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The Office of the Clinical Cent- or plans end administers 
a full range of patient care and other services necessary in the c 
duct of effective cl" search at NIH. Among the mo3t important 
of these functions is the maintenance of an environment in which 
advanced, academically-oriented medical t a superior brand of 
clinical care; and productive medical research may be e:cp?cted to 
flourish . Equally important in an era of extremely rapid advances in 
medical and hospital practice is the role of this office in helping 
NIH fulfill its responsibility for widely disseminating new knowledge, 
techniques and viewpoints to medical and related disciplines 

Substantial achievement was made in all of these domains during 
the past year. 

Admission and Occupancy Data 

After 5 years of carefully planned expansion, total activation 
of patient care and clinical research physical facilities was achieved 
in the second half of 1957- Thus, 1958 represented the first full 
calendar year in which maximum utilization of the Clinical Center's 
$16 beds and associated patient care services could be attempted » 
The number of inpatients admitted totalled 3»300 during 1958, an 
increase of 59b over the preceding year. The daily average census 
was 38U as against 329 in 1957, while the average length of stay was 
Uh as compared to Ul days in the previous year,, The occupancy rate 
for the year was 7h% of available beds as against 66* for 1957^ 

Approximately 12,000 patients have been admitted since the 
Clinical Center opened in July 1953 <= Ea h of these patients was 
referred by a physician and each was accepted because his medical 
background and current condition suited the rather precise require- 
ments of one or more studies being conducted in the Clinical Center. 

US6 2 


To obtain this m 

receivo and 

a im- 

about one in ton 

ratio of 
to best e 

the diagncsos accepted arc particu 

nesses such as cancer, heart abnormalltier. , rhe 

respirat:. :js, epilepsy, and severe js. 

In several of these conditions ther 

properly referred and qualified patients. 

However, in some other catopori' 
disease or conditions Is so 

cooperation of ins throughout th 

ra needs arc to be met. Thus, with f the 

Clin decided 

to intensify nationwide efforts to apprize ;sion, 

methods and diagnostic needs of th< 

in a substanti ed volume of 

the Union and a 10 per 
from outside I Bethesda are 

land, and Virpinia), 

The following table prov d occup.v 

data for the five calendar years I ;as been 

Jn operation. 

Admission and Occupancy Data — Calendar Year 

Total Number of Admissions 

Percentage From Other Than 
Immediate Area 

Total Patient Days 

Daily Average Number of Patients 

Average Length of Stay 

Percentage of Bed Occupancy 

Outpatient Visits (includes admis- 
sion examinations and follow-up 
research examinations) 






































21*, 5U1 

Bach year a number of c oia 

i duties these »• 

itients must ■ at 

leave to pur 

7 one or mon* year? 

•vldency 16 8 

medic ,a 

These ^ sped;. 


Diagnoetlc Radiol 

Internal Medicine 


Lar tecnt activities 

of form** 

replies to -t was revealed U 

time tea Iraost as many mc <ools, 

unlverel! »; 27 were lr. completing their 

medical specialty trr Lae medical practice; and 

16 were engaged in various combinations of 
private practice - 

Obvious .ntrlbution to r- ind teaching 
these young men will make in the years ahead cannot be assessed pre- 
cisely. However, it is certain that their contribution will be vastly 
greater because of the experience received in the 2 or 3 years spent 
at Bethesda, 

Medical Board 

The Medical Board for 1958 had a new chairman and four other 
new members- There were a number of changes in appointments to the 
various committees of the Medical Board „ 

There was the addition of a Steering Committee to the Standing 
Committees of the Medical Board and incorporation within its functions 
those formerly assigned to the Pharmacy, Nutrition, and Nursing Com- 
mittees, The Steering Committee considers problems and policies 
pertaining to patient care not specifically delegated to any other 

Office of the Director Page h 

Standing Committer f the Medical 

Board Director or 

Associat - tors of the 

Institutes T ^unh problems and 
presents ] od cal 8oard 

The leen active- aga, n this 

year prir.> Iving normal 


The added to the Organization 

and By Laws Locunv 

Ap/ JtandardSj 

xndards for 
im h 

Ann- airmen for Standing 

Com Article V, Section 6, A 

anc B 

ri.ttee to the 
and incorpora- 
tic i'ormorly assigned to 

the Pharmacy Committees, Article V, 

Sect -tion 

^mmittees are renumbered, 
Tht ant becomes 

becomes item 9 

Amt al Orders," 

the word 
n e: ence between the second 

a sentence at the 

end i 

ling more exc 
asearch Committee of 
the Medi 6, C» paragraph no 6. 

Le II, item 3, to include 
responsibil rector, Clinical Center =, 

Amendment i . iules and Regu- 
lations, ftule~No U y cord Standards, Inpatients" 
to include cervi« ination on all female in- 
patients over ag< 

Office of the Director Page 5 

Amendment No, 13 ,, Revision of Article V, Section 6, no„ 6„, 
Clinical Research Committee, and Article VII, Rules and 
Regulations, Rule No- 13, Normal Volunteers, to permit 
review at Institute level of procedures utilizing normal 
volunteer subjects which involve oaly withdrawal of blood 
or collection of urine. 

Panel of Consultants 

During the year 1958, there were 1,29? visits by medical 
care consultants to the Clinical Center, Six medical care consultants 
have been added to the roster, the total nurtoer now being I6I4 

The system for calling medical care consultants has been 
satisfactory, Institute staff members responsible for covering a 
particular specialty are designated as panel chiefs „ If a staff 
member wishes the services of a medical care consultant he makes 
known his desire to the responsible panel chief who then makes 
arrangements for securing the services of the consultant desired* 
In this manner a consultant can be used by more than one Institute 
during one visit to the Clinical Center. It is felt that the 
consultant program, as a whole, has worked out very well. 

Visitor Program 

The Clinical Center has materially increased its participation 
in the processes by which scientists from all over the world exchange 
first-nand information concerning medical research problems^ tech- 
niques, findings, and trends Management of the official NIH Visitor 
Program was transferred from the Division of Research Services to 
the Office of the Clinical Center Director on January 1, 1958. 

