2312) 



IV. IMPLEMENTATION OF THE COMPREHENSIVE PLAN: 



Implementation of the Comprehensive Plan for Analysis and Interpretation of Collabora- 
tive Perinatal Project Data Is carried out through teams of researchers, headed In each 
of the ten primary areas by a member of the Professional Staff Consultants Section of 
the Perinatal Research Branch. On each team, there is a member of the Office of 
Biometry staff, who participates fully In the development of the analysis. In addition, 
two members of the Section for Production of Data Analysis, Perinatal Research Branch, 
are assigned to each primary area to facilitate data processing. The assignments are 
as follows: 



PERINATAL RESEARCH 
PRIMARY DATA ANALYSIS AREAS BRANCH OFFICE OF BIOMETRY 



Cerebral Palsy 

Mental Retardation 

Communicative Disorders 

Visual Abnormality 

Convulsive Disorders 

Learning Disorders 

Minimal Brain Dysfunction 

Congenital Malformations 

Birthweight-Gestctional Age 
Relationships (Prematurity) 

Neuropathology, General 
Pathology and Placentology 



Dr, K. B. Nelson 

Dr. S. H. Broman 

Dr. P. J. LaBenz 

Dr. R. Feinberg 

Dr. K. B. Nelson 

Dr. S. H. Broman 

Dr. P. L. Nichols 



Dr. J. H. Ellenberg 
Dr. P. W. Shaughnessy 
Mr. D. Rubinstein 
Miss E. C. Jackson 
Dr. J. H. Ellenberg 
Dr. P. W. Shaughnessy 
Dr. T. C. Chen 



Dr. N.C.Myrianthopoulos Mr. D. Rubinstein 
Dr. B. H. Williams Miss E. C. Jackson 



Dr. J. S. Drage 



Dr. T, C. Chen 



In each of the ten primary areas, a program plan has been developed and approved 
v/hich expands in detail on the summary statements in the Comprehensive Plan and 
gives a detailed approach to the analysis and identifies major components. Milestone 
charts have been designed to set goals and to aid in the measurement of progress. 
Written monthly reports are being prepared to record progress and identify problem 
areas. In order to facilitate the primary data analyses and/or complete major efforts 
well underv/ay, additional analyses are to be completed in the follov/ing secondary 
areas: 

Toxemia (See Contract Narrative, Contract NOl-NS-3-2320) 

Maternal Infection during pregnancy (See report by Infectious Diseases 
Branch, C&FR, NINDS) 

Labor and Delivery (See Individual Project Report - 

Serial No. NDS (CF)-71 PR/PSC 1904) 

Neonatal Hyperbilirubinemia (See Individual Project Report - 
Serial No. NDS (CF)-74 PR/OC 2112) 

Maternal anesthesia-analgesia during labor and delivery (To be developed) 

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Four- Year IQ (See Individual Project Report - 

Serial No. NDS (CF)-72 PR/PSC 1968) 

Physical Growth and Development (Birth to Seven Years) (See Contract 
Narrative, Contract NOl-NS-2-2320) 

Twins (To be developed) 

Genetic and Socio-economic Factors (See Individual Project Reports - 

Serial No. NDS (CF)-67 PR/PSC 1514, NDS (CF)-70 PR/PSC 
1857, NDS (CF)-71 PR/PSC 1910) 

Drugs taken during pregnancy (See Contract Narrative, Contract NOl-NS-2- 
2322) 



V. SUMMARYOF WORK IN PROGRESS: 



In the cerebral palsy area, consistent definitions and criteria have been established. 
Records of all children with definite or suspect diagnosis at seven years have been 
reviewed and reclassified according to these established criteria. Interval histories 
have been examined to separate "acquired" from "congenital" cerebral palsy. There 
are about 200 cases of cerebral palsy In the study or about 1 In 200 births. This figure 
Includes milder cases than are usually reported, but provides a fuller spectrum of motor 
defects than has heretofore been available. Many children showing definite cerebral 
palsy diagnosis at one year, do not show such deficits at seven years but remain at high 
risk for mental retardation. Isolating variables which permit prediction may be possi- 
ble; e.g., few children who could pull to standing by one year showed significant 
motor deficits at age seven. Work has progressed on the planning of a demographic 
analysis; maternal, obstetric and early pediatric variables have been selected and 
defined and a data analysis request to relate these factors to the development of the 
child Is nearing completion. 

Children who are mentally retarded (I.Q.'s under 70 as determined at age seven years) 
form a heterogeneous group and tbey have been subdivided into four major categories: 
severe retardation (I.Q.under 50) with and without signs of central nervous system 
damage, and mild retardation (I.Q. between 50 and 69) with and without such signs. 
Preliminary findings indicate that the following conditions are related to a low intel- 
lectual functioning at age seven: Low socioeconomic status of the family, high hous- 
ing density, low maternal Intelligence, less parental care, less advanced mental and 
motor development at eight months, poor language development at age three, lower 
psychometric test performance at age four, and smaller physical measurements during 
the first seven years of life. 

A major decision in the Speech, Language and Hearing area during FY 1974 was to 
arrange for the analysis and interpretation of the data under contract. Efforts to se- 
cure a contract are underway. Since collection of data in this area will not be com- 

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plete until the end of this fiscal year, the use of the contract mechanism is expected 
to expedite data analysis and relieve part of the burden from the Perinatal Research 
Branch data production staff in the coming fiscal year. Tabulations of available 3- 
year data have been made by items and item combinations. Correlation matricies 
were constructed to explore intratest relationships at age 3, and relationships with 30 
selected 8-year, perinatal and other variables. Tabulations were also run on 8-year 
data presently available. Crude estimates of prevalence of disorders have been made, 
and potential key variables for construction of indices have been tentatively selected. 
Specifications have been drawn for conducting detailed analyses. These efforts are 
now aimed at preparing the data for the contractor. 

In the visual abnormality area, frequency distributions of findings on a sample of the 
seven-year visual screening tests have been obtained, and a review and classification 
of some 4,000 ophthalmologist consultants' reports have been undertaken. (When the 
screening test is failed, ophthalmologic consultation is obtained.) Preliminary evi- 
dence suggests that current prevalence figures regarding color vision impairment, 
20/20 visual acuity, amblyopia, and ocular muscle imbalance may need to be revised. 

In the convulsive disorders area, criteria for specific diagnosis have been established 
and 710 records reviewed of children thought to have convulsive disorders. There Is 
some overlap with the cerebral palsy cohorts. The review of cases will continue and 
should near completion by the end of this fiscal year. Variables to be used in analy- 
sis have been selected and defined. 

Children with learning disorders have been identified. A working definition of learn- 
ing disorders for Collaborative Perinatal Project children has been derived by identi- 
fying those children who are one year below grade expectancy in reading or spelling 
on the Wide Range Achievement Test and who have an I.Q. of 90 or above on the 
Weschler Intelligence Scale. 

Progress in the area of minimal brain dysfunction was toward the delineation of the 
syndrome from a list of 25 symptoms in four major areas: behavioral, cognitive and 
perceptual motor, academic, and physical. Several hundred children have been 
identified who have symptoms in two or more of the major areas. Risk of symptoms in 
one area is increased if there are symptoms in another area, but this association is not 
strong since most children only have symptoms in a single area. 

Work has continued to progress on the comprehensive analysis of congenital malforma- 
tions. The analysis is in 11 parts. Those partsdealingwith theepidemioiogyof congenital 
malformationshave been completed and reports are being written or are in press. Only a third 
of malformations were diagnosed at birth. Except for three minor malformations, there 
were no significant racial differences in incidences. Twins have significantly more 
malformations than singletons, but the difference is wholly contributed by monozygotic 
twins. Multiple birth, pregnancy complications, male birth, and maternal diabetes 
were positively correlated with increased risk of malformations, whereas maternal 
weight gain was negatively correlated. With respect to diabetes, the risk is doubled; 
the effect seems to be correlated with the severity of the disease and not with insulin 

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intake. Genetic studies, blood type studies, a study of clinical significance, devel- 
opmental and survival studies, studies of maternal factors, and chromosome studies are 
all either underv/ay or in the design stage. 

Progress in the area of birthweight-gestational age includes the generation of an exten- 
sive set of tabulations showing distributions, frequencies, and rates of birthweight- 
gestational age groups and their relationship with secondary parameters such as pla- 
cental weight, maternal education, socioeconomic index, gestation at registration, 
year of birth, institution, body length at birth, and head circumference at birth. 
Frequency tables showing neonatal death and stillborn rates and survival figures have 
been generated. These tables are under intensive analysis and results will provide 
essential input to other analysis areas. 

Studies in the area of pathology are being undertaken by two separate contracts, one 
for neuropathology and one for the general and placental pathology. The neuropathol- 
ogy contractor will prepare a catalogue of gross brain abnormalities, plot fetal brain 
weight against gestational age, and analyze the events of pregnancy, labor and de- 
livery for those brains deviating from the projected growth curve. Intra- and extra- 
utero growth rates will be compared. Intra-cranial hemorrhage, telencephalic leuco- 
encephalopathy, and cerebral necrosis will be studied in detail and related to possible 
risk factors. The general and placental pathology contractor will review deaths and 
classify associated factors. Demographic characteristics, maternal conditions and 
events of pregnancy, labor and delivery will be related to deaths. Organ growth rates 
will be analyzed and related to maternal conditions as contributory factors. Placental 
abnormalities will be classified and related to demographic characteristics, maternal 
conditions, and events of pregnancy, labor and delivery. Placental weight and placen- 
tal pathologv will be analyzed with respect to birthweight-gestational age classifica- 
tions and pregnancy outcome. 

The study of "Toxemia of Pregnancy" was undertaken through contract in Fiscal Year 
1974. The objective is to establish a precisely-defined classification by a time-trend 
analysis of the data pertinent to clinical diagnosis: blood pressure, proteinuria, and 
edema. Contributory factors for each toxemia classification will be examined. Dur- 
ing the first year of this contract, the time-trend analysis of blood pressures will be 
completed. 

The neonatal hyperbilirubinemia study was implemented during Fiscal Year 1974. This 
study has been designed to assess the relationships of intermediate (10-20 mg.%) levels 
of serum bilirubin on subsequent neurological and mental development of CPP children. 

A study of the relationship between intelligence at four years of age, as measured by 
the Stanford-Binet Intelligence Scale, and a variety of factors related to pregnancy, 
delivery, postnatal status, and environment, was completed during Fiscal Year 1974. 
A substantial monograph has been developed and has received outside review. The 
manuscript has undergone revision and publication is anticipated in six to nine months. 

Detailed analysis of physical growth data on CPP children to seven years of age is under- 
way. Four anthropometric variables are under consideration: weight, height, head 

6 z 



circumference, and chest circumference. Analyses are dimensional, incremenfal, and 
bivariate and the analysis will be by race, sex, and socio-economic groupings. A 
contractor is working on the bivariate analysis. Based on these findings, an expanded 
contract analysis will be developed. 

The file of family linkage groups was updated and revised during Fiscal Year 1974. 
The recreated file is in an improved format to facilitate familial studies on CPP 
mothers and children. This file will be an essential resource for the analysis of gene- 
tic factors in other study areas. The file has been used to estimate genetic components 
in obstetrical variables, both continuous and discrete. Work continues on the genetics 
of intellectual and motor performance and additional papers are being prepared. 

The study of the effects of maternal drug ingestion on perinatal deaths and congenital 
malformations continued during Fiscal Year 1974 under contract. The study will des- 
cribe drug utilization patterns during pregnancy and provide for a rapid evaluation of 
drug teratogenicity through the use of a screening device to calculate relative risk. 
Other relationships are being explored and a number of papers have been published. 



VI. CONTRACT DEVELOPMENT: 

A contract is being developed for an extensive analysis of speech, language and hear- 
ing data. The specific aims of the proposed contract are 1) to identify a small number 
of key speech, language and hearing variables, including indices which will serve to 
describe communicative ability or status of the subject tested; 2) to explore the associa- 
tion between speech, language, and hearing variables, and other Collaborative Peri™ 
natal Project variables, in order to establish etiological and predictive relationships 
where they exist; and 3) to prepare a final manuscript in publishable form, which re- 
ports on the samples and methods, results, and their interpretation and potential appli- 
cation. 

The correlation of minor chromosomal variants with congenital malformation is also be- 
ing developed as a contract project. This project will utilize data collected by five 
of the Collaborative Perinatal Project institutions, which are participating in a chro- 
mosomal study of minor variants. The collection of the chromosomal material was 
sponsored by the National Institute of Child Health and Human Development. 



VII. OTHER FUNCTIONS: 



The Perinatal Research Branch continues to process incoming research protocols, per- 
form automatic data processing, and manage the phase-out of data collection contracts. 



7z 



CONTRACT NARRATIVE 
Perinatal Research Branch — Professional Staff Consultants Section 
July 1, 1973 through June 30, 197^ 

UNIVERSITY OF MICHIGAN (NOl-NS-1-2390) 

Previous Contract No (NIH-NINDS 71-2390) 

Title: Genetic typing in children in NIH Perinatal Project 

Contractor's Project Director : H.G. Gershowitz, Ph.D. 

Current Annual Level : None 

Objectives : To obtain approximately 5,000 blood specimens from children 
born in the Collaborative Perinatal Project, separate red cells from serum 
and freeze both in liquid nitrogen for future studies. 

Current Status of Contract: This contract has been tenoinated. 



CONTRACT NAERATIVE 

Perinatal Research Branch — Professional Staff Consultajits Section 

July 1, 1973 through June 30, 197^ 

UNIVERSITY OF WISCONSIN : (NOl-NS-2-2302) 

Previous Contract Number : (NINDS-NIH 72-2303) 

Title : Genetic-odontometric study of pre and neonatal growth 

Contractor's Project Director : Richard Oshorne, Ph.D. 

Current Annual Level : $59,988.00 

Objectives : To correlate the incidence and prevalence of dental and facial 
abnormalities with neiirological defects, congenital abnormalities and other 
disorders of childhood. 

Methodology : Der.tal casts and photographs of teeth will be obtained on about 
2500 Collaborative Study children who are schediiled for routine seven and 
eight year examinations at selected participating institutions, and on 
children rescheduled because of known complications during their prenatal 
and neonatal periods of study. 

Major Findings : During the first year of study about 22 percent of children 
were found to have dental abnormalities. 

Significance to the Program : The study of dental abnormalities is expected 
to yield an index of stress or abnormality during prenatal development. 

Proposed Course of Contract : Collection of casts and film on 2217 children 
has been completed. The contractor is now proceeding with morphological 
evaluation of the data. The contractor has performed a preliminary analysis 
of data on the first 675 children to develop variables and methodology for 
analysis. 

Because of r'.elays in instrumentation the morphological e-alaation and final 
anaiysiE will not be completed within the contracx j.eriod, i.e. by May 31, 197^ < 
Howevei=, since it is anticipated that there will be a considerable unexpended 
balance at the end of the contract period, the possibility is considered of 
extending the contract without allocation of further funds to enable the 
contractor to complete the analysis and prepare a final report. 



11:: 



CONTRACT NARRATIVE 
Perinatal Research Branch, C&FR, NIMDS 
Office of the Chief 
July 1, 1973 throiigh June 30, 197^ 

UNIVERSITY OF MICHIGAN (NOl-NS-2-2320) 

Previous Contract N;mber : NIH-NINDS-72-2320 

Title : Bivariant Physical Growth 

Contractor's Project Director : Stanley M. Garn, Ph.D. 

Ciirrent Annual Level : $l6,900 

Objectives : 

a. Merge per-capita income and socio-economic index data per individual 
with anthropometric data on tapes and; after checking, convert to 
MIDAS for the most economical approach to b. through e. below. 

b. For k anthropometric variables, (A, weight; B, height; C, head circum- 
ference; D, chest circumference) by 2 sexes, by 2 races by 3-U economic 
groupings, compute new percentiles for at least 2 ages to see how the 
percentiles and therefore the medians are affected, within a race and 
between races at comparable economic groupings and between economic 
groupings . 

c. For k anthropometric variables and a single later age, for 2 sexes, 
by 2 or more races, compute new mini bivariant tables. A by B, 

A by C, A by D, B by C, B by D, C by D, for each of 3-^+ economic groupings 
and thus ascertain the extent to which income effects size percentiles A 
by size intervals B by constant age. 

d. For a set of ages, including the ages used in b. and c. compute the 
increments (deltas) for the same k dimensional variables given in b. and 
o. and percentiles for the deltas, by sex, race, and 3-^ so.jio-economic 
groupings to ascertain: the effect of income, income/needs or socio- 
economic index on deltas, within races and for comparable groupings, 
between races. 

e. Using a special MIDAS program, convert the low-income skewed sample into 
a more nearly national probability sample, by weighing, so that one set 

of "norms" can be developed that will not be simply limited to the original 
lower-income wei5;hted distributions. Specifically, percentiles (see b. 
above) and increments (see d. above). 

Major Findings : Detailed analysis of PRE physical growtli data (dimensional., 
incremental, and bivariant) chow the need for a) excluding impj'obable, miscoded, 
or mispunched values, b) meaningful arrangement by PRB SE Index and other 
measures of social class, c) corrected and imcorrected birth weight groupings, 

13 z 



Contract Number: NOl-NS-2-2320 

and d) PRB SE Index in developing bivariant tables. That is, achievement of 
the above objectives indicates the need for a more comprehensive analysis of 
the physical growth data. 

Co-urse of Contract : September 28, 1973 through September 27, 197U, 



14 



CONTRACT NARRATIVE 

Perinai-al Research Branch, C&FR, NINDS 

Office of the Chief 

July 1, 1973 through June 30, 1974 

JOHNS HOPKINS UNIVERSITY (NOl-NS-2-2321) 

Title: Ancillary Studies 

Contractor's Project Director : Janet B. Hardy, M.D. 

Current Annual Level: $51,600 

Objectives: 

Using NINDS Collaborative Perinatal Project data and data unique to the Johns Hopkins 
Collaborative Perinatal Study, the contractor will conduct studies in the following areas: 

a. Perinatal infections. 

b. Problems in communication. 

c. Growth and development of the child in an inner-city population. 

d. The psychological, biological ana environmental factors relating to I.Q. 

e. The etiology and characterization of seizures. 

f. The effect of maternal smoking on the fetus. 

g. The toxicity of low levels of bilirubin. 

Course of Contract : June 28, 1972 through June 27, 1975. 

Publications: Hardy, J. B.: Birth weight and subsequent physical and intellectual devel- 
opment. N. Engl. J. Med. 289:973-974, 1973. 

Hardy, J. B. and Mel I its, E. D.: Does maternal smoking during pregnancy h /e a long 
term effect on the child? Lancet. 2:1332-1336,1972, 

Hardy, J, B.: Clinical end developmental aspects oF congen'to! rjbeiJQ. Arch. Otolaryn- 
gol. 98:230-236, 1973. 

Wolff, S. M,: The ocular manifestations of congeni'-al ruLelia, A prospective study of 328 
cases of congenital rubella. J. Pediatr. Ophthalmol. 10:101-141, 1973. 

Welcher, D. W., Wessei, K. W., Mellits, E. D. and Hardy, J. &.: The Bender-Gestalt 
test as an indicator of neurological impairment in voung "inner city" children. Percept. 
Mot. Skills. 38:899-910, 1974, 

Hardy, J. 3.: Congenital rubella. In Ryan, S. J., Jr. (Ed.): Eye and Systemic Disease . 
New York, Grune & Stratton, In press. 



I ^^^ 



CONTRACT NARRATIVE 
Perinatal Research Branch, C&FR, NINDS 
Office of the Chief 
July 1, 1973 through June 30, 1974 

BOSTON UNIVERSITY (N01-NS-2-2322) 

Title: Maternal Drug Ingestion and Fetal Abnormalities 

Contractor's Project Director: Dennis Slone, M.D. 

Current Annual Level: $165,000 

Objectives: 

Using NINDS Collaborative Perinatal Project data, the objective of the contract is to 
conduct a research program on maternal drug ingestion and fetal abnormalities. The 
study will describe maternal drug utilization patterns, provide for a rapid evaluation of 
drug teratogenicity, report on the relationship between Dilantin and selected congenital 
malformations, investigate the possible teratogenic effects of virus vaccines, and investi- 
gate possible associations between dnjgs and congenital cardiac malformations. 

Course of Contract: June 28, 1972 through June 27, 1975 

Publications: Heinonen, O. P., Shapiro, S., Monson, R. R., Hartz, S. C, Rosenberg, 
L. and Slone, D.: Immunization during pregnancy against poliomyelitis and influenza 
in relation to childhood malignancy. Int. J. Epidemiol. 2:229-235, 1973. 

Monson, R. R., Rosenberg, L., Hartz, S. C, Shapiro, S,, Heinonen, O. P. and Slone, 
D.: Diphenylhydantoin and selected congenital malformations. N. Engl. J. Med. 
289:1049-1052, 1973. 

Slone, D., Heinonen, O. P., Monson, R. R., Shapiro, S., Hartz, S. C. and Rosenberg, 
L.: Maternal drug exposure and fetal abnormalities. Clin. Pharmacol. Thar. 14:648- 
653, 1973. 

The Boston Collaborative Drug Surveillance Program (Maruscript prepared by Olii P. 
Heinonen): Diethylstilbestrol in pregnnncy. Frequency of exposure and usage patterns. 
Cancer. 31:573-577, 1973. 



17 z 



CONTRACT NARRATIVE 
Perinatal Research Branch, C8JR, NINDS 
Office of the Chief 
July 1, 1973 through June 30, 1974 

PENNSYLVANIA STATE UNIVERSITY, M.S. HERSHEY MEDICAL CENTER (NOl-NS-3-231 1) 

Title : Analysis of General Pathology and Placental Pathology of the 
Collaborative Perinatal Project Data. 

Contractor's Project Director : Richard L. Naeye, M.D. 

Current Annual Level: $136,500. 

Objectives: 

This contract is in two parts. One part concerns the general pathology and the other part 
the placental patholog)'. 

The Objective of the genera! pathology analysis is to review and classify deaths as to the 
cause of death, factors contributing to death, and other associated factors. The patholo- 
gist is making an assessment of morphologic diagnoses using Collaborative Perinatal Project 
(CPP) protocols, magnetic data tapes and histologic tissues and slides. An analysis will be 
completed in the following areas: 1) demographic characteristics (age, race, marital sta- 
tus and others deemed pertinent to a pathology-oriented analysis) of women to develop 
possible associations with stillbirth, neonatal, and later death; and other classifications of 
specific pathological or anatomical findings deemed relevant to demographic characteris- 
tics. 2) maternal conditions and events of pregnancy, labor and delivery to test for asso- 
ciations with stillbirth, neonatal, and later deaths, and with specific pathological and 
anatomical findings or groups of findings thought by the pathologist to need exploration. 
For neonatal and later deaths, date obtained on the child prior to death will be examined. 
Such analysis will include specific conditions; such as, hyaline membrane dis^'.se, eryihro- 
blastosis, congenital malformations, and sudden unexplainable death. 3) organ growth 
rates, inck'ding the quantitative extent to which a ,ariety of reported maternal conditions 
and events way modify or contribute to altered organ growth ratos. 

The Objective of the placental pathology analysis is to rftviev and classify the various 
placental findings as reported on study proiocols (Paiti-1 a^d f'oth-2), by using the proto- 
cols, magnetic data tapes, and placental microscopic slides and tissues. Through such a 
review, the pathologist will be expected to define and create additional classifications of 
placental pathology. Analyses v/i 1 1 be completed on 1) demo^raph'c characteristics and ■ 
maternal conditions and events of pregnancy, labor, and delivery by the classical and in- 
novative pathological and anatomic categories established by the pathologist; and 2) birth- 
weight-gestation classifications by placenta! weight, placental pathology and pregnancy 
outcome. 

Course of Contract: June 1, 1973 through June 30, 1976 

19 z 



CONTRACT NARRATIVE 

Perinatal Research Branch, C";FR, NINDS 

Office of i-he Chief 

July 1, 1973 through June 30, 1974 

CHILDREN'S HOSPITAL MEDICAL CENTER, BOSTON, MASSACHUSETTS (N01-NS-3--2312) 

Title: Combined Neuropathologic and Epidemiologic Study 

Contractor's Project Director: Floyd H. Gilles, M.D. 

Current Annual Level: $206,150 

Objectives: The contract will analyze the neuropathology collection of the Collaborative 
Perinatal Project (CPP). A safe, readily accessible site for the neuropathology collection 
will be created to receive the material formerly housed in the Perinatal Research Branch, 
Cc.FR, NINDS, Bethesda, Maryland. An estimate of the quality of the materia! and a 
catalogue of gross brain abnormalities will be prepared. Plots of fetal brain weight of 
grossly normal brains against estimated gestational age, utilizing a Gompertz function, 
will be madCo For brains lying beyond one and/or two standard deviations from the growth 
curve projected by the model, an analysis will be made of events of pregnancy, labor, and 
delivery, A comparison will be made of rate of brain weight acquisition in utero to rate of 
brain weight acquisition after birth as a function of total (gestational plus survival) age, A 
study will be made of intracranial hemorrhage including topography of hemorrhage (ventri- 
cular, within plexus; ventricular, external to plexus; ganglionic eminence' extraventricular 
within brain; arachnoidal; extra-arachnoldal), time of age at death, incidence of venus 
thrombi, incidence of Galenic or sinus thrombi, incidence of venous dilatation, and eval- 
uation of such risk factors as pulmonary disease, cardiac diseases, liver and spleen weight, 
fetal experiences, and body and brain weight. A study will be done on the risk factors 
associated with perinatal telencephalic leucoencephalopathy to test the following hypothe- 
ses: 1) infants with perinatal telencephalic leucoencephalopathy differ from those without 
it in regard to (a) immunoglobins and (b) maternal immunoglobins; and 2) infants with peri- 
natal telencephalic leucoencephalopathy differ from those without in regard to thymus 
weight and morphology. A study of cerebral necrosis is to be completed which would in- 
clude criteria of necrosis in the perinatal brain, neuronal necrosis (relative maturity of 
neuronal aggregate; relative immaturify of neuronal aggregate, topography; modes of re- 
pair), white matter necrosis (necrotic foci; topography; modes of repair) and an evaluation 
of selected risk factors in relation to pobclassification of neuronal and white matter necrosis. 

Course of Contract: June 1, 1973 to June 30, 1976. 



21 z 



CONTRACT NARRATIVE 

Perinatal Research Branch, C&FR, NINDS 

Office of the Chief 

July 1, 1973 through June 30, 1974 

BETH ISRAEL HOSPITAL, BOSTON, MASSACHUSETTS (NOl-NS-3-2320) 

Title : A Dynamic Time Trend Analysis of Collaborative Perinatal Project Data on 
Patterns of Blood Pressure, Edema and Proteinuria (Toxemia) as Related to 
Pregnancy Outcomes. 

Contractor's Project Director: Emanuel A. Friedman, M.D. 

Current Annual Level: $165,000 

Objectives: 

The objective of the "Toxemia of Pregnancy" contract is to establish a precisely-defined 
toxemia classification by an analysis and interpretation of Collaborative Perinatal Project 
(CPP) data pertinent to the clinical diagnosis of toxemia: blood pressure, proteinuria and 
edema. The objective of the CPP is to provide leads to the etiologies of neurological and 
mental abnormalities of children througn a study of the conditions and events of pregnancy, 
labor and delivery. Toxemia \s one major abnormality of pregnancy, which is known to 
have an increased risk to the fetus of death or premature birth with attendant risk to the 
survivors. However, toxemia has been vaguely and inconsistently defined and this research 
effort is intended to classify the varying degrees of toxemia so that each precisely-defined 
degree of toxemia can be assessed for a possible etiological relationship with neurological 
and mental abnormality of the child. 

An estimated 5% to 10% of CPP women have manifested one or more symptoms of toxemia. 
Extensive data on systolic and di,Jstolic pressure, varying degrees of proteinuria and degrees 
of edema have been collected on CPP women. 

The retecrch as detaiied in the work scope plan contains a dynamic tir.ie-related trend 
anolysis cf blood pressi re, edema, and proteinuria; and intorc.or'-elaticns of the preceding 
faclors. The dynainic tlme'»re!at&d trenc' analysis for each of the factors would include the 
determination of patterns over the course of pregnancy for individual gravidas, categorizing 
the trend patterns, establishing relationships to maternal and pregnancy outcome factors, 
and defining the relative risk effect for each trend pattern. Another major analysis would 
be muliiple regression analysis of contributory factors for each of the toxemia classifica- 
tions. 

Course of Contract: June 25, 1973 through January 31^ 1976 



23? 



Serial No. NDS (CF)-73 PR/OC 2052 

1. Per?nai-al Research Branch 

2. Office of the Chief 
3o Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The First Year of Life 

Previous Serial Number: Same 

Principal Investigators: Joseph S. Drage, M.D., Chief, Perinatal Research Branch, 

C&FR, NINDS 
Janet B. Hardy, M.D., Associate Professor of Pediatrics, 

The Johns Hopkins University 
Esther Jackson, Head, Section on Applied Statistics, 

Office of Biometry, C&FR, NINDS 

Other Investigators: None 

Cooperating Units: The Johns Hopkins University; Perinatal Research Branch, C&FR, N!NDS| 

Office of Biometry, C&FR, NINDS 



Man Years: 




Total: 


,02 


Professional: 


.01 


Other: 


.01 



Project Description: 

Objectives: "The rirst Year of Life" is planned as a volume to report on tSe frequency 
distribution of a number of findings reported on Collaborative Perinatal Project children 
during the first year of their lives. It will include information on birthweight-gestation 
distributions, bilirubin levels^ age at hospital discharge^ and distributions of various 
pathological findings detected during '■tie nursery stay ana during the first year of life. 
Of particular interest will be information regarding brain abnormality as detected during 
the nursery period. This volume is intended to serve as a general description of the 
Collaborative Perinatal Project children during their first year of life and as a reference 
document for further in-depth studieso A draft of the manuscript, tables, and graphs is 
nearing completion* 

Honors and Awards: None 

Publications: None 

25 z 



Serial No. NDS (CF)-74 PR/OC 2112 

1. Perinatal Research Branch 

2. Office of the Chief 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Meonatal Hyperbilirubinemia 

Previous Serial Number: None 

Principal Investigators: Joseph S. Drage, M.D., Chief, Perinatal Research Branch, 

C&FR, NINDS 
Janet B. Hardy, M.D., Associate Professor of Pediatrics, 

The Johns Hopkins University 
Esther Jackson, Head, Section on Applied Statistics, 

Office of Biometry, C&FR, NINDS 

Other investigators: None 

Cooperating Units: The Johns Hopkins University; Perinatal Research Branch, Cc.FR, NINDS; 

Office of Biometry, C&FR, NINDS 

Man Years: 

Total: .02 

Professional: .01 
Other: .01 

Project Description: 



Objectives: The neonatal hyperbilirubinemia study has been designed !o assess the rela- 
tionship of intermediate levels of serum bilirubin on the subsequent neurological and men- 
tal Jevelopment of Collaborative Perinatnl Project children. There has been increasing 
concern that neonatal serum bilirubin levels between 10-20 ng.% may be damaging to the 
central nervous system; not in the classicol sense of "kerricterjs" associated Vi^ith levels 
above 20 mg.%, but rather damaging in more subtle yet clinically significant ways. Neo- 
nates are to be studied in five birthweight-gestationol age categories, by three socio- 
economic classes, for a varieiy of outcome measures; incluci'ng mental and motor assess- 
ments at 8 months of age, and a spectrum of neurological findings at age one year, which 
will include motor performance, reflexes, tone, abnormal movements, eye findings, and 
the over-all neurological classification of normal, suspect, or abnormal. 

Honors and Awards: None 

Publications: None 27 z 



Serral No. NDS (CF)-63 PR/PSC 1163 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: An Investigation into the Relationship Between Congenital Heart 
and Great Vessel Anomalies and Selected Maternal Factors as 
Recorded in the Collaborative Perinatal Research Projects 

Previous Serial Number: Same 

Principal Investigator: Lenore Bajda^ M.D., PRB, NINDS 

Other Investigators: None 

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 vari- 
ables and congenital heart-great vessel anomalies. 

Collaborative Study records received from onset of the project (January l'^59 through De- 
cember 1964) provided for this investigation !12 live and stillbirth congenital heart cases 
in a population of approximately 38,000. Only cases with a specific diagnosis supported 
by catherizution, surgery, or autopsy were used for a st'.tistical analysis oi relc^ionships 
between nioterna! factors and pregnancy outcome. Maternal paramerers used included age, 
gravidity, parity, prior pregnancy outcome, general health, blood group, smoking history, 
and viral antibody status. 

Preliminary analysis indicated that there was a definite preponderance of mothers over 30 
in the C-V Study grjup. Controlling for race, and removing cases with chromosomal v^ber™ 
rations, 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 systerr.ic disease complications and prior pregnancy loss among the 

29 z 



Serial No. NDS (CF)-63 PR/PSC 1163 

mothers of the cardiacs. A breakdown of these factors for greater specificity is pending 
completion of data processing of the entire study sample. The consequent increase in 
study sample size should support earlier findings and perhaps provide further clues for 
identifying etiological relationships. 

The urgent priority of processing the entire Collaborative Project study data hos temporar- 
ily delayed further work on this study. 

L 

Honors and Awards: None 
Publications: None 



30 z 



Serial No. NDS (CF)-63 PR/PSC 1181| 

1. Perinatal Research Branch 

2. Professional Staff Consiiltants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^+ 

Project Title: Population Dynamics of Tay-Sachs Disease and other 
Sphingolipidoses 

Previous Serial Number: Same 

Principal Investigators: Dr. N.C. Myrianthopoulos , PRE, NINDS 

Other Investigators: Dr. Stanley Aronson, Brown University 

Cooperating Units : None 

Man Years : 

Total: .05 
Professional: .05 
Other: .00 

Project Description : 

Objectives : To confirm experimentally the epidemiologic finding that the 
selective advantage of the Jewish TSD heterozygote is due to possible 
protection of the heterozygote from tuberculosis. 

Methodology : Experiments are in progress to measure the rate of growth of 
the mycobacterium tuberculosis in media with and without hexosaminidase A, 
and the rate of infection by the mycobacterium of tissues with e.nr' :ri.thouL 
lipid accumulation. 

M ajor findings : None 

Honors and Awards : None 

Publications: None 



31 z 



Serial No. NDS (CF)-65 PR/PSC 127^ 

1. Perinatal Research Branch 

2. Professional Staff Constiltants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Indiridual Project Report 
Jiily 1, 1973 through June 30, 197^ 

Project Title: Genetic Bases of Neonatal Reflexes 

Previous Serial Number : Same 

Principal Investigators: Dr. A.F. Waylor, PRB, NINDS 

Other Investigators: Dr. W.Co Myrianthopoulos , PRB, NINDS 

Cooperating Units: None 

Man Years : 

Total: .00 
Professional: .00 
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 frequenices of losses of suck, rooting, 
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, ant 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 



33 



qz 



Serial No. NDS (CF)-65 PR/PSC 12T6 

1. Perinatal Research Branch 

2. Professional Staff Consultant 

Section 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 



Project Title: 



Sequential Aspects of Occurrence of Spontaneous Abortion in 
Family Histories 



Previous Serial Number: Same 

Principal Investigators: Dr, A.F. Naylor, PRB, NINDS 

Other Investigators: Dr, Dorothy Warburton, College of Physicians and 

Surgeons of Colimibia University 

Cooperating Units: None 

Man Years : 

Total: .25 
Professional: .20 
Others: .05 

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 -co compen- 
sate for unsuccesful pregnancies. Also conditional risks have bee^i estimated. 

Proposed cour^oe: A manuscript arguing that age effects arc real but laying 
emphasis en the decided importance of parity effects has been accepted for 
pubxicati.on. Dr. Warourton is to bogin a draft of the second manuscript which 
will deal with conditional risks of spontaneous abortion. 



Honors and Awards : None 

Publications: Naylor, A.F.: Sequential aspects of spcntaneous abortion I. 
Maternal age, parity and pregnancy compensation art:.fact. Soc. Biol. 2l(2) 
press, 197^. 



in 



35 z 



Serial No. NDS (CF)-66 PR/PSC 1338 

1 . Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The Association of Mental Subnormal ity with Head Circumfer- 
ence, Congenita! Malformations, and Other Conditions of the 
Newborn Term Infant 

Previous Serial Number; Same 

Principal Investigator: Lenore Bajda, M.D., PRB, NINDS 

Other Investigators: None 

Cooperating Units: Office of Biomotry 

Man Years: 

Total: 

Professional: 
Other: 

Project Description: 

This study was designed to observe relationships between head size and other physical 
features at birth, ot one year and at the 4 year Psychological examination of term live 
single birth children in the Collaborative Perinatal Research Projects It was anticipated 
thct this study would function as follow-up to the Nelson &. Deutschbergt-r paper which 
used a small sample of the CRP population, (''Head Size at One Year as a Predictor of 
Four- Year IQ" (Develop. Med. Child Neurol. 12: 487-495, 1970). However, action 
on this study is in abeyance due to the urgent priority cf processing the entire Collabora- 
tive Project data for task force analysis. 

Honors and Awards: None 

Publications: None 



37Z 



Serial No. NDS (CF)-67 PR/PSC 1510 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: The Study of Maternal Effects in the Productior; of 
Congenital Malformations 

During FY 197U this project was incorporated with Serial No. NDS (CF)-7^ 

er/psc 2109. 



39 z 



Serial No. NDS (CF)-67 PR/PSC 151^ 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Record Linkage of Relatives Registered in the Collahorative 
Study 

Previous Serial Number: Same 

Principal Investigators: Dr. A.F. Naylor, PRE, NINDS 

Other Investigators: Dr. N.C. Myrianthopoulos , PRE, NIEDS 

Cooperating Units : None 

Man Years : 

Total: .25 
Professional: .25 
Other : . 00 

Project Description : 

Objectives : To identify all relatives of gravidae registered in the Collabora- 
tive Study. 

Methodology : Keypunched reports of relatives have been edited for many errors 
and the NINDS serial numbers of the registrants have been condensed into non- 
redundant family groupings. Data records from a previously created version of 
the Variable File (Varfile) have been selected and resequenced to correspond 
to the family groupings. Thus, quite usable genetically grouped data can now 
be accessed. Systematic errors in renumbering non-Core NINDS numbers and 
some suspiciously large family groups have been reviewed and corrected. 

Proposed course : Review will be completed of certain ambiguous relation- 
ships (e.g., "aunt" or "niece",) and of NINDS n-mibers suspected of 
transcription error. The file, when recreated, will be very free of 
errors and will be in form improved to facilitate familial studies on 
both Project mothers and children. 

Honors and Awards : None 

Publications: None 



h±z 



Serial No. HDS (CF)-6T PR/PSC 1515 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through Jiine 30, 197^ 

Project Title: Rh Hemolytic Disease in Negro and White Infants^ 

Previous Serial Numher: Same 

Principal Investigators: Dr. A.F. Naylor, PRE, NIWDS 

Other Investigators: None 

Cooperating Units: None 

Man Years : 

Total: .00 
Professional: .00 
Other: .00, 

Project Description : 

Objectives : To confirm a report that high Rh antibody levels have smaller 
morbid effects in Negro than in white babies , although this is not true for 
ABO antibodies. 

Major findings : Preliminary and indirect confirmation has been obtained, from 
a small data sample under study in this Section, for reports in the literature 
that high Rh antibody titers are not as highly associated with serious morbidity 
in Negroes as in whites. 

Prop ose d course : The existence of the Var.i able Data File will make possible 
the easy execution of the required data processing. 

Honors and Awards : None 

Publications: None 



^3z 



Serial No. NDS (CF)-67 PR/PSC 15l6 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Size of Placenta in Relation to Mother-Fetus Antigenic 
Difference 

Previous Serial Number: Same 

Principal Investigators: Dr. Dorothy Warturton, College of Physicians and 

Sirrgeons of Columhia University 
Dr. A.F. Naylor, PRE, NINDS' 

Other Investigators: Dr. G. Nicholas Rogentine, Immunology Branch, NCI 

Dr. Robert Cefalo, Obstetrics, National Naval Medical 

Center 

Ms. Doris Mahoney, PRB, NINDS 

Cooperating Units: Obstetrics Department, National Naval Medical Center 

Man Years : 

Total: .00 
Professional: .00 
Other: .00 

Project Description : 

Objectives ; To investigate the hypothesis published by Billington in 1964, 
that sensitization of the mother during pregnancy against paternal antigens 
leads to non-p'ithological placental hypertrophy and increased birthweight in 
succeeding pregnancies. 

Major findi nf;s: Analysis of a large semple of study data has given convincing 
support to the hypothesis. A paper reporting the evidence has been published. 

P roposed course : The arrangements for a laboratory study, described in the 
1969-70 Annual Report have been active and hk maternal sera have been tested 
against paternal leucocytes. Logistic complications which cause poor data 
return for effort have stopped collection of new cases. The accumulated data 
will soon be reviewed for publishability. 

Honors and Awards : None 

Publicationij : None 

45 z 



Serial No. NDS (CF)- 69 PE/PSC 1?^ 

1. Perinatal Research Branchy, NIKDS 

2. Section on Professional Staff 
Consultants 

3. Bethesda^ Maryland 

PHS-WIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Neonatal Polycythemia: I. A Manifestation of Chronic 
Injury Daring Distress 

Previous vSerial Number: Same 

Principal Investigator: Miles M. Weinberger^ M.D.^ Greenbrae, California 

Other Investigators: Arthur Oleinick, M. D., Portland Oregon 

John A. Churchill;, M.D.;, Harper Hospital^, Detroit, Mich. 

Cooperating Units: None 



This project has been discontinued due to the departure of the senior 
investigator. 



kT z 



Serial No. NDS (CF)-69 PR/PSC 175-^ 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Growth and Intellectual Development of Children from 
Interracial Matings 

Previous Serial Number: Serial No. NDS (CF)-69 PR/BS 175^ 

Principal Investigators: Dr. Lee Willerman, University of Texas at Austin 

Other Investigators: Dr. Alfred F. Naylor, PRB, NINDS 

Dr. Ntinos C. Myrianthopoulos , PRB, NINDS 

Cooperating Units: Department of Psychology, The University of 

Texas at Austin 

Man Years: 

Total: .35 
Professional: .30 
Other: ,05 

Project Description : 

Objectives : Results of analyses of growth and intellectual development have 
been prepared for separate publication. 

The conclusions reached in an initial publication in Science that analyses of 
CPP interracial for IQ data indicate that race of mother as a postnatal environ- 
mental indicator accounts for much of the black-white IQ have been strengthened 
in a paper published this year. Analyses of Bayley Mental and Ilotor Scores, 
taken at 8 months , when maternal social influences will have had little effect 
show no differences between children of black/white and white/black matings. 
Also a luore con zincing demonstration has been made of lack of bias in genetic 
or socio-economic factors affecting foirr year IQ on the paternal side. 