During the fiscal year 1958 almost 10 g 000 visitors, mostly 
professional and pre-professional, were registered in the official 
program By contrast about 2,300 were registered in 1953, the year 
the Clinical Center ooenedc Data for the 10~year period 19k9-1958 
indicates this trend in greater detail: 


19149 . . 

1950 . . 

1951 . . 

1952 . . 



1956 . . 




Of tho jitors rag if 

' if ic w 
daily at N 

35 o corded i ind 30 of these 

required aseic he Clinical Center o 


! services pr nter 

is Institute clinical prorraras, one of the roost 
of the proceedings of the semi-monthly Combined 
nces. These proceedings are distributed 
[H staff and to over 250 NIH medical consultants 
)or of these proceedings are also prepared for 
publication and submitted to various medical and scientific Journals 
each year During 195$ the following appeared in the Annals of 
Internal Medicine: 

Among the 
staff to the i 
important is editi} 
Clinical Gtaff Cor 
regularly to the 
in the area. A m 

"Primary Aldosteronism" 
"Factitious Diseases" 

"Acromegaly, Jejunal Ulcers, and 
Hypersecretion of Gastric Juice" 

- Vol, W, N 0o 3, March 1958 

- Vole U8, No. 6, June 1958 

- Vole U9, No. 3, Sept. 1958 

- Vol. U9, No. 6, Dec. 1958 

"Waldenstrom^ Macroglobulinemia" is scheduled for publication 
in the March 1959 issue of the Annals of Internal Medicine, and four 
other conferences have been selected and are in preparation for this 
journal during the coming year. 

Office of the Director 

A Manual for Metabolic 
607), a new JiO pp. printed publication, ma !■ 
the first of i I cationa nlanned 

which engaged 

ibes the -ept for metaboV 

reeea , and includes a detailed - 

cedures required studies. It t of 

many inquiries about the technical 

■ on 

• 58 a nev 3ystem was begun of 

Stafi st of 3,000 ptajn 

licated an intorest 

Efforts were also made to stimulate I v>a physicians 

to attend Clinic Conferences ;\ge 

were riven wide distribution, and two confer. i the ov 

ing for the convenience of physicians Leo. 

A comprehensive >ecial report or. Institutes of Health 

and Clinical and a guest editorial on "Tr r a Car- 

in Medical Research" appeared in the May 1958 issue of Resident 

Preadmission Services 

An important function of the Office of the Director is to pr 
vlde a central service to all the Institutes for receiving, recording, 
analyzing, routing, and following up on letters of patient referral 
from physicians. All inquiries from high officials of the Government 
and Members of Congress concerning admissions and the patient care 
program are also handled by this office and the Director has NIH-wide 
responsibility for policy and content review of the replies made to 
all such inquiries 

Since its establishment in 1957, the Preadmission Service Unit 
has made real progress toward achieving more consistent, efficient, 
and comprehensive management of these important professional and 
special communications,, The volume of the Unit's 1958 workload can 
be seen in the following data: 

Office of tho : Pg 


Inpatient lotters , . . . 3,108 

Outpatient comspondonce pre: . 1,7^3 

Physician referrals 

Acknowledgement letters •••••••••• 

Rejection letters ..... ..... 

Congressional contacts 

Telephone calls ....... » 102 

Letters answered . lhO 






The Pctient 
eetabli. 7. Dur olendar year it 

continued to cixry out ita primary 



2. To provide a patient library service. 

orvicc procrnm. The 
American Red Croaa Gray Lcdlea i 
to carry ou 

charged eea of t 


In addition, the adminiatrative management o- 
Volunteer program was placed in thia 


The divcrsional rrogror. -a 

of the recreation leader and includes spsrta, both active and pass 
in nature, vith spectator parti cipati or., and a broad program of social 
activities. In the selection of activities it is essential that the 
patient have medical approval to inaure compatible ity with the 
Individual tolerance levels and the patientE roll in his reoearch study. 

The first half of the 1958 calendar year i resented a trying 
situation in that several vacancies occurred in the sports and social 
phases of the program. However, by the end of the summer these 
vacancies were filled and except for a period of orientation many 
phases of both activities produced a very effective program. 

Patient Activities Section p age 2 

Mr. Levis Wills, was employeed as a Recreation Supervisor to 
plan and coordinate the sports activities and Miss Mary Ann Porterfield 
was employeed to cover responsibilities in the social field and 
responsibility in the development of a program for children. 

These activities are carried out in the lUth floor gymnasium, 
assembly hall, and solarium for the general patient population; in 
the 5th floor solarium for seizure patients, and in the Uth floor 
solarium for normal volunteer patients. 

The patients and normal volunteers are provided supervision 
to cover their daily activities. Each morning between 10:00 and 12:00 a.m. 
the gymnasium is reserved for the Mental Health Institute. We do not 
assume direct responsibility for the supervision in their recreation 
program. However, frequently they request our assistance and in most 
cases we extend this courtesy. 

The gymnasium is open daily between 1:00 and 2:00 p.m. for 
patients needing specific supervision in the NINDB. Between 2:00 and 
3:00 p.m. all medically cleared patients from other institutes 
participate in a general interest program. The normal volunteers 
participate in activities between 3:00 and 5:00 p.m. and this time is 
divided into two phases. Between 3:00 and 4:00 p.m. the volunteers 
participate in general interest activities and between 4:00 and 5:00 p.m. 
the male volunteers devote their time to basketball. Daily between 
5:00 and 7:00 p.m. the Recreation and Welfare Association has the gym 
reserved for the employees. 

Tentative plans include sport activities outside of the 
Clinical Center when weather permits. Since the golf course at 
Glenbrook will be inactivated, due to construction, it is important 
that consideration be given to an area for developing a playground. 

The social phase of the recreation program involves parties, 
spectators activities (live shows, games, motion pictures," dramatics, 
hobbies and arts and crafts etc). In addition we continue to schedule 
a cr^ft program Monday nights for the normal volunteer. However the 
title of this program is misleading since the work involves the use 
of the hands and mind. It is true on occasions the volunteer 
participates in a craft activity but it also includes lectures on 
subjects of general interest. The fundamental reason for this type 
of program is to broaden the intellectual scope of the normal volunteer. 
For instance, one evening Dr. Chapman lectured on drug addiction, 
another evening there was a demonstration in the use of linoleum 
blocks; in this particular instance the volunteer was permitted to 
carve linoleum blocks for his individual use. Other topics of interest 
will be engraving, silk screen process, art and sculpturing. The 
lecture series will include such topics as Public Health Service in 

Patient Activities Section Page 3 

Foreign Countries, Iimigration Laws, Missel Program and similar topics 
that will stimulate the general interest. 

The Childrens Program remains in its infancy. Time has been 
set aside for units housing children . We shall, study the situation 
more intensely and make every effort to outline a constructive program 
that will be effective for the children of the Clinical Center. 

Gray Ladies 

The Volunteer Program of the American Bed Cross Gray Lady 
Service provided by the Montgomery County Chapter, has had a successful 
and productive year. Increased interest has been shown in the Children's 
Program witb games, reading and entertaining or helping the children 
with their studies, taking the children for walks either out of doors 
or on the sun deck. 

One of the problems reported by the Gray Lady Service is their 
room rental service. They state it is the most frustrating, aggravating 
and irritating service they attempt to fill and they feel they are 
doing a most unsatisfactory Job. 

Statistics show that an average of 85 Gray Ladies per week have 
contributed 10,000 hours of service for 1958. Progress has been made 
in intergrating the Gray Lady Service into the following areas: 
rehabilitation, recreation, chapel, volunteer training program and 
patient library. Special emphasis must be placed on the time and effort 
the Gray Ladies contributed to the nnn »pi Christmas Program. 

Patients' Library 

Program goals developments and trends including projected 
plans and future work. The program goals listed in the Summary Report, 
1957, represent the Patients' Library's goals for 1958, with the 
following additions: 

A. Improvement of quality of library service through... 

1. Greater emphasis on reader's guidance program for 
individual patients. 

2. Development of program to familiarize patients 
(seizure, mental and other as needed) with the 
library and its resources. The n^ is not only 
to open new interests to them here, but to help 
them recognize the library as a place they can 
turn for entertainment, education etc. after 
discharged from the Clinical Center. 

Patient Activities Section Page h 

Progress Made 

1. Regular, direct service to patients by trained librarians; 
the two librarians now give weekly book cart service to Ik 
nursing units and substitute for the three library Gray 
Ladies on the 6 nursing units assigned to them, when they are 
absent. In general, the mental health patients are reached 
through group reading sessions and when they visit the library. 

2. Group reading and discussion programs have been conducted 
weekly during 1958 in the following institutes: NIMH, 

A weekly story hour for children from all units is held 
in the Occupational Therapy Music Room. Stories are read 
or told to individual children or small groups on the units 
during regular book cart visits or when requested by the 
nursing staff. 

3. Active membership of librarians on the care team on each unit: 

Each librarian concentrates on an institute at a time, attending 
two or three meetings a week, getting acquainted with staff, 
patients and work of the units and coordinating library services 
with other activities of the patients. 

Invariably attendance at meetings results in a marked increased 
in the number of patients referred to the library for special 
attention by the medical staff, aB well as in a noticeable 
improvement in the service the library is able to provide. 
In the case of seizure patients and other with difficult 
behavior characteristics, understanding of these traits and 
knowledge of the special interests and background of the 
patients and helpful in personal contacts with them. 

U. Greatly improved service to eye patients (NINDB) and other 
blind patients throughout the Clinical Center through special 
service by one of the librarians to these patients. 

5. Satisfactory routine for sterilization of library books 
loaned to patients in isolation. After a long period of 
testing,, the Sanitary Engineer notified us that the 
Cryotherm apparatus is satisfactory for this purpose and 
does not greatly harm the books. The books are placed in 
Polyethylen bags on the unit, taken directly to the 
sterilisation ovens and then returned to the library. 

Patient Activities Section Page 5 

6. Successful, week- long celebration of National Library- 
Week, March 17 to 21, resulted in increased use of and 
interest in the library, and in several requests for 
additional library service. The cooperation of 
personnel throughout the NIH in helping publicize the 
program and in taking in the group meetings and story 
hours was noteworthy. 

3. Major problems incurred in the past and problems anticipated 
in the near future. 

Our major problems continue to lack of regular channels 
of communication with other departments serving patients 
in the Clinical Center, lack of necessary space, and 
shortage of personnel necessary to meet additio n al 
requests for library service and to develop the library 
program beyond the rather basic services now offered. 

Normal Volunteer Program 

The administrative management of the Normal Volunteer Program was 
placed in this section to maintain financial, personnel and activity 
records; insure observance of the basic rules governing utilization in 
research studies; provide a continuing program of social recreation 
and educational activities; and maintain liaison between the two agencies 
under contract supplying the Volunteer subjects. 

Since its inception in I95U when 17 Volunteers contributed 1278 
days of service we show a steady increase. In 1958* 85 Volunteers 
contributed 9331 days of service. 

Gee graphic ally the Volunteers arrive from over half of the 
United States and occasionally from foreign countries. However the 
greater percentage are from Pennsylvania, Michigan, Illinois and Indiana. 
The average age for 1958 was 2U.5 years however, this includes several 
of the older Volunteers that participated in specific projects demanding 
subjects in this age group. When these subjects are excluded the 
average age decreases to 21 years. 

There have been interesting strides toward our ultimate goal 
in reaching supply over demand. Volunteers discharged in 1958 show 
a 9$ increase over that of 1957 • We feel that in the near future we 
will have reached the optimum number of normal subjects our total 
census at the Clinical Center can support. 

The average age, male and female, since the inception of thiB 
program is 21.2 years. During this year we have recruited and admitted 
subjects from the Brethren Home for the aged for specific studies in 
the NHI. One Volunteer was 80 years young. Obviously then, the average 
will be elevated. 