The physical development data have been re-analyzed to compare all properly 
selected CPP interracial and monoracial matings. Hospital variation and other 
background factors were corrected for by multivariate ''"egression. At birth 
children born to white mothers, whether by white or black fathers, are very 
similar in weigh"G and length. Monoracial blacks are definitely smaller and 
Interracials with black mothers may be intermediate in cize (small numbers 
cloud the issue}. At four months interracials with white mothers fall 
behind in weight (but catch up after one year), perhaps because of social 
stresses in the household. A paper has been subaitted for publication. 

h9z 



Serial No. NDS (CF)-69 PR/PSC 175^ 

This Project was inadvertently omitted last year and is still in progress. 

Honors and Awards : None 

Publications : Willerman, L. , Naylor, A.F., Myrianthopoulos, N.C.: 

Intellectual DeA'elopment of Children from Interracial 
Matings: Performance in Infancy and at k Years. 
Behav. Genet. , 197^, Vol. k, pp. 83-90. 



50 z 



Serial No. NDS (CF)-70 PR/PSC 1850 

1. Perinatal Research Branch 

2. Professional Staff Consultants 
Section 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: The Clinical Signs of Toxemia of Pregnancy and Their 
Association with the Outcome of Pregnancy 

During FY 1974 this project was incorporated with Contract Narrative 
Number: NOl-NS-3-2320 . 



51Z 



Serial No. NDS (CF)-TO PR/PSC l857 

1. Perinatal Research Branch 

2. Professional Staff Consultant 

Section 

3. Bet has da, Maryland 



PHS-NIH 
Individual Project Report 
Jiily 1, 1973 through June 30, 197^ 



Project Title: The Genetics of Intellectual and Motor Performance 

Previous Serial Number: Same 

Principal Investigators: Sarah H. Eroman, Ph . D . , PRB-NINDS 

Other Investigators: N. C. Myrianthopoulos, Ph.D., PRB-NINDS 

V. E. Anderson, Ph.D., University of Minnesota 
Paul L. Nichols, Ph.D., PRB-NINDS 

Cooperating Units: The Dight Institute for Hiiman Genetics, University of 

Minnesota 

Man Years : 

Total: .Ok 
Professional: .02 
Others: .02 

Project Description: 

Qhjectives: The objectives of the study are to assess the contribution of 
genetics to the variance of behavioral measures, particularly intellectual 
and motor performance, by correlating scores and measixrements of twin, 
sibling and half-sibling pairs. The results of the psychological tests 
given at ag^s erght months, four years and seven years have been anaJ.yzed. 
As a physical parallel, correlations for height, weight and heaa 
circumference at several ages have been computed. 

Analyse? and the p-^eparation of additonal papers are continuing. 

Honors and Awards: None 

Publications: 



Nichols, F. L. : The effects of heredity and environment on intelligen::e test 
performance in k and '( year old white and Negro sibling pairs. Doctoral 
dissertation, University of Minnesota, 1970. 



53 z 



Serial No. NDS(CF)-TO PR/PSC l85T 



Myrianthopoulos , N. C, Nichols, P. L., Bromarx, S. H., and Anderson, V. E.: 
Intellectual development of a prospectively studied population of twins. 
In Proceedings of the Fourth International Congress of Human Genetics , Paris , 
6-11 September 1971. Amsterdam, Excerpta Medica. 

Nichols, P. L. and Anderson, V. E.: Intellectual performance, race and 
socioeconomic status. Soc . Biol . , December 1973, in press. ^ 

Nichols, P. L. and Broman, S. H.: Familial resemblance in infant mental 
development. Dev. Psychol . , May 197^, in press. 



5h z 



Serial No. NDS (CF)-71 PR/PSC 1904 

1 . Perinat-al Research Branch 

2. Professional Staff Consultant-s 
Section 

3. Bethesdc, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The Study of Labor and Delivery 

Previous Serial Number: NDS (CF)-71 PR/OB 1904 

Principal Investigator: Primarily a Task Force Investigation 

Dr. Vincent Tricomi, Chairman 

Other Investigators: Emanuel A. Friedman, M.D., Harvard University Medical 

School, Boston, Massachusetts 
Luke Gillespie, M.D., Boston Lying-in Hospital, 

Boston, Massachusetts 
Marvin Green, M.D., New York Medical College, New York, 

New York 
Esther Jackson, Office of Biometry, NINDS 
Schuyler G. Kohl, M.D., State University of New York, 

Downstate Medical Center, Brooklyn, New York 
Kenneth R. NIswander, M.D,, University of California 

at Davis, California 
David Rubinstein, Office of Biometry, NINDS 
Vincent Tricomi, M.D., Brooklyn-Cumberland Medical 

Center, Brooklyn, New York 

Cooperating Units: All Institutions participating In the Collaborative Project 

Man Yeors 

Total: 0.2 

Professional: 0.1 
Other: 0.1 

Project Description: 

This study has two objective;;; 

A. To determine the influence of specific !abor and delivery factors on the 
fetus In terms of immediate outcome and later neurologic and psychologic 
development. 

55 z 



Serial No. NDS (CF)-71 PR/PSC 1904 

B. To develop a reference standard of actuarial, obstetric, and labor and delivery 
features that will result in optimal outcome. 

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. FRIEDMAN 
has demonstrated in a long series of publications that these associations do not necessarily 
reflect the underlying physiology or pathology of uterine activity. He devised a method- 
ology 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 dyscoor- 
dination 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 (Pediatric-Neurology, 
Pathology, Physical Growth and Development, etc.) as variables for their respective 
special studies. 

Methodology: 

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 v/ith incomplete data on labor and delivery in the Study record will be 
identified ciid their general characteristics (race, age, parity, sex of fetus) and outcomes 
will be tabulated separately. 

Initial computer printouts have been obtained and are in the process of being analyzed. 
Further work in this area is waiting for a toxemia classification being determined under 
contract. 

Honors and Awards: None 

Publications: Friednian, E. A. and Kroll, B. H.: Computer analysis of labour progression. 
J. Obstet. Gynaacol. Br. Commonw. 76:100-1079, 1969. 

Friedman, E. A. and Kroll, B. H.: Computer analysis of labor progression. II. Distribu- 
tion of data and limits of normal. J. Reprod. Med. 6:43-48, 1971. 

Friedman, E. A. and Kroll, B. H.: Computer analysis of labor progression. III. Pattern 
variations by parity. J. Reprod. Med. 6:63-67, 1971. 

Friedman, E. A. and Kroll, B. H.: Computer analysis of labor progression. IV. Diag- 
nosis of secondary arrest of dilatation. J. Reprod. Med. 7:176-178, 1971. 

56 z 



Serial No. NDS (CF)-71 PR/PSC 1904 

Friedman, E. A. and Kroll, B. H. : Computer analysis of labor progression. V. Effects 
of fetal presentation and position. J. Reprod. Med. 8:117-121, 1972. 



37 z 



Serial No. EDS (CF)-71 PR/PSC 1907 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Congenital malformations in the Collaborative Study 
I. Epidemiologic survey in singletons 

During FY 197^ this project was incorporated with Serial Wo. NDS (CF)- 7^ 

pr/psc 2109. 



59: 



Serial No. NDS (CF)-Tl PR/PSC 1908 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: A chromosomal study of children in the Collahorative Study 

During FY 197^ this project was incorporated with Serial No. NDS (CF)- 74 

pr/psc 2109. 



61 z 



Serial Wo. NDS (CF)-71 PR/PSC 1910 

1. Perinatal Research Branch 

2. Professional Staff Consultant 

Section 

3. Bcthesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Genetics of Obstetric Variables and the Role of Maternal 

Factors in the Determination of Intelligence and Neurological 
Performance 

Previous Serial Number: Same 

Principal Investigators: Dr. A.F. Naylor, PRE, NINDS 

Dr. N.C. Myrianthopoulos, PRE, NINDS 

Other Investigators: Dr. D. Warburton, College of Physicians and Surgeons 

of Columbia University 

Cooperating Units: Department of Hun-an Genetics and Development, Colimbia 

University 

Man Years : 

Total: M 
Professional: .hO 
Others: .05 



Project Descrition: 

Objectives : To analyze the variation of obstetric and gynecological factors 
into heritable and non-heritable com.ponents. 

Maj or find i ngs : The completed Record Linitage files hare been used to estimate 
genetic con:pcnents in obstetric variables, both continuous and discrete: age 
at menarche, lengths of stages of labor, gestation time, diagonal conjugate, 
anemia, toxemia, albuminuria, hemorrhage, abnormal preoentation, abruptio 
placentae, fetal death and nine others. A manuscript detailing the resixlts 
has been sent to a journal for possible publication. 

Honors and Awards: None 

Publications : None 



63 z 



Serial No. NDS (CF)-Tl PR/PSC 1913 

1. Perinatal Research Branch 

2. Professional Staff Consultant 

Section 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through Jujie 30, 197^^ 



Project Title: The Clinical Significance of Generalized Petechiae at Birth 

Previous Serial Number: Same 

Principal Investigators: Luz A. Froehlich, M.D., NIAID 

Jean G. Oliver, M.A., PRB, NINDS 

Other Investigators: Jean S. Nemore, R.N., B.A., PRB, NINDS 

Cooperating Units: All Collaborating Institutions 

Man Years : 



Total: 


0.0 


Professional: 


0.0 


Others : 


0.0 


Current Status: 





Evaluation of selected variables did not show any significant differences 
between cases with significant petechiae and matched controls with respect 
to speech, language and hearing findings. Other variables should be 
considered before another evaluation is made. However, eight-year data 
were not yet available for a significant number of cases at the time of the 
first evaluation. Therefore, this project has been temporarily suspended 
until 8-year data collection is completed and in house. 

Honors and Awards: None 

Publications : None 



65 z 



Serial No. NDS (CF)-T2 PR/PSC 1968 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Preschool IQ: Prenatal and Early Developmental Correlates 

Previous Project Title: Prenatal and Postnatal Factors Associated with IQ 

at Age Four 

Principal Investigators: Sarah H. Broman, Ph.D., PRB-NINDS 

Paul L. Nichols, Ph.D., PRB-NINDS 
Wallace A. Kennedy, Ph.D., Florida State University 

Cooperating Unit: Florida State University 

Man Years : ■ ■ 

Total: 1.3 
Professional 1.0 
Other : . 3 

Project Description: 

Objectives : The relationship between intelligence at age four as 
measured by the Stanford-Einet Intelligence Scale and a variety of factors 
related to pregnancy, delivery, postnatal status and social environment 
was examined among 26,760 white and Negro childrn enrolled in the 
Collaborative Perinatal Project. 

Method : The plan of analysis of these data was exploratory, beginning with 
an identification of antecedents related to IQ. The i-esults of a variable 
screen in which 169 prenatal, postnatal and environmental factors were 
correlated with IQ are presented. Th'? 83 variables found to be signi- 
ficantly related to IQ are defined and their distributions in the sample 
are given. Mean IQs for different levels of each pvecactor variable are 
displayed within race, sex, and social class controls. Significant main 
effects and interactions are discussed. The combined effects of the 
antecedent variables were examined in a multiple regre£.32on model, and also 
in a discriminant function model with subjects divided into low and normal 
IQ groups. Methodological issues discussed include the selection of an 
outcome variable at four years, the problem of non-linear relationships 
between antecedent variables and IQ, and the effect; of combining data from 
different study centers. 



67 z 



Serial No. NDS (CF)-T2 PR/PSC I968 

Major Findings: All predictor variables accounted for only 25 to 28 percent 
of the variance in IQ among whites and 15 to 17 percent among Negroes, The 
prenatal variables alone accounted for 20 percent of the IQ variance 
among whites , and eight to 11 percent among Negroes , with maternal 
education and the socioeconomic index contributing the largest proportion 
of explained variance not only in this first analysis but also when 
neonatal and childhood variables were added. The estimated effects 
of maternal education and the SEI on IQ were greater in the white than in 
the Negro samples. The neonatal variables accounted for only an additional 
one to two percent of the variance, while the infajicy and childhood 
variables added four to six percent. 

Prior to birth the most consistent predictor of mental retardation at age 
four was maternal education. When variables from the neonatal period were 
included a diagnosis of brain abnormality, size of the neonate (head 
circumference or length), maternal education and socioeconomic index of 
the family were significantly associated with retardation. After status 
during the first year of life and size at four years were considered, the 
factors most highly related to retardation were delayed motor development 
at one year, mental and motor test scores at eight months, maternal 
education, and relatively rare conditions associated with neurological 
impairment during the first year of life. 

Proposed Com'se: This study has been completed. Publication plans are 
in progress. 

Honors and Awards: None 

Publications: 



Broman, S. H., Nichols, P. L., and Kennedy, W. A.: Precursors of Low IQ 
in Young Children. In Proceedings of the BOth Annual Convention of the 
American Psychological Association, Honolulu, Hawaii, September 1-8, 1972, 
pp. 77-78. 



68 2 



Serial No. NDS (CF)-T2 PR/PSC I969 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: A Study of genetic markers in children of the Collaborative 
study 

Previous Serial Number: Same 

Principal Investigators: Dr. N.C. Myrianthopoulos, PRE, NINDS 

Dr. Henry Gershowitz, University of Michigan 

Other Investigators: None 

Cooperating Units: Collaborating Institutions at Boston, Buffalo, Memphis 

and Oregon 

This study has been discontinued because the contract expired. 






Serial No, NDS (CF)-72 PR/PSC 1971 

1. Perinatal Research Branch 

2. Professional Staff Consultant 

Section 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 



Project Title: Biosocial factors Associated with Commvinicative Disorders 
and Competence in Children 

Previous Serial Number: Same and incorporating Serial No. NDS (CF)-72 

PR/PSC 1970 

Principal Investigators: Paul J. LaBenz, Sc.D., PRE, NINDS 

Other Investigators: Leo G. Doerfler, Ph.D., Pittsburgh Eye & Ear Hospital 

Harry M. Beirne, M.D. , Children's Hospital, Buffalo 
Luke Gillespie, M.D., Children's Medical Center, Boston 
John V. Irwin, Ph.D., Memphis State University 
Frank M. Lassman, Ph.D., University of Minnesota 
Mrs. Jean G. Oliver, PRB, NINDS 
Mr. David Rubinstein, OB, NINDS 
Earl D. Schubert, Ph.D., Stanford University 
Warren Torgerson, Ph.D., Johns Hopkins University 
Peter Workman, Ph.D., University of Massachusetts 

Cooperating Units: All institutions participating in the Collaborative 

Project 

Man Years : ' 

Total: • 1.3 
Professional: 1.2 
Others: .1 

Project Description: 

Speech, language and hearing (SLH) data have beer, collected on about 15,000 
children at age 3-years, and additional data are being collected at age 
6~years. Analysis is being planned to achieve four main objectives: 

1. Estimation of the incidence of SLH deficits relative to de.uographic 
characteristics of the s'cudy population; 

2. Selection of the most meaningfuJ and reliable SLH variables f.nd indices 
to provide an economical descriptor of commimicative behavior; 

3. Discovery and delineation of relationships between SLH and perinatal 
variables, especially those relating to neurology; 

71 z 



Serial No. NDS (CF)-72 PR/PSC 1971 

k. Improvement of SLH methodology for clinical and research applications. 
Method: 

Data are to he tabulated variously in order to assess the influence of test 
procedures and demographic factors upon the variance of scores, and to enable 
estimates of the incidence of communicative disorders as defined operationally. 
Factor analysis techniques will be applied to derive a minimal set of indices, 
or a single composite index, which can serve to describe communicative status. 
Correlation methods will be used to establish relationships with variables 
of interest. These variables will be drawn from documented areas relating 
to family history, maternal pregnancy and delivery, neonatal and subsequent 
examinations, intercurrent health events, and measured physical and mental 
characteristics . 

Current Status : 



Tabulations of the available 3-year data were made by items and by combinations 
of items, scored according to protocol and by revised criteria. Alternative 
methods of scoring were found not to contribute materially to improved descrip- 
tion of communicative performance. Correlation matrices were constructed to 
explore intratest relationships at the 3-year level, as well as relationships 
with 30 selected 8-year, perinatal, and other veiriables. Several clusters 
of associations were discerned. Tab\ilations were run on the 8-year data 
presently available. Data collection will be completed at the end of the 
cvurrent fiscal year. Crude estimates of the prevalence of certain communi- 
cative deficits were derived from both the 3-year and the 8-year data. 
Tentative selection was made of potential key variables for indices, and a 
strategy was developed for constructing indices relating to several aspects 
of commiinicative performance. Specifications have been drawn for conducting 
detailed analyses of the SLH data. Efforts are underway to arrange for the 
analysis and interpretation of the data extramurally under contract. 

Publications: None 

Honors and Awards: None 



72 2 



Serial No. ffl3S (CF)-T2 PR/PSC 1972 

1. Perinatal Research Branch 

2. Professional Staff Consultant 

Section 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 



Project Title: Relationships between Spontaneous Speech and Other Test 
Results at Eight Years of Age 

Previous Serial Number: Same 

Principal Investigators: Jean G. Oliver, M.A., PRB-NINDS 

Other Investigators: Pa\il J. LaBenz, Sc.D., PRB-NINDS 

Cooperating Units : All Collaborating Institutions 

Man Years: 

Total: .08 
Professional: .05 
Others: .03 

Project Description; 

The study of spontaneous speech can provide useful insights into communicative 
skills, and appears to be a valid index of expressive language proficiency. 
Samples of spontaneous speech, elicited by visual stimuli (pictures) are 
being tape-recorded opportimistically at quality control visits where 8-year 
old children are retested. These samples are later transcribed for analysis. 
Performances are rated for comparison with data derived from the standard 
speech, langua/je and hearing battery. 

Method: 



A tentative set of rating scales has been devired to enable evaluation of 
several aspects of spontaneous speech output. 'iTiese include the length of 
the speech sample as well as the content in terms of quality, organization, 
elaboration and grammatical structure. Relationships between spontaneous 
connected speech and a number of traditional measures of communicative 
ability are to be explored using correlation methods. 

Current Status : 



Continuation of individual project with eight-year spontaneous language 
samples. Samples are completed as of June 30, 197^ • Following vhich 



73 z 



Serial No. WDS (CF)-72 PR/PSC 1972 



analysis will continue. 
Publications: None 
Honors and Awards: None 



7Uz 



Serial No. NDS (CF)-73 PR/PSC 2053 

1 . Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The Natural History of Congenital Toxoplasmosis: Case 
Studies from the Collaborative Perinatal Research Project 

Previous Serial Number: None 

Principal Investigator: Lenore Bajda, M.D., PRB, NINDS 

Other Investigators: Dr. John L. Sever, ID, NINDS 

M. R. Gilkeson, ID, NINDS 

Cooperating Units: Infectious Diseases Branch, NINDS 

Man Years: 

Total: .50 

Professional: .50 
Other: .00 

Project Description: 

In searching Collaborative Perinatal Research Project data for indicators of fetol risk to 
maternal Toxoplasma infection this study used a subgroup cf47 gravidae wii markedly 
elevated titres or four-fold rises in Toxo antibody and their 94 control gravidae provided 
from on initial study population of 23,000. (Sever, J. L. Toxop' jsrt'osis: Serological 
and Clinical Studies — Serial No. NDS (CF)-65 PR/ID 1270). Al! pertinent data in the 
Collaborative Research Project central file protocols we'e analyzed for statistically signi- 
ficant finding?. Pediatric data included al! recorded inedicc! information thru the 7 year 
Neurology examination as well as global !Q scores of t'^e 4 jnd 7 year Psychological 
examination. 

Major Findings: Maternal findings included: a) a const&tit exposure to Toxoplasmosis 
throughout the child-bearing years, b) higher exposures among Negroes and women born 
in Puerto Rico, and c) no correlation wrth urboti vs. rural life. Addirional Findings re- 
quiring further support included prior pregnancy failures within 1 year to IMP being greater 
than in rhe controls (8.5 to 3.2%) and the observation that a frequent use of sulfa drugs in 
the treatment of urinary tract infection during the pregnancy (17%) may have a sianificant 
effect on the spectrum of pregnancy outcome. Pediatric findings at a seven year endpoint 

added to the previously published one-year findings the additional obs va.i'ons of a high 

75 z 



Serial No. NDS (CF)-73 PR/PSC 2053 

rate of "undefined" abnormallHes in the case group compared to controls and a high rate 
of low IQs (24 compared to 17%). The latter finding may become less significant on ad- 
justment for birthweight and degree of maternal education. The implication of the as yet 
unknown and difficult to measure effect of drug treatment for coexistant medical problems 
in the Toxoplasma-infected mother demands further study. 

Results of this project were presented at a Symposium on Toxoplasmosis during the Xlllth 
Congress of the Medical Women's international Association, Paris, France, September 3- 
8, 1972, Publication is pending. 

Honors and Awards: None 

Rjbllcations: None 



76 



Serial No. NDS (CF)-T3 PR/PSC 205^+ 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 
3= Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197'-^ 

Project Title: Congenital malformations in the Collaborative Study. 

II. Epidemiology of congenital malformations in multiple 
births . 

During FY 197^ this project was incorporated with Serial Wo. NDS (CF)-7i+ 

pr/psc 2109. 



772 



Serial No. NDS (CF)-T3 PR/PSC 2055 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 19lh 

Project Title: Congenital malformations in the Collaborative St-udy. 
III. Study of the effects of medical, genetic and 
socioeconomic factors in the occurrence of congenital 
malf ormat ions 

During FY 197^ this project was incorporated with Serial No. HDS (CF)- jh 

pr/psc 2109. 



7'9; 



Serial Wo. NDS (CF)-73 PR/PSC 2056 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^+ 

Project Title: Congenital malformations in the Collaborative Study. 
IV. Maternal diabetes and congenitail malformations. 

During FY 197^ this project was incorporated with Serial No. NDS (CF)-7l+ 

pr/psc 2109. 



61 z 



Serial No. M)S (CF)-73 PR/PSC 2057 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. B^-thesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197U 

Project Title: Seizures with Fever, Beginning in the First Ye'ar of Life 

During FY 197^^ this project was incorporated with Serial Wo. EDS (CF)-73 
PR/PSC 2058. 



83 



Serial No. NDS (CF)-73 PR/PSC 2058 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Convulsive Disorders Data Analysis Group 

Previous Title: Data Analysis Group on Convulsive Disorders 

Previous Serial Number: Same 

Principal Investigators: Karin B. Nelson, M.D. , PRB, NIWDS 

Jonas Ellenberg, Ph.D., OB, NINDS 

Other Investigators: John Freeman, M.D., John Hopkins 

Gregory Ferriss, M.D., Louisiana State University 

Kenton Holden, M.D. , Child Development Study, Providence, 

Rhode Island 
Marianne Schuelein, M.D., Georgetown University 

Cooperating Units : All Collaborating Institutions 

Man Years: 

Total: 3.1 
Professional: l.k 
Other : 1.7 

Project Description : 

I. Objectives 

A. Etiology 

We will examine maternal characteristics, conditions of pregnancy, 
labor, delivery f.nd the neonatal period, and illnesses and injuries of early 
childhood for their association with seizure disgnoses. Our aim is to identify 
points at which preventive eff jrts could be directed. 

B. Prediction 

Once a seizure has occurred, what is the likelihood of the develop- 
ment of a chronic epileptic state? What other characteristics are to be 
anticipated? The clinically important question of prognosis requires that we 
explore for fsctcrs or clusters of factors which allow reliaDle prediction on 
the basis of fiven clinical characteristics. 



35 



Serial No. IDS (CF)-73 PR/PSC 2058 

C. Clustering of handicaps 

There are a number of different convulsive disorders of varying 
gravity and frequency. In some, no other handicap need be anticipated while 
in others deficits are often multiple. To examine and quantify these assoc- 
iations of handicaps by specific diagnosis is a third objective of this study 
area. 

D. Frequency ' 

For some seizure disorders, little is known of frequency, and CPP 
data will make a real contribution in stating the observed occurrences of 
different convulsive disorders of early childhood in a population of known 
size. Although some reservations are appropriate in generalizing from Study 
frequency data because owe population was not initially selected as a sample 
of the American population, still our figures will offer the best information 
yet available as to frequency. 

II. Methodology 

A. Case review 

With the help of a group of expert consultants, we have established 
criteria for specific diagnoses of various seizure states. We have then 
examined approximately 710 records of children in the Study thought to have 
convulsive disorders. There is an overlap of I96 cases with the CP case 
material. We are at present approximately half way through the abstraction 
and classification of seizure records, and the reduction of this clinical 
information to a codable format. ■ 

B. Analysis 

The basic proced\ires to be used in the study of seizure disorders 
are similar to the CP analysis:* first, a demographic exploration of the dis- 
tribution of specific seiz\ire diagnoses by ins-citution, year of birth, sex, 
race, and so on. Then we will explore the association betwee.. seizure 
diagnoses aiid anteceden"". characteristics of the mother, cf obstetrical events, 
of the neon;ital couise, and intervening illnesses and in.pxcies. Then the most 
prominent antecedent factors will be exanu.ned to attempt ^o assess the inde- 
pendent contribu"uion of each. The role of f ani ,y hi^.zcr/ and of malformations 
will b3 examined. Pathological infomiation will be discussed when available. 

The variables to be used in these analyses ha"«e been selected and 
defined in the forms necessary for the studies, "n some cases, dichotomies 
are to be used, and are baseO. upon available frequency analysis and other 
inforination within the study. 

D^s. Freemaxi ajid Holden are participating in the study of neonatal 
seizures. Dr. Gregory Ferriss and Dr. Mariajine Schuelein have assisted in 
classification of seizure cases. 

*Serial No. NLl(CF)-73 PR/PSC 2059 0^ 

00 z 



Serial No. NDS (CF)-T3 PR/PSC 2058 

III. Major findings 

No analyses have yet been imdertaken on t>ie data on convulsive disorders. 

IV. Significance 

The convulsive disorders are a common and socially costly medical problem. 
It is estimated that about k million Americans have some form of epilepsy. 
The cost of the epilepsies in direct welfare payments and in medical expenses - 
ignoring other costs — has been calculated at more than $H billion per year. 

As indicated in Objectives, above, we hope to generate medically useful 
information relevant to this important problem. The direction of future 
programs of prevention could be significantly influenced by information obtain- 
able in this Study. 

V. Proposed course 

It is planned to complete chart review, then to analyse the data as 
outlined under Methodology, above. 

Honors and Awards: None 

Publications : None 



87z 



Serial No. NDS (CF)-73 PR/PSC 2059 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197*+ 

Project Title: Cerebral Palsy Data Analysis Group 

Previous Serial Number: Same 

Principal Investigators: Karin B.Nelson, M.D., PRB, NINDS 

Jonas Ellenberg, Ph.D., OB, NINDS 

Other Investigators: Robert Groover, M.D., Mayo Foiondation Rochester, Minn. 

Alan Leviton, M.D., Children's Medical Center, Boston, 

Mass. 
William Clark, Jr., M.D. , University of Oregon Medical 

School, Portland Ore. 

Cooperating Units : None 

Man Years : 

Total: 3.5 
Professional: 1.6 
Other : 1.9 

Project Description : 

I. Objectives 

A. Etiology 

Oujc first responsibility in this study area is to investigate and 
interpret the interwoven factors antecedent to CP diagnoses, in an attempt to 
cast light on the etiology of CP. We examine maternal, obstetric, social, and 
neonatal conditions, and events of early childhood, and attempt to weigh their 
assocation with CP outcomes. Our aim is to identify areas in which preventive 
efforts might most usefully be concentrated. 

B. Prediction 

Given a child with suspect or definite abnormalities, the central 
question posed by parents and physician is, Wliat may we expect? We are there- 
fore exploring for fe.ctors or clusters of factors which will aid in prediction 
of the subsequent course of affected Study children. Preliminary data suggest 
that useful information may be expected in this area. 

89z 



Serial No. NDS (CF)-T3 PR/PSC 2059 

The emergence of intensive means of intervention in neonatal 
nurseries has increased the requirement for reliable indices for prediction 
of later neurological status of young infants. Neonatologists need to know 
what the neurological outcome of their intervention., may be, and they need 
as rapid as feasible a turn-around time. Oiur findings may well have relevant 
applications to this need. 

C . Clustering of handicaps 

To plan optimal medical and education management for involved 
children, and to plan the facilities in which such children receive care, it 
is necessary to know what other handicaps may be anticipated in children with 
CP. The nature, number, and severity of associated deficits will therefore 
be examined in Study children. 

D. Frequency data 

The CPP was not designed for estimates of frequency which could be 
generalized to the population at large; still, the CPP data will represent the 
best information available for estimates of the frequency of certain conditions 
in a population of known size. We will therefore examine the relative frequency 
of different types of CP as they have appeared in Study children. We will note 
as well the occurrence of such non-CP motor deficits as, e.g., the acute cere- 
bellar ataxias, as observed in our sample. 

II. Methodology 

A. Case review 

We have, with the help of a group of expert consultants, settled 
upon consistent definitions and criteria for CP outcomes. We have then 
examined the records of more than 8l5 children for CP diagnoses, including 
all children with definite and suspect CP diagnoses at seven years, and most 
of those with definite diagnoses at one year. We have reclassified th°^3 
children according to the consistent standard definitions agreed upon. 

We have examined the interval histories of all the CP car>es in order 
to seysA-ite out those children whose motor deficits appear to have followed 
distinct illnesses or injuries of the early years, in order to exclude these 
cases from test fo"' obstetrical antecedents. Acquired CP is also of substantial 
interest in its own right. The risk factors for acquired deficits are of 
medical and social concern, and may possibly lend themselves to preventive 
efforts more readily than congenital defects. 

B. Analysis 

A "dump" of the variable file on CP cased was run so that we could 
exclude from further consideration all prenatal, obstetri::al, and early 
pediatric conditions which appeared very rarely in our case material. 



90 z 



Serial Wo. HDS (CF)-73 PR/PSC 2059 

We are now planning an analysis to see how these cases are distributed 
by such demographic factors as sex, institution, race, year of birth, birth 
weight and gestational length. From these demop;raphic analyses we will seek 
basic information which will allow us to identify any factors so critical as 
to require inclusion as control variables in the next phase, the analysis of 
antecedents. 

Maternal, obstetric and early pediatric variables have been defined, 
and a data analysis request nearly completed, to examine the role of these 
factors upon the development of the infant. The approach decided upon for this 
phase is an examination of relative risk: we examine the occtirrence of, for 
example, abruptio placentae among children who do and children who do not have 
CP at 7 years. Comparing the frequencies by chi squares, we get a two x two 
table which enables us to slate that abruptio placentae raises the risk of CP 
by a factor of (say) 3- By a further manipulation of the data, we are able to 
state what proportion of the CP cases can be accounted for by abruptio placentae. 
To take another exam^jle, we might find that first trimester rubella raises the 
risk of microcephaly by a factor of 20, and that of all cases of microcephaly, 
5^ appear to be accounted for by first trimester rubella. This expression of 
relative risk and attributable risk appear to satisfy our need for a direct 
and straightforward expression of the information sought: what risks do each 
of these antecedents entail? 

The last phase of this study will take the most prominent risk factors 
identified and submit these to multivariate analysis, to try to assess the in- 
dependent contribution of each. 

III. Preliminary findings to date: 

A. First, an approximate figure of 1 CP case in 200 births. 

B. At one year, two relatively common forms of CP, spastic diplegia and 
quadriplegia, are 3 to !+ times as common among Negroes as among whites. No 
such difference by race exists at T years. We are doing what we ca.. to try to 
understand what may account for this difference at one year. 

C. Many childre.n who appeared to warrant definite cerebral palsy diagnoses 
at one ye-^r, do not show handicapping motor deficits when seven years old. 
These children remained at relatively high risk for menta] retardation, however. 
It appears possible to isolate certain factor'^ which permit prediction of 
whether CP diagnoses at one year will be lost or retained. For instance, for 
spastic diplegia and quadriplegia, of children who coijld pull to standing by 

one year of age, very few showed significant motor deficits by the age of 7- 
Other predictors also appear useful. 

IV. Significance: 

Approximately 750^000 persons in the United States are \ictinis of cerebral 
palsy; the milder forms of CP are more frequent still. As these individuals 
are afflicted from earliest childhood, the loss in economic terms, and in social 

91 z 



Serial No. NDS (CF)-73 PR/PSC 2059 

and other costs, is immense. Many are lifelong dependents of their families 
and the State. 

Our aims, as expressed in Objectives (above) are to identify areas in 
which preventive efforts may be effectively directed to improve clinical 
prognostication, to examine clustering of handicaps, and to estimate relative 
frequency of CP conditions. Useful information in these area may help to 
indicate fruitful directions for both medical and social policy decisions. 
Considering the magnitude of the problem, advances against this chronic handi- 
capping neiorological condition of childhood would seem strongly compatible 
with Institute objectives. 

V. Future Course: 

Continued effort on data analysis as outlined in Methodology, above, will 
proceed in the next year. In any time available while, awaiting machine 
analyses, investigators will undertake hand tabiilation on specific diagnotic 
categories, in pursuance of the stated Objectives. 

Honors and Awards : None 

Publications: None 



92 z 



Serial No. EDS (CF)--73 PR/PSC 2060 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Birthweight-Gestational Age 

Previous Serial Number: Same and incorporating Serial No. NjJS (CF)-73 PR/P 2066 

Principal Investigators: Dr. Bill H. Williams, PRB, NINDS 

Other Investigators: Dr. Janet Hardy, Johns Hopkins 

Dr. Joseph S. Drage, PRB, NINDS- 

Dr. Kinne D. McCabe, PRB, NINDS 

Ms. Esther Jackson, OB, NINDS 

Mr. William Weiss, OB, NINDS 

Dr. E. David Mellits, Johns Hopkins 

Cooperating Units : Johns Hopkins University 

Man Years: 

Total: 0.50 
Professional: 0.25 
Other: 0.25 

Project Description : 

During the past year a set of tables have been generated which show basic 
information (distributions, frequencies, rates) of birthweight-gestational 
age groups and their relationships with secondary parameters such as placental 
weight, maternal education, socioeconomic index, gestation at registration, 
year of birth, institution, body length at birth, head circumference at birth. 
Frequency tables , neonatal death and stiZ Iborn rates , meajis , standard 
deviations, survival figures, and horizontal and vertical percentage table 
have been generated. These variables ai'e to be related to antecedent factors 
(prenatal) and later outcomes (particularly neiirological development). 
Empirical Risk Tables for babies in certain birthweight-gestational age 
categories will be developed and a Birthweight Index will be generated which 
will be of some predictive value in relating antecedent factors to birthweight- 
gestational age outcores. Plans are underway to incorporate the entire body 
of information generated in the form of a monograph which would be of ure to 
clinicians. 

Honors and Awards : None 

Publications : Noae 

93z 



Serial No. NDS (CF)-73 PR/PSC 206i 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. P^thesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Physical Growth and Development Task Force 

During FY 197^ this project was incorporated with Contract Narrative Number: 
NOl-NS-2-2320. 



95 z 



Serial No. NDS (CF)-73 PR/PSC 2062 

1. Perinatal Research Branch 

2. Professional Staff Consultant 

Section 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^^ 



Project Title: Minimal Brain Dysfunction 

Previous Serial Number: Same 

Principal Investigators: Paul L. Nichols, Ph.D., PRB, NINDS 

Other Investigators: Ta-chaun Chen, Ph.D., OB, NINDS 

Cooperating Units: None .; ■ 

Man Years : 

Total: .85 ■ 
Professional: .75 
Others: .10 



Project Description: 

Specific diagnoses of minimal brain dysfunction have not been made 
for CPP children, but there is information available related to the most 
frequently cited symptoms (e.g., hyperactivity, learning difficulties, and 
equivocal neurological signs). This study will attempt to develop MBD 
criteria from the available CPP data, characterize MBD children in terms of 
demographic, psychological, and physical variables, and relate MBD or its 
components or individual symptoms to socioeconomic, maternal, neonatal, 
perinatal , and otner antecedent variables . 

A major effort has been made to define symptoms in terms of relevant 
variables from the 7-year psychological and neurological examinations; 
a list of 25 symptoms in h major areas has been .-compiled: behavioral, 
cognitive and perceptual-motor, academic, and phsycial. 

Associations will be exajnined among these symptoms. In selected 
samples, U-year and 8-year data will also be included. A listing of 
cases grouped by symptom combinations will be examined to furtner 
group symptom patterns and to identify cases for chart review, 
possibly by outside clinical consultants. In addition, combinations 
of outcome variables will be derived using statistical procedures 
of increasir<5 degrees of complexity. Cluster and factor analytic 
techniques will be employed. 

97 z 



SeriaO. No. NDS (CF)-T3 PR/PSC 2062 



MBD children vill be characterized in terms of demographic variables, 
IQ, and other psychological variables, and compart i to "normal" children. 
These comparisons will help determine whether control groups shoiild be 
composed of laj*ge random samples, smaller matched groups, or both. 

Antecedent variables to be investigated will be selected from among 
family characteristics, maternal characteristics, the prenatal period, labor 
and delivery variables, the neonatal period, infancy and childfiood charac- 
teristics, and intercurrent events. It is expected that a miinimum of 300 
antecedent variables will be examined. The selection and definition 
of many of these variables will be in collaboration with investigators in 
other primary areas. For the purposes of preliminary analyses and screening, 
variables from all periods will be treated together. 

Antecedent variables will be examined in combination using multiple 
regression or discrimir.ant function techniques where appropriate. The ability 
of variables from different periods (prenatal, neonatal, etc.) to differentiate 
between affected and "normal" children will be investigated. 

Based on test data already received, the following preliminary observations 
are possible: (l) Several hundred children have been identified who have 
symptoms in two, three, or even all four of the major areas described above. 
(2) The risk of problems in one area is increased between kofa and 100^ if a 
child has problems in one of the other four areas. That is, there is a 
significant association among the symptoms in the four areas. (3) Most 
children, however, who have problems in one area (e.g., physical) do not have 
problems in the other areas (e.g., behavioral or academic). In other words, 
the association among symptoms is not a strong one. 

Honors and Awards : None 

Publications: None 



98 



Project Title: 



Serial No. NDS(CF)-Ti+ PR/PSC 2106 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Developmental Factors Associated with Mental I^etardation 
at Age Seven 



Previous Project Title: None 
Previous Serial Number: None 



Principal Investigators ; 

Cooperating Unit: None 
Man Years : 



Sarah H. Broman, Ph.D., PRB-NINDS 
Paul L. Nichols, Ph.D., PRB-NINDS 
Peter W. Shaughnessy, Ph.D., OB-NINDS 



Total 1.2 

Professional 1 
Other .2 

Project Description: 

Objectives : Mental retardation is a serious social problem. Approximately 
three percent of the population is affected according to the conventional 
criterion of intelligence t^st scores under 70. Data collected in the CPP 
are being analysed to determine the primary and contributing roles of 
biological, environmental and familial factors in mental retardation in a 
biracial popuJ.ation of itO,000 seven year old children followed since the 
prenatal period. In this popxilation, approximate!,/ two percent of the 
vl'ito nample and fiA^e percent of the Negi'C sample nave IQs under 70. The 
identification of early signs of mental retardation will facilitate pre- 
vention, diagnosis and treatment, and will add substantially to knowledge 
in this area which has been largely derived f ro; i small retrospective 
studies of institutionalized retardates, The objectives of thi'i study are 
to determine the degree of association between mental i-etardation to age seven 
and the following classes of variables: 

l) Biological factors 



a) Complications of pregnancy and delivery 

b) Adverse neonatal conditions 



99; 



Serial No. NDS (CF)-?^ PR/PSC 2106 

c) Neurological and general medical status at one and 
seven years 

d) Childhood diseases and accidents 

e) Physical growth rates 

2) Socio-environmental factors 

a) Socioeconomic status and social mobility of family 

b) Parental education 

c) Family composition 

d) Geographic area of residence 

3) Familial factors: Qcciirrence of retardation in relatives 

a) Monoi:ygotic and dyzygotic twins 

b) Full and half-sibs 

c) Other relatives 

k) Cognitive development 

a) Mental and motor development at eight months 

b) IQ scores, fine and gross motor development and 
conceptual development at age four 

c) Conceptual, visual -motor and behavioral development 
at age seven 

Method; In this study, mental retardation is defined as an IQ of TO or 
less on the Weschler Intelligence Scale for Children, or, for the relatively 
few children who could not be tested according to study protocol, 
equivalent IQs from other tests or reliable clinical Judgements of re- 
tardation. This latter soixrce of classification applies mainly to severely 
retarded children who have been institutionalized. 

Since children with IQs under JO form a heterogeneous group, they will be 
subdivided into four major categories consisting of those with severe 
retarciation (IQ under 50) with and without signs of central nervous system 
damage, and those with mild retardation (IQ between 50 and 69) with and 
without such signs. Specific neiirological diagnoses, such as cereb".'al 
spastic paresis, seizure states, Down's syndrome (mongolism) and head trauma, 
will be obtained from the neurological examination given at age seven. 
Ths four groups will be further subdivided by ethnicity (white, Negro 
and Puerto R\can) and, where the nvunter of cases ir r-uffic3 ent , by sex. 
Comparison groups will be composed of cnildren with IQs in the borderline, 
average and superior ranges. These groups will also be checked for signs 
of central nervous system damage and will be subdivided by ethnicity and 
sex. Factors that are associated with performance in the average and 
superior groups, and are most usefuJ. in differentiating these groups from 
the mentally retarded, will be examined. 

Major Findings: Preliminary findings indicate that the following conditions 
are related to e low level of intellectual functioning at age seven: Low 

100 z 



Serial No. NDS (CF)-?^ PR/pSC 2106 

socioeconomic status of the family which includes income, and occupation and 
education of the parents, high housing density, low maternal intelligence 
test scores, less prenatal care, less advanced mental and motor development 
at 8 months , poorer language development at age three , lower psychometric 
test performance at age four, and smaller physical measurements during the 
first 7 years of life. A large sample, soon to be available, is necessary 
to detect relationships between mental retardation and relatively infrequent 
events, such as serious medical complications in the prenatal, neonatal and 
early childhood periods. 

Proposed Couxse: Plans for the analysis of CPP data on mental retardation 
fall into a preliminary phase, a pilot phase and a main study phase. The 
current pilot phase consists of an examination of associations between IQ 
level (retarded, average and above-average) and 59^ variables from the 
prenatal, socio-environmental, medical and psychological histories of the 
30,000 children whose records are now on tape. This phase is expected to be 
completed in November of 197^ • The main study phase, in which analyses 
will be made on the total popiilation of approximately 1+0,000 children, 
should be completed by December of 1975? leaving approximately 6 months for 
the preparation of a final manuscript presenting the findings. 