Calendar Year 1958 



Serial No. 


The patients tell the Chaplains that the most Important feature of 
the Clinical Center to them is the personal attention and care that they 
receive. To be treated as persons, rather than diseases, means even more 
to them than the superior physical and technical resources of this research 
hospital. It is to help the patient as a person to find meaning and courage 
in the face of chronic illness and death that chaplaino are members of the 

To find meaning and courage for one's life is an individual matter. 
To be of help requires that the chaplains relate to the patients individually. 
The number of patients per chaplain is, therefore, a matter of major importance. 
Experience here and elsewhei'e shows that one chaplain can minister to the in- 
dividual needs of no more than 50-70 patients. That is what the Protestant 
and Catholic chaplains are now doing here. The Catholic Chaplain, who was 
made a full- time member of the staff this year, covers adequately the 60-70 
catholic patients. However, about 250 of the 300 Protestant patients are 
not being covered. There Is adequate provision for a part-time Jewish 
Chaplain for the 20-30 Jewish patients. 

To minister to the individual spiritual needs of all the Protestant 
patients requires four more Protestant Chaplains. Last year's annual report 
stated this to be the moot urgent need of the Chaplains Department. It still 
is. Not only does this lack endanger the important morale factor in the 
patlentG hospitalization, it also is an embarrassment, when the churches end 
hoGpitals of the country expect the Clinical Center to provide leadership in 
this field as it does in all toe rest. 

Ministry to Patients 

Initial lntervievs 















. Pre-Oper. 



Chapel Services 






Holy Ccnsiunion 






Patient Referrals 




77 1 * 

Social Workers 










Staff Relation* 





Staff Mtgs. 


Lectures, etc, 

■ 19 









Conference a 



Correspondence 752 


737 brs. 


vith Chaplains 6l 

Study & Research 739 hrs, 


From January through June 1958, chaplaincy aervice for Catholic 
patients was given on a consultant basin as it had been since the opening 
of the Clinical Center in July, 1953. This service was provided by St. 
Jane deChantal Church in Betheada. As was pointed out in previous reports, 
only the absolute necessities of a Chaplaincy Program could be provided 
by this type of service. 

Toward the end of June, 1958 the Catholic Chaplain was appointed to 
the staff of the Clinical Center on a full-time basis. Being here full-time 
affords the Chcplain time not only to provide more public services, such as 
daily Mass and special services as the seasons indicate, but also more tima 
to devote to the needs and care of the individual patients. Also it allows 
him time to participate in the programs of and work more closely with the 
other departments dealing with patient care, such departments as Rursing, 
Social Service, and Patient Activity. Furthermore, the Chaplain being here 
full-time is more available and can be of greater assistance to personnel, 
who request consultations with him. 

The average Catholic patient census is between TO and 75 patients. 
This number has proven to be the nrniTnum number of patients one Chaplain 
can provide efficient personalized service for in this type of hospital. 
This became quite evident during the months of August and September when 
the Catholic patient census averaged between 90 and 100 patients. 


Divine Worship Service (Mass) for patients and personnel daily 
(two services on all Sundays and Holydays) 

Special religious services daily during months of May and October. 
Special religious services as scheduled during seasons of Advent and Lent. 
Sacraments odministered to patients daily. 
Participation in Orientation program of all new Nursing and Social 

Service personnel. 
Participation in Gray Lady Training Program. 
Participation in programs of Patient Activity Department. 
All Patients visited on a regular schedule for counselling. 
All Patients visited each Saturday for confession. 
Seriously-ill patients visited every day. 
Weekly consultations with families and patients. 
Consultations with Staff regarding welfare and rehabilitation of 

patients . 
Consultations with personnel on request. 
On call for emergency service twenty-four hours a day. 


Jeviah chaplaincy service is on a contract basis providing for 
twelve visits per month. Rabbi Joseph M. Brandriss has been serving 
as Jeviah Chaplain since November 2k, 195^. Approximately 8$ of the 
patients admitted to the Clinical Center are Jewish. 


Sabbath services on Fridays aad special service for the various 
major and minor Holidays of the Jewish calendar, including some socials 
after the celebrations. 

Visits to patients. 

Counseling of patients and families. 

Religious education when requested and/or needed. 

On call for emergency aer/ice twenty-four hours a day. 

Calendar lear 2S$Q 




Serial Number 

Major Goals 

The Employee Health Service acts ao the focal point for all natter* 
relating to the health of employees Its primary goal continues to be the 
maintenance of the health of all employees at the highest possible level 
through the provision of accepted preventive health measures such as 
Immunizationa, periodic X«*rays, health advice, guidance and referral, and 
periodic physical exandnationa In meeting these health needs of employees, 
the research program can be expected to benefit as a result of decreased 
absenteeism and a higher level of effective performanceo The control of 
occupational disease hazards and the prevention of occupational injuries 
and diseases through the provision of periodic physical examinations, 
laboratoiy determinations^ and surveys of hazardous work areas will always 
be a ma.1or goal of the Employee Health programo 

Continued efforts will be made to improve procedures for the manage- 
ment of injuries and illness occurring on the reservation in spite of the 
preventive medical programo The development of the research potential of 
occupational medicine, and in particular the contribution the Employee 
Health Service can make to the research program of NIH, will continue to 
receive our interest,. 