Honors and Awards : None 

Publications: None 



101 z 



Serial No. NDS (CF)-74 PR/PSC 2107 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The study of visual abnormalities in the Collaborative Study 

Previous Serial Number: None 

Principal investigator: Dr. Richard Feinberg, PRB, NINDS 

Other Investigators: Ms. Esther Jackson, OB, NINDS 

Richard J. Apell, O.D., Gesell Institute of Child Development, 
Nev/ Haven, Connecticut 

William R. Baldvi'in, O.D., Ph. D., Massachusetts College of 
Optometry, Boston, Massachusetts 

Richard E. Hoover, M.D., Baltimore, Maryland 

Robert P. Kling, M.D., Georgetown University Hospital, Wash- 
ington, D. C. 

Milton A. Whitcomb, Ph. D., National Academy of Sciences, 
Washington, D. C. 

Sheila Z. Wood, O.D., Washington, D. C. 

Francis A. Young,. Ph. D., Washington State University, Pullman, 
Washington 

Cooperating Units: None 

Man Years: 

Total: 1.2 

Professional: 1.2 
Others: 0.2 

Project Description: 

Objectives: To determine the extent to v/hich genetir, matarnal, obstetric, pediatric and 
environmental factors produce eye and visual abnormalities in the Study children; to assess 
the relative frequency of such anomalies, and, to study the concomitance of visual abnor- 
malities with other sensory and motor neurologica! and systemic disorders. 



1037. 



Serial No. NDS (CF)-74 PR/PSC 210? 

Methods Employed: A reference cohort of visual defecl^ is defined for which the specified 
outcome will be tabulated. Outcome variables have been selected. Initial computer 
printouts have been obtained and are in process of being analyzed. 

Major Findings: At this stage, no major findings can be reported. 

Proposed Course: This will include: the analysis between visual abnormalities and outcome 
variables; anecdotal treatment based on case histories of unusual visual abnormalities; spe- 
cial studies of high-incidence disorders and of concordance in siblings and twins. 

Honors and Awards: None 

Publications: None 



lOi+2 



Serial No. NDS (CF)-T1+ PR/PSC 2108 

1. Perinatal Research Branch 

2. Professional Staff Consiiltants 

Section 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Developmental Factors Associated with Learning Disorders 
at Age Seven 

Previous Project Title: None 

Previous Serial Number: None 

Principal Investigators: Sarah H. Broman, Ph.D., PRB-NINDS 

Paul L. Nichols, Ph.D., PRB-NINDS 
Peter W. Shaughnessy, Ph.D., OB-NINDS 

Cooperating Unit: None 

Man Years : 

Total 1.2 

Professional 1 

Other .2 ' ' 

Project Description : 

Objectives: Children with normal intelligence and poor school performance, 
particularly in reading, present significant problems in etiology and 
the development of effective remedial techniques. Longitudinal data 
collected in the CPP on a biracial population of U0,000 seven year old 
children permit a study to be made of early events, beginning in the prenatal 
period, which differentiate between children with learning disorders in the 
first and second grades and those without a significant discrepcJicy betveen 
intellectual ability and school perform nice. The accurate identification of 
precursors of behavior patterns iden^ified as learning disorders will 
facilitate prevention, early diagnosis and treatment. The objectives of 
this stuciy are to determine the degree of association between learning 
disorders at age seven and the following classes of variables: 

l) Biological factors . - 

a) Complications of pregnancy and delivery 

b) Matarnal drug intake during pregnancy 



105 z 



Serial Wo. WDS (CF)-'jk PR/psC 2108 

c) Adverse neonatal conditions and early abnormal 
neurological signs 

d) Neurological and general medical status at one and 
seven years 

e) Childhood diseases and accidents 

f) Physical growth rates 

2) Socio-environmental factors 

a) Socioeconomic status and social mobility of family 

b) Parental education 

c) Maternal intelligence level 

d) Family size and composition 

e) Geographic area of residence 

3) Familial factors: occiirrence of learning disorders in relatives 

a) Monozygotic and dyzygotic twins 

b) Full and half-sibs 

k) Cognitive and behavioral factors 

a) Mental and motor development and behavior ratings at 
eight months 

b) IQ scores, fine and gross motor development, concept 
formation and behavior ratings at age four 

c) Verbal and performance IQ scores, conceptual and visual- 
motor development and behavior ratings at age seven 

Method: In this study, a child is considered to have a learning disorder if 
he has an IQ of 90 or above on the Weschler Intelligence Scale for children 
and is more than one yeeir below grade expectancy (actual grade placement) 
in reading or spelling on the Wide Range Achievement Test. Children with 
learning disorders will be compared with average and above-average ach"" overs 1 
order to isolate early signs and current characteristics. Comparisons will 
be made within ethnic (white, Negro and Puerto Rican), sex and social class 
subgroups. It is recogniz;ed that this study is limited by the absence of 
any follow-up on the academic performance of the CPP children beyond age 
seven. 

Major Findings: Very preliminary findings, because o ' the, small samples now 
available, indicate that when compared with control groups matcned for IQ, 
children with learning disorders had families with lower socio-economic index 
scores (differences seemed to stem mainly from the pare;:ts' occupation), 
mothers of higher parity (whites) and mothers who were more often unmarried 
during pregnancy (Negroes). Among Negroes more of these children were 
rated as dysmature at birth. At eight months thej were foujid to have a 
shorter dui'ation of response and a less intense social response. At seven 



106 z 



Serial Ro. NDS (CF)-?^ PR/PSC 2108 
years they were rated as less cooperative, more hostile and as lacking 
in self-confidence. Among Negroes, children with learning disorders 
had a significantly lower vocabulary subtest score on the Wechsler 
Intelligence Scale for Children. With an increased sample, it is 
expected that many more characteristics of chij.dren with low academic 
performance relative to their ability will be identified. 

Proposed Course: Plans for the anlaysis of GPP data on learning disorders 
fall into a preliminary phase, a pilot phase and a main study phase. The 
current pilot phase consists of an examination of significant differences 
between the learning disorder groups and their controls, matched for IQ, 
on 59^ variables from the prenatal, socio-environmental, medical and 
psychological histories of the children whose records are now on tape. 
This phase is expected to be completed in November of 191k. The main study 
phase, in which analyses will be made on children drawn from the total 
study population of approximately U0,000 should be completed by December 
of 1975, leaving approximately 6 months for the preparation of a manuscript 
presenting the findings. 

Honors and Awards: None 

Publications: None 



107 z 



Serial No. NDS {CF)-'jk PR/PSC 2109 

1. Perinatal Research Branch 

2. Professional Staff Consiiltants 

Section 

3. Bethesda, Maryland 



Project Title: 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Comprehensive Analysis and Interpretation of the 
Collaborative Perinatal Project Data on Congenital 
Malformations 



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(CF)-67 PR/PSC 1510, 

(CF)-71 PR/PSC 1907, 

(CF)-71 PR/PSC 1908, 

(CF)-73 PR/PSC 205^^, 

(CF)-73 PR/PSC 2055, 

(CF)-73 PR/PSC 2056 



Principal Investigators: Dr. N.C. Myrianthopoulos , PRE, NINDS 



Other Investigators; 



Dr. A.F. Nctylor, PRE, NINDS 

Mr. Dan White, PRE, NINDS 

Ms. Ann Coates , PRE, NINDS 

Mr. David Rubinstein, OE, NINDS 

Dr. C.S. Chung, University of Ha'jaii 

Dr. Herbert Lubs, University of Colorado 

Dr. Marie-Louise Lubs, University of Colorado 

Dr. David Smith, University of Washington 



Cooperating Units; 



Office of Biometry, NINDS 

Department of Pediatrics, University of Colorado 

Department of Public Health, University of Hawaii 



Man Years : ,• ■ 

Total: 2.U0 
Professional: I.80 
Othe-: 0.60 

Project Description : 

Objectives : To thoroughly study the epidemiologic characteristics of congenital 
malformations in singletons and twins; to assess and interpret the influence 
of maternal, socioeconomic, neonatal, medical and other environmental factors 
on the occurrence of congenital malformations ; to determine the :.'isk of 
familial occurrence and to elucidate the role of genetic factors and the 
mode of inheritance of certain malformations; to determine the severity 
and clinical significance of congenital malformations and their assoc- 
iations with neurological,, psycliological and sensory handicaps; and to 



109 z 



Serial No. EDS (CF)-T^ PR/PSC 2109 

assess the long-range effects of malformations on s\irvival, growth and 
development . 

Methodology : A congenital malformation is defined as a gross physical or 
anatomical developmental anomaly which was present at birth or was detected 
during the first year of life. Malformations have been classified into major 
and minor categories on the basis of their severity, threat to life and cosmetic 
significance. Various epidemiologic and statistical methods are being employed 
in the analysis. 

Current status and major findings : 

I. Epidemiology of congenital malformations in singletons. 

This part has been completed. About 15 percent of single-born children 
had one or more malformations. Half of the malformations were major. Only 
about a third of malforma-tions observed during the first year of life were 
diagnosed at birth. Except for three minor malformations which were more 
frequent among Negroes, there were no significant differences in malformation 
incidences between Negroes and whites. 

A report has been prepared and is now in press . 

II. Epidemiology of congenital malformations in twins. 

The analysis of this part has been completed. Twins have significantly 
more major and minor malformations thaji singletons but the difference is wholly 
contributed by monozygotic twins. The incidence of malformations in dizygotic 
twins is the same as that in singletons. 

A report is now being written. 

III. A study of the effects of medical, genetic and socioeconomic factors in 
the occurrence of congenital malformations. 

The analysis of this part has been completed. Multiple birth, pregnancy 
complications (mostly through hydramnios) and male birth vere positively 
correlated with increased risk in major malformations, whereas maternal weight 
gain was negatively* correlated with major mal format i on3 . Maternal iiabete.^ 
di)j:'ing pregnancy was significantly correlated with single or multiple major 
malformations. This finding is further explored in Pa;-t B'. 

IV. Special s-nalysis of the effects of diabetes in the mother or. the occurrence 
of congenital malformations in the offspring. 

The analysis for this part has been completed. The risk of having a 
malformed child in mothers wiLh continuous diabetes, is doubled compared to 
that of non-diabetic mothers, with regard to major malforiraticns and increased 
significantly with regard to minor malformations. The effect seems to be 



110 



Serial No. NDS {CF)-'jk PR/PSC 2109 

associated, with the severity of the disease and not with the intake of 
insulin. 

A report is now being written. 

V. Genetic studies of congenital malformations. 

This part is in progress. The studies have been designed to derive 
empiric risk figures of repeating a malformation when it has onpe appeared in 
a family; to identify familial aggregations of specific malformations; and to 
clarify the mode of inheritance in identified familial aggregations. 

Specifications for data production have been written and submitted. 

VI. Study of associations of ABO and Rh blood types with congenital 
malformations . 

This study has been designed to confirm earlier suggestions of association 
of blood groups with congenital malformations, based on small samples. Speci- 
fications for data production are now being written. 

VII. Study of the clinical significance of miaor malformations. 

VII. Longitudinal study of- development, morbidity and survival of children 
with malformations. 

These two studies are now being designed in collaboration with Dr. David 
Smith of the University of Seattle, Washington, and Dr. C.S. Chung of the 
University of Hawaii. The objective of Part VII is to establish which minor 
malformations are worthwhile detecting and why, and which ones can be ignored 
or considered as normal variants. Part VIII is a continuation of Part VII 
utilizing the 7-year data on malformations. 

IX. Study of the effects of maternal factors in the production of .ongenital 
malformations . 

A pilot study haf. shown that maternal factors myy play a role in the 

causj-tion of some malformations. An extensive study utilizing all available 

malformation data through age 7 years is being planned and will be designed 
when the 7-year data become available . 

X. Study of the effects of major and minor malformations on outcome at 
7 years. 

This part will investigate the effects of major and minor malformations 
on neurological outcome, psychological test performance, and speech, language 
and hearing performance. The study will be designed when the 7-year data 
become available. 



Ill z 



Serial No. NDS (CF)-7i+ PR/PSC 2109 

XI. Correlation of minor chromosomal variants with congenital malformations. 

This part will te carried out as a contract operation with Dr. Herbert 
Lubs, University of Colorado, Denver, as principal investigator. The study 
will utilize data which have been developed by five Collaborating Perinatal 
Project institutions which are currently participating in the Denver-based 
chromosomal study of minor variants. 

The administrative mechanism for contract award has been initiated. 

Honors and Awards : None 

Publications : 

Myrianthopoulos, N.C.: Age: risk of seizures in infants. In: The Epidemiology 
of Epilepsy, M. Alter and W.A. Hauser, Eds. U.S. Department of Health, 
Education and Welfare, Public Health Service Publication No. (NIH) 73-390 
(NINDS Monograph No. lU ) . D.C., U.S. Government Printing Office, 1972, 
79-81. 

Myrinathopoulos , N.C.: Maternal factors influencing risk of epilepsy. In: 

The Epidemiology of Epilepsy, M. Alter and W.A. Hauser, Eds. U.S. Department 

of Health, Education and Welfare, Public Health Service Publication No. (NIH) 

73-390 (NINDS Monograph No. lU). D.C., U.S. Government Printing Office, 
1972, 103-107. 

Myrianthopoulos, N.C. and Chung, C.S.: Epidemiologic survey of congenital 

malformations in singletons. Report from the Collaborative Perinatal Project. 
National Foundation, Orig. Art. Series , 197^ » in press. 

Woolf, CM. and Myrianthopoulos, N.C: Polydactyly in American Negroes and 
whites. Am. J. Hum. Genet. 25:397-^0^1, 1973- 



112 z 



Serial No. NDS (CF)-7i+ PR/PSC 2110 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 



PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 



Project Title: 



Delayed Motor Development at One Year: Antecedents 
and Intellectual and Neurological Outcomes 



Previous Project Title: None 

Previous Serial Number: None 

Principal Investigators: Sarah H. Broman, Ph.D., PRB-NINDS 

Karin B. Nelson, M.D., PRB-NINDS 

Cooperating Unit : None 

Man Years : 

Total .02 
Pr o f e s s i onal . 01 
Other .01 

Project Description ; 

Objectives : A diagnosis of delayed motor development at one year made a 
significant independent contribution to IQ variance at age four, and 
was the best discriminator between retarded and normal children at 
that age. The purpose of this study is to define this condition more 
precisely in terms of specific development levels, and to examine some 
of its antecedents and longer-range outcomes at age seven. Factors to 
be investigated include maturity at birth, physical growth, presence 
of neurological and other medical diagnoses at one and at seven years, 
socioeconomic status and education of the parents, and IQ at age seven. 

Proposed Course ; Requests for the necessary computer printouts have 
been made. It is expected that this study will be completed within the 
next fiscal year. 

Honors and Awards ; None 

Publications: Broman, S. H., Nichols, P. L., and Kenn^jdy, W. A.: 
Precursors of Low IQ in Young Children. In Proceedings of the 80th 
Annual Convention of the American Psychological Association, Honolulu, 
Hawaii, Septe;.;ber 1-8, 1972-5 PP- 'i'7-78. 



113 z 



Serial No. EDS (CF)-Ti+ PR/PSC 2111 

1. Perinatal Research Branch 

2. Professional Staff Consultants 

Section 

3. Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1, 19T3 through June 30, 19lh 

Project Title: A Centennial Bibliography of Huntington's Chorea 

Previous Serial Number: None 

Principal Investigators: Dr. N.C. Myrianthopoulos , PRB, NINDS 

Dr. G.W. Bruyn, University of Utrecht, The Netherlands 
Dr. F. Baro, University of Louvain, Belgitun 

Other Investigators: None 

Cooperating Units: World Federation of Neurology Research Group on 

Huntington's Chorea 

Man Years: 

Total: O.itO ■ 

Professional: 0.10 
Other: 0.30 

Project Description : 

Objectives : To prepare a complete bibliography of Huntington's chorea from 
the first publication on the disease in 18T2 up to and including the papers 
read at the Huntington Centennial Symposium, in Columbus, Ohio, March, 1972. 

Methods employed : Sources for the bibliography were private collec.' l.ons of 
reprints , special bibliographies prepared by Medlars and searches of publi- 
cations in the National Library of Medicine of the United St;\tes, the Library 
of the Royal Society o^ Medicine, London, the Library of the Faculty of 
Medicine, Sorboune , Paris and the libraries of over 500 ffi'=dical associations 
and institutes throughout the world. Many ai-ticle& were translated and all 
were read. References were classified by >ear of publication, author, subject 
and country of origin. 

Currsnt status : The project has been completed. 

Honors and Awards : None 

Publications : Bruyn, G.W. , Baro, F. and Myrianthopoulos, N.C: A Centennial 
Bibliography of Huntington's Chorea 1872-1972 . Leuven , Leuven 
University Press, 197^, 31*+ pp. 

11^3 z 



Serial No. NDS (CF)- 66 PR/P 13^5 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, fferyland 



PHS-WIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Influencing Factors in Sudden Unexpected Death 

During FY '7^ this project was incorporated with Contract Niimber: 
WOl-IS-3-2311. 



117 z 



Serial Wo. WDS (CF)-70 PR/P 1859 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 197^4 

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 

This project is discontinued due to the departure of the principal 
investigator. 



119 z 



Serial Wo. NDS (CF)-72 PR/P 1973 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, IVforyland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Renal Glomerular Development in Infants of Diabetic 
Mothers 

Previous Serial Number: Same 

Principal Investigators: Toshio Fujikura^ M.D. 

Other Investigators: Shirley Driscoll, M.D. 

Cooperating Units: Boston Hospital for Women 

Lying-in Division 
221 Longwood Avenue 
Boston^ Massachusetts 02115 

This project, is discontinued due to the departure of the principal 
investigator. 



121: 



Serial No. NDS (CF)-72 PR/P 1974 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1^ 1973 through June 30^, 1974 

Project Title: The Significance of Meconium Staining in Neonatal Deaths 

Previous Serial Number: Same 

Principal Investigators: Toshio Fujikura^ M.D. 

Other Investigators: None 

Cooperating Units: All Collaborating Institutions 

This project is discontinued due to the departure of the principal 
investigator. 



123 z 



Serial Wo. NDS (CF)-72 PR/P 1975 

1. Perinatal Research Branch 

2. Section on Pathology 
3- Bethesda, Maryland 

PHS-WIH 
Individual Project Report 
July 1, 1973 through June 30, 197J+ 

Project Title: Organ Weight Change of Infants with Pulmonary Hyaline 
Membranes 

During FY '7^ this project was incorporated with Contract Number: 
WOl-WS-3-2311. 



125 z 



Serial No. NDS (CF)-72 PR/P 1976 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: The Effects of Profound Hypothermia in Newborn Mice: 
Prolonged circulatory arrest without cerebral, damage 

Previous Serial Number: Same 

Principal Investigators: Toshio Fujikura^ M.D. 

Other Investigators : None 

Cooperating Units : None 

This project is discontinued due to the departure of the principal 
investigator. 



127 z 



Serial No. NDS (CF)-73 PR/P 2064 
1. Perinatal Research Branch 
2„ Section on Pathology 
3« Bethesda, Maryland 

PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 1974 

Project Title: Blood Flow Study in the Intervillous Space: 

Sickling of maternal erythrocytes in the placenta 

Previous Serial Number: None 

Principal Investigators: Toshio Fujikura, M.D. 

Cooperating Units: All Collaborating Institutions 

This project is discontinued due to the departure of the principal 
investigator. 



129 z 



Serial No. WDS (CF)-73 PR/P 2065 

1. Perinatal Research Branch 

2. Section on Pathology 

3. Bethesda^ IVferyland 

PHS-MH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: Placental Transmission of Sickling Erythrocytes from 
Mother to Fetus 

Previous Serial Number : None 

Principal Investigators: Toshio Fujikura, M.D. 

Other Investigators: None 

Cooperating Units: All Collaborating Institutions 

This project is discontinued due to the departure of the principal 
Investigator. 



131 z 



Serial Wo. NDS (CF)-73 PR/p 2066 

1. Perinatal Research Branch 

2. Section on Pathology 
3- Bethesda, Iferyland. 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 197^ 

Project Title: The Influence of Birth Rate on Neonatal Mortality by 
Birth Weight, Gestational Age, Race and Sex 

During FY '7^ this project was incorporated with Serial No. NDS (CF)-73 
pr/psc 2060. 



133' 



Serial No. NDS-63 OD/OPE 1144 

1. Office of the Director 

2. Office of Planning & Evaluation 

3. Bethesda, Md, 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: 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., OPE, NINDS 

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, Md. 
Blanche L. Vincent, S.N.O., Greensboro, N.D, 
Zula C. Meekham, B.S.N. , PRB, NINDS 
Rose R. Tortoiilla, PRB, NINDS 

Cooperating Units: Georgetown University Hospital, Retarded Children's 

Clinic, Selected Maternity Hospitals and Physicians in 
Metropolitan Washington 

Man Years: 

Total: .50 

Professional: .30 

Other: .20 

P roject Description : 

Ob jectives : Design an instrument for the conduct of a retrospective study of 
seizures, cerebral palsy, mental retardation and other neuro ogical and sen- 
sory disorc'.ers of infancy and childhood in order to test certain basic and 
important iiypothescs concerning the occurrence of neurological damage. 

Methods Employed : Recognized damaged outcomes of pre£,nancy, especially 
mental retardation, have been studied and related to defined perinatal or 
postnatal events which are involved in the biological oi psycho-sociological 
mechanism underlying the following hypotheses: (1) anoxia, (2) toxic influences 
on the brain, (3) metabolic influences, (4) trauina to the head, (5) infection 
of the brain, (6) dehydration of the child, (7) genetic or familial patterns, 
(8) socio-economic status, (9) prematirity, and (10) nuLrition. 

Current Status : A paper on mental retardation is in review. 
Honors and Awards: None 
Publications: None 

135 z 



Serial No. NDS-63 OD/OPE 1146 

1. Office of the Director 

2. Office of Planning & Evaluation 

3. Bethesda, Maryland 

PHS-NIH 
Individual Project Report 
July 1, 1973 through June 30, 1974 

Project Title: Public Health Implications Study of Perinatal Mortality in 

the Collaborative Study and in the Collaborative Study Cities, 

Previous Serial Number: Same 

Principal Investigators: Z. A. Shakhashiri, M.D., OPE, 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 information for the United States and cities. 
The respective state or city health departments 
provided natality and mortality data for the Project. 



Man Years; 




Total: 


.20 


Professional; 


.10 


Otbar: 


.10 


Proiect Description: 




Obiectives: To evaluate 


feta 



Study population and of the cities from which that population is drawn, with 
the aim of comparing the two populations, 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 were sought for the study cities frop either 
the state or city health departments, whichever had jurisdiction for these 
records. Th3 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 birth weight, length of gestation, race and sex, and plurality 
for the years 1959 through 1966. Corresponding data were compiled by 
institution ror the PRE study population. The state or city health depr.rt- 
ments furnished either completed tabulation? or raw data to be tabulated. 

Considerable effort has been expended to create a usable data Tile of the 
external data being obtained in connection with this study. The aim is to 



1 ? 



'-37z 



Serial No. NDS-63 OD/OPE 1146 

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. 

Current Status : A manuscript is in preparation describing birth weight, 
fetal, neonatal and infant mortalities during 1959-1966 for the COLR popula- 
tion and the populations of the cities where the COLR institutions are 
located. Separate and comparative trends will be outlined and comparatively 
interpreted. 

Honors and Awards: None 

Publications: None 



138 



AmrUAL REPORT 

July 1, 1973 through June 30, 197^ 

SPECIAL PROGRAMS BRANCH 
Collaborative and Field Research 
National Institute of Neurological Diseases and Stroke 
National Institutes of Health 



The Special Programs Branch of C&FR, NINDS was established in July 1971 by 
moving the management of the Neurological Information Network and PL I480 
contracts out of the Office of the Director, NINDS. Since then, the Branch 
has had responsibility for: 

1. The development, conduct, and direction of programs for 
scientific and technical information exchange relevant 
to the neurologic and sensory sciences on a national 
and international basis. 

2. Coordination of collaborative research and development 
studies with Institute goals and objectives carried 

out in foreign laboratories with U.S. Government support 
under Public Law (PL) U80. 

3. Development and conduct of national and international 
directed research programs of relevance to the Institute's 
goals and in response to Institute program needs and 
priorities as identified by the Director, NINDS and the 
Associate Director, C&FR, NINDS. 

As of June 30, 197^, the Special Programs Branch was abolished. In March, 
Dr. Bering left his position as Head of the Branch to go into private 
medical practice. Mr. Weissberg was appointed Acting Head for the remaining 
time. 

Much of the in-house activity during the year was associated with the 
Institute's decision to phase out the Neurological Information Network (NIN). 
The NIN consists of the central office in the Special Programs Branch, the 
Brain Information Service ;BIS) at UCLA, the Information Center for Hearing, 
Speech, and Disorders of Human Communications (ICHS&DHC) at the Johns Hopkins 
University Medical School, the Clinical Neurology Information Center (CNIC) 
at the University of Nebraska, and the Cerebrovascular Disease Alerting 
Service (CVD) at the Mayo Foundation. 

The decision to do away with the NIN was made in July 1973 based in large 
measure on the limited NINDS budget for research contracts. In a com- 
petition for research dollars, the information activities were accorded a 
low priority. The NINDS Science Information Program Advisory Committee 
(SIPAC) was informed of this decision and asked for comment. The Committee 



aa 



reaffirmed its "belief in the need for and value of the NIN information services 
but, in the light of the NINDS decision, suggested each center he given an 
additional contract year to allow an opportunity to seek other funding sources. 
The Institute accepted and Implemented this recommei iation. 

Currently, each of the major centers has all of its products on a fee-for- 
service hasis. This income is insufficient to maintain operations, hut the 
centers are looking elsewhere for additional support. If such support is 
not obtained, the Center at Johns Hopkins will close by October 30, 197^j 
and the Center at UCLA will close by December 31, 197^- The Clinical 
Neurology Information Center at the University of Nebraska is expected to 
be funded in September 197^ for at least one full contract year. The Mayo 
Clinic was funded in May 197^ for a full contract year. 

The publication "Parkinson's Disease and Related Disorders," is a recurring 
bibliography from the NLM data base that has been distributed at no charge 
by the NINDS. In May 197'+? by arrangements with the National Technical 
Information Service (NTIf. ) it is being distributed at a cost of twenty 
dollars ($20.00) per yea:r to subscribers. This shifts the cost of producing 
this bulletin from WINDS to the user, resulting in savings to the Institute 
of approximately $^+,000.00 per year. 

Over this past year, the Institute has done much to shift the cost of and 
responsibility for maintaining science information products from itself to 
the scientific community. The Neurological Information Network was main- 
tained by NINDS for the decade from 196^+ to 197^- It was born at a time 
when there was considerable interest in exploring and furthering various 
avenues of scientist-to-scientist communication. Its demise is attribut- 
able in large measure to the reduced f-unding at the Institute which has 
occasioned a reexamination of priorities. During the decade of its exist- 
ence, the NIN supported major centers at an annual cost of about $300 to 
$it00 thousand apiece, with a total yearly budget of about $900 thousand. 
For most years, the cost of the NIF represented approximately one percent 
of the total Institute budget. At its height, the NIN annually provided 
products and services to more than 10,000 members of the community served 
by the Institute. The result of instituting charges for products and 
services has resulted in a fifty percent reduction in subscribei s to the 
irformat?on bulletins. Although each center is p-ursuing rigorously the 
addition of new subscribers, that source of income alone will not be 
eno\agh to allow the centers to survive. 

The results of two evaluation studies, one on the Parkinson's Disease and 
Related Disorders bulletin and the other on thci Biogenic Amines bulletin 
from UCLA, were completed this past year. The evaluation pointed up some 
interesting points. The users of the Biogenic Amines bulletin very much 
valued the inclusion of authors' addresses with each citation. This 
bulletin was on a fee-for-subscription basis and of the fifty percent of 
former subscribers who elected not to continue when a charge was instituted, 
sixty percent continued to read it, obtaining it from others or from their 
library. Forty percent of the respondents to the survey on Parkinson's 
Disease and Related Disorders claimed that its use had prevented duplication 



2aa 



of research in some fashion. The studies were done by Prof. F.W. Lancaster 
and are available from the National Technical Information Service (NTIS). 

The Branch completed work on a directory. Information Resources for the 
Neurosciences . This 90 page compilation was distributed to all members of 
the Society for Neuroscience as well as to many others. It is available 
from the Superintendent of Documents at a price of $1.25. 

The Clinical Information Center at Nebraska has been underway for about 
two years and they are already beginning to draw considerable notice from 
the clinical community. They have a National Advisory Committee drawn from 
the leaders in neurology and neurosurgery. They have produced an innovative 
new alerting bulletin called "Concise Neurological Reviews" in which papers 
on the same subject are grouped together and the current claim of each paper 
listed and referenced. The citations are then given as a separate section. 
Reading this bulletin allows one to get a feel for current work going on in 
various areas. If more information is required the reference is there. 
The mailing list for the publication is now over 2500. This is a true in- 
novation in the field of dissemination and scientific information. 

The Central Office of the Ne-urological Information Network in the 
Special Programs Branch has continued to produce Parkinson's Disease 
and Related Disorders , an alerting bulletin of relevant citations which 
comes out monthly. 

Mrs. Lunin of the Center at Johns Hopkins University was requested by the 
Pan American Health Organization to serve as a consultant for the information 
center for speech and hearing which was established in Caracas, Venezuela, 
by the Latin American Federation of Speech and Hearing Societies. This 
Center is backed by the Venezuelan Government. 

As a result of her visit, the Information Center for Hearing, Speech, and 
Disorders of H\mian Communication at the Johns Hopkins University will 
provide bibliographic searches (at cost) and current alerting bulletins. 
The Center in Caracas will serve all Latin America. The Information Center 
in Baltimore will also provide guidance on other projects that the Center 
in Caracas wishes to pursue. The Center in Caracas can provide information 
to the Center in Baltimore concerning research, facilities, and services in 
Latin America. 

The Special Programs Branch is also responsible for the Special Foreign 
Currenty (PL ^480) Program of the NINDS. This will be described in a 
separate section. 

Projects terminated in FY 7^ were: 

1. Evaluation of the Neurosurgical Biblio-Index, 
Serial No. NDS (CF) -72 SP 20lH 
This was a one-time effort that was completed. 



3 aa 



Information Sources for the Neurosciences 
Serial No. NDS (CF) -72 SP 20li3 

This was an effort leading to a publication which 
terminated with the publication. 

Cerebrovascular Disease Abstracts On Line 
Serial No. NDS (CF) -72 SP 20i;l+ 

This project terminated because of lack of funds 
to institute a large-scale on-line system. 



4 aa. 



Special Report 
Foreign Currency Credit Award Programs 
(PL lt80) 

The foreign currency credit award or PL U80 Program is a program of research 
supported by United States owned foreign currencies which are not convertihle 
to dollars. It is authorized by the Agricultural Trade Development Systems 
Act, 195^;, Public Law ^80, subsection IQl+K, which permits the use of foreign 
currencies, "to conduct and support scientific activities overseas, includ- 
ing programs and projects of scientific cooperation between the United States 
and other countries such as coordinated research against diseases common to 
all mankind, that are unique to individual regions of the globe, and promote 
and support programs of medical scientific research." 

At present, NINDS has 30 ongoing projects with a dollar value of about $900,000. 
There is one project in Egypt, nine in India, two in Israel, six in Poland, 
and twelve in Yugoslavia. Funds are available in Burma, Pakistan, and Tunisia, 
but we have no projects in these countries. The scientific content of the 
PL i;80 research is complementary to other WINDS research and covers both clini- 
cal and laboratory studies. 

Each of the PL 1|80 projects must have a U.S. scientist to serve as a collaborat- 
ing "Project Officer" who works with the foreign scientist in varying degrees. 
The role of the U.S. project officer varies from that of an active collaborator 
participating in the research to that of an informed sponsor. However, the 
project officer must each year review the expenditures and the progress report 
and advise the Institute whether the progress has been satisfactory. Of the 
30 projects, ten have project officers who are in either the NINDS intramural 
or C&FR programs. The other 20 project officers are in universities throughout 
the country. 

It has always been difficult to find scientists who are capable of doing re- 
search even in countries that are fairly well developed. The former NIH 
international fellows who have trained in this country and are working in the 
areas of interest to NINDS in their nati/e lands have been contacted to see 
if they can develop suitable projects. This has met with some Success, and 
many of oui* developing projects are from former fellowship holders. 

In Y'ugoslavid. the involvement of other U.S. agencl'^s in the PL 1+8C program 
has resulted in a gross overplanning for the use of available funds. A Joint 
U.S. -Yugoslavian committee has worked out an extension of the program with 
contributions from both the United States and Yugoslavian sndes. Available 
funds for the Yugoslavian activities have been used up. New projects must 
be done on the basis o." U.S. dollars matched by Yugoslav diua->"s. No great 
interest in doin^ this has been shown. 

There is an enormous amo'jnt of money in Egypt available for PL i+80 research 
projects, but r>here is difficulty in finding qualified people. The number 
one priority hvjalth problem is schistosomiasis, followed by infectious diseases 
(including meningitis). Egyptian researchers are interested in the neuro- 
logical d'seases, particularly accidents and injuries to the CNS. However, 

5aa 



the facilities are very poor and modern equipment in the laboratories is 
scarce. Clinicians and scientists who have trained outside of the cotmtry 
are well trained and capable. They are limited "by the lack of facilities 
and financial support. Also, there is very little infrastructure of techni- 
cians or facilities for repair and maintenance of equipment. This is a 
reflection of the fact that there is only about 1|0^ literacy in the country. 

In Poland, the funds available for use have been sharply curtailed. The 
amount available for projects on neurological and psychiatric disorders 
effectively limits the number of possible projects to a very few. In light 
of this, we are forced to examine carefully the proposals and to assign 
priority rankings for the Institute and to collaborate with NIMH in an overall 
ranking for both areas . 

The PL 1;80 program in Israel is almost totally phased out because all funds 
have been used. The only projects are ones which have been previously funded 
and are still continuing. There are no additional funds for Israel. 

The FINDS PL i+80 projects are as follows: 



Egzel 



India 


01-011-1 




01-015-1 




01-016-1 




OI-O2I+-I 




01-03i+-l 




Ol-Oi+O-1 




Ol-Ol+l-l 




01-099-1 




01-109-1 


Israel 


Ob-015-l 
06-028-1 


Poland 


O5-CO2-I 



03-OO6-I Venoms of Poisonous Snakes 



Nutritional Disorders of Nervous Systems 
Neural Mechanisms in Regulation of Feeding 

Behavior 
Neurologic Investigations of Cerebrovascular 

Disorders 
Collaborative Neurologic Studies 
Neurotoxins from Plan Materials 
Polypeptide Constituents of Snake Venoms 
Effect of Protein Malnutrition on Brain 

Metabolism 
Causes and Treatment of Non- Traumatic 

Paraplegia 
Uptake of Drugs 

Synthesis of Sphingolipids 
Molecular Biology and Immunology of 
Trachoma Agents 



Characteristics of Muscle Disease 
05-00l;-l Biology and Pathology of the Neuron and the 

Glia 
05-013-1 Biology and Histochemistry of Gliomas 
O5-O27-I Biochemistry of Myelination and De-Jiyelination 

(No additional funds) 
O5-O28-I Anoxemic Damage of the Immature Nervous System 
05-035-1 Brain Fine Structure in Rabbits with Hereditary 

Tremor 



6 aa 



Yugoslavia 02-003-1 Neural Correlates of Behavior 

02-006-1 Histogenesis of the Forebrain in the 

Human Embryo 
02-008-1 Function of Cholinesterases 
02-015-1 Metabolism. Release and Uptake of 

5-Hyd.roxytryptamine 
02-023-1 Vestibular Influences on Cerebral Activity 
02-020-1 Function of Biosynthesis of Ribosomes 
02-025-1 Neuropathology of Close Brain Injury 
02-027-1 Molecular Weight of Acetylcholinesterase 
02-023-1 Sialidase Action on Complex Molecular 

Aggregates 
02-0^2-1 Brain Survival in Anoxia and Hypothermia 
02-0'!l-1 Synaptic Transmission in Brain 
02-090-1 Gene Transcription and Gene Expression in 

Sea Urchin Embryos 

Project ( 01-015-1 ) -s concerned with the central and autonomic control system 
modulating alimentary receptor activities and regulation of feeding behavior. 
Project (02-003-1) is a large program to understand the function of the 
nervous system by studying physiological and biochemical correlates of behav- 
ior. In conjunction with this study is the development of a major research 
source, the International Brain Research Laboratory in Yugoslavia. 

Five projects are basically neurochemical or metabolism chemistry studies, but 
they are generally unrelated. Project (02-0l+2-l) is a study of the tolerance 
limiting role of metabolic events occurring in the brain at conditions of 
anoxia induced at different levels of hypthenia down to zero degrees Centi- 
grade. Project (05-027-1) is a study of the role of neuroglia cells in the 
myelination and derayeli nation with emphasis on the enzyme chemistry of the 
processes. This relates to human demyelinating diseases such as multiple 
sclerosis. Project 05-OOU-l is somewhat similar and is a study of the glia 
of the nervous system and their function in diseases of the brain. Partic- 
ular attention will be given here to the intracellular oxidation rf^ductior 
enzymes. Project (02-032-1) deals with the maintenance of the suri'ace 
characteristics of membrane systems and the enzyme actions required to main- 
tain these membrane systems. The control and regulation oi protein synthesis 
in Violopical systems is the focus of Project (02-020-1). 

There is one r.europathological study, Project (05-035-1) » which is a study of 
the fine structure of the brains of rabbits with h^Breditary tremor. This 
tremor is similar to that seen in Parkinson's disease and this study may have 
some bearing on that disease. 

There are six studies which relate to the neurological diseases of children. 
One of these , Project ( 06-015-1 ), is purely biochemical in natui'e; to synthe- 
size the sphingolipids of the brain. This has been important because these 
substances have been used to elucidate the enzymatic defects of sach diseases 
as Tay-Sachs disease. Another study, Project (01-02^-1), is both biochemical 
and clinical and deals with the enzymatic processes involved in the sulfation 
and desulfation of glycosaminoglycans and cerebroside sulfate. These 
biochemical mechanisms have been studies in specific metabolic liForders of 



7a 



aa 



( 



children. Using material from these patients the scientists working on this ^ 
project have identified specific enzyme defects. There are two neuropatho- 
logical studies relating to childhood. Project (05-006-1) is a study of the 
histogenesis of the forebrain in the human embryo, in which material from 
products of abortions is used. The other, Project (05-028-1), is a 
neuropathological study of the anoxia damage of the immature nervous system 
in the brains of newborns and infants who succumb to early anoxia. There 
are two studies that are related to the nutrition of the brain. Project 
(Ol-Oi+1-l) studies the effect of protein malnutrition on the brain, metabo- 
lism, and development. Project (Ol-Oll-l) is a general study of the 
nutritional disorders of the nervous system particularly those defects 
associated with malnutrition, the malabsorption syndrome, and ^jeripheral 
neuropathies of uncertain etiology. 

There are two projects relating to trauma of the nervous system. Project 
(02-025-1) is a neuropathological study of closed brain injuries to deter- 
mine the topographic distribution and histopathology of closed head injuries. 
Project ( 01-099-1 ) is a study of the role of arterial venous malformations 
of the spinal cord which cause paraplegia and other myopathies of unspecified 
origin . 

Project (OI-OI6-I) is a study of clinical angiographic and anatomic aspects 
of cerebrovascular disease in India. Project (05-002-1) is an extensive 
multiple disciplinary program for the study of muscle disease in Poland. 
Project ( 05-013-1) is a study of the biology and histochemistry of gliomas 
using tissue culture and immunochemical methods. Project (02-023-1) on the 
vestibular influences on normal and abnormal cerebral electric rhythms to see 
how these vestibular inputs contribute to the spread of cerebral electric 
rhythms and to correlate these with clinical problems. Project (02-090-1) 
is a new study on gene transcription and gene expression in Sea Urchin 
Embryos . 

These projects are concerned with naturally occurring neurotoxins. Of these. 
Project (03-OO6-I) is concerned with obtaining material of the poisonous 
animals such as snakes from the Middle East, North Africa, and as far South as 
the Union of South Africa. Biochemical and physiological studies will be 
carried out on the purified toxins which will also be available to "he investi- 
gators in the United States and antisera wil'l be developed. Project (Ol-Ol+O-l) 
is a similar study being carried out in India, studying the poisonous snakes 
of India. Tne thirc. Project, (Ol-034-l), is on neurotoxins from plant 
materials, and is focusing on toxicity of B-diamincpiopionic acid (ODPA) 
which is extracted from the seeds of Lathyrus Sativus in India. This will 
be studied in connection with the devexopment of human lathyrism. 

There are five projects concerned with synaptic transmitters. Project 
( 02-027-1) is to purify and characterize acetylcholinesterase and to deter- 
mine its molecular we?.ght and other chemical characteristics. Project 
(02-OO8-I) is to determine the role of the acetylcholinesterases in the 
acetylcholine system transmission conduction of excitation. There are two 
others which are somewhat related in the study of the metabolism release of 
5-hydroxytryptamine and its function as a transmitter in the brain. Projects 
(02-015-1 and 02-OUl-i). These four are in Yugoslavia. The fifth project, 

8aa 



in India, Project (Ol-109-l), is a study of factors which alter the uptake 
and retention in various tissues of catecholamines and drugs which affect 
their action. The drugs studied are primarily the catecholamines, catechola- 
mine depletors, and other drugs which either mimic the effect of catecholamine 
or block it. 



9aa 



< 



Contract Narrative 
Special Programs Branch, C&FH, UIITOS 
Fiscal Year 197^ 



Cerebrovascular Disease Abstracting and Retrieval Service at the 
Mayo Foundation (PH ^3-66-933) 

Title : Cerebrovasciilar Disease Abstracting Service 

t. 

Contractor's Project Director : Robert G. Siekert, M.D. 

Current Annual Level : $25,000.00 

Objectives : To provide a bibliographic alerting service of nev literature 
on cerebrovascular disease by preparing abstracts of appropriate articles 
for publication in Stroke - A Journal of Cerebral Circulation . 

Methods : The abstracts are from pertinent articles culled from a review of 
about li+8 serial journals. Either the author's abstract is used or a new 
one is prepared if the author abstract is too long or otherwise unsuitable. 
They are then given to the editors of Stroke for incorporation in the 
joiirnal. 

Major Accomplishments : The journal Stroke carries in each of its bimonthly 
issues approximately 100 abstracts prepared under this contract. 