Major Progress 

Progress for a program, of the type and calibre the Employee Health 
Service is pursuing^ is at beat difficult to evaluate because of the many 
intangibles involved, However, over the years, we are convinced that most 
employees at all levels of management are beginning to accept the concept 
and understand the value of preventive medicine in an occupational setting,, 

Greater awareness of the importance of controlling occupational 
disease hazards in the environment has occurred during the year<, Part of 
this awareness has resulted from Joint activities of participating members 
of the Environmental Health Advisory Group — Plant Safety, Sanitary Engineer- 
ing and Employee Health, ^^ 

An effort was made during the year to deal more effectively with 
actional and mental health problems We hop© to increase our efforts along 
these lines during the next year, realizing that 60 to 80$ of ineffective 
performance in industry i 3 due to emotional problems* We can think of no 
area that needs our concerted efforts more than this one 

Major Progress ( Continued ) 

In collaboration with many KJR scientists k our support of and 
participation in clinical research has increased during the part years 
and as in the past B we stand ready to assist any NIH scientist in the 
proper utilization of the employee population In suitable research 
projects o 

Major Problems 

—*■! i nr i ■m il m wjm m 

Added responsibilities placed on our clerical staff warrant a 
thorough review and revision of grade structures applicable to themo 

Space continues to be a problem and we are hopeful that progress is 
being made in obtaining additional space for the Bnployee Health Service 
in the area of B2"A level now occupied by the Laboratory Aids Drancho 

Selected Statistics 

TUm 122L 22£ 122 

Total Visits .000. •<..»•,• 28,011* 28,616 23*339 29,959 
Visits for Occupational Injuries 

and Diseases • ••o«»o*«*« 2,600 2,896 2„960 2 S 633 

Immunizations ooo«oo««oo o 8,23u 8j,760 3^009 2 8 lil3 

Pre~empo Physical Scamlnationa 000 1*828 l p 637 1&915 1 8 351 

Laboratory Examinations „ . « » 12,868* 6,528 5,9l<5 6 C U75 

Referral* to Personal Physician*, „ 1,631 1*1*5U 1*353 1*681 

* Revised system of accounting which now includes all procedureso 

Serxal Mo CC~12 

lo Clinical Canter 

2 Employee Health Service 

3o Bldg ID, Rmo B2«A06 


Individual Project Report 

Calender Tear 1S58 

Pert Ao 

Project Titlei Reactions to Influenza Vims Vaccine 

Principal Investigaton John M» Lynch, M.D. 

Other Investigators* Oexmain Lo Schndt, M.D., Frances Mo Daly, RoN. 

Cooperating Unites DBS 

Men Tearst 

Totalt 1/5 

Professional! 1/5 

Project Descriptiont 


Meaningful,* statistically significant statistics on the incidence 
of adverse reactions following the administration of influenza 
vaccine have been in demand for some time«> With the inauguration 
of large scale influenza immunization programs last year* infor- 
mation on reactions was urgently needed to predict the possible 
impact on large groups of individualse The impression that re= 
actions to influenza virus vaccine might be more disrupting to an 
employee population than the actual influenza needs confirmation 
or correctiono 

Methods Baployed t 

Machine tabulation and statistical analysis of answers obtained 
by questionaire on all employees receiving influenza virus vaccine 
during 1957 - 1556 and 1956-1959 seasons are currently being 
undertaken o 

Pat ient Material * 

The relatively stable e significantly large enough cooperative NIH 
otrolcyee population was considered the ideal resource for con- 
ducting this clinical research,. The vaccine was offered on a 


Patlent Material : (Continued) 

voluntary basis and 2021 employees participated in 1957-1958 
and 2675 participated in 1958-1959o 

Maj or Findin gs t 

■ ^w m mi i m K ill 

Contrary to our expectations, there was a relatively low incidence 
of systemic reactions reportedo Local reactions, while seemingly 
high, were apparently minor in significanceo Absenteeism varied 
from 2 to IS of those employees who reported reactionso Detailed 
findings and discussion are currently being incorporated into a 
paper which is being prepared for publication* Summary of pre- 
lindnary findings is attached. 

Significance to the Clinical Center and NIH t 

Perhaps the greatest significance of this study is the denonstration 
of the clinical research potential of occupational Kedicine Proper 
utilization of the vast pool of clinical roaearch material found in 
the NIH employee population is a major goal of the Employee Health 

Specifically, this study appears to allay fears that reactions to 
Influenza Virus vaccine will seriously disrupt normal operations 
and productivity of an organization, the majority of whose employees 
received vaccine during a relatively short time* 

Proposed Course of Project « 

Publication of findings is planned as soon as possible,. Reaction 
data will be accumulated in future years following immunization 
programs and will be compared to previous findings » 



% of individuals 

reporting reactions « 

1st. Inoculation 

2nd Inoculation 





Any Systendo Reatioa 




Fever, Unspecified 



Fever* under 100° F 


0o3 5°3 

08 2 3 

Fever, orer 100° *" 




12 oh 











Any Local Reaction 






26 o3 

17 oO 



22 o$ 






Other Reaction 




Ho Reaction 




Individuals Reporting Lost Tine 

86 (l*oW) 
158 (lo8 days, 

28 (2 o 0Jt) 

Total Days of Lost Time 


Sh (1 6 days, 

Number Vaccinated 




Number Reporting 

H589 (SW) 

2088 (78%) 

1381 (68%) 

Calendar Yea 




Program Goals : 

1. To provide competent casework service to the patients 

QT>d their families in order to alleviate such financial and emotional 
stress as would hinder their full participation in the research and 
treatment pro g ram of the Clinical Center. 

2. To offer appropriate consultation about the social and 
emotional factors related to illness to other members of the multi- 
disciplinary team in order to facilitate the individual patient's 
adjustment in the research setting and to formulate Clinical Center 
policy with regard for the total welfare of patients. 

3. To participate in research undertaken in the Clinical 
Center frcm the standpoint of social, cultural, economic, and emotion- 
al aspects of illness by using such social work methods as history- 
taking, interviewing and ongoing observations in the Center and at heme. 

k. To utilize the social worker's knowledge of the ccoounity in botb 
the local and national sense for the continued medical social, treatment of 
our patients; and to engage in comnunlty and other professional projects 
concerned with Improved Bocial planning for the chronically ill person. 

5. To participate In interdisciplinary training programs in 
order to work toward optimum communication and utilization of pro- 
fessional knowledge among the disciplines for the advancement of the 
research treatment goals. 

6. To provide a staff education program aimed toward constant 
improvement In the quality of casework at the Clinical Center and 
toward furthering the staff's knowledge of basic research methods 
requisite to research participation. 

Progress ; 

Staffing . For the first time since the opening of the Clinical 
Center the staff of this Department is complete. This has been accomplished 
by employing two medical social workers en a part-time basis (three days 
a week) and bringing on one new graduate whom we have accepted responsi- 
bility for supervising for several years in the hope that we can develop 
new staff more adequately now that the crucial service areas are coveredo 
It is hoped that the part-time staff members can be replaced by full-time 
personnel, out because of the crucial shortage of medical social workers 
it is not known when this can be accomplished. 

icaal sad all 

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Serial ii~6~ 

During 195? the Anesthesiology Department fulfilled its 
primary obligations of providing for the clinical management of 
patients undergoing therapeutic surgical procedures and diagnostic 
studies requiring anesthesia, and for the conduct of inhalation 
therapy of all ty->es for patients in the Clinical Center, Research 
activities included the work of one of our staff members who served 
as principal investigator in the Anesthesia Research Laboratory and 
our oarticipation directly or indirectly in various programs of the 
surgical services of the several institutes with which we work 

Our full-time staff was composed of the followinp personnel 
(based on the yearly average): 

Clinical Anesthesiologists - 2\ 
Research Anesthesiologists 
Nurse Anesthetists 
Anesthesia Technician - 1 
Inhalation Theraoists - 2\ 

The services of outside consultants were utilised collectively 
on an average of three visits per week to make up staff deficiencies * 
In addition the services of a research anesthesiologist employed 
by the Surgical Neurology Branch of NINDB were available two-thirds 
of the year, his clinical anesthesia duties being limited to the 
major operations performed by the Surgical Neurology Brancho (Sh cases) 

I„ ANESTHESIOLOOICAL PROCEDURES. The administration of anesthesia 
was carried out in ^.176 instances and special procedures such as nerve 
blocks and consultations were done 110 times, The length of time 
required to complete surgical procedures at the Clinical Center is 
comparatively long: 

5l6 operations, or hh% t had anesthesia longer than 3 hours 
2U0 operations, or 2056, had anesthesia longer than 5 hours 
135 operations, or 11^5$, had anesthesia longer than 6 hours 

It can be seen that a relatively high proportion of the ooerations 
done required more than three hours' anesthesia time and that over 
one out of five of our cases was maintained under anesthesia for 
periods in excess of five hours, one out of ten for six or more hcura, 
Included among the lengthy procedures was the management of ->atients 

Anesthesiology Department 

undergoing open cardiotomy with the use of an extracorporeal 
circulation apoaratus in which the heart and lungs were bypassedj 
CO such operations were done since February 195c\ Also, there* 
were 36 patients who had their body temperatures lowered to permit 
special tyoes of surgery,, 23 for correction of defects of the heart 
and great vessels and 10 during intracranial operations. In other 
words one out of ten of all the anesthesias were for patients 
undergoing complex surpery in which adjunctive techniques, extra~ 
corporeal circuit or hypothermia, were used* Other lengthy procedure© 
in which extensive resections were done for cancer and craniotomies 
for localisation and extirpation of epileptogenic foci required the 
administration of anesthesia over unusually long periods-, All of 
these lonper procedures necessitated the assignment of at least two 
anesthetists to handle the technical details and to nrovide for the 
problem of human fatigue which is bound to develon when one oerson 
stays at the anesthesia controls and monitors vital sims hour in 
and hour out. 

As noted in last year's report, 37% of all anesthesias were 
concerned with diagnostic procedures performed in locations outside 
the Main Surgical Suiter This figure serves to emphasize the 
investigative nature of the work being done in this institution., 

The tabic below indicates the institutes responsible for the 
surgical cases with whom we work Only four of the seven institutes 
have surgical staffs. As may be expected the majority of surgical 
work (9256) is done by NHI, NCI, NINDB, and N3DR 

Administration of Anesthesia. 1958 













18 o5 



11 06 











= 160 o 0g 

Anesthesiology Department 

our staff of two or more inhalation therapists were responsible 
for the setting up and carrying out of various forms of inhalation 
treatments. These treatments are tabulated below: 

Oxygen tent therapy , 9,220 hours 

High humidity tent therapy „ „ <- • . ■> « 17,285 houro 
Gxyper by nasal catheter or mask , » , - 15,883 houro 
Aerosol therapy n «.*...o.o.. 6,367 treatments 
Intermittent Dositive preesure 

breathing with aerosol <..o«««oo U*697 treatments 
Respirator use -. . « , . . • 307 hours 

Resuscitation (artificial respiration) ( „ 261j hours 
Miscellaneous special treatments 1,329 hours 

Administration of oxygen combined 

with other gases 380 hours 

A continuing program for the education cf new oersdnnel in 
regard to the use of standard oxygen and suction devices, which 
are installed in the patients' rooms, was carried out. A total of 
152 registered nurses, 25 oractical nurses, 5li attendants, and 9 
clerks attended teaching sessions which were held weekly , Special 
demonstration sessions were held at the request of nurses from NIA3D 
on the use of the croupette and two sessions were held for the nurses 
engaged in postoperative care of patients from NHI- 

IIIc RESEARCH ACTIVITIES . The Anesthesia Research Laboratory, 
under the direction of Dr f . J- W Severinghaus of our staff, working 
with physical facilities provided by NHI, was in operation during 
the first part of the year- Dr* Severinghaus left to accept an 
appointment at the University of California in June, The work of 
this laboratory was primarily concerned with the development of an 
apparatus to permit rapid and accurate analysis of oxygen and 
carbon dioxide tensions in gas blood or in any liquid mixture, Aa 
the result of experimental studies, Dr.. Severinghaus and his associate^ 
Mr, A, F e Bradley, designed, developed and perfected methods cf 
operation for an improved polarographic call oxygen electrode and 
a carbon dioxide electrode together with associated curvettes, bath 
and circuitry, to produce a functional integrated analyzer for these 
gases, A model was exhibited at the Federation of American Societies 
for Experimental Biology. This work is fully described in a brochure 
which was attached to our monthly report for the month cf April, 1958c 