Significance to Biomedical Research : For several years, these abstracts were 
offered to a small group of researchers on an informal basis. As more and 
more scientists heard about this service, the demand for the abstracts 
became so great that a way of general distribution was needed. The publica- 
tion of these abstracts as a special section in the journal Stroke makes 
the service available to all those who would find it useful. 

Proposed Course of the Contract : This program is under the surveillance of 
an WINDS project officer and its performance is reviewed periodically. 



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Contract Narrative 
Special Programs Branch, C&FR, NINDS 
Fiscal Year 197^+ 



Clinical Neurology Information Center (NIH-NINDS-72-2300) 

Title: The operation of a Clinical Neurology Information Center 

Contractor's Project Director : Walter J. Friedlander, M.D. 

Current Annual Level : $122,3U1|.00 

Objectives : To develop, as part of the NINDS Information Center Network, a 
specialized Information Center on Clinical Neurology. This Center -will he 
an international focal point for information relating to those diseases of 
interest to NINDS, especially information relating to diagnosis, treatment 
and prevention of diseases of the train and central nervous system. The 
Center will produce reviews of various clinical prohlems of interest to the 
Government, bringing together the relevant clinical knowledge as it applies 
to the problem. These reviews may focus on an entire disease problem as a 
whole, or on any distinct part cf a disease entity. The Center will iden- 
tify sources of information relevant to clinical neurological problems, 
including indexing services, abstracting services, periodical journals, 
books, monographs, etc. 

Major Accomplishments : In its second year of operation, the Center is obtain- 
ing from leading clinical specialists critical reviews. The manuscripts 
completed are the following: 

1) Developmental Dyslexia: Neurological Aspects 

2) Minor Hemisphere Syndrome 

3) Evaluation of Speech Therapy in Acquired Aphasia 
h) Clinical Evaluation of Neuropsychological Tests 

These will be published in a single volume by Raven Press. 

The Center's most innovative product and one that has been received enthusi- 
astically by the approximately 2500 scientists who receiA'-e it is the 
Concise Clinical Neurology Review . This publicat'' on emphasizes a one 
sentence (terse) abstract of each paper in a cluster of terse st"teraents on 
a single topic. A number of these topics are covered in a single issue of 
the bulletin. The bibliographic citations are referenced by a number and 
appear together in a second part of the bulletin. Approximately 2ii0 papers 
are selected for inclusion each month, based on a review of 69"-: serials. 
This publication seems to fulfill a need not otherwise met in neurology It 
is produced once e^Terj two weeks. 

Another accomplishment was the publication and distribution of a state-of-the- 
art review on Epilepsy. This is available at the cost of $1.50. Over 500 
copies have been sold. 



llaa 



Proposed Coiirse of the Contract : This program is under the surveillance of 
an WINDS project officer and its performance is under continued review. 



12aa 



Contract Narrative 

Special Programs Branch, C&FE, NINDS 

Fiscal Year 197^ 

The Brain Information Service at UCLA (NIH-NI]TOS-70-2063) 

Title: Operation of Specialized Information Center on 
Nonclinical Neurosciences 

t. 

Contractor's Project Director : Michael Chase, Ph.D. 

Cxirrent Annual Level : $32^,000.00 

Objectives : To operate, as a part of the National Institutes of Neurological 
Diseases and Stroke Weiirological Information Network, a specialized informa- 
tion center for the nonclinical neurosciences to serve as a national focal 
point for the identification, collection, storage, retrieval, analysis, 
repackaging, and dissemination of information. The major thrust of this 
information center shall be information analysis products and services. 
The center will identify and locate all information items and services 
pertaining to its subject area. The center will use the identified and 
stored information to make available comprehensive information service 
primarily to the scientists supported by NINDS, secondarily to the remainder 
of the biomedical community throughout the country, and, when possible, 
internationally. These services shall include: a) current awareness 
services; b) bibliographic services; c) translation of foreign language 
articles when essential; d) answering scientific questions relating to 
the subject area; e) promotion and preparation of reviews and other special 
articles for the general as well as specialized segments of the biomedical 
community, organizing and supporting workshops on specialized topics in the 
brain sciences and publishing their proceedings where necessary; f) f-ornish 
information about ongoing research; g) preparation of a publication series 
in the brain sciences. 

Major Accomplishments: The Brain Information Service reached a level where 
rele'/art scientific information was sent yearly to approximai:ely 10,000 
scientists. This was prior to fee charges for all products. This coming 
year will indicate changes due to these charges. The current alerting 
bulletins produced are: 

1. Sleep Bulletin and Sleep Review are monthly bulletins with 
2071 subscribers. These bulletins have proved so useful 
that the Association for the Psychophysiological Study of 
Sleep has requested that all members have copies of the 
bulletin and have made subscription to them a requisite 
for membership in the association. As of January, 1973? 
a subscription charge of $15 per day was instituted. The 
fee this year was raised to $2k per year. 



13aa 



2. Biogenic Amines and Ueiirotransmitters in the Nervous Systems ; 
an annotated iDibliography which appears semi-monthly and 
contains several one-page reviews or abstracts in depth. 
Distributed to 1000 at a fee of $2l+ per year. 

3. Neuroendocrine Control Mechanism ; Hypothalmic-Pituitary- 
Gonadal System - an annotated semi-monthly with several 
one page reviews or abstracts in depth. Distributed at 

a fee of $23 per year. 

k. Memory and Learning - Animal Studies : a monthly listing of 
published studies. Distributed for a fee of $21 per yt.ar. 

5. Memo of Current Books in the Brain Sciences : comes out 

monthly and is annotated. Distributed for a fee of $l8 per 
year. 

The Index to Current (EEG) Literature is prepared by the BIS as a quarterly 
supplement to the jo\irnal Electroencephalography and Clinical Neurophysiology . 
The cost of printing and distribution is borne by the EEG Society. 

The Sleep Research series. Volume 2, 19T3 , contains expanded abstracts of 
the year's Association for the Psychophysiological Study of Sleep meeting, 
current claims of research findings and all pertinent bibliographic 
information for the year. 

The Brain Information Service is placing more emphasis on the preparation 
of research review papers in specific areas of scientific endeavor. A 
manuscript prepared this year is Neural Systems and Circuitry Concerned 
with Sleep-Vaking Behavior by Morgan, P.J. and Stern, W.C. . 

The Conference Report Series has been continued. These reports have been 
in great demand with as many as 3500 requests for a single report. Three 
new reports produced this period are: 

1. Society for Neuroscience 3rd A_inual Meeting 

2. 7th Annual Winter Conference on Brain and Behavior 

3. Cellular Mechanisms Subserving Changes in Neuronal Activity, 
Other products being completed are' 

1. Translation of a Soviet Research Report on the Auditory System, 
by Alt 'man 

2. Updated Review on Synopses by Bubis and Sandbank 

3. A C\jmulative Bibliography of Electrical Recordings in the 
CNS, 1973- 



lUaa 



Significance to Biomedical Research : The Brain Information Service performs 
an essential service to the "biomedical community "by distilling and repackag- 
ing all new information items within its area of responsibility. An 
indication that this program is becoming central to the advancement of research 
in the basic brain sciences is the formal relationship being developed with 
the BIS by the Society for Weurosciences in which the Society looks to depend 
more and more on the BIS as a resoixrce to the neuroscience community. The 
BIS mailing list numbers more than l6,000 scientists. 

Proposed Course of the Contract : This program is under the surveillance of 
the NINDS project officer and its performance is reviewed periodically. 
Continued funding for this program will be greatly reduced. 



15aa 



Contract Narrative 

Special Programs Branch, C&FR, NIIIDS 

Fiscal Year 197^ 

The Johns Hopkins University (NIH-71-228l) 

Title : Operation of an Information Center on Human Commimi cations 

Contractor's Project Director : John E. Bordley, M.D. 

Current Annual Level : $250,000.00 

0"b.1ectives : To operate as a part of the National Institute of Neurological 
Diseases and Stroke Information Center Network, a specialized information 
center on speech, hearing and human communication to serve as a national 
focal point for the identification, collection, storage, retrieval, analysis, 
repackaging, and dissemination of information. The major thrust of this 
information center shall be information analysis products and services. 
The center will identify and locate all information items and services 
pertaining to its subject area. In addition, th<= center will U3e the 
identified and stored information to provide comprehensive information 
service to the biomedical commimity throughout the country and, when pos- 
sible, internationally. -These services shall include: a) current awareness 
services; b) bibliographic services; c) translation of foreign language 
articles when essential; d) answering scientific questions relating to 
subject area; e) promotion and preparation of reviews and other special 
articles for the general as well as specialized segments of the biomedical 
community; f ) furnish information about ongoing research and g) develop 
new methods of information analysis and dissemination. 

Major Accomplishments : Cxirrent Citations on Communications Disorders is the 
major current awareness product of the Center, having 'jirculation of 
1,200. This bulletin is divided and distributed in two parts: (l) Hearing 
and Balance, and (2) Language, Speech, and Voice- These bulletins provide 
up-to-date coverage of articles, editorials, chapters of books, small 
ronogrpphs. unpublished papers, reports, etc. in the fiexd of the com- 
municative sciences. The audience includes scientists, clinicians, educators, 
administrators and students. 

Biblioprofiles are comprehensive bibliographies on selected subjects that 
include a short state-of-the art digest by a scientist expert in the subject 
area. These products are available for a nominal charge that covers the 
cost of reproduction and mailing. The following is a list of Biblioprofiles 
that have been produced this year. 

Completed and/or Printed 

No. 8 Etiology and Pathology of Deafness 
No. 9 Speech Development 
No. 10 Acquired Aphasia 



16 



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Prepared BiTpliographies are produced periodically and are comprehensive 
bibliographies on selected topics. These had been available free but are 
now available for a nominal charge that covers reproduction and mailing. 
Those that have been produced this year were: 

Communication Disorders Related to Cerebral Palsy 

Language of the Deaf 

Speech Education of the Deaf 

Communication Disorders in Psychoses with Special Emphasis 

on Autism 
Hearing Screening Tests 
Language and Learning Disabilities 
Apraxia 
Speech and J.anguage Disorders Associated with Genetic and 

Metabolic Abnormalities 

In Preparation 

Gross Anatomy of the Ear 

Congenital Anomalies of the Ossicular Chain 

Ear Protectors 

Facial Nerve Tumors 

Voice Prints 

Significance to Biomedical Research : The Information Center for Hearing, 
Speech and Disorders of Human Commionication performs a valuable service to 
the biomedical community by distilling and repackaging all new information 
items covering the many disciplines in its area of responsibility. Special 
attention is given to the information needs of those whose programs are 
supported by NINDS funds. 

Proposed Course of the Contract : This program is monitored by the WINDS 
project officer and its performance is reviewed periodically. Continued 
funding for this program will be greatly reduced and contingent on its 
ability to be relatively self sufficient. 



173^ 



PHS-NIH 

Individual Project Report 

July 1, 1973 through June 30, 197^ 



Serial No. NDS(LF)-72 SP 201+2 
Special Programs Branch 
Office of the Chief 
Bethesda, Maryland 



Project Title: Parkinson's Disease and Related Disorders 

Previous Serial Number: Same 

Principal Investigator: Mr. Alfred Weissherg 

Cooperating Units: Bibliographic Services, National Library of Medicine 

Man Years: 

Total: O.UO 
Professional: 0.20 
Other: 0.20 

Project Description: 

Objectives : The monthly bulletin, Parkinson's Disease and Related Disorders : 
Citations from the Literature is a concise bibliography of papers from the 
current literature of interest to those in the subject field. This current 
awareness service is intended to help researchers and clinicians maintain 
knowledge of developments in the field of Parkinsonism. It was sent to a 
user group of approximately 1000 located in more than 50 countries. In 
May 197^ it was placed on a paid subscription basis and the circulation 
has yet to be determined. 

Methods Employed : The bulletin is produced monthly by a special search of 
the newly produced tapes of the National Library of Medicine MEDLARS system. 
The NLM provides camera ready copy composed by the computer and it is 
sent to the National Technical Information Service for printing and dis- 
tribution at a fee of $20 per year. 

Significance to Biomedical Research : Current awareness bulletins help keep 
researchers and clinicians up-to-date on developments in their subject, 
saving them many hours of library work, reducing the likelihood of dupli:;a- 
tion of research, and providing new knowledge to the clinicians at an 
early date. The value of the Parkinsonism bulletin to the neurological 
community is attested to by its enthusiastic acceptance. 

Proposed Course of the Project : The effect of placing the bulletin on a 
fee basis has to be determined. The results of this charge should be 
available by the end of the year. 

Honors and Awards: None 



Publications: 



None 



19aa 



ANNUAL REPORT 

Associate Director's Report 

July 1, 1973 through June 30, 1974 

Extramural Programs 

National Institute of Neurological Diseases and Stroke 

Extramural Programs, NINDS, have been characterized in FY 1974 by (1) an 
increase in the number and proportion of funded research grants; (2) the 
initiation of a targeted effort on neural growth and regeneration; (3) an 
evaluation of the program project-clinical research center mechanism; 
(4) completion of the NINDS Report on Brain and Aggressive (Violent) 
Behavior; and (5) launching of the NINDS "new" individual research fellow- 
ship program. In addition, the release of FY 1973 funds for research grants, 
training grants and fellcvships required an intense and special effort in 
the development and implementation of guidelines and procedures for the 
appropriate and meaningful investment of these funds. 

Research Programs - As documented in the body of this Report, both the 
targeted and investigator-initiated grant-supported research efforts of the 
Institute have moved ahead successfully. The research efforts in spinal 
cord injury, neural growth and regeneration, and virologic-immunologic 
disorders of the nervous system have increased significantly; research on 
stroke, multiple sclerosis, epilepsy, neuromuscular disorders and disorders 
of special senses including hearing have maintained previous impetus; research 
on head injury and neoplasms of the CNS continues to decline. The balance 
between fundamental and clinical research remains unchanged, with a large 
proportion of fundamental research easily identifiable with the Institute's 
clinical research goals; a small proportion is basic to the overall effort 
but difficult to assign to a specific area. 

The individual, investigator-initiated research grant continues to be 
the core mechanism of NINDS research grant support. However, the continuing 
growth of the program project and clinical research center programs has 
caused concern both of staff and the National Advisory Council. An analysis 
of the growth of these programs, the alternatives available and the consequences 
of each alternative has resulted in the Institute continuing present guide- 
lines but with increased monitoring of their administrative and scientific 
impact . 

Following established NINDS practices for the use of available funds in 
the last quarter, a large proportion of the released FY 1973 "impounded" 
research grant funds were awarded in support of meritorious projects for two 
years. This mechanism provided a brief but meaningful period of support and 
avoided the establishment of commitments against the President's FY 1975 
budget which does not take into account the release of these funds. 

Research Manpower Programs - The NINDS "new" institutional fellowship 
program was launched successfully in FY 1974. The number oi fellowships 
awarded in each area was in agreement with the NINDS plan presented and 
approved previously. As was anticipated, the number of applications in 
neurovirology and neuroimmunology was below the level of anticipated national 

i 

1 bb 



need in these areas; the number of applications in neurophysiology and J 
neuroanatomy was above that level. With the expectation that the NINDS 
institutional fellowship program will be initiated in FY 1975, the recruit- 
ment and training impetus needed in neurovirology and neuroimmunology should 
become available. The Research Career Development Award and Teacher- 
Investigator Award Programs also will be reopened for new applications in 
FY 1975. 



With the release of "impounded" FY 1973 training funds and the FY 1974 
apportionment, the "old" training programs have been continued under previous 
guidelines. This continuation is an extension of the program's "phaseout" 
but under the policies and at the funding levels of previous years. 

Administration and Organization - The "informal" reorganization of the 
Branches and Units of Extramural Programs has been accomplished and functioned 
successfully during FY 1974. Administrative steps have been initiated to 
formalize this organization. There is an obvious need for staff scientist- 
administrators with clinical background in neurology and otolaryngology to 
complement our pre-clinical staff. 3ecause of the characteristics of the 
job and the salary level, it has been extremely difficult to recruit highly 
qualified candidates. However, a candidate for each position (neurology, 
otolaryngology) has now been identified and discussions are in progress with 
the expectation that they may join our staff next year. 

Because of the continued growth and complexity of the Extramural Programs, 
in FY 1974 there has been a continuation of steps to increase the delegation 
of authorities to staff. This has increased the responsibilities for planning 
and operational decision-making for the Acting Deputy, the Acting Branch 
Chiefs and the Administrative Officer, EP. In parallel with this, operational 
sections on a "program" basis within each branch have been proposed. 



i 



bb 



AMUAL REPORT 
July 1, 1973 J through June 30, 197^ 
Extramural Programs 
National Institute of Neurological Diseases and Stroke 

TABLE OF CONTENTS 

Subject Page 

Introduction 1 

Fundamental Research in the Neurological Sciences 

Introduction 5 

Nerve Membrane 6 

Neurotransmitters ' 8 

Neuromuscular Junction 10 

Parkinson's Disease 12 

Viruses and Neurological Disorders 19 

Neuromuscular Diseases 30 

Nerve Gro-w-th Factors 32 

Regeneration in the Central Nervous System 33 

Cerebrovascular Disorders 35 

Trauma (Head Injury) 39 

Convulsive Disorders k3 

Multiple Sclerosis kj 

Communicative Sciences 

Sensory Process 53 

Hearing and Deafness '^k 

Middle Ear 55 

Inner Ear 57 

Cochlea 58 

Hearing Disorders 60 

Cochlear Prosthesis 60 

Comparative Hearing -• 62 

Speech and Language 63 

Olfaction 65 

Taste 67 

Research Training 69 



cc 



Annual Report 
July 1, 1973 through June 30, 197^ 
Extramural Programs 
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 extensive 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 metabolically 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 addition- 
al billions comprising the nervous system, work together to make the human body 
an effective and coordinated living organism. 

The economic burden of the neurological, sensory, and communicative 
disorders 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 conse- 
quences of brain injury 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 neiu'-omuscular apparatus, the most familiar of which are 
stroke, head and spinal cord injury, epilepsy, multiple sclerosis, parkinsonism, 
muscular dystrophy, cerebral palsy, aphasia, brain tumors, and otosclerosis. 
These disorders 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 and training programs of the National Institute of 
Neurological Diseases and Stroke are focussed on the identification, stimulation, 
and support of essential research problems aimed at the improved diagnosis, 
treatment, and prevention of disorders of the nervous system, the neuromuscular 
apparatus, the ear, and human conmiunication. They include disorders of the 
young (cerebral palsy, epilepsy, learning disabilities) of adulthood (head and 
spinal cord injury, multiple sclerosis, brain tumors,) and of the aged (stroke, 
parkinsonism, otosclerosis). Tlie administrative instruments used bo accomplish 
these purposes include research projects, research program projects, clinical 
research centers, outpatient clinical research projects, specialized research 
centers, research career awards, research career development awards, teacher- 
investigator awards, institutional research fellowship awards and individual 
research fellowship awards. 

Injuries to the spinal cord are occurring with increasing frequency. 
Once the spinal cord degenerates in the area of injury, paralysis always devel- 
ops. In March 1971;. the NANDS Council recommended approval of plans to support 
a limited number of Acute Spinal Cord Injury Clinical Research Centers. 



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Each Center's plan would include investigation, development and evaluation 
of improved methods of emergency treatment, rapid transport-ation, diagnostic 
techniques, medical-surgical repair, and the training of required professional 
scientific and technical personnel. Operation of one of these Centers would 
be a major undertaking for even a large medical institution. Therefore, 
modest funds were provided on a competitive basis for feasibility studies at 
six insxitutions to enable them to determine if they could develop the 
requirem.ents for a full scale Center. Five of them applied for support for a 
full scale Center, but only three could be supported with the funds available 
at that time. This year an additional Center was funded, making a total of 
four. It is e>rpected that each fully developed Center will ultimately con- 
tribute important information on the prevention of degeneration of the spinal 
cord and on the restoration of function in the injured spinal cord. Close 
liaison with other government and non- government agencies with responsibilities 
in this area is being maintained. 

In June 1972 in the face of what appeared to be a favorable fiscal 
situation and with knowledge of an urgent need, the NAITOS Council recommended 
the establishment of a program of Stroke Acute Care Research Units (SACRU). 
Grants not to exceed $75,000 per year would be awarded to develop Institutional 
focal points for (l) increased attention to the development or improvement of 
methods of prevention, diagnosis and treatment of the acute stroke; (2) devel- 
opment and evaluation of treatment methods and, (3) related training of 
professional and scientific personnel in this area. Twenty-six SACRU appli- 
cations were received and I8 were recommended for approval by the initial 
review comi^iittee and by the Council in March 1973- In view of the limited 
resources at that time, however, the Council felt that only 10 SACRU could be 
financed without crippling other equally urgent programs. One additional 
Unit was funded this year, making a total of 11. It is hoped that the SACRU 
program will ultimately be a broad one to improve the prevention, diagnosis 
and treatment of the acute stroke episode at many medical institutions. 

The development and use of biomedical methods for the treatment of 
behavioral disorders recently has generated discussion in the scientific 
community and on the part of the public about issues of efficacy and safety 
and about appropriate criteria for their use on hurnans. The issues have 
become particularly sensitive with the use of psychosurgical methods for 
the treatment of uncontrollable violence and rage behavior. At the request 
of DKEW, illMDS invited 48 distinguished leaders in basic and clinical 
research to .join in the preparation of a report based on four workshops: 
(1) neuroanatomical and neurophysiologic studies; (2) neurochemical, endocrine, 
pharmacological, and genetic studies; (3) behavioral studies; and (k) clinical 
studies- A summary of the Report on the Biomedical Research Aspects of Brain 
and Aggressive Violent Behavior has been published in the Archives of Neurology, 
Vol. 30, Ja.nua.ry 19lk. The full Report has been forwarded to m.EM. 

A National Advisory Commission on Multiple Sclerosis was appointed in 
February 1973^ by the Secretary, DHEW, in accordance with an act of Congress 
(P.L. 92-563). The charge to the Commission was: "To determine the most 
effective means of finding the cause of and cures and treatments for multiple 
sclerosis." In February 197^^ the Commission submitted its report, "Report 
and Recommendations of the National Advisory Commission on Multiple Sclerosis," 



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containing a series of recommendations on funding research, on budget planning, 
and on the management of research on multiple sclerosis. 

The following Table shows the number of research grant applications 
considered by the Council at its June meetings in each of the last five years. 

JUNE '70 JIME '71 JUNE '72 JUNE '73 JIME '7^ 

Number 339 352 398 k6j ij-28 

The number of applications has increased nearly 30 percent. This is attributed 
primarily to the effectiveness of the research training programs of the Insti- 
tute in which the output of fully trained investigators has only in recent 
years reached its full potential. It is tempting to suggest that the modest 
decrease in the number of requests from FY 1973 to FY 197^ is due to the 
proposed phasing out of the training program in 1973- Another factor may be 
that, because available funds have in the past permitted the funding of such 
a siiiall proportion of approved proposals, many investigators have been 
discouraged from applying. 

Tlie following Table shows the number of grants awarded and the total 
amounts of funds expended (in millions) each year for the past five years. 

FY '70 FY '71 FY '72 FY '73 FY '7^ 

Number 1,267 1,256 1,280 1,056* 1,065 

Dollars $48.8 $53-6 $6U.2 $62. 1|* $70.0 

■^Exclusive of impounded funds 

The largest change was in FY '70 when all research on vision was 
transferred to form the National Eye Institute. In FY 1971 and 1972 the number 
of projects remained essentially the same even though there was some increase 
in funds. This is due primarily to the increased cost of doing research. 
If the increasing cost of research continues, it will not be possible even 
to maintain the present program without appreciable increases in available 
funds . 

For many years the Extramural Programs were organized on the basis of 
a Research Grants Branch and a Training Grants and Awards Branch. During 
the past year the activities have been reorganized along more functional 
programmatic lines consisting of a Regular Programs Branch and a Special 
Programs Branch. The primary responsibility of the former is the planning, 
development and operation of the investigator- initiated research grant 
programs. The primary responsibility of the Special Programs Branch is the 
planning, development and operation of the targeted grant programs including 
program projects, clinical research centers, special projects, training 
programs and manpower studies. Tliis reorganization has been implemented with 
the approval of the Acting Director, NINDS, pending final action by the 
Director, NIH. 



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The reorganization and the uncertainty about the "old" research 
training program have resulted in some changes in the scientific staff. 
However, with the addition of three staff scientists this year, two in the 
Special Programs Branch and one in the Regular Programs Branch, the profession- 
al staff is fairly well stabilized. There is still some question in regard to 
future staff needs in view of the fact that the IS^k appropriation included 
funds for the "old" research training program which presumably was to have 
been phased out. 

Tlrie numbers of grants and ttie amoiants of funds in the various disorder 
categoi'ies are shown in Appendix E. 



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FUNDAMENTAL RESEARCH IN THE NEUROLOGICAL SCIENCES 

Introduction 

The history of science shows clearly that improvements in the treat- 
ment of disorders are largely dependent on sound basic research. However, the 
great importance of fundamental studies on neurological, sensory, and commun- 
icative components frequently is not adequately emphasized because their 
immediate application to clinical problems is still not entirely clear. 

This Introduction is usually devoted to illustrating why basic 
research on synapses, neurotransmitters, nerve membranes, etc. is so important 
to the ultimate prevention and treatment of neurological and communicative 
disorders. At this time, however, what might be considered a less biased 
point of view may be of interest. Therefore, part of an article in Science , 
December 28, 1973? page 1331? is quoted below. 

"Neurobiology is often cited as molecular 
biology's successor as the center of the 
,,;•.;, most challenging unsolved problems of 
biology. Understanding brain and nerve 
function requires, among other things, 
understanding how neurotransmitters 
interact with receptors on their target 
cells and thus transmit signals between . .- . 

• .. .. .•,;•,-.;■■ ,v neurons. Similar studies are being , .-.: .-. 

,.;•: performed with drugs. For example, -.-.:, . 
,,,■;■.: .. Solomon Snyder and Candace Pert at Johns ,. .. . : .■• 

.,-,..._ ..... . ■..■.,--, .,,-., , Hopkins University School of Medicine, :-. ■;■-.■; ,. 

... ..,-..•. -■ Baltimore, Maryland, demonstrated that .■...■ 

,. ;,. opiates bind to specific receptors in rat ' ■.-. ■ ... ' ■ . 

• .■,,.,, ..•-.,. ;,...,.-;. and human brain. Their investigations . ,_ ^ ■.-.•':■ 

-.--.-,.- ■ should aid in unraveling the still unknown -. '"■ • .. " ' -■ 
mechanism of opiate action. Moreover, the 
binding assay should facilitate design and • ■. '■■ ■ 
testing of opiate analogs useful for alle- 
.. »- ....,., . ^.. -.viating pain and of antagonists useful for :': 
.: ■.. '-. .•.,.. treating addiction to opiates such as heroin. ...- ;-j. i--- 

. .-..■,■.•.■...- .v-v" The function of the nervous system is also 
... . "• ,,..",..,^^ . being investigated at a higher level of - ■ 
■•. .- ■■■■.■■■ ..:-.," organization- -that of the whole animal. A , - 
._.. ,..; ....:■,,,.,.■. ..,. current approach to analyzing the factors . .-■■ 

^..,..,.,,..,, . .-..%..- that influence behavior fuses the disciplines ■■..- 
..- .; of genetics, physiology, and biochemistry. 

......... For example, Seymour Benrer and his colleagues .■=■.■ . ,• 

.:._ •^.■■r',.:- .■ at the California Institute of Technology, ' ■ 
. ;,_• .■:■■..•: Pasedena, are investigating fruit fly mutants 

that exhibit abnormal behavior. These researchers 
... _ ., have traced in their mutants the physiological 
.,..,-. defects responsible for such altered behavior as 

inability to respond to light. Since they can 



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"teach" fruitflies to perforin simple tasks, 
such analysis may be applicable to more com- 
plex activities, including learning, if the 
investigators can develop mutants unable to 
learn . 

Mem.branes are a recurrent theme in many of the 
most active research areas, and membrane research 
itself is an active area of investigation. Not 
only do membranes play an essential role in im- 
mune responses and the transmission of nerves 
impulses, but also in such critical functions as 
energy transformations in plant and animal cells. 
One of the concepts now stimulating much research 
is membrane fluidity- -that the components of cell 
membranes can and do move, and that these move- 
ments may be correlated with the physiological 
state of the cell." 

Many other areas of important basic research could be described. 
A nerve growth factor has been discovered which in extremely low concentrations, 
causes a reiaarkable growth in nerve fibers. Many efforts are being made to 
modify the shape or chemical composition of drug and chemical molecules in 
subtle ways to try to make them mnre effective in therapy, to reduce their 
toxicity, and to develop chemical tools to elucidate cellular functions and 
structures. Examples are new and better anesthetics, drugs for the treatment 
of epilepsy and parkinsonism, and chemicals for analytical tools to study nerve 
membrane structure and other neurophysiological problems. These important 
approaches involve such sophisticated techniques as x-ray crystallography, 
electron paramagnetic resonance spectroscopy, and nuclear magnetic resonance 
spectroscopy. On the basis of such highly specialised infonnation on the 
atomic relationships and the electron activity in the effective molecules, it 
will be possible to synthesize other compoimds that are more effective or 
■I'Thich play a crucial role in studies on how present drugs work. 

Nerve Membrane 

The excitable membrane consists of two functional components. One 
is activated electrically by depolarization and cannot be activated by 
specific chemicals like synaptic transmitters. The other can be activated 
only by transmitters and not by electrical stimulation. The postsynaptic 
membranes of single neurons are known to contain both components closely 
packed in the molecular complex of protein and phospholipid. It is well 
known that chemical activation induces a change in membrane potential which 
in turn influences the activity of the electrically excitable co-nponent. 
In other words it seems that the voltage change produced by chemical 
activation is what ultimately affects the electrical activity. However, it 
is still uncertain whether the chemical activation itself, without voltage 
change, has any effect on the activity of the neighboring excitable component. 

The production of action potentials (nerve impulses) in nerve fibers 
can now be interpreted as being due to a specific increase in the permeability 



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of the nerve membrane to sodium and potassium. The nerve membrane also plays 
a crucial role in neurotransmission at the synapse and, in association with 
the muscle membrane, at the neuromuscular junction. Therefore, detailed 
knowledge of how these membranes function is of the utmost importance. 

One team of investigators is studying the reaction of acetylcholine 
(ACh) with its receptor at the neuromuscular junction. It appears that 
the sise of the pore associated with the receptor must be very large, since 
the conductance changes little as the predominant external cation is changed 
from lithium to sodiiuii to rubidium. The anion seems to have no effect on 
the conductance or equilibrium potential. 

Increasing the viscosity (decreasing the diffusion rate of ACh) 
slows the kinetics of the falling (decay) phase of the miniature end plate 
potential. This supports the idea that the kinetics reflect the ACh 
concentration near the receptor. It cannot be that the concentration of 
conducting receptor lags behind that of the local ACh concentration. These 
studies are significant because they bear on the question of how quickly 
ACh binds to the receptor and then causes an increase in conductance through 
the m.embrane. Tiiis information will lead to a rational approach to the 
treatment of neuromuscular diseases and other synaptic disorders. 

This group has also studied the characterization and isolation 
of the receptor for tetrodotoxin (TTX) in crab, frog, and mammalian nerve 
m.embranes . Treatment of crab fibers with carbodiimide inhibited the binding 
of TTX and suggested that blockage of a carboxyl group might be responsible. 
In the case of frog sciatic nerve, depression of the compound action potential 
by TTX is very rapid and is completely and rapidly reversed by washing with 
saline. Efforts have been ::iade to characterize the TTX receptor in nerve 
membranes from bovine cerebellum. Results suggest that blockage of sulf- 
hydryl groups by reduction-alkylation or N- ethyl-male imide does not affect 
the TTK receptor. On the other hand, protein denaturation in guanidine 
hydrochloride or urea irreversibly eliminates all binding. Further preliminary 
purification of the receptor has been completed with the use of selected 
detergents. These experiments provide further evidence regarding the nature 
and distribution of receptor sites in the neuron membrane involved in the 
control of cation permeability. This information is essential to an under- 
standing of how the nerve functions and of the causes of its disorders. 

It is well known that calcium plays an important role in the 
permeability of the nerve membrane and consequently, in the conductance of 
nerves, that is, the transmission of an impulse along the nerve. However, 
the exact mechanism is still unknown. Tlierefore, one laboratoiy has been 
studying the influence of external calcium concentration on aeetylcholine- 
induced conductance changes and on receptor inactivation in snail neurons. 

Lowering the external calcium from 7 to 1.75 mM consistently 
decreased the acetylcholine- induced conductance as much as 53 percent. This 
result, in comparison with the work of other investigators, suggests that 
in snail neurons calcium ions may supply the nerve membrane f/ith fixed 
positive charges that regulate its permeability to chloride. 



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To study receptor inactivation, acetylcholine (ACh) was applied to 
the neuron twice only four to 12 seconds apart. This pulse interval was 
chosen "because it was sufficiently long to elicit a second response and yet 
too short to allow complete recovery of the receptors from the effect of 
the first application. The membrane conductance change elicited by the 
second ACh application was used to estimate the influence of test solutions 
on receptor inactivation. It was found that lowering the external calcium 
concentration consistently increased the degree of receptor inactivation. 
It was previously shown that at the endplate the rate of receptor inactivation 
increases when the external calcium concentration is raised. The fact that 
the same change in external calcium that leads to a decrease in the ACh- induced 
conductance also leads to an increase in receptor inactivation has led to the 
hypothesis that a reduction of the fixed membrane charges that allow cations 
or anions to permeate through the membrane facilitates the transition of the 
conductance mechanism to an inactive state. 

Neurotransmitters 

Any controlled movement or other activity of the body invol\^es the 
participation of at least two or three nerves, and usually many more. The 
connections between the nerves across the synapses are carried out by neuro- 
transmitters. It is obvious, therefore, that neurotransmitters are very 
important and any disruption of their function may be disastrous. 

One investigator is concentrating on the identification of synaptic 
transmitter compounds and the mechanisms by which nerve cells regulate their 
accumulation. It has been shown that gamma-aminobutyric acid (GABA) is the 
inhibitory transmitter at the lobster neuromuscular junction. GABA was 
shown to be the most active inhibitory substance present in extracts of the 
lobster nervous system and it was the only physiologically active compound 
selectively localized in inhibitory neurons. The only excitatory substance 
found was glutamate which ■was present in about equal amounts in excitatory 
and inhibitory axons. The concentration of GABA was about O.IM in inhibitory 
and O.OOIM in excitatory axons. 

It remained to be demonstrated that GAM was actually released when 
an inhibitory neuron was stimulated. This was done and it was found that 
the amount released was proportional to the number of stim.uli applied. Like 
the release of chemical transmitters at other synapses, it depended on the 
presence of calcium in the bathing fluid. Numerous details in regard to 
GABA are still unresolved, such as the storage form, release mechanism, 
isolation and nature of receptor sites, etc. 

Since an active uptake m.echanism exists for GABA in lobster nerve- 
miuscle preparations, the amounts collected (l-^l- X lOl-^ moles/impulse) are 
only an overflow of this uptake. Recently it ' is been possible to localize 
the site of uptake. Certain aldehyde fixatives will covalently bond amino 
acids to tissue components. With this method, using h3 labelled GABA, the 
site of uptake ira,s shown to be the connective tissue cells and the Schwann 
cells that surround nerve and muscle in the preparation. Physiological 
studies of the role of this mechanism are being continued as are studies of 
other neurotransm.itters, both inhibitory and excitatory. 



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Synaptic transmission is mediated by specific chemicals, so-called 
"transmitters." After being released from the presynaptic terminals, they 
combine with receptor molecules localized on the surface of the postsynaptic 
membrane. Whether their effects are excitatory or inhibitory to the post- 
synaptic neurons is solely determined by the ionic specificity of each 
receptor. For example, acetylcholine (ACh) induces either excitatory or 
inhibitory responses at the postsynaptic membrane, depending on the type 
of receptor. 

Studies in one laboratory have shovn that the excitatory receptor 
(D-type) induces a permeability increase mainly to sodium ions, whereas the 
inhibitory receptor (H-type) induces a permeability increase exclusively 
to chloride ions when the receptors are activated. It was discovered that 
certain proteolytic enzymes, such as carboxypeptidase B and leucine- 
aminopeptidase, can selectively block either the excitatory or inhibitory 
tj'pe of ACh-receptor. This observation suggested that a C-terminal lysine 
or arginine molecule is a critical element of the inhibitory receptor 
whereas an N-terminal leucine or alanine is a crucial component of the 
excitatory receptor. 

Further studies were made on the effects of carefully selected 
compounds on the responses of the excitatory (D-type) receptors and the 
inhibitory receptors (H-type) to ACh. Pyrodoxal 5 '-phosphate (PLP) is a 
well known coenzyme of the vitamin Eg group. The postsynaptic activity 
of the ACh receptor was rapidly depressed by a 10-20 minute exposure to 
1 mM PLP. The effect was specific to the H-type receptor without affecting 
the D-type receptor. As mentioned above, carboxypeptidase B also depresses 
the activity of the H-type receptor and not the D-type, the effect being 
consistent with that of PLP. These findings suggest that the beta-amino 
groups of lysyi residues, including C-terminal lysine are present near the 
reactive site of the H-type receptor. 

The chemical p-Nitrothiophenol (NTP) was known to form stable 
bonds with myosin A, the amino acid composition of which suggests that NTP 
makes a bond with a carboxyl side-chain of glutamate or aspartate residue in 
the myosin A. Although the structure of postsynaptic receptors may be quite 
different from that of myosin A, a comparative study of the reactive sites 
in many enzymatic structures m.ay lead to the elucidation of basic receptor 
mechanismis such as ionic requirement, conformational change, and triggering 
of the permeability increases. It was found that the expuaure of the 
receptor membranes to IniM NTP markedly depressed the excitatoiy response of 
the D-type (excitatory) receptor without affecting the activity of the H-type 
receptor. Previously it was shown that the D-type receptor membrane becomes 
permeable to many cations (Wa, Mg, Ca, K) when activated. The present 
results with NTP suggest that D-type (excitatory) receptors include glutamyl 
or aspartyl residues In the vicinity of the reactive sites which may 
participate in the cationlc movement across the receptor membrane. 

This work Illustrates especially well how very sophisticated 
approaches are being utilized to analyze at the molecular level the structure 
and composition of the excitatory and inhibitory receptor sites in the 
postsynaptic nerve membrane. Studies like this will ultimately lead to an 



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understanding of hov the highly coinplex systera of excitatory and inhibitory 
responses work together to make the billions of individual synapses function 
smoothly. 

IvTeuromuscular Junction 



'■''cause the neixroir.uscular junction is a special kind of "synapse," 
studies on it not only throw light on nerve-muscle relationships, but may 
also result in the delineation of basic principles applicable to synapses 
ill .'general. One investigator has studied whether some nerves other than 
the normal inctor nerve will innervate a skeletal muscle. This problem was 
attac;:ed by an ingenious method of transplanting the frog's own muscle to 
the lyrnph space underlying the skin to which the ends of various nerves 
could be implanted. 

With this procedure it was found that skin sensory nerves and 
post-ganglionic sympathetic nerves show no evidence of innervating skeletal 
m.uscle, or of maintaining it in any way. Tlie muscle atrophies and develops 
acetylcholine sensitJ.vity as if it were totally denervated. On the other 
hand, preganglionic autonomic nerves (gastric, splancnic) are quite capable 
of for:ning functional sytiapses. A totally denervated muscle atrophies to 
about 20-25 percent of its original weight in about six months. A muscle 
Innervated by a preganglionic nerve atrophies only to about 40-50 percent 
of its original weight. With a somatic motor nerve the atrophy is even less, 
only 6O-80 percent of the original weight. 

In the case of vagal innervation, the number of muscle fibers 
decreases, but innervated fibers appear as large and healthy as control 
fibers. With splancnic Innervation the number of fibers decreases slightly, 
and the remaining fibers all look approzliriately equally atrophied despite 
innervation. It appears, tlierefore, that the gastric vagus is better able 
than the splancnic to maintain healthy m.uscle fibers. 

In m.ost cases not all muscle fibers become re-innervated by the 
autonomic preganglionics. Vagus -Innervated muscles produced an average of 
only hk piercent of the maximum tension produced by the controls and contained 
only approximately half of the number of muscle fibers in the control. 
Splancnic innervation is usually even less successful. I^aximum tetanus tension 
is typically only 10-20 percent of the direct-stimulus value. However, the 
number of fibers is not reduced much. Thus, either some of the fj.bers are 
rem.aining unlnnervated, or the synapses are not sufficiently effective to 
produce threshold stimulation in a large fraction of the fibers. 

There are distinct differences in synaptic pharmacology/ between 
somatic motor nerve innervated and autonomic re innervated m.uscle. In the 
latter there is no evidence of chollnesterase at the new neuromuscular 
.junctions. Apparently the autonomic preganglionic nerves are incapable of 
inducing the necessary synthesis or concentration of the enzyme at their 
synapses. Both vagus and splancnic evoked excitatory junction potentials 
are blocked by hexamethonlum at one-tenth the concentration recessary to 
block control synapses. Cujrare apparently is equally effective in both cases. 



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Denervation of the transplanted muscle leads to a generalized 
sensitivity to acetylcholine (ACh) just as it does in denervated muscles 
left in the leg. Reinnervation by somatic motor nerves, the vagus or the 
splancnic all lead to a similar reduction in the sensitivity of the muscle 
to ACh except at the sites of the new nerve endings. 

Wien a somatic motor nerve is implanted into a muscle already 
innervated a few weeks or months earlier by a splancnic nerve, the somatic 
nerve can mostly displace the splancnic. Individual muscle fibers were 
seen at intermediate stages in which they were innervated by both nerves, 
with different junctions on the same fiber showing the differences typical 
of the two nerve types. It is not clear whether the somatic nerve can 
completely displace the splancnic. Evidence thus far indicates that the 
splancnic nerve endings are simply forced into a small part of the muscle 
which they continue to innervate successfully. 



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PARKII'iS on ' S DIS5ASE 

Parkinson's disease is a progressive neuroloyjcal discraer of un..i'Owr: 
canse affectin,;^ certain brain areas responsible for the control and re<7alat;'on 
of movement. Estiinates of prevalence rani^e from one case per 1,00'': to one per 
200. Pritnarily a disorder of rr.iiJdle are or later, its prevalei"/:e is tiioo.'-ht 
to be increasir'.f: witli 1:he iri'-irease in the averare life spar. 

Vp.e onset of iMarl^insonisr-: is ultimately charao-teri::fed t^y bro^or, 
rigidity, and bent posture. Intel]. l,^ence is usual]-y unaffected. In tue r'uily 
developed disorder tne L'ace :.'.-:.. ms t e ■-. •aracteristic " . .h.g' " vlL. a ar ; stare 
comin,'" from unllinki nr eyes. The nationt ofter! sits notionless , rarely crossiiv 
his le,f';s or foldjny his arins . 