One of our staff, Dr„ G R, Christenson, directly participated 
in a project sponsored by NIDR, namely s the investigation of physiologie 
responses of ambulatory dental patients during general anesthesia - 
This study calls for the simultaneous recording of electrocardiograms 
electroencephalogram, blood Dressure, and oulse rates during the 
administration of anesthesia to oatients undergoing surgery in a dental 

Anesthesiology Department 

chair- The project required the services of three people, an 
anesthesiologist, nurse anesthetist, and anesthesia technician 
one half day each week The project description is being submitted 
by the principal investigator, Drv Driscoll, from HIDRo 

We also collaborated with Dr- Kenneth Hall, Research 
Neuro-Aneethesiologist of NINDB- Dr Hall was concerned with the 
laboratory and clinical investigation of a new anesthetic drug p 
Fluothane, and mixtures of this substance with ethyl ether. After 
preliminary studies had shown that Fluothane was a valuable and 
relatively safe potent anesthetic agent with non-explosive properties, 
we began to make increasing use of it for maintenance of anesthesia 
in situations where the characteristics of fluothane could be used 
to the best advantage „ Fluothane was used in a total of P8 anesthesias 

IVo MAJOR PROBLEMS ENCOUNTERED, Cnce a^ain, our outstanding 
problem was provision cf sufficient anesthesiolopists for the ever 
increasing surgical schedules- We began the year with two vacancies 
in our quota of four staff anesthesiologists „ Only one ot these 
positions was filled in mid July, In addition, we lost the part-time 
services cf our Research Anesthesiologist in June Our deficiencies 
were covered by employing consultants on a more or less per diem 
basis, Three, and later four, consultants reserved a day or two a 
week to work with us,- We made full utilization of the abilities of 
our nurse anesthetists who now have had several years of experience 
at the Clinical Center , Nurse anesthetists working in collaboration 
with an anesthesiologist managed the anesthetic care of patients 
undergoing all types of procedures t Anesthesia technicians gave 
valuable assistance and helped to alleviate the shortage of professional 
personnel. Every conceivable avenue for the recruitment of anesthesiolo- 
gists was explored including attempts to interest anesthetists in 
Great Britain (durinp visits by the undersigned to England and Scotland,,) 

The problem cf fluctuations in the surgical schedules, resulting 
in the peaking of workloads during the middle days of the week, was 
less troublesome this year. Progressive increase in the number of 
procedures requiring anesthesia plus the shortage of anesthetist 
personnel on certain days caused the surgeons to schedule procedures 
at times when ancillary personnel (surgical nurses, x-ray technicians, 
and anesthetists) were available rather than at times when it might 
have been most convenient from their personal standpoint- 

We are still faced with the continuing problem of obtaining 
adequate civil service grades commensurate with the duties performed 
by our inhalation therapists and our anesthesia technicians o Current 
classifications greatly hemper our efforts to recruit and retain 
intelligent personnel capable of administering inhalation therapy 
treatments or acting as technical assistants to anesthesiologists, 

Anesthesiology Department 

The problems described above are inherent not only in the 
economic situation today which does not allow us to compete on 
an even basis with the opportunities which exist outside government 
service but to some extent are related to the particular circumstances 
which attach themselves to patient care activities in the Clinical 
Center .- 

V, CHANGES AND IMPROVEMENTS MADE, There were no outstanding 
changes during the year- Improvements consisted of applying the 
best technical know-how and skill in the anesthesiological management 
of human beinps whose life and safety are in our hands durinp the 
most crucial periods of their lives r. To this end we sought to provide 
the physical facilities and professional knowledge necessary to carry 
out this objective. We are gratified to state that morbidity and 
mortality due to anesthesia was essentially nil. 


1, Electrodes for Blood ^2 an{ * PCO2 Determinations 

(Sevoringhaus and Bradley, Jour„ of Applied Physiology, 
Vol,, 13, November 1956) 

2 Inflatable Plastic Tub for Hypothermia (Hebert and 
Merzig, Anesthesiology, Volo 19, March April 1958) 

3o Anesthetic Management of Children Undergoing Cardiac 
Catheterization and Cardio-angiography (Hebert, 
Christenson, Clark) presented at annual meeting of 
Southern Society of Anesthesiologists, April 26, 1958, 
Gatlinburg, Tenn- 

Staff members attended a number of national meetings for the 
dual purpose of keeping up with advances in cur specialty and of 
seeking out anesthesiologists in training centers who might be 
candidates for our staff at later da tea „ The undersigned attended 
the World Conpress on Cardiology at Brussels, September, and following 
this meeting, went on to visit hospitals and clinics in Great Britain, 
Stockholm, Copenhagen, and Helsinki.- 

C„ L, Hebert, Mo TJ7 





Program Goals: 

The primary objective of the Diagnostic X-ray Department is to provide 
a complete diagnostic radiological service for the seven Institutes of the 
National Institutes of Health. Other objectives include the operation of a 
diagnostic radioactive iodine thyroid clinic and the provision of X-ray faci- 
lities and service for research studies involving animal radiography. 

Developments and Trends : 

As indicated in previous report, emphasis was placed upon radiation 
protection within the Department during 1958 as a result of publicity and 
increased awareness of the problem. Added filtration, better collimators, 
and lead shielding were measures used to accomplish this objective. 

In February 1958, the Diagnostic X-ray Department assumed the responsi- 
bility for diagnostic radioactive iodine studies. This new function was under- 
taken at the request of the National Cancer Institute. 

A complete residency training program in radiology was inaugurated by 
the Department on July 1, 1957. One new resident will be accepted annually 
with a maximum total of four residents during any one year. 

Plans were developed for structural changes in the patient reception 
area to permit the performance of all radioactive isotope work within the 
Department in this new facility. Assistance was also provided in the planning 
of radiological facilities for the new surgical wing. 

The trend toward a greater proportion of special radiographic examin- 
ations continued during the fiscal year 19ii9o Retrograde ileo-ureterograms 
| were developed, more sectional laminography was consummated and there was 
an increase in the utilization of animal radiographic facilities. 

Another trend which became increasingly apparent during the final six 
II months of the calendar year 1958 was the increase in workload of the Department. 
Over 2,000 more examinations were completed! during the months of July through 
December 1958 than for the some period of the preceding year. 