Standard treati^ent in thr- r)ast consisted of I'ihysical tliorapy, a 
variety of irurrs, and siu7.":ery. The treatments of choice are -uow tt'ala.ic 
surrery or a highly efToctive fort:; of repJ.acement therap.y mah.iny use cf lar;'i 
cral doses uL' L-Dopa. Although L-Dopa induces remarkable :'rn)ri"'ve' ent :in !::ost 
parkinsonism patients, it iias very anploasant side ef j ects ac I sn-j-' ^atiui l,s 
do not respond. Therefore, a stall letter treatment must oe I'lU;.! and the 
':reatest lon*3 ranre contribution of tiie researcti on L-Dopa may ;o tuat, as 
the first direct lead to the real- cause of par::insoi;isn', it pri v'i les an 
appi'oach to the 'ievelopinent of a ;a'j,ch i';ore efi/ecLlvo i\,r' \ . i' Li '•I'an.y . Jespiue 
many recent aeveloptdents, evidence i/iaJ.caLes tiiat tue i^ever i.iel, :i.>i.ls l'I' tnerapy 
have not substantially increased life e:>7pectancy. 

Eecei'tly numerous biochei';ical abfor'-alities ir. Par; . i.noi 'i 's diseaso 
have been found. Most of these findinifs are related to a decrear.e '<<' dopari'ine 
concentration in the parkinsonian brain. Most of the l.'ioihie' :ical clian;-es in 
dopa;tiiiie metabolism iii tne par!:ii,sonian Drain (;an be duplicate'..! by [iia;:in;^ a 
lesion in the substantia nirra in animals. The chan^jes in a>-etylcnolir:e 
metabolisiii. in i^rivinson's disease, reflected ;.',/ the liypersensitivit.y to 
clioliners'^ic st j.mul.ation and therapeutic ii.iproveinent with ant i-hiolinerric dru,;';s, 
also j.s secondary to the loss of ni,-jro-striatal neurons. Tlie rlc, -en oration of 
tne substantia nit-^ra cells and certain other neuror.s appears to be the prirary 
changes in this disease. Most investigators agree tliat it is only the Jicnroi'.s 
which contain melanin (e. g. substantia nigra, locus coeruleus, dorsa]. nucleus 
of the vagus, etc.) which degenerate in F&rkinson's disease. This suggests 
tiiat there is some peculiarity of the neiuroinelan J n-cun.tainirig cells of the 
central riervous syste:'' which predispose the.m to degenerati on in. i^arkinsonis.: . 
Investigation of the structure, :".ode of s;>Aithesis and possible function of 
neurone Ian in is one important approacl-i to tlio etloloj^y and path.o.'^enesis of 
idiopathic Parkinson's disease. Attempts are being iiade to leteriiane if there 
is arty biochemical inter-relationship between substantia nigra n^eJ.ani satioti 
and the af'^iorr^ialities in catecholamine n'etaboli.sm ■■"'■ t^"'s iisi'rhjr. 

L-Dora acts as a t'leraneutic a'-ei'.t ii' farki '^so;- 's .fisease nrosn-ably 
by increasing the concentration, of dopatidne in the basal -ar'^d.ia. L-Dopa 
therapy aJ.S'.^ increases dopamine levels in the iiypotlia.la.'-us and '"edian e';!ine!;ce 
whicii i:ay alter the secretion i.l.' ro.leasing factcirs which control pituitary 
hormona.1 secretion. InvestP-atioiis of Vie effect oV L-Dopa on bypot!ialam.i>.'- 
pituitary hormonal control may be important in t.be early detectioo o.r adverse 



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side effects of long term L-Dopa therapy. 

A group of researchers has found that of fifty-one parkinsonian 
patients who have been on long-term L-Dopa therapy, six developed "start 
hesitation," a difficulty in initiating walking, as a side effect. This 
symptom can also occur in untreated Parkinson's disease. In some untreated 
parkinsonian patients, rigidity is present and the gait is usually slow with 
short shuffling steps, whereas in some L-Dopa-treated patients who develop 
start hesitation, the muscle tone is either normal or decreased and the gait 
is normal or minimally impaired once the patient starts walking. These 
investigations showed that patients with start hesitation had a significantly 
better response to L-Dopa therapy than the remainder of the L-Dopa- treated 
patients. 

An abnormality in dopamine metabolism has been well documented in 
Parkinson's disease and implicated in Huntington's chorea. Both diseases 
are extrapyramidal disorders resulting from anatomical and functional abnor- 
malities of the basal ganglia, which contain the highest concentration of 
dopamine in the brain. In Parkinson's disease, the levels of dopa.mine in the 
brain are decreased. L-Dopa, the liranediate precursor of dupamine, is the most 
effective therapeutic agent for Parkinson's disease, presum.ably due to an 
increase in brain dopamine. These investigators recently obtained an IND from, 
the FDA and university approval to study the biochemical and clinical effects 
of gairana-hydroxybutyrate (gHB) in patients with extrapyramidal disorders. 
Much of the initial investigation has been designed to determine absorption 
of the drug, plasma disappearance rates, toxicity, optimal rlose, etc. Initial 
indications are that chronic treatment with GHB appears to he safe and well 
tolerated by patients and the investigators are encouraged by preliminary 
experience to undertake a thorough, well controlled evaluation of this prom.ising 
drug. In addition, GHB will be given a clinical trial in other neurological 
disorders such as Dystonia Musculormn Deformans, Spasmotic Torticollis and 
Tardive Dyskinesias. 

Tlie aim of studies in another laboratory is to develop an understanding 
and prevention of such unwanted effects of L-Dopa therapy as drug- induced 
psychotic behavior and involuntary movements. The hope is to separate these 
actions of L-Dopa from the therapeutic action against Parkinson's disease. 
It is also hoped that these studies and similar ones to be performed on psy- 
chotic patients will provide new forms of theraj^y for natiurally occurring 
psychoses as well as those induced by L-Dopa. 

Tlie research focus is to attempt to control psychotic behavior and 
dyskinesia with cholinergic drugs such as physostigmine, oxotremorine or 
poperidine while at the same time not interfering witli the control of F^rkin- 
sonian symptoms by L-Dopa. It is felt that the h;;rp'^''~''^sis that Parkinson's 
disease results from a loss of inhibition of p striatal cholinergic neiuron 
by a nigrostriatal dopaminergic neuron is overly simple and th_e work, will 
be continued without such a hypothetical model. 

Tiie preclinical work includes toxicological studies of the agents 
used and tests of the agents on various animal models. These models include 
modification of the ability of L-Dopa to induce turning behavior (toward the 



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side of the lesion) in caudectomlaed mice a.nd actions of cholinergic drugs 
on caudectoinized liilce. Since cholinergic drugs modify the behavior of mice 
intoxicated by L-Dopa, other intoxications will also be studied such as those 
of LSD and amphetamine. 

Clinical studies to date have tested only physostigmine and it has 
been e-stab] ished that relatively large doses of this compound can be tolerated 
by Parkinson's patients, provided that dosage is increased gradually and 
probanthine is given. Tlie initial results on four patients have been promising. 
Garrulity and hyperactivity of patients on L-Dopa has been decreased while 
affecting the control of parkinsonian symptom's relatively little. No e-<rperience 
has yet been gained with the treatment of natural psychoses. 

Clinically;, attempts are being made to control troublesome side effects 
of L-Dopa which greatly limit its therapeutic effectiveness. If these side 
effects can be alleviated without producing significant deterioration of the 
therapeutic action on parkinsonism, it will be of great clinical benefit. 
Improvement with the administration of physostigmine to those with mental 
abnonnalities secondary to L-Dopa has been demonstrated. Other investigators 
have demonstrated the effectiveness of physostigmine in treating toxic psychoses 
and in decreasing the involuntary movements of Huntington's chorea. Anti- 
cholinergic drugs do not produce abnormal involuntary movements or increase 
those produced by L-Dopa. However, although not identical, there is some 
similarity between the mental effects of L-Dopa and those of anticholinergic 
toxicity, and the latter can potentiate the former. Thus, there is a certain 
rationale from clinical e:>rperience for the use of cholinergic agents to 
alleviate both the mental effects and the abnormal involuntary movements 
produced by L-Dopa. Fluctuations in the therapeutic response constitute 
another troublesome aspect of therapy with L-Dopa. Preliminary studies on 
the effect of low protein diets plus MK 486 in smoothing out this response 
are very encoiu-aging. 

Studies by another investigator are aimed at correlating biochemical 
(biogenic amines) parameters with neiurological abnormalities associated with 
extrapyramidal disorders. In these studies monkeys with siorgically induced 
lesions in specific brain areas who ez-diibit abnormal movements are used as an 
e;<perimental model. Tiie effect of the lesions on putative neurotransmitters is 
being evaluated. Histological changes resulting from the destruction of various 
anatomic pathways are to be correlated with biochemical changes. The clinical 
studies involve an evaluation of the effectiveness of L-Dopa therapy and an 
investigation of the fate of L-Dopa in patients undergoing treatment for 
extrapyramidal disorders. 

Developm.ent of a primate model of parkinsonism producing tremor and 
hypokinesis, has been an important feature of this project. The administration 
of L-Dopa in combination with 1-alpha-methyl-dopa hydrazine (MK-486), a dopa- 
decarbonylase inhibitor that acts peripherally, resulted in a transient 
disappearance of tremor in the extremities contralateral to the lesion with 
a concomitant development of involuntary movement. Trivastal (ET-495), an 
agent that stim.ulates dopamdne receptors, had an effect similar to that of 
L-Dopa. Trivastal relieved the tremor and induced involuntary movements for 
a longer time than L-Dopa. 2-Er-alpha-ergocryptine (CB 15^), another agent 



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that stlniulaten (lopamJnc receptors, also lias tremor relieving and involuntary 
movement induGiii,q; activities. 

Tlie effects of actJvation ami bl(x-l;ade of dopamiiie receptors on 
dopamine synthesis in striatal slices sliow that apomorphine, an agent that 
stimulahcs dopamine receptors, inhibits the Ijiosynthesis of -^ '•'i^-dopamine 
frr.m J ^ '- 1.;/ r-osine more effectively In striatal slices tlian tyrosine hydroxylase 
activity ^^n vitr o. Tlie effective inhibition of I'-^H-dopamine biosynthesis in 
striatal slices by apomorphljie '-(.uld eitlier be -iue to its accumulation in the 
dopami]Te containing neurons and subsequent inhibition of tyrosine hydroxylase 
or to dopamine receptor stimulating activity of the drug resulting in a 
feedback control of dopamine biosynthesis. In order to study further the 
effects of activation or blockade of dopamine receptors on presynaptic 
dopamine synthesis the effects of Trivastal and of Halo^eridol on ^C-dopamine 
syntiiesis from ^^C-tyrosine in striatal slices is being investigated. Trivastal 
stimulates dopamine receptors, liut unlike apomorphine. does not contain a 
catechol group and does not inhibit tyrosine hydroxylase activity in vitro. 
The administration of Trivastal results in an inhibition while administration 
of Haloperidol results in the stimulation of -'-^C-dopamine synthesis in 
striatal slices. Treatment of rats with Haloperidol antagonizes the Trivastal- 
indi.iced. inhibition of I'+C-dopamine synthesis. These results support the idea 
that a receptor mediated feedback exists which controls the rate of dopamine 
synthesis. 

Tlie findings that agents which stimulate or block dopamine receptors 
af f e" t the rate of striatal dopamine synthesis and the recently presented 
evidence that a dopamine-sensitive adenylate cyclase may be the receptor for 

of the effects of c-AMP 
obtained from various 
regions of the brain. The results of these studies have shown that dibutyryl 
cyclic Af4P (dJb-cAI4P) stimulates tlie synthesis of ^^C-dopamine in striatal 
slices as well as in other regions of the brain (i.e. median eminence, cortex). 
In separate e:qjeriTnents it was establlslied that the cyclic nu'^leotide and not 
the butyryl moiety of dB-cAMP is effective in the stimulation of -^^C-dopamine 
synthesis. Tlie dB-cAJjlP- induced stimulation of ^^C-dopamine synthesis was not 
abolished by blocking the dopamine re'.;eptors with Haloperidol or by stimulating 
the dopamine receptors with Trivastal. However, this phenomenon may be dose 
dependent since the results of preliminary studies indicate that the stimulation 
of C-dopamine synthesis by low conr^entrations of 'TB-cAi4P (5x10~5m) j_c; less 
effective in striatal slices obtained from Haloperidol. 

Wiile a large amount of information has been obtained on the effect 
of dopa on tlie Krain, little or nrjthlng is known about tlie transport of this 
compound, arid its derivatives between brain and blood, or about tiie mechanism 
of renal excretion. Mslng the techniques of tubular mlcrofusior and also 
a newer technique or subcapsular microinfusirjn the possibility rf tubular 
so'-retion of levodopa and Its derivatives is being investigated in another 
laboratory. '■Jornpetition and inhibition of these transport processes are 
being examined by using related amino acids and alsr^ rrobenacid. Tlie ventric- 
uloclsternal perfusion technique of I^jjpenheimer will be applied to study 
the transport of these cr.mj^ounds In the brain. Inulin and labeled drug will 
',.e a'i-J_e'l t-"- the nerfnsiriri c.rd i.iti fin an'] tlie rate of clearance of the drug from 



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dopamine in mamraallan brain prompted tlie investigation 
on I'+C- dopamine synthesis from I'^C-tyrosine in slices c 



the cerebrospinal fluid will be studied by analyzing the perfusate. Arain, 
related compounds, inhibitors;, and also dopa decarboxylase inhibitors will 
be added to determine their effect on these transport processes. 

The rationale of another study is to help understand the behavioral 
side effects of L-Dopa therapy in human disease. L-Dopa administration is 
known not only to increase dopamine levels in the brain but also to result 
in a release of serotonin and then an inhibition of its synthesis. Tiie 
dopamine increase may be noted in serotonergic neurons and may act as a 
"false transmitter." Studies are proposed to deteimiine whether the behavioral 
effects of L-Dopa on mouse locomotor activity is centrally or peripherally 
mediated, and whether it is specifically related to L-Dopa conversion to 
catecholamine or to interference with other transmitters such as serotonin. 
Studies will be perfonned of the turnover of catecholamines both in peripheral 
tissue, such as heart, and centrally following chronic L-Dopa therapy. 
Inhibitors such as methysergid and cyproheptadine may help implicate serotonin 
receptors because of their known action as receptor blockers. Of interest 
are the preliminary data which suggest that both L- and D-Dopa m.ay produce 
behavioral depression whereas only L-Dopa causes stimulation. Since D-Dopa 
is not converted to dopamine, a mechanism such as altered serotonin content 
and turnover might be a more reasonable explanation of the induced behavioral 
alteration. Other preliminary results suggest that the L-Dopa effect is 
centrally rather than peripherally mediated. Studies are being carried out with 
5,6-hydroxytryptophan to enhance the stimulant effect of L-Dopa and reduce 
the depression. Assays of catecholamines, serotonin, and their metabolites will 
be conducted not only on whole brain but on selected regions such as cortex, 
caudate, hypothalam'os , cerebellum, and medulla. 

The objective of another research project is to provide a basis for 
testing the speculation that the administration of L-Dopa in the form of a 
metal chelate complex might reduce the tendency for e:ctracerebral decarbo:-:y- 
lation and facilitate the transport of intact L-Dopa into the brain. An 
e>5)loratoiy investigation of the L-Dopa chelates of Cu(ll), Sn(ll), and 
MG(II) is being undertaken with a view to devise an effective system, for the 
optimal transport of intact L-Dopa to the brain. Tlie stoichiom.etry, 
therm.odynamics, and solubility of the chelates in aqueous and nonaqueous 
solvents are to be investigated by potentiometric, spectroscopic, and other 
physiochemical methods. Radiolabeled L-dopa chelates are to be studied in 
anim.al experiments to determine transport into the brain. 

The large patient population of Parkinsonism and allied diseases 
accumulated over the past 10 years by one laboratory continues to be documented 
and studied with emphasis on the following: defining the effects of pharma- 
cological agents on the natural history of the disease; testing the therapeutic 
efficiency of newly developed agents; monitoring of side effects and toxic 
reactions of presently employed treatment pro.-'-r'ms; and searching for new 
insights as to the etiogenesis and pathogenesis of these disorders. Central 
to this program are the developments regarding the effects of mionamines 
particularly dopam.ine on the symptoiTis of basal ganglia disorders. Tliese 
investigators have been deeply involved in the application of these new 
findings from, a diagnostic and therapeutic viewpoint as well as their implica- 
tions as to the causation of these disorders. Essentially this program is 



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continuing docjmentation of cases of F&rkinson's Disease and other types 
of Parkinsonism. In accord with a specially designed protocol, including a 
rating scale for Parkinson signs and symptoms, patients are evaluated 
at regular intervals. These data have been useful in delineating the natural 
evolution of the F&rkinson process and determining the effects of various 
treatment programs. This patient population also acts as a resource for 
exploring biochemical abnormalities in CSF and other body fluids which may 
exist in this disorder. In this regard, emphasis at present is on the 
various monamines, their metabolites and enzymes as well as the essential 
amino acids. Additionally, autopsy material is periodically being derived 
and correlative clinical and pathological studies carried out. The long 
term assessment of the therapeutic effectiveness of levodopa in Parkinsonism 
is being investigated. The large patient population under treatment will 
continue to be examined at regular intervals as to the state of their 
F&,rkinsonism, evidence of side reactions and/or toxic effects from the use 
of this agent. Here again, specific studies to identify metabolites of 
levodopa in CSF are in progress with a view to determining correlations 
with beneficial as well as side effects. When available, autopsy material 
is obtained and studied to determine the effects of administration of levo- 
dopa on the pathological Parkinson process in brain as well as its effects 
on other organs . 

The assessment of new pharmacological agents or other modes of 
therapy are being explored. As a result of the remarkable effects of levodopa, 
renewed interest in developing other pharmacological agents has occurred. 
Over the past three years, some 15 such agents have been investigated. Of a 
promising nature are two which will undergo trial. One, an analogue of 
oxytremorine, which has been shown to have overall anti-I^rkinson activity; 
the other, a piperazine compound which appears to have anti-tremorigenic 
activity. 

There are two major routes of dopa metabolism. The first involves 
decarboxylation to dopamine and subsequent catabolism of dopamine to 0-methyl- 
ated deaminated, and beta-hydroxylated products. The second major route of 
catabolism involves transamination processes to pyruvic and lactic acid 
derivatives. 

It has been suggested by one investigator that the prolonged time 
course of the therapeutic effect of L-dopa results from the accumulation of 
one of its metabolites, 3-0-methyl dopa, which is slowly de-0-methylated to 
dopa and subsequently decarboxylated to dopamine. A correlation between 
neurologic status, psychiatric status, and metabolism of L-dopa is being sought. 
A population of patients with Parkinsonism are placed on a standardized regimen 
of therapy with L-dopa. These patients are evaluated in great detail with 
regard toiheir neurologic status, psychiatric status and levels of urinary 
dopamine and dopamine metabolites in the drug-free state. The major finding 
thus far in this study of l6 patients with primary Parkinsonism is that the 
pretreatment levels of each of the two metabolites of dopamine, dihydroxy- 
phenylacetic acid (DOPAC) and homovanillic acid (HVA), are specifically 
correlated with the severity of the neurologic disease or with a specific 
indication of mental status. DOPAC varies with the severity of the neurologic 
disease, and HVA, with the psychiatric status. The primary objective of this 



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study is to confirm tbf=se observations and to extend the findings to patients 
other than those with primary Parkinsonism. Additional correlations will be 
examined between the metabolites and the therapeutic as well as the adverse 
effects of the drug in an attempt to clarify the numerous problems and to 
lend additional data to evaluate the several theories which have been put 
forth to •explain, the clinical observations. 

A strong relationship between the sulfate conjugation of a major 
metabolite of L-dopa (3.^^-dihydroxyphenylacetic acid, DOPAC) and the severity 
of I^,rkinsonism is suggested. It has been found that administration of large 
am.ounts of L-dopa to patients results in saturation of the mechanism for 
sulfate conjugation of L-dopa metabolites. It was, therefore, of considerable 
interest to explore the biochemica.1 detenninants of the sulfujylation process. 
The ejrperimental approach used here is a comparative study of the sulfate- 
conjugating systems in the liver and brain which may participate in the 
metabolism of L-dopa or its metabolites in the intact manmal. Tlie program 
is directed to determine the relative capacities of the liver and brain to 
generate sulfate esters of L-dopa and its metabolites, and to examine the 
ability of dopa m.etabolites to saturate these systems, also to compare the 
etiLiyme system.s of the brain and liver which generate the sulfate conjugates 
of L-dopa and/or its metabolites with those reported to catalyze the pro- 
duction of sulfate derivatives of other substances (e.g., phenol, serotonin, 
estrone) . 

Small concentric hyaline spherules are a distinctive feature of 
the form of Parkinsonism known as I^ralysis Agitans. Tliese round structures, 
known as Lewy bodies, occur in various pigm.ented neurons throughout the brain 
stem (e.g., substantia nigra and locus coeruleus). They occur most frequently 
in idiopathic Paralysis Agitans, less frequently in the postencephalitic 
variety, and small nLunbers of them are also encountered in 5-20'^ of "control" 
autopsies. Wien examined by electron microscopy, the bodies are composed 
of filamentous structures which are more densely packed in the central portion 
of the spherule. The precise biochemical nature of Lewj^ bodies is not known. 
Efforts are being made to collect these bodies in greater number for chemical 
and m,orphological analysis. The answer would be of interest in relationship 
to the problem of parkinsonism, even though the specific nature of this 
relationship is not evident at the present time. 



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VIRUSES MP lilEUROLOGICAL DISORDERS 

Earlier workers in virology suspected the possibility that some 
viruses might remain dormant in the body for a prolonged period before pro- 
ducing overt disease and that others may be responsible for persistent and/or 
recurrent infection. Most of the clinical and experimental viruses are 
identifiable by a relatively short incubation period lasting only days or at 
the most a few weeks, with rapid proliferation of virus prior to the acute 
phase of disease followed by the disappearance of virus resulting in an 
increase in specific antibody that is supposed to provide protection against 
future infection with the same virus- Opposed to rapid onset of viral 
disease, virus infections with a prolonged incubation period and slow initiation 
of pathological processes may be responsible for many chronic and degenerative 
neurological disorders of man. Added evidence to the concept of "slow viruses" 
was gained when veterinary virologist Sigurdsson identified three infectious 
diseases that affected native Icelandic sheep not observed anywhere else pre- 
viously. Two were neurologic and one a pulmionary disease. All had a protracted 
incubation period which resulted in the animals' death. Since then several 
other slow viruses both in animals and man have been found which specifically 
attack the nervous system. This report in general is related to these neuro- 
tropic viruses. 

Methods most commonly used for the detection and characterization of 
a viral agent in cell cultures include observations for cytopathic effects, 
hemagglutination, hemadsorption, interference, fluorescence and electron 
microscopies, antigen detection, immune adherence, enzyme induction, neucleic 
acid homology, radioactive labeling of nucleic acid precursors in nutrient 
media, reverse transcriptase detection, cell fusion, providing "helper" viruses, 
radiation, co-cultivation and immunofluorescence. 

It is generally agreed that latent agents are those which can be 
detected more or less regularly by available techniques and which can persist 
in the organs of animals in the absence of symptoms. Masked (occult, hidden) 
agents, on the other hand, are those which are suspected as being present but 
which cannot be demonstrated directly by available techniques. The pie sent 
distinction between masked and latent viruses, therefore, is conditional, and 
is dependent upon the sensitivity of available detection techniques. This 
leads to confusion, because some viruses now considered "masked" may be fully 
infectious, but be present in such small quantities or in such a physical state 
that present methods are inadequate for their demonstration. If an effective 
detection technique should becom-e available or if existing techniques should 
be made more sensitive, then these viruses would be detected and therefore 
designated as "latent." 

It is apparent, therefore, that there are two prime considerations 
in evaluating viral masking latency: (l) the methods available ".'or detection, 
with special reference to their sensitivity and (2) the basic nature of the 
virus-host relationship. The two considerations can hardly be separated in 
studies of masked and latent viruses. Although some insight has been obtained 
concerning the possible mechanisms of defectiveness in certain viruses, the 
approaches to unmasking "hidden" (or "occult") animal viruses have been largely 
empirical. Little is really known about why certain experimental conditions 



19 



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favor virus unmasking, or the exact relationship between m/L viruses and host 
cells. A few fundamental studies of this type have "been done. For example, 
adenoviruses can be isolated from adenoid tissues which are cultivated 
in vitro but homogenates of similar original tissues usually contain no 
detectable virus. There is some reason to believe, however, that fully 
Infectious virus is present in such tissues, but not in readily detectable 
concentrations. Work with adenoassociated satellite virus and distemper virus 
suggests that these two viruses can exist in a truly masked form and are com- 
pletely lost if the viability of the infected cell is lost. Human diseases 
wnich have been studied extensively to determine whether viruses are in any 
way associated include: Kuru, subacute sclerosing panencephalitis, progressive 
multifocal leuioencephalopathy and Jakob- Creutzfeldt disease. Convincing 
evidence is accumulating from transmission studies in higher primates, in 
vitro cultivation of brain tissues and electron microscopy of affected tissues, 
that m/l viruses are etiologically associated with each of these diseases. 
The recent reports on the isolation of new papovaviruses in humans, particularly 
in the brain, have revived interest in the possible role cf this class of 
viruses in chronic human disease. 

Using some of the techniques, referred to above, a program to study 
explant cell cultures from human and simian brains is conducted with an attem.pt 
to determine the presence or absence of masked and latent viruses which might 
be suspected in viral etiology. It is hypothesised that if a group of brain 
tumors with common antigen(s) can be identified, these might be candidates to 
examine for a viral etiology. It is hoped to develop a serological test 
which will detect putative antibody responses to such antigen(s) in the tumor- 
bearing patients. If such antibodies exist and can be detected, then larger 
groups of normal subjects and patients would be tested. 

Criteria of the responses of the experimental animal to viral 
infection are established by careful m.onitoring of the physiologic behavior 
of the CNS following inoculation with tissue extracts and whole cells derived 
from brains of patients with chronic encephalopathies. 'Tlie early events of 
the experimental diseases in animals have been detected by electroencephalo- 
graphy because the EEG record becomes abnormal before any other signs of 
diseases appear. These techniques are being perfected for clinical application. 
However, at present they permit the selection of diseased animals for study 
d^uring 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 
CWS disease produced in animals may not be identical with the disease of the 
patient from whom the tissue was obtained. A dramatic exam.ple of species 
difference in host response is the effect of sim.ian virus B. In man it induces 
a disease that is severe, and usually fatal, whereas for monkeys this viri.is 
is innocuous. 

Subacute sclerosing panencephalitis (f)SPE) is a degenerative 
neurologic disease of children and young adults. It is characteri::ed by pro- 
gressive mental and motor deterioration, myoclonic jerks, and com.a. Tlie 
patients become severely emaciated and die from intercLurrent infections. The 
diagnosis established during the incipient stages often shows a personality 
disorder or m.ental retardation and the EEG shows slowing and dysrhythmia. 



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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 animal, producing symptoms typical of SSPE in the animal, has pro- 
vided an important new lead in isolating and understanding the causative agent 
in SSPE. During the last few years, evidence of a relationship between SSPS 
and measles virus has been established. ¥ith the help of electron microscopy, 
tubular structures have been seen in the brain with SSIE which resemble the 
nucleocapsides of measles virus. Further investigation showed a high or rising 
titer of m.easles 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 SSEE. These patients had no history of clinical measles, but had 
received live measles virus vaccine. Some had rubeola once in their lifetime. 
Several characteristics common to both measles virus and SSPE have been found. 
They include antigenic properties, host-range, cytopathogenic effects, tem- 
perature 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 neuroadapted strain of measles 
virus or a distinct but antigenically related virus? Is a second "helper" or 
"co-virus" necessary to produce disease? If such 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 usually 
high serum measles antibody levels noted in most SSPE patients a response to 
viral antigen in the 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 CNS? 

Six SSPE strains have been isolaisd and examined in sufficient 
detail. They were found to be more neurovirulent for newborn hamsters and less 
reactive with neutralizing antibody than classical wild or vaccine strains of 
measles virus. Data suggest that the isolate "SSPE (l)" is also poorly reactive 
with antibody. Consistent possession of increased neurotropism and decreased 
reactivity among the SSPE viruses examined suggested that they were variants of 
measles virus of a similar nature. This hypothesis was supported by finding 
among plaque isolates from both wild and vaccine strains of measles virus 
variants which resemble SSPE viruses in neurovirulence and reactivity with 
antibody. It seems likely that the SSPE strains emerge as variants from a 
measles virus population under selective pressures in the patient. Although 
these variants may simply represent the most persistexit viral genome, it is 
possible that those properties which they were found to possess in common, or 
some associated viral characteristic, allow such viruses to produce SSPE. 

At present there are no known markers of wild strain or vaccine 
strain measles virus that allow determination of which of these strains might 
have given rise to a particular SSPE strain. Attempts to distinguish between 
wild or vaccine strains as a source of SSPE virus will be further complicated 



21 cc 



V;.- Sr.F"^ isolates prooably being highly selected variants of a virus population 
whicli originally infected the patient. Tlie exact relationship of SSPE virus 
to i.easles virus ray be defined only after detaile.i biochemical and genetic 
studies. Reports liave appeared which describe some chemical an.i p'nysical 
properties of ir.easles virv.s R'd/i a;.' of ?l"'\ a;'d inicleocapsi.is from measles 
virus l..'----te.i cells. 'I'lie co:;;parisons of virus specified RNA's in measles 
ir.fect^ '-T's viti- t>'ose in :"SP^- vii'us infected cells have reveale-.l tlius far 
O'-'y quantitative difi"erences among six distinct species of RIM- Tlie search 
for ii.inor qualitative differences between measles and SSPE virus specified 
RIjA of each species has net l:eeii co:;;pleted. Studies are undeCTvay compariufj 
,:ieasles ano SSPE viruses in terms of virion WW and peptides, as well as virus 
specifiet pepti-es of infected ceills. Mutants of measles and SSPE virus have 
beeji isolate! v: ic';\ are te;!'.perature sensitive and which have otlier markers 
tb^at wi?! serv-:^ ■')■■. both bioche;!iical a;-i genetic investigations. 

Altlit.u.-di the etiolo;;^ of itiultiple sclerosis is onscure, epidemiologic 
evblence has suggested a coimnon exposure factor, m.ost likely a virus, in those 
a;'-'^ '.jte ■ wit;, this most prevalent nuraar^ 'lemyelinating disease. Studies of 
viral causes of deniyelination have, i- ti e past, taben two general rlirections: 
( 1 ) in vi vo an: ±^ vitro stuiies of animal viruses and ( ";) attempts to isolate 
virus T'ro!!! Imir.an lisoases such as ;,-,iltiple sclerosis, Schilder's disease and 
progressive -..lultif O'^al leukoenceplialopathy (H'-IL), a disease in which papova 
virus-like particles has been observed by electron microscopy- Ihe study o.l 
a. -imal viruses causing demyelinabion lias been limited by the paucity of 
experi. ental riodels. Work has ceiitered on two known viruses, JHM. a neurotropic 
strain c. ::;ouse i:epatit:"s viru.s, an.; progressive multifocal leul;oencephalopathy. 

d'lli virus-induced demyelination in mice appears to be the ii;ost 
practical model to study. li; 19^9, 'Jheever and co-workers isolated a murine 
virus causing a lisseminated encephalomiyelitis with extensive destruction 
of myelin, '[he virus entitled J!iI4, also caused liver disease and su^^sequently 
has been grouped vath the :r:c-.-..se liepatitis viruses (MIP/) designated MrfV-i)-. 'Llie 
demyelinating lesions produce: .■) acute infections with the neutrotropic strain 
of r;cu.se liepatitis virus were fcun:J to be dependent on duse of virus, age of 
mice, ani the route of inoculation, kluorescent antibody studies showed that 
the antigen was inost prominent in cells of the white matter and immunosuppression 
studies gave no indication that there was an irraniuiopatMologic mechanism in the 
demyelination. Electro:: jcicroscopic studies were initiated on animals after 
the acute phas'- at a time when virus couLl no longer be recovered but when there 
was still an active irflammatory respc^nse. Initial studies in tliese animals 
as lor g as one year after the acute infe^.-ticn have siiowrj the presence of 
coronavirus viriors in oligodendroc\d:es and what appears to be continuing 
:1 e my e 1 i I : at i c n . 

i'rom trie star/ipoint of hu::ar: disease, atter.pts tc tra:.smit ]:iultiple 
sclerosis and Schil-;er's disease tc experii/ieiital aniinals has, taus far, been 
unsuccessful. In vitro stu.dies, although pcc^rly documented, liave also deen 
futile, ircgressive -multifocal leul'oencephalopathy (Fi-lL), the only hmnan 
de-r/elinating disease clearly acsociateM with a virus has been of great interest, 
lliis subacute process, usually occurring ir: patients with incompetence of their 
irmune response, has been characterir.ed by multifocal neurologic deficits. 
Patients have no fever or ceredrc spinal fluid abnor\"alities and tk:e coujrse is 



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progressive, eiidinio; in death. Multiple foci of demyelination surrounded 
by oli^AOdell.-■roglia have been found in the brain. Many of these cells have 
inclusion bodies containing papovavirus-like particles. Attempts to isolate 
virus remained unsuccessful, until recently when a virus which appeared 
unrelatea to previously recognized papovavirus was isolated as an SV40-like 
agent frc-n two cases of FML. The nucleic acids of the two SV40-like viruses 
isolaxc; ; f'rom patients with FML have been compared with the DM of laboratory 
strains of SVUo. Tlie DM of these isolates are both covalently closed circu- 
lar duplexes with a sedimentation rate the same as SV4o DM. Tlie denatured 
DNA hybridized with denatured SV^O DM at the same rate has homologous DM. 
■i^ilien digested with Hemophilus influenza restriction endonuclease, the DM of 
both the SV40-IML viruses yielded 11 fragments but in each case there were 
two fragments which differed from the classic strains of SV40, and these 
two fragments from the SV^+O-EML viruses differed from one another. 

Biological characterization of the SV40-like viruses isolated from 
PI^IL were also carried out. Both showed remarkable stability to chloroform, 
pH 3 and heating at 50°C for 120 minutes. Both viruses were shown to grow 
efficiently in BSC-1 and primary African green monkey kidney cells although 
SV^O-H'IL 2 virus showed more efficient replication, and the viruses differed 
in their plaquing characteristics. Both agents caused transformation of 
rodent cells, and in Wi-35 human cells there was apparent serai-permissive 
infection with the production of infectious virus but also cell transformation. 
Serum and kinetic neutralization tests showed complete cross reaction of SV^O 
virus. Tumor, "T", antigens appeared identical to SV^O. No tumors were in- 
duced in mice inoculated with the viruses, but tumors were produced in hamsters 
resem.bling the tumors induced by classic strains of SV4o virus. 

These studies have given further infonnation on the mechanisms by 
which virus may produce demyelinating disease both in experimental animals 
and man and represent some detailed characterisation of the papovaviruses 
associated with progressive multifocal leukoencephalopathy, the first chronic 
hmnan demyelinating disease demonstrated to be of viral etiology. 

Progressive multifocal leukoencephalopathy is caused by papova 
viruses, but their exact identity and significance in producing demyelination 
still require clarification. A strain has been isolated for which antibodies 
were detected in 60'^3 of randomly collected human sera of all ages. This strain 
is distinct from, polyoma and 3V^0 viruses. Tumors have been successfully 
induced in the brain of hamsters after intracerebral inoculation of this 
h'oman papova virus. Tlrie research plan now includes; morphologic identification 
of the DM in this and other papova viruses by means of the Kleinschmidt 
spreading technics identification of viral antigens by electron microscopy; 
isolation and characterization of papova viruses derived from other cases of 
P!"IL; morphologic studies on the formation of brain tumors induced in hamsters 
by ?naman papova virus; and comparative studies of polyoma, SV^O, and papova 
viruses from other laooratories. 

Transmissible mink encephalopathy (TI'IE) is akin to scrapie, kuru, 
and Creutzfeldt-Jakob disease, i.e., the noninflammatory, subacute spongiform 
viral encephalopathies. These diseases develop over a period of months and years. 
The infectious agents are still unidentified, but their properties are distinct 



23 



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from conventional viruses. A study is being undertaken inclulinf: sequential, 
morphologic study in hainsters ana monkeys inoculated with Tf-Ti brain suspensionn 
in order to determine the localization and precise ultras true tural alteratioi.s 
that develop in neirrons, particularly in synaptic regions, months before th.e 
onset of clinical s.y7Tptoms. Tlie proposed TMl-; stuny is important in that the 
disease mimics hiiman '.'reutzfeldt-Jakob disease. Thus far, all ultras tr'/ic tural 
studies have aealt with the advanced stages of the disease in jnan and animal, 
hut the elusive agent could conceivably be visualize-! earlier. T'-'ie changes 
in the plasma membranes that appear to initiate the spongiform alteratior.s 
in ne'orons warrant in depth study, for speculations concerning abnormal mem- 
branes are supported by biochemical data on the infer; ticjs ag=nt. 

Herpes simplex viruses, tjrpF- 1 ana ^, are the ''^ause C'l' life- 
threatening encephalitis in the United States today. Tlie real frequency and 
clinical spectrum of disease is not fully known. Studies are li;'iited by th.e 
current necessity of isolating herpes simplex virus (KSY) fromi the brain ir. 
order to make a definitive diagnosis. In prelimiriary studies T^assivo 
hemagglutinating antibodies (p1-!A) in cerebrospinal fluid ]:av.; ; cnj •'^.unl early 
during the courses of several patients with liSV encephalitis arid w.l uii herpes 
virus hominis encephalitis, 'fliis PHA-''S?" technique will be evaluaten as an 
imjriediately available diagnostic test for ^'SV encephalitis, derebrcspinal 
fluid PPIA will be correlated with isolation, of -.'■irus at -rain biopsy and 
with measures of conventional and complement requli-ii g !'Outi'ali/ing avtibodies 
in sera and CSF during various neurological conditions. Diagnostic studies 
will be augmented by _in vivo and in vitro studies of the pharnacology of 
potentially useful antiviral agents in e:cperimental models a;;;i ii: man. An 
in vitro assay of antiviral activity in body fluids due to adi:iinistered Irug 
which is not affected by interferon or homologous seriuri an.tiboiy has been 
developed which includes concentrations of idoxuridine in seru::., urine and 
cerebrospinal fluid of patients with suspected diagnoses of ''erpes virus 
hominis encephalitis. Minimal inhibitory concentrations cf agent vs strains 
of HSV will be correlated with am.ounts of drug in blood, spiral fluid, brair. 
and urine after therapy. Possible antiviral syrieTiZy of several antiviral 
drugs in combination will be assayed. These data will guide ratlc al therapy 
in man. 

A number of viruses iiave been i:, plicated in the etiology of multiple 
sclerosis. It is surprising that vaccinia infection, almost universally 
distributed, has net been sought as a possible agent. However, no studies 
on va.ccinia studies of multiple sclerosis have been 'cne with the ex-.-eption. 
of electron m.icroscope searches for elem.entar;,'" particles which have been 
unsuccessful. It Is clear that, if vaccinia antibodies are founu in a 
significant number of spinal fluiis cf multiple sclerosis victi-.s and in no 
other chronic neiurologic disease, this ii.ight be of some diagnostic use for 
neurologists faced with early states of th.e disoruer, such as optic ueur-itis, 
which may or may not progress v.o the full-grr'-- pictiure of miulticle sclerosis. 
More important, if vaccinia can be related to e'^iology, a significant red'.ictioi 
in incidei'.ce of routine vaccination, :iiay lead to a reduction In this lisease- 
A study is oelng conducted on the frequency of the presence of r_eutrali ;in.g 
an/tibodies against vaccinia virus in thie spinal fluids of victims of ■ -iltiple 
sclerosis and of other patients suffering from chronic brain s;,a;ircr:es - Rre- 
liminai-y evidem^e suggests that only multiple sclerosis patients "nave sucli 






antibodies in spinal fluid. In order to relate the possible etiologic sig- 
nificance of this finding to the disease itself, it is proposed to do fluores- 
cent antigen studies on brains of multiple sclerosis victims using specific 
antibodies against fo'or soluble antigens of vaccinia (WP, S, HS, HL). 

Visna is the only model of slow, virus-induced demyelinating 
disease in animals, but work on this infection has been limited because the 
disease has only been produced in Icelandic sheep. Previous attem.pts to 
adapt visna virus to other breeds of sheep or other animals have not been 
successful. Because of the importance of this model to study demyelination 
relative to multiple sclerosis, further attempts have been made to adapt the 
visna virus. Preliminary studies have been carried out using the rapid serial 
passage of visna virus in fetal sheep. Intracerebral inoculations have been 
carried out by laporotomy at 50 days gestation in fetal lambs. Lambs were 
sacrificed at weekly intervals thereafter for virus titration on sheep testicle 
tissue cultures and for explants of brain tissue. Preliminary, but encouraging, 
results have been obtained indicating there is definite replication of visna 
virus in the brains of American sheep. 

Visna was originally recognized in Iceland as a naturally occuring 
fetal central nervous system disease of sheep and was described by Sigurdsson 
as one of the prototypes of slow infection. The causal agent is now classified 
as an oncorna virus, based on its physical, biochemical, and biological 
properties. Following intracerebral inoculation of susceptible Icelandic sheep, 
a persistent infection develops followed by a late appearance of neutralizing 
antibodies. Clinical disease appears in 6 months to years after inoculation, 
and is characterized by a severe lymphocytic infiltration of leptomeminges and 
choroid plexus, with marked perivascular cuffing and eventual destruction of 
white matter. 

This study postulates that the disease is immunopathological in 
nature and will attempt to elucidate (a) the cellular sites of virus 
replication; (b) the role of the immune response (antibody and cell-mediated) 
in disease production; and (c) the mechanisms of virus persistence. Approaches 
will include (a) a sequential study of infection in Icelandic sheep; (b) the 
effect of immunosuppression, using antilymphoid serum of cyclophosphamide; 
(c) the effect of active immunization before and after inoculation; (d) the 
correlation of various measures of antibody and cell-mediated immunity with 
disease progression; (e) a search for evidence of an auto- immune response 
against myelin antigens; and (f) the influence of age of inoculation upon 
the course of infection. 

The overall goal of one laboratory is to investigate the biological 
properties of neurotropic viruses and their interactions with neural tissue at 
the cellular level. 

The specific goals are to analyze the nucleic acids and proteins of 
visna virus, to define the ultrastructure of the internal nucleic acid component 
of visna virus, to examine the brains of patients with post-encephalitic 
Parkinson's disease for the presence of influenza virus antigen and to study 
experimental murine influenza virus encephalitis with reference to the role 
defective autointerfering virus has on the course and outcome of disease. 



25 



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60-70S visna virus RNA extracted from purified virus was found to 
be composed of a heterogeneous population of molecules with two predominant 
forms, long, single filaments and branched strands. The mean length of the 
two forms ivas 6.6um. The shortest single filament measur^ed 1.0 um. the 
longest 9-3 urn- The width of these strands was 96 A. Visna virus RNA has 
been analyzed by electrophoresis- Although these experiments are still in a 
preliiuirary staj^e^, they indicate that the major viral KNA moities can be 
distinctly resolved in such gels and that the electrophoretic mobility of 
the major nucleic acid species is similar to that erctracted from MA tumor 
viruses. Preliminary experiments have been conducted to determine if the 
tissues of normal sheep or the tissues of iiaedi or visna-afflicted sheep 
contain nucleic acids which share sequence homology with the 3H-DM formed 
by the visna or maedi virus polymerase reaction.^ Both MA and DM have been 
extracted from sheep tissues, hybridized to the ~'H-DWA product and analyzed 
on hydroxylapatite columns. These studies are now in progress and no specific 
conclusions can yet be drai-m. 

Analysis of the proteins of visna virus indicated that visna virions 
have 11 to lU polypeptides, including at least two glycoproteins. Treatment 
of the virus particles with the proteolytic enzyme, broraelein, resulted in 
the removal of most of the glycoproteins as well as some of the non- carbohydrate 
containing polypeptides. It iTas concluded that in the number and general 
pattern of their mobility in gels, the proteins of visna virus resemble the 
proteins of the RNA tumor viruses. Studies of the amino acid composition of 
visna virions have shown that they do not contain any unusual amino acids. 
Tlie virus particle appears to contain small quantities of the polyamines, 
spermine and spermidine. 

Influenza virus antigen has been demonstrated in brain sections of 
patients with post-encephalitic Fiarhinson's disease by immuno-f luorescence 
staining. Sections of midbrain and hj'pothalam.us from 6 patients with well- 
documented post-encephalitic Parkinson's disease and 5 patients with idiopathic 
parkinsonism were studied. Using the direct immunofluorescence technique, 
sections were stained with f luorescein-conjugated gamma globulin against the 
neurotropic influenza A virus strains (iWS and ¥S-N), other unadapted influenza 
virus A strains (PRc, Japan, Swine), influenza B (Lee), measles virus and herpes 
simplex virus. Specific fluorescence in the nuclei of cells in the substantia 
nigra and hypothalamus was seen only in sections prepared from post-encephalitic 
Parkinson's disease cases after exposure to gamma globulin against the two 
neiirotropic influenza A strains. The specificity of the staining reaction was 
demonstrated by the inhibition of fluorescence after pre-treatment with 
unconjugated gamma globulin against either the IJ¥S or WS-W strains. No 
inhibition i-ra.s observed, however, after pre-treatment with gamma globulin 
against any other type A or B influenza virus strain. Specific fluorescence 
■'.■ras not seen in brain sections cf patients with idiopathic Parkinson's disease. ^ 
These findings indicate that antigenic sites capable of binding to gamma ' 
globulin directed against the NWS and ¥S-N strains of influenza virus exist 
in the midbrain and hypothalamus of patients who suffered from encephalitis 
lethargica and later developed post-encephalitic parkinsonism. Furtheimiore, 
they suggest that post-encephalitic parkinsonism may be etiologically distinct 
from idiopathic Parkinson's disease and that there is a link between influenza 
virijis and the post-encephalitic form of the disease. 

26 cc " 



The results obtained in these investigations may help to provide 
new approaches and techniques which will be of use in demonstrating that 
certain human subacute or chronic neurological diseases are due to viruses. 
They may help to explain the ways in which viruses cause destruction of nerve 
cells, define the factors which limit the progression of viral encephalitis, 
and point out similarities between viruses which cause slow infections of 
the nervous system and viruses which cause tumor formation. 

The objectives of another investigation are to understand the 
intricacies surrounding the replication of visna virus nucleic acid species 
and how these synthetic processes relate to the ability of this slow virus 
to persist in susceptible host tissue until overt clinical symptoms appear. 
The fonnation of visna virus DM intermediates designated as "DM provirus" 
will be studied in cultures of sheep choroid plexus cells, and the presence 
of these virus-specific DMs will be detected by testing the DM fraction of 
infected cells for its capacity to stimulate reassociation kinetics of 
denatured DM generated in vitro by reverse transcriptase or by testing for 
its integration within "network" DM formed by Britten-Kohn repetitive 
sequences under conditions of self -annealing. The aim in these experiments 
is to deteiTiiine whether visna "provirus DM" occupies a specific site on 
a host chromosome and associates with a given size class of cellular DM. 
The effect of various inhibitors of protein and nucleic acid synthesis on 
the formation and integration of these visna DM species will also be examined. 

Other major projects included in the research plan are molecular 
investigations of viral-specific nucleic acids and proteins in diseased tissue 
of sheep infected with visna virus and a search for resident gene products of 
covert slow virus genomes in human autopsy speciments of brain and other organs 
from victims of multiple sclerosis. Tlie latter tests will focus on demonstra- 
ting viral nucleic acids having sequences homologous to visna virus by re- 
association kinetics of visna transcriptase product, by DM-KM hybridization 
studies with labeled visna RNA and by use of complementary "minus" strand DM 
generated by the visna transcriptase for implicating the presence of viral 
MA transcripts in cell fractions. 

Ultrastructural investigation of viral infections involving the 
central nervous system of experimental animals and man is being conducted 
in another laboratory. Emphasis is being placed on disorders in which a 
viral infection may be associated with demyellnation or with a teratogenic 
effect. Human biopsy material from chronic degenerative disorders of the 
CMS of suspected viral etiology will also be screened electron microscopically 
for detection of a possible agent. Specifically, cerebellar maLformation 
induced by the Kilham rat virus, and the effect of the blue tongue virus on 
the cerebral cortical development will be examined. The study of the dem.yelin- 
ating disease induced in mice with the JHM strain of mouse hepatitis virus 
will focus on two specific questions, namely, the mechanism of demyellnation 
and the role of the immune response in demyellnation, and the source of cells 
involved in remyelination. Human studies will consist of a continued attempt 
to determ.ine the occurence and frequency of viral isolates in progressive 
multifocal leukoencephalopathy, and electron microscopic monitoring of 
cultures derived from patients with known and suspected viral disease of the 
nervous system. 



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The long-term objective of the rnultiple sclerosis (l-IS) virology 
program is to isolate infectious agents from tissue obtained from patients 
with multiple sclerosis and to characterize them and determine their relation- 
ship to the disease. The rationale for a virologic approach to the study of 
multiple sclerosis is based on the fact that some clinical feat'ores are common 
to both multiple sclerosis and the known slow virus diseases. Serologic and 
epidemiologic inf orrraticn incriminate a viral agent as the cause of multiple 
sclerosis. The recent isolation of a virus (6/9^+) fromi the brains of two 
patients with multiple sclerosis lends f'jrther support to the viral etiology 
concept. The multidisciplinary approach to the LIS problem (clinical, 
virologieal, immunological), com.bined with the newer laboratory techniques 
(cell fusion, use of halogenated pyriraidines, imraunosuppression) to activate 
viruses in tissue culture and in ezperimental animals, offers a better chance 
for the discover;/ of a virus than l:as previously existed. 

Multiple sclerosis fits the three criteria establishe i by Sigurdsson 
for slow virus diseases (a) after infection, a period of irany months or years 
without signs of disease; (b) after clinical signs appear, a proi;racte;i coiur-se, 
usually ending in serious inrpairment or death; (c) infeotii'r: l:!;it-;i lo a. 
single host species and lesions limited to a single organ system. Tlr.is , IK 
is probably acquired but not m.anifested during childhood, runs a protracted 
coujTse, seems to be species specific for man and attacks ii:itially only the 
CTIS. 

Som.e of the laiowii slow virus diseases of m.an (kur'i, Jakob- 
Creutzfeldt and SSPE) and animals (scrapie, visna, transmissible mink en- 
cephalopathy and Aleutian disease of mink) have been phcwn to be transm.issible 
to susceptible hosts. Although several studies have been conducted to induce 
or propagate these virus --' h MS-tissue erctraots, the efforts so far have 
rem.ained unsuccessful. ^ -- one other hand inoculation of animals with cells or 
tissue extracts obtained from cases of SSPE, progressive m.ultifocal leukoenceph- 
alopathy, kuru, Jakob -Creutzfe Id t, mink encephalopathy, scrapie and visna 
resulted in the production of disease. Only with two viruses, SSPE and H^IL, 
\Tas it possible to demonstrate and characterize a virus or its components inside 
CIjS cells. With tissue obtained from. Jakob- Creut^feldt cases, it has been 
possible to relate the presence of a virus to m.orphological transformation in 
both the original brain cultures and in cells derived from, cultui'es exposed to 
the original brain cells. 

Recently, after brain cells from two cases of i-'S were fused with 
indicator cells, an agent referred to as the 6/^k virus was isolated. This 
agent has not been propagated in embryonatea hens eggs. Hemagglution-inhibition 
tests with egg-grown 6/9k virus permitted it to be differentiated from both 
the HA-2 and Sendai agents; two prototj'pes cf para influence -/ir-.xses to which 
G/^k is related. 'Jsing physic ochemical technique;j sucn as purification, 
determination of m.olecular weight, sedimenta-^" - co-efficient, analysis of 
roiA species and electrophoretically-distinguisha^le protein migration, o/9k 
virus has been fui-ther studied and compared with tlie related Sendai virus. Tlie 
6/9h virus inoculated intracerebrally into newborri Sj^ian hiamsters caused 
severe clinical and pathological syndrome. On the other 'aand, :LA---: virus 
produced severe hydrocephalus but failed tc cause the marked wasting character- 
istic of 6/9U. 



-O cc 



It has been pointed out earlier that in two chronic CWS aiseases^ 
visna and PML^ a link has been suggested between slow virus infection and viral 
oncogenesis. Tlie rationale in searching for a similar correlation in IC is 
supported by the observation that the 6/94 agent is different antigenically, 
biochemically and biologically from more than one of the parainfluenza I 
prototype viruses and argues against it being a laboratory contaminant. Also^ 
the presence of nucleocapsids in brain cells of one I'lS case before fijsicn, 
the appearance of virions upon fusion of these cells with both indicator 
cells (CV-l and WI38) and lack of virions and nucleocapsids in non-MS brain 
"fused cultures" argue against the indicator cells being the source of virus. 
Tlae isolation of a similar (6/9^) virus from the second case of I'lS lends further 
strength to the possible association of this agent with multiple sclerosis. 

Brain tissues obtained at the time of biopsy or autopsy are being 
studied by light and electron microscopy in order to document and define any 
moarphologic alterations and to search for viruses in tissue sections from 
patients with, multiple sclerosis. These studies also include electron 
microscopy of tissue culture cells derived from nervous system of patients with 
MS, EM of supernatant fluid from cell cultures to search for viral 
particles, morphological characterization of virions, morphological analysis 
of tissue to define ijnitype cultures and morphological characterization of 
6/94 virus to augment the viral isolation studies and to define virus nucleo- 
capsici in cells prior to fusion and before other tests for the presence of 
virus could be considered positive. 

In general these programs are designed to achieve the following 
objectives (a) to obtain a viable MS tissue; (b) to investigate a variety 
of environmental parameters in search for the optimum conditions for the 
growth of human central nervous tissue in cell culture and their characteri- 
zation; (c) to continue attempts to isolate, rescue and identify virus(es) 
from tissue cultured cells using standard and newer virologic techniques; 
(d) to further characterise the 6/9^ virus, and (e) to study immunological 
reactions in MS patients. 



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NEUROMUSCULAR DISEASES 

The muscular dystrophies (MD) and myasthenia gravis (MG) constitute 
the most prevalent neuromuscular diseases affecting approximately 200,000 (>1D) 
and 30,000 (MG) individuals in the United States alone. The predominent 
symptom is muscular weakness. However, while MG is attributable to a failure 
of neurochemical transmission at the myoneural junction, the various dystrophies 
are primarily genetic in origin and are variously attributed to deficiencies 
in muscle proteins, muscle membranes, nerves, capillary circulation, and 
endocrine regulation. 

Myasthenia gravis results from a defect in transmission across 
the neuromuscular junction probably due to either the inadequate release of 
acetylcholine from the nerve terminals or from a decreased sensitivity of 
the motor endplate to the transmitter. The size of the miniature endplate 
potentials arising from the spontaneous continual release of acetylcholine 
from the nerve endings is greatly reduced. There is also evidence that MG 
may be an autoimmune disease resulting in antibody interference with neuro- 
muscular transmission. Drug therapy in MG is based either upon increasing the 
available acetylcholine at the junction or upon the suppression of the auto- 
immune mechanism. Anticholinesterase agents are used to decrease the rate of 
destruction of the neurohumor at the endplate and ACTH and corticosteroids 
are presumably effective through suppression of the immune process. The 
theophylline derivative, oxtriphylline, has proved effective in patients 
refractory to anticholinesterases. 

Muscular dystrophy includes a group of diseases characterized by 
progressive wasting of skeletal muscle, primarily in children 2-6 years of age. 
The condition is usually inherited, may be sex-linked (Duchenne type), trans- 
mitted as a dominent trait (facio-scapulo-humeral) or may demonstrate autosomal 
recessive inheritance (limb-girdle type). A varient MD of late onset may 
occur in both men and women after the age of thirty. The "white, fast" 
muscle fibers are primarily affected while the "red, slow" fibers are 
relatively resistant. In the case of the Duchenne type, patients and 70 per- 
cent of female carriers usually show high levels of serum creatine phospho- 
kinase, demonstrate excessive synthesis of collagen by isolated polyribosomes 
of muscle, and reveal a larger than normal percentage of red blood cells with 
abnormal membranes under the scanning electron microscope. There is no accepted 
treatment for MD other than physical therapy although unconfirmed reports 
suggest that the administration of high oxygen, glucagon, or coenzyme 0. are 
beneficial in the dystrophic mouse. 

Recent research on MG has centered about the development of a sensi- 
tive in vitro method for the evaluation of potential pharmacological agents 
and their mechanisms of action. Biopsies of guinea pig intercostal muscle 
have been effectively employed for this purpose. Parallel experiments with 
biopsies of human intercostal muscle have served both to demonstrate the 
potential clinical effectiveness of certain drugs and to devise therapy for 
individual patients who are refractory to anticholinesterases or are overly 



30 



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sensitive to certain agents. The preparation allows for an extensive electro- 
physiological analysis of miniature endplate potentials, frequency and 
amplitude, an analysis of endplate potentials, quantal content of acetylcholine, 
equilibrium potential, muscle fiber membrane resistance and capacitance, and 
receptor density. The facilitory action of oxtriphylline at the myoneural 
junction (and probably its clinical effectiveness) is related to its properties 
of increasing acetylcholine depolarization of muscle, increasing endplate 
potential, decreasing muscle membrane resistance and slightly depolarizing 
the postsynaptic membrane. 

The clinical evaluation of germine monoacetate (a derivative of the 
veratrum alkaloids) has continued to produce promising data suggesting that 
this agent uiay also eventually be available for therapy. It should be noted 
that the anticholinesterases, the theophylline derivatives, the veratrum 
alkaloid derivatives and the adrenocorticosteroids , each owe their clinical 
effectiveness to a different physiological mechanism. This is especially 
important in MG where individual responses to drugs are often unpredictable. 

Research in the area of muscular dystrophy is largely concentrated 
on tissue chemistry. One of the primary objectives has been to determine 
whether the myosin protein of human dystrophic muscle is intrinsically 
abnormal. Extensive studies of the enzymatic and contractile behavior of 
this protein have produced divergent results with reports that these reactions 
are normal, abnormal, and normal under certain optimal laboratory conditions 
but abnormal otherwise. The activities of other enzymes such as "alkaline" 
RNAse II have been found to be specifically increased in dystrophic mouse 
muscle. Investigations on human muscle have sho^m that the abnormalities 
observed in protein synthesis differ for each form of dystrophy, confirming 
that the dystrophies are inherited as separate diseases caused by defects in 
different gene products. 

Although Duchenne's Muscular Dystrophy has been associated with 
elevated levels of serum creatine phosphokinase, recent reports indicate that 
the activities of at least four other enzymes are similarly increased at birth, 
continue to rise for the first two years, and gradually decline to normal 
levels at the terminal stages (15 to 20 years of age). A limited preliminary 
clinical trial with diethylstilbesterol has resulted in reduced serum enzyme 
levels and apparently has arrested the expected deterioration of muscle 
strength. Finally, it may be noted that by a combination of techniques, such 
as light and electron microscopy, analysis of serum enzyme levels and protein 
synthesis by muscle biopsy tissue, it is possible that all or most female 
carriers of the Duchenne type of dystrophy can be identified and thus event- 
ually provide for effective genetic counseling. 



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NERVE GROWTIi FACTORS 

Nerve growth factors (NGF) have been isolated from mouse salivary 
glands, tumors, and snake venoms, and may be present in the serum of patients 
with neuroblastoma and disseminated neurofibromatosis (von Recklinghausen's 
disease) . In vivo , the administration of this material results in the en- 
largement of spinal and sympathetic ganglia. The addition of NGF to tissue 
cultures of embryonic ganglia of chick, mouse or human, stimulates extensive 
outgrowth of neurites. 

The normal physiological role of NGF in the body is not kno^^m, nor 
is its detailed mode of action. However, it has been ascertained that NGF 
is an "insulin- like" protein that can be fractioned into a aumber of poly- 
peptides of varying biological activity and can be reconstituted from its 
fragments. NGF may be one of several related growth promoting substances, 
since other proteins which stimulate the elaboration of epithelial tissue 
have also been isolated and partially characterized. However, the administra- 
tion of specific antiserum to NGF into newborn mice causes degeneration of 
sympathetic nerves (producing "immunosympathectomized" animals) and suggests 
that NGF does function physiologically. 

NGF might be of great potential value in promoting regeneration in 
the central nervous system. However, NGF stimulation has thus far been limited 
to ganglion cells and neurons in the central nervous system of sympathetic 
origin. No preparation has been shown to promote the growth of neuropile 
or tracts of the spinal cord. 

Much of the current research in this area is concerned with the 
isolation, structure, and chemistry of NGF derived from different sources, 
and the relation of these factors to biological activity. The mechanism of 
action of the protein on target cells is also under investigation. Recent 
reports indicate that stimulation of growth is not mediated via cyclic AMP, 
and that NGF need only bind to the cell surface to exert its actions. It 
is estimated that there are approximately 20,000 binding sites per cell. 
Of potential importance is the observation that the glial cells of the ganglia 
may constitute the normal physiological source of NGF for the neighboring 
neurons . 



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REGENERATION IN THE CENTRAL NERVOUS SYSTEM 

Regeneration of nerve cells in the severed mammalian spinal cord 
leading to a restitution of function has not been reproducibly demonstrated. 
Outgrowth of fibers has been shown to be abortive, presumably because of 
mechanical blockage due to glial or connective tissue proliferation. Recently 
there has been a revival of interest in this subject because of a number of 
advances in neurobiology. It has been shown that nerve fibers may grow con- 
tinuously throughout life, or at least material synthesized in the cell body is 
constantly transported along the fiber. Recent electron microscopic investigation 
of synaptogenesis in the CNS indicates that these connections may be continuously 
"remodeled." It has also been noted that lesions in the CNS can stimulate 
extensive collateral sprouting of axons which potentially could establish 
functional connections. Successful regeneration of the severed spinal cord 
has been observed in the goldfish. Also, the extensive development of tissue 
culture methods for nerve cells, ganglia, spinal cord, and cortex has made 
available a variety of materials which might be implanted into the damaged 
spinal cord. 

Much of the current research in this area is concerned with the 
nature and extent of collateral sprouting that occurs after experimental 
lesions are made in the CNS. The results are extremely variable and probably 
reflect different growth potentials in different regions of the CNS. For 
example, in the hippocampus, depending upon the fibers involved, sprouting may 
be minimal or elaborate and new synapses may form, become functional and 
eventually degenerate. Sprouting stimulated by severing the mammalian spinal 
cord usually does not cross the transection extensively, but results in the 
formation of sjmapses on the proximal side of the lesion. There are several 
possible reasons to account for this behavior aside from the formation of 
impenetrable scar tissue. The establishment of a synapse a small distance 
laterally may inhibit further growth of the sprout or the regenerating axon 
(contact inhibition) . Cell bodies which become partially deaf ferentiated after 
the lesion may attract the growing sprout; the nearest cells with available 
surface for contact are on the proximal side of the lesion. A variety of 
experimental procedures have been proposed to encourage the regenerating fibers 
to grow across the transection. These include the administration of a number of 
drugs which may suppress glial proliferation, possibly via inhibition of the 
body's immune defense mechanism. Radiation of the severed cord to inhibit 
^ glial growth is also being attempted. 

The specificity of new synapses established by the sprouting fibers 
is also under investigation. Aside from the viability of new connections, the 
consequences of aberrant synapses on behavior are being evaluated. In the case 
of the regenerating olfactory nerve, electrophysiological, anatomical and 
behavioral parameters can be utilized to assess the reorganization of the 
olfactory bulb. This system lends itself to detailed analysis because of the 
exquisite sensitivity of the olfactory epithelium to odors. Regeneration in 
the nervous system of several invertebrates is also under study. 



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Finally, it may be noted that rat spinal cord segments, maintained 
in tissue culture, can be successfully grown in the spinal subdural space, 
eventually becoming vascularized, but remaining isolated from the intact 
spinal cord. This preparation has not successfully produced anatomical 
connections between the two parts of the transected cord, but significantly, 
has not been rejected by the immune defense system of the host animal. 



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CEREBROVASCULAR DISORDERS 

Beyond the age of forty, the greatest killers of Americans are 
cancer, heart disease and stroke. In the United States it has been estimated 
that about two million citizens have cerebrovascular disease with an approximate 
death rate of 200,000 per annum. Research on the pathophysiology of cerebro- 
vascular disease over the past twelve years has been intensified, primarily 
through the leadership and fiscal support of the NINDS. The support level 
has risen steadily through the years to $8,900,000 in FY' 74 (43 research 
grants, 15 clinical research centers, and 11 stroke acute care research units.) 

it has long been known that the mature brain cannot be deprived of 
oxygen or its chemical substrate glucose for very long periods of time. 
Consciousness is lost when the arterial p0« has been reduced to 30 Tor or 
when the cerebral blood flow (CBF) has been decreased by 50 percent. There- 
fore hypoxemia and ischemia share equally in the genesis of brain anoxia. 

Studies on the physiological factors controlling cerebral blood 
flow (CBF) in animals have demonstrated that cerebral vessels seem to respond 
either directly to pH changes, viz. effect on vessel walls or via a sensor at 
some distant point, acting via a neural pathway. Additional data from the 
same research center have also shown that CBF remains unchanged when the 
blood pH is altered, provided the pH change is not produced by changes in 
pC02 . Therefore CBF is independent of blood bicarbonate alterations. The 
investigators concluded that the two important factors controlling CBF are 
extracellular Hf concentration and extracellular pCOo . 

Further studies in the same center on regional cerebral blood 
flow (rCBF) after stimulation of the cervical sympathetic nerves in dogs 
have reinforced earlier studies, substantiating the fact that rCBF is 
diminished following sympathetic stimulation. The effect was reversed when 
the animals were hypotensive and the effects were greater in the cortical 
areas as opposed to the subcortical white matter. 

Work in another center on acid-base and CBF changes in cat brains 
during seizure activity yield data which demonstrated that the CBF increased 
within a few seconds of seizure onset. The CBF increase was maximum when the 
blood pressure increase was maximum. Coincident with the increase in CBF, 
there was an increase in brain pH. The pH fell and returned to baseline 
levels in three to four minutes. Autoregulation of CBF to pressure was 
impaired for several minutes after each seizure. These data do not seem to 
support the concept that cortical perivascular acidosis is responsible for 
the dilatation of cerebral vessels during seizures. In addition, they imply 
that during seizures CBF is equal to, or occasionally in excess of the 
metabolic requirements of the brain, and that the onset of electrical 
evidence of seizure activity is frequently accompanied by cortical alkalosis. 

Patients with orthostatic hypotension were given five percent carbon 
dioxide in air and cerebral blood flows were recorded before, during, and 
after the carbon dioxide exposures. A majority of the patients studied 



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gave evidence of idopathic autonomic insufficiency (lAI) . CBF remained 
constant during norepinephrine induced hypertension. Apparently autoregula- 
tion was preserved but tilting reduced the perfusion pressure below the base 
line pressure for autoregulation. These results would seem to imply that in 
man the autonomic nervous system normally has an effect on the regulation of 
CBF, responding to changes in perfusion pressure but not to metabolic 
stimulation. 

Work in another stroke center on CBF changes in animals, as 
affected by the introduction of 70 ,(. of autogenous fresh femoral arterial 
blood into the subarachnoid space of the contralateral hemisphere, demonstrated 
a decrease in CBF in the ipsilateral hemisphere. This late change (30 minutes 
to 4 hours after injection) in vascular tone was thought to be due to a 
neuronally transmitted "diachisis." 

Investigators using the technique of unilateral sympathetic stimula- 
tion in anesthetized dogs found that CBF on the stimulated side was reduced 
by 80 percent but total brain blood flow as only decreased by 5 percent. 
Additional studies with tritiated water, where the average upLjku of brain 
water per gram was calculated, tended to reinforce the concept that CBF is 
heterogenous . 

Investigations carried out by another group to further our under- 
standing of the mechanism of irreversible cerebral damage by hypoxia or 
ischemia have shoi\na that mitochondrial oxygen requirements are critically 
dependent on the rate of electron flow, plus the presence of a phosphate 
acceptor, generally verifying the supposition that mitochondrial oxygen 
requirements are proportional to the "turnover number" of cytochromes. 
Tissues of lower respiration rates will be satisfied by oxygen gradients. 
Additional experiments by this group on anoxia and anoxic depolarization in 
the intact animal suggest that cells close to capillaries tend to become 
hypoxic later than cells distant from capillaries and may be influenced by 
potassium diffusing from distant cells. 

Animals with injected autogenous fresh arterial blood (see above), 
in addition to the blood flow changes, also gave evidence that the metabolism 
of ATP and glycogen in the treated hemisphere showed signficant decreases 
while AMP, pyruvate, lactate and the pyruvate-lactate ratios were significantly 
elevated. Metabolic derangements still existed 24 hours following the 
hemorrhagic insult. In addition, the NADH/NADf ratio was also at the lower 
limit of normal range. Brain stem norepinephrine was increased, dopamine 
decreased, and serotonin levels unchanged. 

Preliminary data from another stroke center has sho^^m precipitous 
depletions of catecholamines (epinephrine and 'norepinephrine) in cats with 
experimentally occluded middle cerebral arteries. Sham operated animals 
gave evidence of decreased catecholamines but less than half of that recorded 
in the experimental animals. 

Over the past few years there has been an intense interest in the 
mechanisms of thrombus formation and its possible relationship to the genesis 
of thrombotic strokes. Hageman factor activation has been shown by some 



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investigators to result in the activation of the intrinsic coagulation system 
and also in the generation of bradykinin. Bradykinin in minute amounts 
produces hypotension, increases blood flow and microvascular permeability, 
influences smooth muscle contraction, incites pain and stimulates leukotaxis. 
More recent observations from this stroke center have established that collagen, 
a constituent of the subendothelium of blood vessels, activates the Hageman 
factor and thereby initiates coagulation. These studies have demonstrated a 
structural and chemical specificity for the activation and binding of plasma 
Kallikrein-like activity by the collagen of the endothelium. It is therefore 
suggested that a plasma Kallikrein system may function as a surface-bound 
enzyme system. The subendothelial location of collagen suggests that disruption 
of the vascular endothelium may produce a local activation and concentration 
of collagen-bound Kallikrein-like activity. Collagen induced activation of 
both the coagulation and Kallikrein systems raises the possibility that the 
Kallikrein-kinin vasoactive system may function in both hemostatic and 
thrombotic processes. The activation of these systems by collagen establishes 
yet another interrelationship between coagulation and Kallikrein pathways 
which may have pathophysiologic relevance to various inflammatory human 
disease states. ...... 

An attempt has been made to explain the possible reasons for observed 
differences in mortality and morbidity from stroke through the Nationwide 
Cerebrovascular Disease Mortality and Morbidity Study. Three areas were 
selected--one with high, one with intermediate and one with low mortality 
from stroke; Pueblo, Colorado; Hagerstown, Maryland; and Savannah, Georgia, 
respectively. 

A higher percentage of white men in Savannah had systolic blood 
pressures over 140. The excess was limited primarily to the 140-159 (border- 
line hypertension) group. Mexican-American men in Pueblo had the lowest 
prevalence of systolic hypertension, but the sample was limited to 39 men. 
For the women, the prevalence of systolic hypertension was higher in Hagerstown. 
The pattern for black men was similar to white men. Black women in Pueblo 
had a much lower prevalence of systolic hypertension than women in either 
Hagerstown or Savannah. There was only a small difference in the prevalence 
of systolic hypertension among black and white women in Pueblo. There also 
appeared to be large differences in the sex ratio of the prevalence of 
systolic hypertension. In Hagerstown, the prevalence of systolic hypertension 
was similar in men and women while in Savannah the prevalence of systolic 
hypertension was about twice as high in white men as compared to white women. 

There were no differences in the prevalence of (4th phase) diastolic 
blood pressure over 90 among the men. The black women in Pueblo had a much 
lower prevalence of diastolic hypertension than black women in the other two 
areas . 

There were no consistent differences in the prevalence of elevated 
serum cholesterols among the subjects from the three areas, nor between men 
and women or blacks and whites. This was also true for plasma glucose values. 
White men were less obese in Pueblo than in Savannah or Hagerstown. The 
frequency of cigarette smoking was greater in Savannah than in Hagerstown or 
Pueblo. 



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Non-hypertensive blacks in Savannah had higher concentrations of 
urine sodium but there were no differences in the urine sodiums of hyper- 
tensives from the three areas. The Na+/K+ ratios were also higher for 
hypertensives in all areas and higher for black men in the Savannah area. 
There was no consistent relationship between the Na-f/K+ ratios and the level 
of blood pressure. The striking differences in prevalence of elevated blood 
pressure among black men and women among the areas is further evidence that 
the etiology of hypertension is different among black men and women. 

Aphasia is a frequent complication of stroke and, along with 
hemiplegia, ranks as one of the major rehabilitative problems associated 
with cerebrovascular disease. Studies of auditory reception in patients with 
aphasia demonstrated that injury to the left hemisphere apparently prevents 
phonological analysis of speech sounds. This finding is the same in French 
and English speaking aphasics. The results indicate that phonemic and 
semantic discrimination may vary independently' in aphasia and it appears 
dubious that the phonemic hearing disorders can explain auditory comprehension 
difficulty. 

Animal ablation studies showed that ablation in small steps 
permits recovery from dorso- lateral frontal lobe functions but not of 
orbital frontal functions. This difference has been suggested to be related 
to the phylogenetic age of the respective structures. Once recovered, these 
functions appear to become vulnerable to lesions in widely dispersed regions, 
which would normally have no effect if the dorso -lateral frontal cortex were 
intact. 



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TRAUMA (head DJJUP.Y) 

Investigations in the field of head injury, both clinical and animal, 
are of major interest to the NIWDS which is currently supporting several head 
injury centers as well as numerous related research projects. These studies 
invcl.-e . a'\y disciplines including neurosurgery, neurophysiology, neuropathology, 
anesthesiology, nuclear medicine, and biomedical engineering. 

A fundamental question regarding brain injury is whether patients 
who are admitted to the hospital in coma with signs of brain stem dysfunction 
have sustained an irreversible injury at the moment of impact or shortly 
thereafter, or whether they are comatose because of complications of the 
injujry which are potentially reversible. Hypoxia caused by respiratory 
insufficiency and ischemic brain hypoxia are common complications of acute 
brain injuries and are responsible in part for the continued high mortality 
and morbidity in patients. Consequently, many laboratory and clinical 
investigations are focused on its role in the deterioration of neural function 
in head injury. 

Tliere is much concern with the dynamics of brain injury such as 
alterations in vital signs, the significance of increased intracranial 
press'ore, the status of cerebral blood flow and the interrelationship between 
these two parameters, and the cardiovascular pulmonary complications. Also 
related to cerebral blood flow is the question of autoregulation and its 
im.plication in the head injured patient. Recently the metabolic concomitants 
of brain injury have assumed importance with studies cf the biochemical 
substrates such as lactate, pyruvate, a;lenonosine triphosphate, phosphocreatine, 
etc. 

The treatment of acute brain injuries has been largely empirical. 
A rationale exists for each method therapy, but identification of the patholog- 
ical conditions against which the treatment is directed is difficult in the 
individual patient. Four of the principal methods which have been used to 
treat acute head injuries are hypertonic solutions, hyperventilation, hypo- 
thermia, and steroids. A new therapeutic agent currently being evaluated in 
anim.als is dimethyl sulfoxide. Results from basic studies currently underway 
on the pathophysiology of cerebrospinal fluid formation, on the passage of 
protein through blood vessels, and on the enzymatic control of cortical 
swelling may provide new approaches for the treatment of cerebral edema 
associated with head trauma. 

Clinical evidence has been obtained in one of the head injury centers 
that metabolism miay decrease with cerebral blood flow, not as a manifestation 
of ischemic brain damage, but as a compensatory response to avoid brain damage 
in the face of decreasing blood flow. If metabolism is decreased because of 
ischem.ic brain damage, prognosis is in doubt, and improvement in blood flow 
will not benefit irreversibly damaged neurons. However, if a patient is 
comatose because of a pnysiological decrease in metabolism, he does not 
necessarily have brain damage because there is an appropriate balance between 
metabolism, and flow. If blooo flow can be increased by reduction of edema, 
and increased intracranial pressure, for example, metabolisii' will increase 
also and the patient will improve. Animal experiments designed to test this 



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hypothesis lend support to the clinical observations in that anerobic glycolysis 
did not occur until cerebral blood flow had fallen below ^5fo of normal. How- 
ever, before anerobic glycolysis developed, a marked reduction in brain oxygen 
metabolism (CMROp) and brain glucose metabolism (CMRGi) had occurred demon- 
strating that the decrease in metabolism preceded the derangement of metabolic 
pathirays produced by inadequate delivery of metabolites to the brain. Thus, 
there appears to be a m.echanism whereby decreased cerebral blood flow causes 
a compensatory reduction in brain metabolism of o:-cygen. 

Animal models have provided valuable information but the relevance 
to clinical head injury is not always established. To meet the need for a 
better model, a device is being developed which produces acceleration- 
deceleration head injury in the monkey and is capable of creating a broad 
spectrum of physiological and pathological abnormalities which are predictable 
according to the force of the injury. The injury is produced by a head 
accelerating device which rotates a monkey's head encased in a helmet. 
Measurements are made of systemic vital signs, intracranial pressure, EEG, 
regional cerebral blood flow, and brain metabolism of oxygen and glucose. 
Postmortem findings are then correlated with these measurements. 

Cultured glial cells have been used as an _in vitro model to study 
the effects of dimethyl sulfoxide on brain cells subject to sonic stress. 
This simplistic approach affords a chance to examine one important variable 
in head injury, i.e., the cellular response free of interference by vascular 
and interorgan reactions. Survival of the cells when sonicated in the presence 
of 10'^ DMSO was T8^. The mechanism of this protective action awaits further 
clarification. Glycerol and urea, however, had little or no effect, indicating 
that their usefulness in head injured patients apparently rests on mechanisms 
at a different level. 

Since a great va.ny penetrating injuries of the head result in 
post-traiimatic seizures, a rational approach would be to attempt to prevent 
scar formation at an early stage or to attempt to dissolve it with an antiserum 
after its formation. The success in this latter approach hinges on a clear 
understanding of the pathophysiology of gliosis, since scar formation in the 
brain results from astrocytic proliferation. A brain protein with a molecular 
weight of about 300,000 capable of stimulating the growth and maturation 
of astrocytes under tissue culture conditions has been isolated and partially 
purified. With an antiserum against this protein it may be feasible to bring 
about immunosuppression of gliosis and scar formation. 

The syste mic effects of intracranial trauma are being explored in 
depth in order to provide information which may be of value in the management 
of the head injured patient. In experim.ental head injuries on Fihesus monkeys, 
a variety of changes in the visual evoked responses (VER) have been found. 
These changes seem to be related to multiple fac'.,ors, including hypoxia, 
ischemia, intracranial pressure, etc. With profound hypoxia, the VER disappeared 
in 6-9 minutes and with milder degrees of hypoxia the amplitude remained con- 
stant for 2-3 hours. 

Observations made on nine patients with acute head injury confirmed 
the existence of a significant incidence of impaired phagocy-tosis as reported 



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by other investigators. These studies are being continued with serial 
investigations on these patients at 2, 5 and Ik- days after head inj'jry. 

Observations on acid-base balance after head injury have demonstrated 
that soon after injury most patients have an arterial pH which is slightly 
acidotic. Usually within the first 2h hours a respiratory alkalosis develops. 
Early after injury urinary excretion of hydrogen ion is elevated, but then 
progressively decreased with an increase in urinary bicarbonate. Thus, 
immediately after injury a mixed metabolic acidosis respiratory alkalosis is 
present. With supportive therapy, the metabolic acidosis rem.its and respir- 
atory alkalosis is exposed. 

One of the head injury centers has been interested in quantifying the 
response of cardiac and pulmonary function to head injury and the detailed 
characterization of the manner in which cerebral blood flow (CBF) and oxi- 
dative metabolism (CMRO2) fail after head injury in order to establish a 
more rational basis for treatment. A most important contribution of this 
group has been the development of clinical mass spectrometry for the purpose 
of monitoring cerebral blood flow and oxygen consumption in brain injured 
patients on a continuing basis. This technique is used during a period of 
clinical stability usually a few days after the initial injury. 

Cardiovascular and pulmonary functions, though important to overall 
clinical status, are not considered by these investigators to be fundamental 
causes of persistent and progressive brain dysfunction. Consequently, 
there has been a major shift in the direction of this program towards solving 
in the laboratory regional problems of flow and metabolism centering around 
the hypothesis that flow and metabolism are regulated independently by a 
brain stem mechanism. Another contribution of this group has been the revival 
of the free radical theory of membrane damage and the possibility of therapy 
by antioxidants and free radical scavengers. 

The treatment of acute brain trauma is greatly aided by information 
concerning the extent and location of the injury and the methods available 
to the physician for assessing patient status are now somewhat limited. It 
is extremely important to have available a rapid, safe, atrauinatic and 
repeatable visualization system for obtaining information on brain structures 
to assist in diagnosis and therapy of the severely traumatized brain. 
Differentiation between intracerebral hemorrhage and edema is one of the 
contributions of a study on ultrasonic visualization assessment of brain 
trauma. Hemorrhage can be readily identified as it produces strong echoes. 
Edematous areas seem to produce fewer than normal echoes, and can usually 
be identified with shifts which are visible in the falx and tentorium. It 
seems likely that localization and extent of edema can be identified, and 
the effectiveness of various anti-edema therapies assessed. The demonstration 
that human skull segments interposed betwec-i the ultrasound receiver and 
standard targets do not produce as much distortion as thought has encouraged 
work in this area. Adding simulated scalp and hair does not appear to refract, 
or scatter focused sound beams as much as anticipated. This fact brings the 
realization of transkull brain visualization closer to reality. Visualization 
of autopsied human brains has provided information on the echogram appearance 
of large intracerebral hemorrhages and surgical Interventions. Visualization 



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of patients following intracranial surgei*y suggests that these entities can 
be correctly diagnosed ±n vivo . It appears that several ultrasound scans 
made at intervals, timed to detect sequential changes, will be of more aid 
in the detection and differentiation of small hemorrhages and edema than a 
single scan will. 

It is now established that raised levels of cerebrocortical e;ctra- 
cellular KT"" are associated with astroglial swelling (cerebrocortical edema). 
Moreover, it is suspected that pericapillaiy astroglial swelling in response 
to an ischemic insult of brain occludes capillaries of cerebral cortex and 
further endangers neuronal elements marginally surviving the initial insult. 
At least three enzyme systems which may be responsible for K dependent 
swelling of the cerebral cortex have been investigated. The finding that 
ethacrynic acid inhibits not only all three enzymes but also cortical edema 
formation may have clinical application. 



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COWULSIVE DISORDERS 
History 

CoiiATulsive disorders have been recognized since the beginning of 
recorded history. In spite of this, seizures were for centirries thought to 
be caused by "evil spirits" which possessed the afflicted person. It was 
not until the latter part of the nineteenth century that the investigations 
of John H. Jackson provided a sound basis for the understanding of epileptic 
seizures when he proposed that seizures were caused by "occasional, sudden, 
excessive, rapid and local discharges of gray matter." Increasing sophis- 
tication in the techniques of electroencephalography, neurophysiology and 
electrophyoiology have led to the recognition that epilepsy is not a single 
disease but rather a collective term for a class of chronic convulsive 
disorders whose symptomatology is extremely varied. This variation is 
reflected in the two international epilepsy classifications agreed upon in 
1972, one based upon epilepsy type and one upon seizure type. 

Classification of the Epilepsies 

I. Generalized. 

A. Primary (includes petit mal, grand mal, and hypsarrhythmia) . 

B. Secondary 

C Undetermined 

II. partial (local, focal) (includes Jacksonian, temporal lobe and 
psychomotor). 

Classification of the Seizure Types 

I- Partial (beginning locally). 

A. With elementary symptoms (generally without impairment of 
consciousness); includes: motor (including Jacksonian 
seizures); special sensory or somatosensory; autonomic, 
and compound forms. 

B. With complex symptoms (generally with impairment of con- 
sciousness); includes: impainnent of consciousness only; 
cognitive symptoms; effective symptoms; "psychosensory" 
symptoms; "psychomotor" symptoms (automatisms); and com- 
pound forms. (All of these make up former temporal lobe/ 
psychomotor category. ) 

C. Secondarily generalized. 

II. Generalized. (Bilaterally symmetrical without local onset); 
includes: absences (petit mal); bilateral massive epileptic 
myoclonus; infantile spasms (hypsarrhythmia); clonic, tonic, 



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tonic-clonic (grand mal); atonic and akinetic. 

III. Unilateral (or predominantly). 

IV. Unclassified (due to incomplete data). 

Etiology 

The cause(s) of epilepsy is unknown. Although many epileptics 
have histories of brain damage, drug or alcohol intoxication, nutritional 
deficiencies, fever infectious diseases, brain tumors and abscesses or 
congenital brain malformations, many have no pathology to which epileptic 
seizures can be ascribed. 

Histopathology 

One grantee is studying the histopathology of the hippocampus in 
patients referred for surgical treatment of epilepsy. The most interesting 
group of patients are those in which no pathological lesion can be found 
though there is electrical evidence of its existence. In this group both 
dendrites and associated synapses with darkly stained granular matrices 
were observed together with evidence of ongoing axonal degeneration. It 
was concluded that degeneration hypersensitivity may influence the ictal 
substrate in these cortices. Similarly, another grantee has reported 
that degeneration and astrocytic alterations are frequently present in 
human cortical tissue obtained from epileptics. These findings were similar 
to the changes found in monkeys with altunina induced clinical seizures. 

Diagnos is 

Because of the variety of symptoms associated with epileptic 
seizures, it is difficult in many cases to establish the precise diagnosis, 
particularly in those patients with very mild seizures. One of the main 
diagnostic problems is the interpretation of the electroencephalogram (EEG) . 
Several grantees are investigating the use of computer technology for evalu- 
ating clinical EEGs. One investigator has made maps of the EEG (compressed 
spectral arrays, CSA) and has developed an extensive atlas of no27mal and 
abnormal CSAs. In addition, a large variety of computer programs (EEG Data 
Analysis Systems, EMS) have been developed for use with the PDP-12 computer. 
Another grantee is studying several aspects of normal and abnormal EEG 
activity at rest and in response to visual stimulation, by means of several 
linear and non-linear analog and digital computer processing techniques. 
Specific areas of investigation include the rhythmic waves of alpha wave 
frequency induced by photic stimulation, frequency and phase stability of 
rhythmic alpha activity, and interhemispheric interrelationships. Also 
being studied is the vis ual- oculomotor (i.e., eye-brain-eye) system, with 
particular reference to the EEG and eye-movement (electro-occulographically 
monitored) aspects of processing information containing visual stimuli by 
the brain. 

One project tes developed a program to provide complete, accurate 
detailed clinical histories of epileptic patients. To accomplish this, it 



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was necessary to (l) define seizure types in currently acceptable terms, 
(2) develop a feasible and standard program to gather historical data on a 
wide variety of seizure types, (3) develop a program in which detailed and 
standarized information is obtained on specific types of seizures and, (4) 
develop a program which includes patients with more than one kind of seizure. 

Other investigators are studying the relationship between the KEG 
and seizure by means of a split screen technique which presents the patient 
on one-half of the screen and the EEG on the other with the audio channel 
recording the sequence of events in the auditory sphere. This technique 
has made it possible to accurately exhibit the kind of seizure and its exact 
relationship to the electroencephalographic changes. 

Treatment 

Central to the development of treatment modes for a given disease 
is the availability of animal models of the disease in question. Therefore, 
it is not surprising that many grantees are studying convulsions induced 
in a variety of experimental animals by a variety of methods . Perhaps the 
earliest of these methods originated with the demonstration in I87O that 
excessive electrical stimulation of the brain of animals could produce 
seizixres. Since that time, techniques for the production of seizures by 
electrical stimulation have been refined and, in fact, the use of this 
miodel led to the discoveiy of diphenylhydantoin (Dilantin) in 1938- 
Probably the most useful animal seizure model in recent times has been the 
chemically- induced seizure. In these models, epileptic foci are created 
by direct administration of chronic irritants such as penicillin, alumina, 
etc., to brain tissue. Another model which has become increasingly popular 
is that in which epileptiform activity can be induced by sensory stimulation, 
such as photic stimulation in Senegalese baboons and auditory stimulation 
in rats, mice and rabbits. 

One investigator, using sophisticated electrophysiological techniques, 
has compared the electrical events occurring in single cells from human 
epileptic foci with those described for chronic epilepsy produced by alumina 
in the monkey. The similarity in patterns suggested that these electrical 
events are characteristic of both the human disease and the animal model. 
Another grantee has developed a standarized procedure for producing a penicillin 
epileptogenic focus which repetitively led to EEG after discharges in unanes- 
thetized, immobilized cats. This acute preparation, unlike the chronic models, 
allows a more rapid evaluation of potential antiepileptic drugs. Using this 
acute preparation, the investigator found that standard antiepileptic drugs 
such as diphenylhydantoin and diazepam exerted anticonvulsant effects which 
were dose related. This preparation is currently being evaluated as an 
initial screening model. 

Other grantees are studying the mechanism, by which penicillin 
exerts its epileptogenic effects in chronic preparations. One aspect is 
the paroxysmal depolarization shift (PDS) of cortical neurons, the most 
typical intracellular electrographic manifestation of a potentially epilepto- 
genic lesion, in the penicillin treated cat. The data suggsst that the 
presynaptic terminal axon is depolarized and that this depolarization results 
in the generation of bursts of action potentials originating at axon terminals. 



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Audiogenic seizures are being studied by one grantee "who demonstrated 
a circadian rhythmicity of susceptibility to audiogenic stimulation in genetic- 
ally dete2nnined seizure susceptible rats. Similarly, the administration of 
thiosemicarbazide or 2-methyl-i^-amino-5-hydroxymethyl pyrimidine to genetically 
non-sensitive rats and cats -was capable of inducing reversible audiogenic 
seizure susceptibility. Evidence was also obtained that audiogenic response 
in general is mediated through the inferior colliculus^, while audiogenic 
stimulus bound emotional behavior may be mediated througli extra-lemniscal 
pathways . 

It has been estimated that drug treatment has successfuJl.ly controlled 
seizure activity in approximately seventy-five percent of the patients treated. 
As a result, much effort has gone into (l) the search for new drugs and, (2) 
investigations of the mechanism of action and pharmacology of known drugs. 
One grantee is actively engaged in the synthesis of analogues of 5-phenyl-2, 
ij— oxoz olid inedi one. Another investigator is studying the structure of anti- 
convulsant drugs by X-ray crystallography in an attempt to correlate 
3-dimensional structure with activity. Procyclidine and trihexyphenidyl, 
both anticonvulsant agents, ^^rere shown to possess sterochemical feat^Jires 
similar to those found to be common to diphenyUiydantoin and diazepam (Valium), 
suggesting that these four agents probably function as anticonvulsants by the 
same mechanisms. 

Two projects have studied the mechanism of action of diphenylhydan- 
toin (DPH) and phenobarbital. Both drugs have been found to increase the 
cerebellar Purkinje cell (P-cell) discharge rate concomitant with a decrease 
in penicillin- induced epileptiform activity in cats. Additional evidence was 
also provided that an intact cerebellum was required for the anticonvulsant 
action of DPH, phenobarbital and diazepam. These data add weight to the 
hypothesis that cerebellar inhibitory discharge (via the P-cells) plays an 
important role in the modulation of cerebral cortical excitability, hyper- 
excitability and epileptiform discharge. 

Another project is currently undertaking an extensive evaluation 
of carbamazepine (Tegretol) in humans. Early pharmacokinetics and a pilot 
study have been coiripleted and more extensive pharmacokinetic studies are 
underway. 

In addition to drug treatment, a group of investigators are 
evaluating the use of operant conditioning techniques for the suppression of 
epileptic activity in humans. The techniques involve identifying the rhythm 
from scalp EEG and conditioning to acoustic/visual signals with the purpose 
of enhancing EEG desynchronization. A small number of patienty have 
benefited from operant conditioning of the sensorimotor rhythm and the use 
of this technique will be expended to a larger number of patients. 



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MULTIPLE SCLEROSIS 

Multiple sclerosis (MS) affects approximately a half-million people 
in the United States and is considered to be one of the major neurological 
disorders. The onset of the disease occurs predominately in the 20-40 year 
old group and is usually progressive for the remainder of the life span, 
although intermittent remissions lasting months or years are commonly 
experienced. The symptoms are quite variable, depending on the part of the 
central nervous system affected, but the general pathological findings are 
fairly constant, involving demyelination of white matter and subsequent plaque 
formation. The etiology of the disorder is unknown, although many assume it 
to be eithei viral or immunological. This hypothesis is primarily based on 
information obtained from other demyelinating diseases. However, since myelin 
is a product of neuroglia cell, an aberration of glial metabolism is considered 
to be involved. 

Still unexplained is the unusual geographical distribution of MS, 
The incidence increases significantly with latitude, the highest occurring 
in the north temperate zones of Europe and North America, Adult migrants 
to areas of relatively low incidence, such as Israel, retain the pattern of 
their native region, while children under 15 years of age assume the lower 
risks characteristic of natives of the country to which they have immigrated. 

At one multiple sclerosis research center, efforts toward accumulation 
of basic clinical data on MS have been a continuing activity. Records of all 
the patients are under review as the patients are selected for participation 
in the various research programs. Definititions of categories of MS and a 
standardized examination have been agreed upon. Concern has been for both 
documentation and categorization. The problem-oriented medical record system 
of Weed has been instituted for new patients. This system is particularly 
appropriate in the care of MS patients with numerous medical and social 
problems. A collection of autopsy specimens is under development. The purpose 
of this effort is the rapid retrieval of specimens from MS patients dying 
accidentally or from complications of their MS. Many legal hurdles have been 
overcome, and an alerting system for the investigators is under development. 

Another group of researchers at this center has intensively studied 
the experimental latent herpetic infection which was established in spinal 
ganglia of mice. Mice are inoculated in the rear footpad with herpes simplex 
virus. The virus ascends through the sciatic nerve to the central nervous 
system, utlimately producing posterior paralysis and death in many of the 
animals. Some of the mice undergo a complete clinical recovery. When the 
sacrosciatic spinal ganglia of these latter mice are explanted and placed in 
"organ culture" in vitro , previously latent herpes simplex virv.s is induced, 
and infectious virus is replicated. They have shown that neurons undergo 
productive infection and have provided strong evidence that the virus travels 
in axons. Since it now appears that neurons harbor latent virus, an attempt 
is being made to establish latent infections in a mouse neuroblastoma cell 
line. If this can be accomplished, a system to determine precisely how latent 
infection is established will have been developed. 



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In addition, autoradiographic experiments involving latently infect- 
ed gaiglia and ganglionic transplantation studies which, together, should 
definitively answer these questions, are in progress now. Using viral C-RNS- 
ganglionic cell DNS hybridization procedures, these researchers have been 
unable to detect the viral DNA in latently infected ganglia. Attempts to 
improve this sensitivity and to use in situ hybridization methods are now 
underway. 

These investigators have also been engaged in the study of sera in 
a search for the proteins that are toxic to CNS tissue. They have perfected 
the techniques necessary to grow and maintain myelinated tissue cultures. 
Their studies on the fractionation of the toxic factor in amyotrophic lateral 
sclerosis (ALS) sera are still in the preliminary stages, but the following 
conclusions are probably iustified: (a) the toxic factor is non-dialyzable 
and is, therefore, presumably a protein; (b) it is not inactivated by heating 
to 56-58°C for one hour (not complement dependent); (c) the molecular weight 
is greater than 10^ daltons (determined by Pellicon filtration); (d) it is 
not sedimented from serum by centrifuging at 10^ x g for eight hours (probably 
not a virus); (e) it is present in the cerebrospinal fluid of ALS patients. 
This work on ALS serum is significant, because this is the first demonstration 
of toxicity to living neurons by serum from a degenerative neuronal disease of 
man. This toxic molecule may actually come in contact with the anterior horn 
cells of the ALS patients. It is highly specific to this disease and has not 
been found in the sera of any other patients with neurological diseases. 

Other investigators in this group have been performing histochemical 
studies of experimental allergic encephalomyelitis (EAE) . A new, simple, 
ultrasensitive method for measuring proteinase activity has been developed. 
It has already been used to assay acid proteinase in human brain, since reports 
from several laboratories have implicated this enzyme in processes of demyelina- 
tion. Nerve cell bodies have been found to be remarkably rich in acid protein- 
ase, as compared with axonal, dendritic and glial components. Efforts are 
nov7 under way to find similarly sensitive ways of measuring lipases and 
esterases . 

Two investigators are attempting to determine the antigen which 
provokes the immunologic response to nervous tissue found in MS patients. 
They investigated the response to nervous tissue antigens by blood mono- 
nuclear cells from normal subjects and from patients with MS. With the mono- 
nuclear cell-mediated cytotoxicity test a brisk reaction was found to microbial 
antigens, but no response to nervous tissue was detected. 

Explanations were sought as to why such an energetic response to 
microbial antigens was found, and no response to nervous tissue antigens 
occurred. One possible explanation is that there is no hypersensitivity to 
nervous tissue antigens in MS. However, it is known that a similar problem 
had been encountered in the study of mixed leukocyte reactions. If the cells 
serving as antigen are disrupted, then no reaction occurs. They used a 
cellular antigen-mouse L-929 cells and a particulate mumps skin test antigen 
which could be disrupted. When these maneuvers were performed, the immunologic 
reaction did not occur. It was concluded that a living intact cellular 
antigen is necessary. 



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Another investigator has sought to measure the DNA content of spinal 
cord during the evolution of EAE , It has been found that the DNA content of 
the spinal cord rises significantly during the course of the development ol 
symptoms, and this correlates reasonably well with histologically verified 
increase in lymphocytic infiltration. 

Another group in this research center has been examining the effects 
of demyelinating disease on neurophysio logical m.echanisms of the human central 
nervous system. The approach is to investigate the cortical somatosensory 
evoked response (SER) to trains of stimuli. This work provides insight into 
the m.echanisms of symptom production as well as the natural course of the 
disease. They have worked for several years to develop accurate, reliable 
and sensitive means of assessing central conduction in MS patients and have 
evolved the technique of the SER to trains of stimuli (SERT) , This experi- 
mental technique, along with the described methods of analyses, provides a 
sensitive measure of the underlying physiological deficit in MS: i.e., 
impaired neuronal conduction. 

In another laboratory a program of research continues to utilize 
long-term cultures of mammalian nerve tissues to investigate factors which 
may influence their states of health and disease. The studies include the 
deiryelinating disorders, virus-host cell relationships, and lipid storage 
diseases . 

The ability of EAE serum to produce a total inhibition of oligo- 
dendroglial differentiation and myelin formation has been shown to be 
complement dependent and heat labile. The nature of the myelin inhibitory 
(MI) factor resides in the 7S globulins, as obtained from G=200 Sephadex 
separation, and in the IgG fraction. The time of action of the MI factor 
is under study. By introducing the factor at various periods after initiation 
of culture, it has been determined that inhibition will not occur after the 
tissue has been six days m vitro , i.e, at the beginning of rayelination. A 
parallel set of biochemical determinations utilized S-35 labelled sulphate 
and its incorporation into cerebroside. This is totally inhibited by MI 
factor and is rapidly started once the factor is removed, i.e., disinhibition. 
The anti-glial (demyelination and MI) and anti-neuronal factors are clearly 
separable and are particularly evident in serum from animals inoculated with 
encephalitogenic basic proteins. 

An extensive examination of sera from MS patients and controls is 
in progress. This will determine the incidence of the anti-glial and anti- 
neuronal factors in the various populations. An intensive study of a select 
group of MS patients, who characteristically experience relatively frequent 
exacerbations, will involve monthly sampling of serum over a period of years. 
This will demonstrate the relationship of the immunological factors to the 
clinical state of the patient. In a study paralleling the EAE examinations, 
an electron microscopic investigation of the demyelinating and sclerosing 
effects of MS serum is in progress. 

In another laboratory a study of lipid and protein synthesis has 
been carried out, while the roles of the oligodendroglia and of proteolysis 
in the demyelinative process are being actively pursued. The large increase 

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in the incorporation of (1-1'^C) leucine into protein of myelin from triethly- 
tin (TET) treated rats has been further explored. After about five weeks of 
chronic administration of TET, the incorporation of radioactive leucine into 
myelin purified from incubated spinal cord slices was increased as much as 
5007o over the normal rate. The increased incorporation was inhibited by 
cycloheximide and was seen using thirteen different amino acids as the 
radioactive precursor. In comparing incorporation of radioactive leucine 
into myelin from three different areas of the CNS , the forebrain, brain 
stem, and spinal cord, the increase was far greater in the spinal cord, less 
in the brain stem, and least in the forebrain (860, 162, and 131% of normal, 
respectively) . This finding agrees roughly to the amount of edema observed 
for each of the areas. In the most affected rats, increased turnover of 
myelin protein occurred. 

In an attempt to identify a demyelinating factor, extracts from 
TET-treated spinal cords were prepared and incubated with purified myeline 
and triethyl-tin treated myelin. These extracts did not cause greater-than- 
normal breakdown in either normal or TET myelin, although myelin from TET- 
treated rats appeared to be slightly more susceptible to breakdown in pH 7 
buffer than did normal myelin. Other membrane fractions from the CNS of TET 
rats will be similarly tested. 

The finding that factors exist in normal brain which under certain 
conditions can break down myelin without the intervention of macrophages may 
have implications for many pathologic conditions. Edema often accompanies 
conditions such as those relating to tumors, stroke, encephalitis, and anoxia, 
and spongy degeneration if accompanied by demyelination. An understanding of 
the myelin dissociation occurring in edema may be basic to our understanding 
of the mechanism of demyelination. This myelin "dissociation" described here 
may be related to an initial step in myelin breakdown, with phagocytes clean- 
ing up after the damage has been done. 

In another research center diagnostic enzyme studies have constituted 
an important advance in understanding the sphingolipidoses . Investigators are 
carrying out studies of the development of cholesterol ester hydrolases in 
the myelin sheath. This work is of importance because of the incidence of 
cholesterol esters in brain tissue in pathological conditions. Workers in 
this center have emphasized that progress in the understanding of the neuro- 
cytochemical and neuropathological aspects of human disease depends upon the 
development of further understanding of the detailed chemical structure of 
the normal brain. In order to study brain tissue rationally, it is essential 
that the different cellular types be isolated. These investigacions have 
isolated oligodendroglia from bovine brain which were then fractionated to 
yield a plasma membrane fraction and a myelin fraction which had been attached 
to the cell. The fractions have been extensively characterized. Fetal myelin 
samples also show variations in composition with maturation, but are not 
identical to cell membrane fractions. One interpretation of these data is 
that myelin undergoes compositional differences in both space and time, and 
that there is no simple relationship between myelination processes in immature 
and mature nervous systems. 



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In another laboratory blood samples were obtained from 56 patients 
with MS, 30 with optic neuritis (ON) and 100 controls, and determinations 
made of histocompatability types and of neutralizing and hemagglutinat ion- 
inhibiting antibodies to measles. A statistically increased frequency of 
HL-A type 3 was found in MS patients (42.9%), as compared to controls (23.07o). 
No such increase was found in ON patients. The titers of measles antibodies 
were not significantly different in the three groups. However, HL-A3 
individuals showed higher measles antibody titers than non-HL-A3 individuals. 
The implication is that the increased measles antibody titers found by others 
in MS patients are related to the increased proportion of HL-A3 carriers. 
The relationship of histocompatability type to immunological reactivity in 
general and to other viruses possibly relevant to MS is currently being 
investigated. 

Another group of investigators concerned with immunologic responses 
in demyelinating disease has been using myelinated cultures as a sensitive 
in vitro system to evaluate sera from encephalitic animals. Sera from 
guinea pigs with EAE , induced by various doses of CNS tissue, basic protein 
and M^ tuberculosis H37Rv (heat-killed) in Freund's adjuvant have been 
tested for myelinotoxic activity based on inhibition of myelin development 
in mouse cerebellum cultures. Under these circumstances, approximately 507o 
of the sera from donors sensitized x^7ith CNS tissue-tubercle bacilli combina- 
tions prevented myelin development in culture. 

Antithymocyte sera (ATS) has been reported to be an effective 
suppressant of delayed-type hypersensitivity. These investigators examined 
the influence of rabbit anti-guinea pig thymocyte sera on the induction of 
EAE in guinea pigs. The sera exhibited cytotoxicity _in vitro against fetal 
thymocytes and showed antibody binding by immuno fluorescent staining. In 
guinea pigs, intravenous treatment with ATS around the time of sensitization 
with basic protein in complete Freund's adjuvant with tubercle bacilli almost 
completely prevented EAE. Controls were similarly treated with normal rabbit 
sera, but all were sensitized with basic protein in adjuvant. Physical and 
histological signs of EAE appeared in control animals, although disease in 
the normal rabbit serum treated guinea pigs was delayed one week. In no 
ATS-treated animals were neurologic signs evident, although some shov/ed CNS 
inflammatory cell infiltration. An evaluation of serum treatment following 
sensitization, when physical signs are apparent, is currently in progress. 

Another researcher has been concerned with the development of 
clinical, genetic, biochemical, immunological, virological and morphological 
data on diffuse degenerative brain diseases of unknown or uncertain etiology 
with the aim to establish their etiology and pathogenesis and to develop 
specific therapies. Such data will also permit the delineation of hitherto 
unrecognized nosologic entities. 

Five patients with Batten's disease have been treated for periods 
varying from 6 to 32 months with an antioxidant mixture. The result of this 
treatment was a definite slow-down in the progression of the disease. The 
treatment was made available to a group of investigators at the University of 
Helsinki in Finland, who reported that they observed an improvement in the I.Q, 
of some of their patients. This group is treating 29 patients with this dis- 
order . 



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Biochemical studies on the isolated pigments from the brains of 
patients with Batten's disease have shown that the accumulated material 
is an acidic polymer with a molecular weight of approximately 30,000 and 
efforts are currently underway to characterize this substance biochemically. 



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COMMTMICATIVE SCIENCES 

The crucial nature of the communicative sciences and the aisorders 
which impair normal communication can be assessed when we consider that 
thought, speech and language are closely linked. Language depends upon the 
activity of the cerebral cortex, the last division of the central nervous 
system to developo The speech centers of the brain are located in close 
proximity to the higher association areas. Furthermore, the mastery of 
language proceeds hand in hand with the development of higher forms of 
anticipatory behavior. 

This means that language and speech in particular is bound up 
with the capacity for discrimination, judgement, and concept formation which 
are so essential for thinking in the technical sense. The motor responses 
of speech centers are connected with the vocal apparatus. Conditioning and 
integration in the higher centers ultimately depend upon the sensory impressions 
that reach them through the afferent nerves from the eyes, ears and other 
receptor organs. Likewise, the muscular and glandular responses which are 
set off from the higher areas are mediated through the effector nerves passing 
through the lower centers. Through conditioning and integration, verbal 
habits are so built into each other that they constitute one of the meet 
important parts of the response system of the individual personality. Speech 
and writing are definitely bound up with social stimulation and the response 
to these stimuli. If, then, the very organisation of the mechanism of 
language is built up by social stimuli and corresponding response, and if 
the objects of the conditioning are social-cultural in nature, then it follows 
that the content, the objects of association, will also be social- cultural. 
Therefore, the thought or language response connected with objects in 
experience must likewise be social-cultural in nature. ¥e cannot ignore 
the fact that the essential situations which give rise to language development 
in the person are social-cultural. Even the experience of the material 
world is constantly influenced by the verbalized interpretations furnished us 
by other people. There used to be a long controversy between philosophers 
and psychologists as to whether thought or language came first. Some writers, 
like Max Muller, have held that thought is nothing but a form of inner 
speech. John Watson held that thought was sub- vocal speech. Today, such 
extreme views are no longer manifest but there is no doubt as to the inextric- 
able intermingling of thought and language both developmentally and in the 
normally mature functions. 

Sensory Process . 

There are several directions in which substantial progress has been 
made in the current year. The most important is the accumulation of neuro- 
physiological evidence that inner and outer hair cells of the cochlea interact. 
There is strong evidence now that outer hair cells provide an inhibitory input 
to the inner hair cells and that the spiral fioers innervating the outer hair 
cells function as delay lines. Both the inner and outer hair cells appear to 
be excited by the displacement of the basilar membrane toward the scala 
vestibuli. At frequencies below the characteristic frequency, the inputs 
from inner and outer hair cells are approximately in phase and tend to cancel 
each other out. Near the characteristic frequency the spiral fibers produce 
a phase lag of approximately l8o° and remove the inhibitory effect of the 



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outer hair cells. Tliese conclusions are based on single unit responses in the 
auditory nerve of normal and kanamycin treated mongolian gerbils. The experi- 
ments were performed with the help of new surgical techniques developed on 
the project. 

Calculation of low-frequency wave velocity in the cochlea of the 
guinea pig, based on anatomical constants obtained by Fermandez and on the 
elasticity of the scala media measured by von Bekesy, has shown excellent 
agreement with phase data obtained by Kohlloffel with the help of a new 
laser technique. Both von Bekesy's and Kohlloffel 's results were obtained 
post-mortem. Correction of the elasticity values for post-mortem changes 
derived from Kohlloffel 's work brings the theoretical velocity into agreement 
with cochlear time delays measured on live animals by means of cochlear 
microphonics and the Mossbauer technique. These consistencies demonstrate 
that at least one pajrt of the cochlear mechanics is well understood. 

In another piece of theoretical work, it has been shown that most, 
and perhaps all, intensity characteristics of sensory receptor units can be 
described by a mathematical relationship. The relationship is based on a 
power-function transformation, a summation of e:d:rinsic and intrinsic stimulus 
energies, and an eJ5)onential saturation. It appears to be completely 
established in the receptors themselves and is evident in the receptor poten- 
tials. Axonal firing rates produced by these potentials appear to follow the 
same relationship except for a multiplicative constant and an additive thres- 
hold constant. In most receptors, the power function exponent is approximately 
equal to 0.5 with respect to stimulus energy, but gustatory receptors appear 
to follow an exponent of 1.0. 

Hearing and Deafness 

A group of investigators looking into the electrical properties 
of the organ of Corti and the influence of physiological conditions on them 
in order to determine how a vibratory form of energy is turned into a nerve 
impulse by the ear have demonstrated that the spiral capillaries of the 
basilar membrane provide the necessary oxygen to the sensory cells of the 
organ of Corti. Presumably these sensory cells are responsible for the 
electrical potential that can be recorded when the cells are vibrated and 
this electrical response is very dependent upon the available oxygen. However, 
it is not yet kjnoim what the role of this electrical potential plays in 
stimulating the small unmyelinated dendritic nerve endings. Nor are the 
exact sources of this potential l-aiown. They have developed a method for 
visually monitoring, by closed circuit TV, electrode placement in the cells 
and fluid spaces of the organ of Corti. They also now vibrate a small area 
of the basilar membrane by a microprobe placed on the foot of an outer pillar 
cell or on the edge of the basilar membrane. With the use of these techniques | 
they are exploring the various regions of the organ of Corti with micro- 
electrodes to identify the factors that determine and control the generation 
of the nerve impulse. An adequate blood supply is essential and they now 
have the technical capability of combining its observation with electrical 
recording. Others in this group are continuing studies of the changes in 
Corti's organ in experimental ototoxicity, n o i s e- Induced hearing impairment, 
and human presbycusis, particularly on the blood vessels of the inner ear and 



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the pathological i-hanges that occur in them. In gentamicin- intoxicated 
guinea pigs the normal process of devascularization of the superstrial 
portion of the spiral ligament appears to he accelerated. Intervascular 
strands formed by degenerating capillaries and avascular channels (i.e., 
percapillary spaces from which capillaries have disappeared) are frequently 
seen. T^jese changes also appear after prolonged exposure to intense noise, 
in addition to the partial occlusions of suprastrial and basilar membrane 
capillaries that were described. 

In the normal guinea pig the so-called arterio-venous anastomoses 
decending in the spiral ligament behind the stria vascularis appear to 
constitute two distinct sets of capillaries. One set, which is called the 
adstrial capillaries, are closely applied to the basal surface of the stria 
and may be involved in the exchange of fluid with the strial vessels. The 
other set, the poststrial capillaries, apparently supply the spiral ligament. 
Neither set of vessels fulfills the criteria of Chambers and Zweifach for 
anterior venous anastomoses or shunts. Other in this group studied the 
hearing function in old world monkeys and its bearing on the evolution of our 
own hearing and the examination of agents such as noise and antibiotics which 
produce both temporary and permanent hearing loss. Following determination 
of normal hearing thresholds by cperent conditioning, the monkeys were 
subjected to octave bands of noise at 500, 2000 and i|-000 Hz at levels of 100 
and 120 dB SFL for periods up to l60 hours. Temporary and permanent hearing 
loss was evaluated daily after each exposure for a month following the last 
exposure. The animals were sacrificed for examination of the inner ear by 
phase contrast and scanning electron microscopy. Maximum temporaiy and 
permanent hearing loss was found at the center of the exposure band and 
extended as much as an octave above the center. 

Temporary hearing inrpairriient of 40-50 dB occurred following an 
exposure of 100 dB SFL for two hours; no permanent changes were observed. 
At the other extreme, exposures of 120 dB SEL for eight hours daily for 
one month produced temporary hearing loss after the first exposure at and 
above the octave band center and permanent loss of 60-7O dB as measured one 
month after the last exposure. Related histopathology in the cochlea ranged 
from, minimal to fairly excessive hair cell loss. The size of the lesions 
■was less than anticipated on the basis of the threshold changes. Their 
locations were in general agreement with the place principle. 

Middle Ear , 

Another group of investigators is attempting to determine the 
mechanisms involved in middle ear effusions (serous otitis media), to develop 
a battery of practical diagnostic laboratory tests, and to find a suitable 
animal model to test a proposed hypothesis. Preliminary evaluation of the 
middle ear effusions (SOM) revealed a high titer of lysozyme, as high as I80 
times that of the serum. Also, the mucoid type showed a higher titer than 
the serous type effusions. Positive bacterial cultures were obtained in 
approximately one-third the total cases (44) and it is significant that high 
bacterial counts were obtained in 60 percent of the serous type effusions 
while only 20 percent showed high bacterial counts in the mucoid type effusion. 
Immunochemical investigations in monkeys supported the evidence that the normal 



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middle ear mucosa, as veil as the eustachian tube, contained IgG, IgA, IgM 
and IgE-producing cells. However, the quantity of cells and the intensity of 
staining increased considerably after tubal obstruction. While this result I 
cannot be directly translated into clinical SOM, it can nevertheless be 
pointed out that the middle ear mucosa and the tubal epithelium have a local 
immunodefense system. Cytochemical investigations on laborato27y animals 
suggest (a) that the strong acid phosphatase activity is localized in the 
secretory cells of tubal as well as middle ear mucosal epithelium and (b) 
that the coating substances (secreta) of the middle ear mucosa and the tube 
showed a positive reaction with "tricorrrplex flocculation procedure," indicating 
the presence of phospholipid which is the main component of the pulmonary g 
surface tension lowering substance (surfactant). " 

Another research team is attempting to identify causal factors 
underlying pathological manifestations of chronic destructive middle ear 
disease by direct intervention into middle ear structures and towards a more 
thorough search for microbiological agents, particularly viruses. Specific 
hypotheses on the development of middle ear abnormalities were tested by 
a series of experiments in cats, guinea pigs, and monkeys. 

(1) The epithelial-migration hypothesis of cholesteatoma formation 
was tested by transplanting epithelium from the external canal into the middle 
ear or by folding back flaps of the tympanic membrane into the middle ear 
cavity. Development of epithelial growth was infrequent and, except for two 
cases of severe infections, failed to cause serious abnormality of the adjacent 
bullar mucosa. The most striking observation was the ability of the middle 
ear structures to repair themselves and to clear themselves of extraneous 
material. j 

(2) It ■jvas hypothesized that chronic otitis media with patent 
eustachian tube in patients is maintained by a reservoir of diseased tissue 
in the mastoid cells and that this condition was supported by hyperplasia 
of the tissue in the isthmus between the tympanum and the attic. Blocking 
the attic with fascia in six cats, ten guinea pigs, and three monkeys failed 
in more than half the cases brought to post-mortem so far, apparently because 
the tissue failed to "take" and was cleared before it provoked the disease. 

Another team is studying the nature of epidermal cyst formation 
in the middle ear to further elucidate the mechanism by which the cyst causes 
bone resojrption. The first step was to produce epidermal cysts in the middle 
ear of guinea pigs and second to purify guinea pig skin collagenase and induce 
a collagenase antibody in rabbits. A total of 75 animals were observed for 
varying periods up to four months following seven different groups of surgical 
experiments designed to induce middle ear epithelial cysts. From two to ten 
animals survived in each group. In group one ear canal skin was transplanted J 
onto the head of the malleus with and without pre-treatment with inflammatory 
agents. The second group involved rotation of canal skin flaps through a 
marginal perf ormation. The tympanic membrane was split in the third group 
and the canal skin placed between the mucosa and epithelium. The fourth 
group was pre-treated with estrogen injections then talc and vitamin A placed 
at the margin of the tympanic membrane. The fifth group was pre-treated as the 
fourth group and then had a second operation in which the skin was placed 



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against the cochlea. Group six was subjected to placement of canal skin under 
a flap of mucosa over the cochlea. Group seven had explants of the canal skin 
placed between a flap of bone over the bulla and intact middle ear mucosa. 
The results of these experiemtns in which the histology is completed show 
no epithelial cyst formation in animals pre-treated with inflammatory agents. 
A large amount of new bone formation was found in these animals. An epithelial 
cyst was found in one animal in which skin was placed between layers of the 
tympanic membrane. The most promising results so far are with placement of 
canal skin under intact mucosa. Three microscopic cysts were found in eight 
animals as early as six days postoperative. These cysts do in fact resemble 
human cholesteatomas histologically. 

Inner Ear 

In a study of the struct'ore and function of the inner ear one 
investigator found the distribution of short and tall hair cells in the 
chicken to be similar to that of the pigeon. Innervation features are also 
similar in the two species. The major difference is that many short hairs 
in the chicken's cochlea do not have kinocilia. Scanning electron microscopy 
has revealed that the rather massive tectorial membrane of the birds cochlea 
is permeated with open channels. Cavities are present in the under surface 
for insertion of the hair bundles. Guinea pig ears have been stained by 
the Golgi method, and the peripheral innervation pattern studied in the 
cochlea- The innervation pattern of the cochlear nerve fibers seems to differ 
in the basal and upper coils of the guinea pig cochlea. In the basal coil, 
a single fiber usually innervates only one row of outer hair cells, and for 
a distance which does not usually exceed a 100 micron segment of the basilar 
membrane. In the upper coils, single nerve fibers innervate 2 or 3 rows of 
outer hair cel.ls over a longer segment of basilar membrane. It was possible 
to identify bhe nerve fibers as cochlear or as belonging to the efferent- 
olivo- cochlear tract. The olivo- cochlear axons also were studied. Scanning 
electron microscopy was found to be very valuable in a study of the vestibular 
end organs. There seems to be some variation in their length on the ridge. 
The hair bundles of the maculae are different than those of the utricle. 
The sacullar stereo-cilia are shorter so that the kinocilium protrudes for 
about five microns above the other cilia. It has not yet been determined 
if this is a regional or general feature. 

Another investigator is attempting to elucidate the mechanisms 
involved in the release and consumption of chemical energy necessary for the 
transduction processes of the inner ear and for the transmission of auditory 
and vestibular information to the CWS, An important part of this work is the 
study of the effects of vascular insufficiency, ototoxic drugs and noise 
trauma on energy metabolism. Using guines pigs and the chinchillas, the 
rate of change of the major components of the total energy reserve (glycogen, 
glucose, ATP and P-creatine) in ischemia has been determined in the organ 
of Corti, stria vascularis, ganglion spirale and cochlear nerve. Ischemic 
changes are most rapid in the case of P-creatine and (except in the stria 
vascularis) much slower in the case of ATP. In the organ of Corti, ATP levels 
were still at 50fo of the original after 20 minutes of ischemia. The initial 
use rate of high energy phosphate (-»--P) in stria vascularis and organ of Corti 
respectively is 65-8O and 15-21 mmoles of ^wP/kg dry weight/minute. The energy 



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use rates are in striking contrast to the values of the total energy reserve 
in these tissues which are 500-530 and 2k'^ minoles of p/KDW in the organ of 
Corti and stria vascularis respectively. 

The effects of ethacrynic acid administered systemically in dosages 
from 10-50 mg/kg upon EP and high energy phosphates have been studied in 
detail. At high dosage levels the positive EP is rapidly replaced by a 
negative EP of -20 to -kO mY, followed by partial recovery to positive levels. 
At the time of maximum depression of EP, P-creatine levels in the stria are 
unchanged (6-9 mmoles/KDW) and ATP levels are reduced by 20-25/0 {l-lO mm.oles/ 
KD¥) of the original. If the intoxicated ears are subjected to the additional 
stress of total ischemia, the initial rate of change of'^^P in the stria is 
reduced to 20 mmoles/KDW/minute, suggesting a reduction of the metabolic rate 
due to uncoupling of active transport. The ischemic decline rate of the 
predamaged EP is reduced roughly in the proportion to the preischemic EP level. 
If the predamaged EP is <C-20 mV, ischemia produced a rapid reduction to 
negativity. They believe that this negative oxygen sensitive compound of EP 
is an electrogenie potential. This work is relevant to a variety of patho- 
logical states of the ear and the results may help elucidate the etiology of 
Meniere's disease and provide a basis for its treatment. 

Cochlea 

A team of investigators studied cochlear distortion and the role of 
cochlear microphonics (CM) in hearing by investigating the nonlinear cochlear 
processes, the relationship between CM distortion components and corresponding 
single unit activity in the cochlear nerve, and recording of CM and neural 
activity from pathological cochleas in which the active hair cell population 
is spatially restricted. One primary aim has been to deactivate preselected 
segments of the cochlea so that more localized and better controlled recordings 
could be made from, the remaining region. This new method involves the passing 
of very high frequency (6mHz) pulsed current bursts through the organ of 
Corti between intracochlear electrodes placed in scala vestibuli and tympani. 
In studying correlations between CM distortion components and single unit 
discharge patterns, they have demonstrated that time-locking of spikes can 
occur to any combination component of the type nf + mf2 where f]_ and f2 are 
the two input frequencies. Thus, simple periodicities in the spike pattern 
are not sufficient to prove the presence of an actual distortion product 
(energy) in the cochlea. Secondly, they completed an extensive reinvestigation 
of the CM interference phenomenon. It was concluded that the most effective 
interfering frequencies are those just above the best frequency of the 
electrode location. In this very frequency region all cochlear nonlinearities 
are most prominent. Further work was carried out on the CM correlates of 
cubic difference tones. They reaffirmed earlier findings that the CM 
components at 2f]_-f2 have no traveling wave precursors. 

Another group investigating the physiology of auditory and vestib- 
ular systems are attempting to elucidate the functional organization of the 
cochlear nuclear complex and its output pathways and to examine the responses 
of peripheral vestibular neurons to linear accelerations. 'The architecture 
of the trapezoid body (TB) was investigated. The tract is divided into three 
distinct laminae: a dorsal layer of m.edium sized fibers (3-5 micra) a layer 



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of large fibers (8-I5 micra) occupying the middle of the tract, and a ventrally 

located lamina of small fibers. A fiber degeneration study showed that the 

entire tract, with the possible exception of some small fibers, originates 

in the cochlear nuclei. The meditun diameter fibers are tonotopically organized. 

Tlie low best frequencies are represented dorsally, higher best frequencies 

ventrally. Wo tonotopic organization was discernable among the large diameter 

fibers. Both sets of fibers are characterized by narrow excitatory tuning 

curves, irregular steady-state discharge patterns, primary-like responses 

to tone-bursts, monotonic rate- intensity functions and secure phase locking in 

response to low- frequency tonal stimulation. 

Previous work reported on the vestibular nerve concerning the 
response of semicircular-canal neurons to linear acceleration suggested that 
the responses resulted from very small density differences between the cupula 
and endolymph. They now discovered that a thermal gradient of the correct 
magnitude and direction exists in their experimental preparation. Experiments 
are undenray to determine if elimination of the gradient will abolish the 
response. If such is the case, it can be concluded that the semicircular 
canals, though capable of responding to linear accelerations, do not do so 
under normal physiological conditions. In a similar vein, they have found 
that otolith neurons do not respond to even relatively intense angular 
accelerations. The response of otolith neurons to controlled linear acceler- 
ations is being investigated, particularly on input-output relations and on the 
directional sensitivity and response dynamics of units inne2rvating both the 
utriculus and sacculus. The results should help in providing a rational 
basis for the interpretation of clinical disorders. 

Another group studied the electro-anatomy of the cochlea by 
investigating the directional sensitivity of the cochlear hair cells and by 
obtaining neurophysiological and electrophysiological changes of experimentally 
induced endolymphatic hydrops. Temporal patterns of nerve impulses of the 
auditory nerve were examined in anesthetized guinea pigs when the basilar 
membrane was displaced statically by driving the round window with a newly 
designed electromechanical transducer after the helicotrema had been plugged 
with bone wax. The electrical signal to the transducer was a trapezoid wave, 
the duration amplitude and rising/falling time could be controlled indepen- 
dently. Duration ranged from 100 msec to 5 sec. It was found (l) when the 
helicotrema was plugged, the hair cell responses to the trapazoidal displace- 
ment of the round window membrane was larger in the third turn than in the 
basal turn; (2) increase of the amplitude of the displacement of the round 
window membrane resulted in a rebound of the hair-cell responses at the end 
of the stimulus which was greater in the upper turns than the basal turns of 
the cochlea; (3) shortening of the rising time of the trapezoidal motion 
applied to the round window produced transient ringing in the hair cell 
response which was more pronounced in the upper turn; (4) the relationship 
between the amplitude of motion of the driver and magnitude of the hair cell 
responses varied considerably among preparations; (5) about 50fo of the 
auditoiy fibers showed tonic responses to unidirectional development of the 
round window. This group also surgically obliterated the endolyiiiphatic duct 
in guinea pigs. Two to six weeks after surgery, measurement of the EP and 
sodium and~patassium content in the endolymph was performed on I6 hydrops 
animals. CM, ATP, and EP were recorded. Temporal bones were examined 



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histologically. The results were (l) EP recorded in the basal turn showed 
significant reduction in the hydrops animals (63.2 + l6.2 mv) and slight 
increases in Na content in the endolymph were found but were statistically 
nonsignificant; (2) EP recorded in the third turn was also suppressed in 
hydrops animals; (3) in early stages of the endolyrnphatic hydrops, CM 
recorded in the basal turn showed slight loss and CM recorded in the third 
turn showed marked decrease in the maximum output and elevation thresholds 
(h) increase in scala media area varied from 50/^ to 150^ and did not show 
close correlation with EP; (5) histological examination showed atrophy of the 
stria vascularis in only three cases and degeneration of hair cells of the 
upper turns in five cases. Most of the hydrops animals showed no apparent 
neurophysiological changes in the early stage and yet demonstrated functional 
disturbance in the upper turns of the cochlea. 

Hearing Disorders 

In an investigation of otopathology utilizing light microscopy, 
a team of investigators have been highly productive in defining the patholog- 
ical bases for deafness and vertigo. The underlying approach is that an 
understanding of the morphological changes occurring in specific disease 
states is essential to the development of preventative and therapeutic 
measures. The current findings of this group are: (l) Atrophic changes occur 
in the spiral ligament as a function of age, but these changes do not in 
themselves cause hearing loss. Only when the change is so severe as to result 
in rupture of the cochlear duct is hearing loss produced; (2) Blockage of 
the cochlear aqueduct causes no inner ear changes and is compatible with 
normal cochlear functions. (3) Recurring, severe episodes of vertigo may 
be caused by diabetic neuropathy of the vestibular nerves. (k) Small 
asymptomatic schwannoma's may occur on the facial nerve. (5) The morphological 
changes in the temporal bones of seven individuals who suffered from sudden 
deafness strongly indicates that the etiology of this disorder is viral 
labyrinthitis. (6) Moi^hological changes occur in the ossicular points as a 
function of aging. However, these changes induce no functional disorder even 
when the joints are ankylosed. 

Cochlear Prosthesis 

A group of investigators are involved in providing basic infozTnation 
crucial to the development of a prosthesis with potential application to a 
significant portion of the 300,000 totally deaf individuals in the United 
States. In most of these deaf individuals, the acoustic nerve is at least 
partially intact, and the experiments indicate that surviving spiral ganglion 
cells are electrically excitable. Sinrple bipolar stimulating devices have 
been introduced into the cochlea or acoustic nerve in a small number of 
patients. Psychoacoustic experiments have revealed the nature of the sensa- 
tion evoked by such stimulation. Physiological studies in cats have shown 
how the sensation is generated and encoded in the auditory nervous system 
and histopathologic studies have shown that at least most spiral ganglion 
cells survive implantation of a flexible scala tympani electrode. These 
studies have also shown tliat encoding of intelligible speech, if ever possible, 
will require differential, multichannel stimulation of a series of discrete, 
predetermined sectors of the acoustic nerve. The current effort is directed 



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at experiments to determine how and to what extent appropriately restrictive 
sectors of the acoustic nerve might be stimulated from within the scala tympani. 
The effectiveness of stimulation as a function of cochlear place for different 
Implanted stimulating electrodes will "be determined in single unit colliculus 
experiments in which advantage is taken of the binaural response properties 
and cochleaotopic organization of inferior colliculus neurons. A map of the 
response threshold to contralateral electrical stimulation can be derived by 
recording from a series of neurons with best frequencies collectively 
spanning the entire frequency spectrtun (the entire length of the basilar 
partition). Other experiments to be conducted concurrently will provide 
further crucial information about the safety of long tenn implantation and 
stimulation. If stimulation of a small number of appropriately restricted, 
predetennined sectors of the acoustic nerve can be effected, then it should 
be able to provide informxation sufficient for the direct hearing of speech 
for subjects who are now totally deaf. 

Another research team attempting to develop a cochlear prosthesis 
determined, by ui vitro testing of possible electrode materials for electrol- 
ysis longevity, that the best material was iridium, followed by platinum and 
gold. Stimulation electrodes of iridium will last decades even under extrem.e 
current density conditions. PlatinLun does nearly as well at low current 
densities, but does not do nearly as well at high densities, and gold is in 
a fairly distant third place. Stainless steel, tungsten, titanium, tantalum 
and zirconium are all very poor, especially at higher current densities. 
Tlie conducting oxides were also veiy unsatisfactory. 

In regard to electrode fabrication, the trial of the integrated 
circuit microelectrodes has met with some difficulties (in fragility) during 
tissue implantation. Tliough prior experience with these electrodes, vhen 
used to rc-Gord from superficial layers of the cortex, was quite satisfactory, 
problems of fragility arise during deeper penetrations. The investigators 
have now fabricated thicker probes which seem more rugged. If these fail, 
a different substrate material will be tried. They have also succeeded in 
fabricating an insulation material which withstands soaking' in saline for 
prolonged periods. With regard to research on stim.ulation circuiting, since 
it sppeared likely that a satisfactoiy electrode material could be found, 
the investigators have gone ahead with consideration of a means for stimula- 
ting such electrodes within the ear. They have developed a novel scheme 
for driving trans cut aneously an arbitrary number of electrodes using two 
links, an rf link (by coil) providing power and certain clocking cues to an 
internal multiplexer, and an optical data link which instructs the multiplexer 
in the stimulation pulse distribution. The advantage of the optical link is 
its ability to rapidly transmit acoustic information which is sufficient 
for any reasonable number of stimulating electrodes. In its simplest form 
the link is a light-emitting diode in the ear canal coupled to a photo diode 
in the middle ear through an intact tympanic niembrane. A bread-board model of 
this system has been constructed and works well. A hybrid semiconductor 
version of implantable dimensions is under construction and should be finished 
by the Fall of 1973- A single-chip integrated circuit version will be commenced 
as soon as the hybrid version has been tested. 



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The investigators are about to begin another series of inrplants 
and chronic stimulations. Meanwhile they have constructed the necessaary 
stimulating equipment for accurate stimulation and have begun impedance 
measurements on the first batch of implanted platinum- iridium wires. The 
major effort while a^/aiting electrode material has been to further define two 
stimulus variables;, the best location for electrodes, and the electrophysio- 
logical limits of possible information transfer by electrical stimulation when 
compared to equivalent acoustical stimuli. Stimulus "equivalence" has been 
explored for electrodes on the round window, scala tyrnpani, auditory nerve 
via modiolus and stereotaxic posterior fossa placements. In general, the 
nerve electrodes require less current for equivalent responses at the 
inferior colliculus than do the other electrode sites. It has not yet been 
possible to find a systematic difference between acoustic and electrically 
evoked neural activity in the inferior colliculus when the eighth nerve 
electrode is used. Differences must exist which ought to be detectable by 
appropriate stimulus programming and analysis. To date, there is no 
difference in the rate synchrony for the two stimulus modes. The round 
window electrodes seem to produce acoustic "masking" and may possibly detect 
differences in the slow wave responses for "normal" and electrical stimulations 
of t?ie auditory system. 

."Jomparative Hearing 

In an investigation of acoustic perception and echo-location, a 
team of investigators are studying the anatomical and physiological properties 
of the chiropteran auditory system and the behavioral and physiological 
activities on bats actively engaged in echo- location. Various histological 
techniques have been tried in an effort to develop procedures suitable for 
studying the highly specialized cochlea of Pteronotus p. parnellii . To 
date no single technique has been entirely satisfactory but good preparations 
have been obtained by perfusion of the cochlear structures- With these 
techniques, hair cell and ganglion cell populations in different parts of 
the cochlea will be studied, measurements of the large basilar membrane 
thickenings are being made, and in embryonic preparations the development 
of the unusual nerve fiber distribution is being followed. An attempt was 
made to determine neural activity in the densely and sparsely innervated 
portions of the cochlea. W-, (eighth nerve) audiograms have been recorded 
from ten bats with the aid of chronically implanted electrodes. The shape of 
the Bj_ audiogram has proved to be different from that determined for CM 
audiograms in the same preparations. The N^ "on" response audiogram showed 
a broad band of sensitivity which covers the frequencies included in the 
bats' FM signals. It is also sharply tuned to a narrow band of frequencies 
3 to 5 KHz above the CF frequency of the emitted pulse. This indicates that 
the detection of Doppler shifts in the returning CF echoes is important. 
The "off" responses are very sharply tuned and correspond closely in shape and 
frequency sensitivity to the CM audiogram. 

The investigators have recorded CM potentials fraa Pteronotus 
performing landing maneuvers. With the aid of a special period-to-D.C meter, 
they were able to measure precisely the frequencies in the emitted pulses and 
Doppler-shifted echoes. By comparing these frequencies with the bats' own 
audiogram, it has become clear that the bats emit CF signals to which their 

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ears ai-e not very sensitive. However, their flight speed creates Doppler 
shifts in the returning echoes, and their ears are much more sensitive to 
frequencies in the Doppler shifted echoes than the emitted pulses. The 
researchers studied the properties of echoes reflected from the wings of 
flying moths. With the aid of photographic records and the period-to-D.C. 
meter it was clear that the wing-beats of an insect create considerable 
amplitude modulation as well as frequency shifts in the sonar signals. They 
are continuing to obtain evidence that the enlarged median lobe (of Ingvar) in 
Pteronotus is related to their highly specialized sonar system. Neural 
audiograms on unanesthetized preparations have shown that there are strong 
auditory projections to this part of the cerebellum. The audiograms closely 
resemble those recorded at the eighth nerve level. Ablation studies are just 
beginning in an effort to see if this part of the cerebellum is concerned 
with the precise regulation of pulse frequency via laryngeal muscle control 
and whether or not it is also concerned with the fine regulation of pulse 
duration and pulse repetition rate. 

Speech and Language 

In an investigation of cancer of the larynx, every larynx removed 
in toto or in part for cancer is sectioned serially. This technique allows 
the investigators to correlate preoperative diagnostic tests such as 
laryngoscopy, laryngography, cine-laryngography, laminography, etc., with the 
microscopic evidence of cancer within and outside of the larynx. Follow-up 
observations will be related to histological patterns of growth and spread 
within the larynx in the 275 cases now available. Tlie major objective 
of this project is to improve present methods of preserving the functions of 
the larynx and pharynx when these organs must be treated for cancer. Serial 
sections of the larynx and/or hypopharynx are being examined for patterns of 
growth and spread of cancer in specimens which have been removed surgically. 
Thus, information is accumulating on such questions as: (l) Why has radiation 
therapy failed to cure some types of laryngeal cancer and how can it be 
made more generally applicable ? (2) How acciurate are the several systems 
of clinical staging now in use? (3) How can malignant lesions be precisely 
assessed for conservation- type surgical procedure? A study of transglottic 
cancer showed that laryngeal cancer which crosses the ventricle and anterior 
commissure in the vertical direction invades the laryngeal framework in a 
high percentage of cases (t6'/o of ^2 cases studied by serial section). 
Invasion of the laryngeal framework was found to be related to the size of the 
primary lesion. It rarely occurred in lesions below 2 cm, but occurred in 
three-quarters of those cases larger than 3 cm. The degree of differentiation 
of the primary lesion showed no relationship to invasion of the laryngeal 
framework. By contrast, cervical lymph node involvement occurred with 63'y^ 
of the poorly differentiated lesions as compared to 2'kio of these well differ- 
entiated. 

A group of investigators are using cineflu orographic techniques 
to (1) study movements of articulatory structures during speech production 
to obtain descriptive data on variations in the articulatory characteristics 
of various types of speech elements; (2) study the timing characteristics 
of articulatory movements in more detail; (3) utilize the data in the formu- 
lation of general principles which appear to govern the opeiration of the speech 



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articulation process. It was found that, when listeners were asked to identify 
consonants and vowels which had been partially or completely deleted from CV 
syllables by electronic gating, they could identify most consonants and all 
vowels at above chance levels even though the steady-state portions had been 
deleted. They also were able to sort phones into feature categories without 
acoustic segments corresponding to steady states. The most important 
perceptual cues relate to anterior posterior place of articulation of the 
sound. High-speed cineflu orographic films were obtained of three speakers 
producing a constant CVW sequence with a variety of prosodically marked 
boundries falling between the two vowels. It was found that the onset of 
velar lowering for the nasal consonant was consistently delayed in those 
cases where marked junctaT-al boundries were present. The position and 
dynamic properties of /l/ sounds in English were studied in relation to the 
syllabic and m.orphemic position of /l/, vowel context and speaking rate. 
The results indicate that observed articulatory variations can be explained 
by positioning two basic /l/ sounds, a pre-vocalic sound which functions 
as a consonant and a post-vocalic sound which has the characteristic of 
a vowel. The syllabic /l/ appears to be basically a post-vocalic sound 
resulting from changes in the timing of articulatory gestures. 

In the area of aphasia, a group of researchers are attempting to 
develop new measures of aphasic symptom variables. The research is devoted 
to the study of brain function in language and related higher functions in 
humans. These problems arise from the observation of brain injured patients 
with aphasia and similar disorders. Specific areas of study include the 
development of measures of linguistic disturbances in aphasia and the 
investigation of its neurophysiological basis, the investigation of disorders 
of purposeful movement and the study of brain laterality. With regard to 
diagnostic assessment, k'30 children from ages 5 through 12 were tested and 
the data analyzed. Boys were found to have a higher naming vocabulary 
than girls of all ages. Significant differences in sequencing span were found 
as a function of material (letters, digits, objects), modality of presentation 
(oral, visual) and response (oral, pointing), and interactions of modality 
with age. Discriminant analysis of aphasia subscores against clinical 
diagnosis was designed and the data are being analyzed. Agrammatic patients 
were found to be defective in their interpretation of syntactic links between 
words, as in their output. With regard to semantic fields and naming 
disorders, analysis based on 2k aphasics and a non-aphasic control indicates 
differential effects of auditory comprehension level on certain associative 
categories in both latency and number of responses . Poorer responses are 
obtained for words failed in a preliminary nam.ing test. With regard to 
brain laterality, it was found that the rate of learning to recognize words 
presented tachistoscopically did not differ in right or left visual fields. 
Final analysis of results on a study of dichotic ear-storage effects for tonal 
stimuli shows that the left ear is the superior storage channel both when 
S is pre- instructed as to order of report and when instructed after the 
stimuli have been heard. With regard to distiu^bances of purposeful movements, 
forty aphasics, thirty brain injured non-aphasics, and ten elderly control 
subjects have been studied with an ex±ensive test of praxis. Prelinimary 
results indicate that left hemisphere subjects are most impaired, Broca's 
and conduction aphasics are worse on bucco facial movements, and right brain 
damaged are worse on the spatial aspects of movements. The significance of 

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these findings are that aphasia is a frequent by-product of stroke, brain 
tumor, trauma and infection. Diagnostic procedures might be constantly up- 
dated as more is learned about the psychological and neurological bases for 
these language disorders. Aphasia is closely related to language learning 
disabilities of childhood and this investigation has immediate applicability 
to the evaluation and treatment of children. The phenom.enon of brain latera] - 
ality, first observed in aphasia, is of both theoretical and practical 
Interest. 

Another group of investigators are researching behavior modification 
and bio-feedback in an attempt to understand stuttering. One investigation 
is the effect of nonfluency and fluency of contingent and random positive 
and avers ive events on the speech of stutterers. The results tend to support 
the hypothesis that subjects show decreases in nonfluency rate (compared to 
noncontingent blocks) as a result of both contingent money gain (positive) 
and money loss (negative). In a second study the hypothesis that formerly 
neutral stimuli would come to serve as discriminative stim.uli for reduced 
nonfluency through their association with contingent attention is clearly 
supported in some subjects but not others. In a third study, fluency rate 
appeared to increase as a result of contingent events, whether money-gain, 
or money-loss, when compared to blocks of speech in which there was no 
contingent event for individual units of fluency. The significance of the 
results is that they support the general hypothesis that stuttering 
behavior can be approached within an operant learning framework, but that 
it should not be regarded as a simple operant response. Also, the nature 
of the reinforcement contingencies maintaining stuttering in real life 
is complex, probably involving the attention of the listener. 

Olfaction ''-'''' 

A group of investigators are attempting to interrelate various 
electrically recorded responses to the primary neural response and to 
anatomically and behavioral responses. In addition, they are attempting to 
discover physical and chemical properties that are of relevance to the 
stimulating properties of odorants. Catfish taste receptors were shown to be 
extremely responsive to common amino acids. The recording preparation was 
the maxillary barbel nerve isolated surgically in the orbit after removal of 
the eye. Thresholds in some instances were less than 10"-'-^ML-arginine or 
L-valine, thus exceeding olfactory sensitivity for L-asparagine or L- valine. 
However, the technique of touching the surface of the olfactory mucosa 
with the microelectrode may not yield as good a signal- to-nolse ratio as 
recording from a nerve twig. The L-amino acids were always more stimulatory 
to the gustatory receptors than were the corresponding D-forms, as was true 
for the olfactory receptors. The series of gustatory effectiveness at 10" M 
in the channel catfish differed from the olfactory series at lO'Jjvi j_n the 
white catfish. Preliminary indications are that there are species difference 
within a given chemical sense modality. The chronic lesion experiments in 
the bullfrog are continuing. Olfactory nerves were sectioned intracranially 
and some lesioned preparations are now well over a year old. Extensive 
regeneration occurs after simple section of the nerve and normal responses 
are recorded electrically from the nerve and mucosal surface. After 
extirpation of a segment of the nerve there is no reconnection with the 

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olfactory bulb. It has been confirmed that these nerves cells undergo 
complete retrograde degeneration after section of the receptor axons in the 
nerve. Electron microscopic audioradiography with titrated thymidine has 
revealed that basal cells of the olfactory mucosa respond by proliferating 
to form new receptors. These new receptor cells clearly must die when 
functional reconnectlon with the olfactory bulb is not accomplished within 
a few m.onths . However^ it can be deduced from the simple section experiments 
that, if a contingent of primary olfactory nerve fibers finds its way to 
the bulb, then the population of functional receptors can slowly grow as 
Indicated by approach in size of the experimental nerve to that of the control 
nerve. In a study of olfactory regeneration, the nerve section in the 
pigeon is extracranial because of details in the anatomy. Regeneration does 
occur. The most interesting question now is whether behavioral capability 
is recovered. Tlie use of the zinc sulfate solutions topically applied in the 
nose in hamsters to produce reversible anosmia was investigated. Olfactory 
bulb removal in the male has been shown to eliminate mating behavior and zinc 
sulfate treatment was shown to produce that effect temporarily. The recovery 
that was observed was not due to regeneration of olfactory mucosa, but 
rather to relief from inflammation of large portions of the mucosa that were 
not destroyed. Furthermore, the Inhibition of mating was very variable. 
Some animals had to be dosed repeatedly in order to obtain the effect and its 
duration was highly variable. 

A team of researchers analyzing biological odors Improved methods 
to purify and Identify volatile pheromone compounds in hamster vaginal 
secretions. Housing and dietary changes were instituted for both male 
and female colonies. Those modifications increased male performance and 
reliability. The major modification In the bioassay procedure was to move 
the two odor test ports, through which odors are introduced, to the bottom 
of the cage where they are covered with several inches of bedding. This 
was a better approximation to the natural situation where an estrous female 
is resident below ground level in a burrow. Results of mean time in seconds 
spent sniffing at a particular odor source during a four minute test and 
the number of times a particular stimulus was preferred revealed that an 
odor of nonbiologlc origin elicits exploratory sniffing as against a blank 
odor bottle, both in terms of time spent sniffing and number of trials 
preferred. The actual time scores were less than those for the vaginal 
secretion or Its distillate. What was not apparent is the remarkable and 
characteristic difference evidenced by the male in the two odor environments. 
With non-biological odors, males are normally quiet, sleeping or grooming. 
In contrast In the presence of vaginal odors, the male displays Intense 
activity directed almost exclusively to the bottom of the cage. Further 
work on the vomeronasal organ response to odors was achieved by cutting the 
skin of the roof of the mouth and carefully drilling through t}ie hard palate 
until the ventral extent of the vomeronasal organ was uncovered. After 
careful removal of the dura, a fine tipped microplpette is in-produced at an 
angle with a micromanipulator. Conventional amplification and display to 
techniques are used. They have been able to record unit spike activity to 
female hamsters vaginal secretion odors from presumed receptor cells in 
the vomeronasal organ. So far the results are sporadic. Presentation 
methods are being Improved for reliable results. 



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Taste 

A group of investigators studying mechanisms of taste function are 
developing biochemical approaches to study the early events in taste sensation, 
including the initial stimulus -receptor interaction and subsequent alterations 
in levels of a possible "second messenger." Peripheral taste receptors can 
apparently discriminate among taste stimuli, but the interaction between a 
stimulus molecule and a receptor site is weak. Major goals were to develop 
a procedure for obtaining taste bud cells in suspension to develop improved 
assay procedures for measuring properties of taste receptors _in vitro , and 
to explore the possible role of the cyclic AMP system in taste receptors. 
Earlier evidence showed that some taste discrimination of sugars apparently 
occurs at the initial binding step in what has long been regarded as a very weak 
interaction. Biochemical research on taste has relied on relatively crude 
preparations Trom entire taste papillae which contain many other tissue 
components in addition to sensory cells. A procedure has been developed 
to prepare suspensions of taste bud cells using collagenase digestion, 
micro-dissection,, gentle horaogenization, and fractionation of the resulting 
cell suspension on a discontinuous density gradient. The suspensions are 
highly enriched in taste bud cells. Preliminary studies on the catfish taste 
system show that this animal will provide suitable inaterial for more detailed 
biochemical studies. 

It was proposed to explore the possible role of cyclic AMP as a 
"second messenger" in the taste system. Although other recent reports have 
shown the presence of adenyl cyclase in olfactory epithelium and taste papillae, 
no evidence was reported for regulation of activity by chemi al stimuli. 
Evidence has now been obtained that cyclic AMP might mediate taste responses. 
Increased incorporation, in response to sucrose, of l^C precursor Into lUC- 
cycllc AMP in intact bovine taste papillae was found while this incr 
not occur in tongue epithelium. 

Gynemic acid, a specific inhibitor of sweet sensation, was found 
not to act at the binding step for sucrose and it was therefore hypothesized 
to act at a step following the initial binding interaction. The sweet- 
tasting protein monellin was characterized as a carbohydrate-free protein 
of molecular weight 10,700 and isoelectric point 9-3- Its amino acid com- 
position and other chemical characteristics were determined. The significance 
of this research is that taste plays an important role in human nutrition. 
A desire for the sweet taste often leads to eating sucrose containing foods, 
which promote development of dental caries. The sweet-tasting monellin 
offers promise in nutrition, in reducing dental caries and in special low- 
sugar diets. 

Because little is known of the biochemical basis of tasting, it is 
difficult to design taste stimuli for specific purposes, such as flavors 
for special diets for obese or diabetic patients whose caloric intake must 
be controlled. It is also difficult to treat disorders of taste. Tlie current 
studies of biochemistry of taste should help lay a foundation of information 
on the f'jtnctioning of this sensory system. The possibility that the "second 
messenger" cyclic A!4P mediates taste sensation could mean that some of 
the mechanisms in hormone sensitive cells (internal chemoreceptors) and in 

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taste receptions (external chemoreceptors) are fundamentally similar, which 
could provide new experimental approaches and insights. 



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RESEARCH TRAINING 

Fiscal Year 1974 was the initial phase-out year of Federal support 
for NINDS training programs as they have been known to both the NINDS and 
the scientific community. These programs are the Training Grant Program, 
NINDS Special Traineeship Program, Teacher-Investigator Special Traineeship 
Program, Research Career Development Award Program, and the Postdoctoral 
Fellowship Program, 

The phase-out of the programs was announced in a memorandum from the 
Acting Director, NIH, dated February 8, 1973, which detailed the phase-out 
guidelines. In general, the guidelines directed that no funds could be provided 
to support training programs beyond the period of commitment to trainees and 
that only those new trainees could be supported who, prior to January 29, 1973, 
had received a firm institutional commitment to be supported on the training 
grant. Individuals receiving direct support from the NINDS would have their 
periods of recommended support honored and no new awards would be made. 

A major activity in the spring of 1973 had been to establish the 
identity of specific individuals that had firm institutional commitments to 
be supported on training grants prior to January 29 and to establish the 
length of time required for these specific individuals to complete their 
training. With this information available, awards were made for the grant 
year July 1, 1973 through June 30, 1974. The amount of the award was based 
on the number of trainees that would be in the program and a proportional 
amount required to support the training environment. With the exception of 
the "Centers of Excellence," where support was provided for trainees only, 
awards were negotiated 10%. A program that had its full complement of trainees 
received 907o of its recommended support. Regardless of the number of trainees, 
no environmental support was reduced more than 507o of the recommended amount. 

A major activity of the program was to ascertain that the only trainees 
being supported on the training grant, during the budget period beginning 
July 1, 1973, had commitments prior to January 29, 1973. Thus every "Statement 
of Appointment of Trainee" form that was received was checked against the list 
of individuals earlier identified by the program director as having committed 
support. In addition, when continuation applications were mailed to the 
program directors for NINDS support for the budget period beginning July 1, 
1974, an accompanying memo was included with each kit which identified the 
specific trainees who could be supported on the grant. 

The phase-out of the NIH training programs was to have resulted in an 
estimated "savings" of approximately $30 million per year for three years begin- 
ing with Fiscal Year 1974 . During the summer of 1973, the Secretary, DHEW, 
announced a $30 million per year program, for each of three years to support 
an Institutional Research Fellowship Program and an Individual Research Fellow- 
ship Program. The Institutes desiring to participate in these programs were 
required to submit a special justification, for approval by the NIH, for 
disciplines in which institutional grants and individual awards would be made. 



69 cc 



The Institute requested approval and was authorized to make Institutional 
Research Fellowship Awards in developmental neurology, neuro immunology, neuro- 
virology, sensory physiology and biophysics, and minority programs in the 
neurosciences . In addition to these five areas, the Institute requested 
approval and was authorized to make Individual Research Fellowship Awards in 
neurobiology, neurochemistry, neuropharmacology, neuropathology and otopathology, 
clinical investigation, neuroanatomy, neurophysiology, neuroradiobiology, 
audiology and speech pathology. (See appendix A) 

Concerning the new "Weinberger" programs, although the Institute had 
received authority to make Institutional awards in FY 1974, we elected not to 
participate in the program in the initial year. Concerning the Individual 
Research Fellowship Program, the Institute was assigned 373 applications 
which had their initial review in a Division of Research Grants' Study Section 
to which the applications were assigned. Although the Institute anticipates 
funding approximately 200 of these applications, the specific awardees will 
not be selected until sometime in June and hence a breakdown of these awardees 
is not available. 

In Fiscal Year 1974, 59 training programs, which had been supported 
over a number of years, were in their final year of recommended support. Under 
normal circumstances, the directors of these programs would have submitted 
renewal applications by the February 1, June 1, or October 1 deadlines in 1973. 
These applications normally would have been reviewed and would have competed 
for funding at the March 1974 meeting of the National Advisory Neurological 
Diseases and Stroke Council. When the phase-out guidelines were received, 
applications which had been submitted by the February 1 deadline were adminis- 
tratively inactivated and no other renewal applications could be accepted. 

Of the 59 programs in their final year of recommended support, 32 
were extended administratively for one, two, or three years because they 
had commitments to individuals who would not complete their training for one, 
two, or three additional years. Hence, they received an award beginning 
July 1, 1974 to provide for the trainees and they received one or two additional 
years of committed support. Twenty-seven other Program Directors who were not 
committed to trainees received no new money for the budget period beginning 
July 1, 1974. These 27 Program Directors and their training institutions are 
the following: 



GRANT NO. 


PROGRAM DIRECTOR 


5583 


Marshall Allen 


5231 


Ellsworth Alvord 


10,048 


Robert Baker 


5660 


Betty Banker 


5548 


E. C. Brandow 


10,057 


Rosalie Burns 


5667 


William Collins 


5144 


John Daly 


5126 


Robert Feldman 


5641 


Irving Fish 



INSTITUTION 

Medical College of Georgia 
University of Washington 
University of Nebraska 
Case Western Reserve U. 
Albany Medical College 
Medical College of Penna, 
Yale University 
New York University 
Boston University 
New York University 



AREA OF TRAINING 

Neurosurgery 

Neuropathology 

Neurology 

Neuropathology 

Otolaryngology 

Neurology 

Neurosurgery 

Otolaryngology 

Neurology 

Child Neurology 



70 cc 



GRANT NO. 


PROGRAin DIRECTOR 


5315 


G. S. Fitz-Hugh 


5400 


Reinhard Friede 


5721 


Fredie Gargano 


5362 


Frederic Girardeau 


5712 


Nicholas Gonatas 


5286 


William Hardy 


5544 


Gabor Kaley 


5564 


Wolff Kirsch 


10,028 


Sanford Larson 


5596 


Gerard Lehrer 


10,056 


iilliott Mancall 


5618 


Lester Mount 


5557 


Louis Nelson 


5464 


Daniel Pease 


5664 


Charles Poser 


5702 


Robert Ruben 


5620 


Philip Sprinkle 



INSTITUTION 

University of Virginia 
Case Western Reserve U. 
University of Miami 
University of Kansas 
University of Pennsylvania 
Johns Hopkins University 
New York Medical College 
University of Colorado 
Medical College of Wisconsin 
Mt . Sinai School of Medicine 
Hahnemann Medical College 
Columbia University 
Albany Medical College 
UCIA 

University of Vermont 
Albert Einstein 
West Virginia University 



AREA OF TRAINING 

Otolaryngology 

Neuropathology 

Neuroradiology 

Communicative Dis , 

Neuropathology 

Audiology 

Neurophysiology 

Neurosurgery 

Neurosurgery 

Neurochemistry 

Neurology 

Neurosurgery 

Neurosurgery 

Neuroanatomy 

Neurology 

Otorhin olaryngology 

Otolaryngology 



A major decision in the spring of 1974 was the Department's (HEW) 
decision to release Fiscal Year 1973 "impounded" funds for training and 
research. The release of these funds had a major impact on the NINDS training 
activities. The major guideline concerning the use of these funds was that 
they should be used to make new and supplemental awards which would have 
been made had these funds been available when our Fiscal Year 1973 funds 
were being programmed. This resulted in practically all of the training 
grants, except for the "Centers of Excellence," receiving supplemental funds 
and the awarding of grants to support those new programs that we normally 
would have funded in the spring of 1973. Utilizing these impounded funds, we 
made the following number of training grant awards: 



TYPE OF APPLICATION 

Supplemental 
Renewal 
Continuation 
New 



NUMBER OF AWARDS 

196 

10 

1 

5 



AMOUNT 

$ 2,122,239 

635,828 

24,436 

357,362 



Thus 212 awards were made for a total of $3,139,865. 

The following are the new training programs that were funded: 



PROGRAM DIRECTOR 

Leo Abood 
Vahe Amassian 
Melvin Cohen 
Warren Grover 

James Snow 



INSTITUTION 

University of Rochester 

SUNY, Downs tate 

Yale University 

St. Christopher Hosp . (Phila.) 

University of Pennsylvania 



AREA OF TRAINING 

Neurochemi s try 
Neurophysiology 
Neurobiology 
Developmental Child 

Neurology 
Otolaryngology 



71 



cc 



These five programs were given initial budget periods of 16 months, terminating 
June 30, 1975, and two additional years of support at the recommended level. 
Although new programs were funded, the training programs remain in a "phase- 
out" status. When the Institute honors its years of recommended support for 
a program, additional support will not be available. The effect the release 
of impounded funds had on the program was to prolong the phase-out period. 
This resulted from restoring years of recommended support that had adminisrative- 
ly been taken away at those institutions where program directors were not 
committed to trainees as of January 29, 1973. 

Also from impounded funds, the Institute awarded seven research 
Career Development Awards ($148,952), eight Postdoctoral Fellowships ($57,000), 
three Teacher-Investigator Special Traineeships ($60,500), and twelve Special 
Traineeships ($185,671). The 20 individuals who were awarded Postdoctoral 
Fellowships or Special Traineeships had the option of receiving an award from 
the "old" program from impounded funds or being supported through an Individual 
Research Fellowship from Fiscal Year 1974 funds. 

Transfer of Research Career Development Award Program and Teacher-Investigator 
Program to Research 

In November 1973, it was announced that the Research Career Award Program and 
the Research Career Development Award Program would no longer be supported 
as training activities and that they would be continued as research activities, 
to be supported through research funds. The RCDA program, at that time, was 
in a phase-out status. 

On the assumption that support for training programs would be withdrawn 
and in order co protect the Teacher-Investigator Special Traineeships, per- 
mission was given to identify the T-I program as an "Academic Career Award" 
(K-7) rather than as a "Direct Traineeship" (Fll) as initially determined. 

Although funding of these three programs proceeded out of Fiscal Year 
1974 training money, the following approximate amounts will be reprogrammed, 
from the training to the research category, to continue the support of these 
programs in FY 1975: 

PROGRAM AMOUNT NO. OF AWARDS 

Research Career Award Program $ 277,000 9 

Research Career Development 

Award Program 1,450,000 80 

Teacher-Investigator Special 

Traineeship Program 405,000 21 

In April 1974, the NIH announced the continuation of the RCDA Program 
under the old guidelines and the Institute was given permission to announce 
the Teacher-Investigator Program. The "new" T-I Program is different from the 
original Program. The RCDA forms would be used for applying for an award and 
the award is to be made to an applicant institution which submits an applica- 
tion on behalf of a specific candidate. 



cc 



Manpower Evaluation Studies 

In the fall of 1972, the Institute awarded four contracts to conduct 
manpower evaluation studies in neurology, neurosurgery, otolaryngology, and 
speech pathology and audiology. These were awarded to the American Neurological 
Association, the American Association of Neurological Surgeons, the American 
Council on Otolaryngology, and the American Speech and Hearing Association 
respectively. 

In the fall of 1973, these four contracts were extended for an 
additional six months and it is anticipated that final reports will be avail- 
able before the end of the year. 

The Institute, over a period of time, was aware of the need for a 
manpower study in the basic sciences and on June 30, 1973, a contract was 
awarded to the National Research Council of the National Academy of Sciences 
to assess the manpower needs related to the neurology and the communicative 
basic sciences. Dr. 0. Malcolm Ray is the Project Officer. It is anticipated 
that their evaluation will also require about 18 months to complete. 

APPENDICES 

Following are four appendices which show how support for the Institute's 
training programs is divided among the various programs and training areas. 
The numbers and amounts pertain to grants or awards from Fiscal Year 1974 
funds only and do not include awards from Fiscal Year 1973 "impounded" funds 
which were released. 

Appendix A is the anticipated number of grants and the training 
grant funds awarded for FY' 74 according to the Institutes areas of respon- 
sibility. This is a decrease of 10 awards over FY' 73. 

Appendix B is the anticipated number of Special Traineeship Awards 
and the amount for FY' 74 divided among the Institute's areas of responsibility. 
This is a decrease of 54 awards over FY' 73. 

Appendix C is the distribution of 11 Teacher-Investigator Special 
Traineeships among the Institute's areas of responsibility. This is a 
decrease of two awards over FY' 73. 

Appendix D is the anticipated number and the amount awarded for RCAs , 
RCDAs , and Postdoctoral Fellowships in FY' 74, This is a decrease of 11 RCDAs 
and a decrease of 41 Postdoctoral Fellowships over FY' 73. Nine RCAs is a 
decrease of one from FY' 73. 



73 



cc 



APPENDIX A 



Distribution by Scientific Fields, 
of Training Grants Awarded in FY 1974 



FIELD 



Audiology 

Cerebrovascular 

Child Neurology 

Communicative Disorders 

Neuroanatomy 

Neurobiology 

Neurochemis try 

Neurological Sciences 

Neurology 

Neuropathology 

Neuropharmacology 

Neurophysiology 

Neuroradiology 

Neurosurgery 

Neurovirology 

Otolaryngology 

Sensory Physiology 

Speech Pathology 

TOTAL 



NUMBER 

6 

1 
14 

6 

3 

4 

2 

6 
51 
12 

2 
13 

8 

21 

1 
36 

3 

2 



191 



AMOUNT 

$ 305,800 

97,100 

878,100 

461,800 

156,700 

180,100 

102,100 

218,300 

3,880,900 

562,500 

144,800 

680,200 

403,400 

1,034,600 

41,100 

2,544,800 

142,300 

65,400 

$11,000,000 



April 1974 



7^ 



cc 



APPENDIX B 



Distribution, by Scientific Fields, 
of Special Traineeships Awarded in FY 1974 



FIELD 

Child Neurology 

Neurology 

Neuropharmacology 

Neurophysiology 

Neuroradiology 

Neurosurgery 

Neurovirology 



NUMBER 



TOTAL 



18 



AMOUNT 
$ 60,150 

15,200 

26,200 

37,925 

44,760 

72,790 

17,850 

$ 274,875 



April 1974 



75 



cc 



APPENDIX C 

Distribution, by Scientific Fields, 
of Teacher-Investigator Awards Granted in FY 1974 



FIELD NUMBER AMOUNT 

$ 46,000 
20,000 
23,500 
20,000 
23,500 
23,500 
45,000 
24,500 
20,000 



Cerebrovascular 


2 


Immunology 




Neuroanatomy 




Neurobiology 




Neuroendocrine logy 




Neurology 




Neur ophys io logy 




Neurosurgery 




Sensory Physiology 




TOTAL 


11 



April 1974 



$ 246,000 



76 



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APPENDIX E ^ 

RESEARCH GRANTS AWARDED IN FY 1974 BY DISORDER CATEGORY 
(Dollars in Thousands) 

DISORDER CATEGORY NO. AMOUNT % of $ 

— ( 

TOTAL ALL DISORDERS 1,065 59,983 100.0 

1. NEUROLOGICAL DISORDERS 

A. Neurological Disorders of Early Life 

B. Neurological Disorders of Aging 

C. Cerebrovascular Disorders 

D. Convulsive or Related Paroxysmal 
Disorders 

E. Demyelinating or Sclerosing Disorders 

F. Muscular or Neuromuscular 

G. Infectious Diseases 

H. Trauma or Injury 

J. Tumors of Nervous System 

M. Neuroendocrine Studies 

N. Neural Aspects of Learning or 

Behavior 44 2,297 3.3 

P. Nervous System Studies - 

Normal Function 116 5 509 7.9 

TOTAL - NEUROLOGICAL DISORDERS 772 47,659 68.1 



156 


8,976 


12.8 


48 


3,646 


5.2 


54 


6,471 


9.3 


41 


3,051 


4.4 


40 


2,935 


4.2 


113 


5,544 


7.9 


12 


641 


0.9 


61 


4,407 


6.3 


21 


1,070 


1.5 


66 


3,112 


4.4 



78 



cc 



ArPENDIX E (Contd.) 
RESEARCH GRANTS AWARDED IN FY 1974 BY DISORDER CATEGORY 
(Dollars in Thousands) 



DISORDER CATEGORY 



B. Disorders of Speech or Other Higher 
CNS Functions 

C. Disorders of Other Senses 



NO. AMOUNT 7o of $ 



SENSORY AND PERCEPTUAL DISORDERS 

A. Disorders of Hearing or Equilibrium 



112 

27 
133 



8,994 12.9 



2,373 
5,769 



3.4 
8.2 



TOTAL - SENSORY AND PERCEPTUAL DISORDERS 272 17,136 24,5 



3. MULTI -CATEGORICAL 



17 5,101 



7.3 



4 . CONFERENCES 



87 



0.1 



79 cc 



< 



I 

ANNUAL REPORT 

July 1, 1973 through June 30, 197^ 

Extramural Programs 
Data Analysis and Reports 

P National Institute of Neurological Diseases and Stroke 

The Data Analysis and Reports Unit is fully operational with its 
computerized information system which operates, for greater accuracy and 
flexibility, in parallel with the manual system. The data base provides 
generally comparable data for the FY I965 to FY 197^ time span on all re- 
search and training grants, fellowship awards and indirect trainee appoint- 
ments funded by the NINDS Extramural Programs. Considerable update and modi- 
fication of existing programs along with the development of some new programs 
has been accompanied by a documentation of all software, which is nearly 
complete. Considered jointly these processes promote increased efficiency 
and improved effectiveness in responding to user requests and in assuring 
response under a wider range of staffing contingencies. 

The unit, in addition to regularly published data books and fiscal 
year end summary books, has prepared special reports for the Institute's 
three regularly scheduled Advisory Council m.eetings in FY 197^. These reports 
compared the volume of clinical with non-clinical research (March Council), 
analyzed the Program Projects evolution in recent years (November Council) 
and siiinmarized the major extramural grant program trends in recent years 
(June Council). The unit also responded to special requests (typically 10 
or more per week) of the Office of the Director, usually processed through 
the budget office, the program planning and evaluation office, or the in- 
formation office. 

The unit had significant staffing problems during the winter and 
early spring and these have been largely resolved. Therefore, no unusual 
delays or data problems are to be expected. 



81 cc 



4 



AmUAL REPORT 
July 1, 19T3, through June 30, 197U 
Extramural Programs 
Grants Management Section 
National Institute of Neurological Diseases and Stroke 

In addition to normally assigned duties, the Grants Management Office 
during FY 197^ assisted in the Institute's Extramural Grant Funds accounting. 
Records of funds allotted plus obligations and encumberance lists have been 
maintained on a current basis. Fiscal Data maintained in the Grants Ledger 
now reflects a more explicit breakdown of Supplemental and Won-corapeting 
continuation funding. Supplements are now further subdivided between Council 
and Interim awards. Non-competing funding is subdivided between Type 5 regular 
continuation and Type J, change of institution funding. Staff actions are 
maintained in the ledger, (increases, pre-reductions, decreases, unobligated 
balance savings, full and partial deobligations) . This infoimation is avail- 
able on a current basis, and this procedixre will save time at the end of the 
fiscal year as individual grant cards will no longer have to be reviewed. 
The Indirect Cost Management Section is constantly revising indirect costs 
awarded during the fiscal year and extra effort has to be employed in the 
closing weeks of the fiscal year to keep abreast with the fund balances. 

Equipment Accountability submission by the grantees has required staff 
follow-up. Grantee submissions have indicated a lack of understanding of the 
policies as well as improper disposition or insufficient reporting. Grants 
Management Staff has rectified this reporting working with the Scientific 
Administrator and the grantee institution. Program and Management review has 
resulted, in a nujnber of instances, with the grantee continuing the accounta- 
bility for the equipment. 

Impounded Fund Awards required in depth analyses and budget reduction 
schedules for the staff administrator, as many of these grants were adminis- 
tratively withdrawn. 

The Closeout of terminated files proceeded very slowly during the 
present fiscal year. Contributing to this slow production was the prolonged 
illness and eventus.l loss of our Grants Technical Assistant. We are now 
training our Grants Clerk to assume this function. Recent updated internal 
procedures within the file room and the Data Analysis Section hopefully will 
contribute to more expeditious closeout of old files. We have been making 
all reasonable efforts to secure closeout documents, but in some instances 
have been unsuccessful. 

Grants Management has been utilized frequently in t)ie Program Project 
Site Visits. Our office is involved in preparatory fiscal analyses to enable 
the site visit teams and the Institute to more effectively carry out its grant 
management role in the Institute's programs. 

Overall, our staff is still hampered by our heavy workload. We are 
veiy much in need of the services of a Grants Technical Assistant and another 
Grants Clerk to assist the staff. Many management areas not having deadline 
dates are not receiving full attention due to the manpower problem. 

83 cc 



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