Plans and Future Work ; 

It is anticipated brain scanning utilising radioactive iodine will 
be initiated as a diagnostic service within' the Department during the 
fiscal year 1959. Likewise, the thyroid diagnostic clinic activities will 

at will ba or 



j re obta 


- 3 - 

Major Problems Encountered : 

The out9t\i- -rtmentnl problem during the fiscal year 

.3 outside the Service plus t". 

1 Service res: 
'.Hied employees (e.g. fi 
also difficult to procure and retain. 

Financial limitations during the fiscal year prohibited 

equipment, particularly a Kodak X-Omatic u 

Mai: are always to be expected with X-ray 

and, of course, were encountered. The two major diff 
e a major 6hut-down of the Pako film 7 
nit due to cl erosion of the tanks, and mechanical end el« 
'•anced with the Schttnander angiocardiographic 

H onors , Avarda , and Publications : 

The Dc; 2 received an award for her id 

ty locking device used In cor.j 

vKis fonr 
7 Radiology and h 

"ccioty. He t ^ on the 

3 District of Col lical Society 
pay 1 .;t the J r -' 

rs published by 

' on. 

•adlological subjects van by st 


Serial No. CC-35-1 

1. Diagnostic X-ray Oept, 

2. Clinical Center 


Individual Project Report 
Calendar Year 1958 

P»rt i , 

Project Title: An Atlas of Laminographic Anatomy of the Head. 

Principal jator: Marvin L. Daves, M.D. 

William E. Loechel 

Cooperating Units: 

CH (Diag. X-ray) (Also Plant Enginee 

Scientific Reports Branch Pat' 


-jar 1958) 
Total: 1/5 

-ict Description: 

To produce a guide for the interpretation 
of the head. 

as of a formalin fixed head will ! 
f corresponding sections of the head 
Led by labeled illustrations of the cut su_ : 






Serial No, 

The objective of the Dental Department of the Clinical Center is 
to provide a diagnostic and treatment service for the various institutes 
of the National Institutes of Health The aim of the Dental Department 
is to perform a complete dental examination on every patient admitted 
to the Clinical Center. Recommendations for treatment are made to 
the attending physician and dental treatment is rendered to these 
patients when it is requested. 

Last year we cited some obstacles which are inherent in a 
hospital concentrating on research and which makes the administration 
of dental treatment difficult. In one instance of a National Institute of 
Mental Health patient, it was necessary to obtain his confidence by 
first instituting work on his family Finally it was possible to perform 
the much-needed dental work on this patient. We have continued to 
have difficulties with patients on the metabolic studies. Quite often the 
studies are started before necessary dental work is instituted. This 
presents quite a problem because often a patient will have a dental 
emergency during the study, especially when the studies last for two 
or three months. We are also working with the children of low I.Q, 's, 
mongoloids, and patients too young to give sufficient cooperation In 
these cases, it is often necessary to perform their needed dental 
services in the operating room under general anesthesia. 

We have cooperated with the Surgical Section of the National 
Cancer Institute by instituting methods of fixation of the jaw before 
radical surgery, such as jaw resections. This elastic traction or fixa- 
tion has minimized the post operative debility. We are also giving post 
operative treatment to these patients daily. This is their means of 
keeping down inflammation, encouraging the patient to eat and holding 
the jaw in the proper position to help counteract scar contraction. Dr. 
Lloyd has had a continuing project in maxillofacial prosthesis in con- 
junction with treatment of patients in the National Cancer Institute, 
This project is designed to evaluate methods of treatment, design of 
appliances, and to observe the response of patients with surgical 
defects of the face. He is also conducting a similar type of project on 
patients afflicted with congenital deformities such as cleft palate. This 
type of work along with these other cases cited, represents a consider- 
able effort which is not reflected in the monthly reports. 

Page 2 

During the past year, we have obtained the services of a new 
consultant. Dr. Robert Fleming, who has taken an active interest in 
the training of our staff in the principles of Endodontics. 

The cooperative study of patients on the leukemia service of the 
National Cancer Institute has been continued. The oral aspects of 
leukemia are being considered and cared for in a much greater respect 
than previously This is developing into a valuable service. 

Last year we had a serious problem of professional staffing. 
This has been overcome and we are now adequately and fully staffed. 
Our problem which we had last year in non -professional staffing due 
to personality conflicts, has been corrected by resignations 

We are also cooperating with the Division of Radiotherapy in the 
development of newer clinical methods of applying therapy rays. 

The Dental Department has cooperated to a great extent with the 
National Institute of Dental Research in the periodontal, orthodontic, 
and genetic investigations Considerable dental chair-time has been 
necessary to assist in these studies Similar examples of time con- 
suming operations could be mentioned for other institutes such as the 
National Heart Institute. The Dental Department participated in the 
Clinical Research Program of the National Institute of Dental Research 
in several ways; first to define cases, secondly to actually participate 
in studies, and thirdly, as mentioned above, we are contributing 
dental chair-time for maintenance of cases under investigation 

Dr. Swerdlow has continued his work on evaluation of damage to 
human dental pulp by cutting instruments, rotating at various revolu- 
tions per minute. He is receiving recognition for this fine work He 
also has another project studying the effect of traumatic occlusion on the 
supporting tissues of the teeth. 

Work is progressing by the Photographic Department at the 
instance of the Dental Department on an unusual clinical camera to 
photographically document color changes in the gingival tissues in 
sickness and in health. 

The Dental Department is prepared to and did render every type 
of dental treatment found necessary to perform on the patients examined 
Again this year, the number of patients examined, the total number of 
visits, and the dental treatment rendered has increased over previous 
years. The following table is self-explanatory. 

Page 3 

Admissions to 


). 956 







Admissions to 







10, £78 


Admissions to 








There has been a continue d increase over previous years in the 
number of examinations performed, total number of visits, and the 
total number of treatments. Tht: increase parallels the increase in 
the Clinical Center Census. 

The need for an additional dental technician is still apparent. I 
am sure that it will be necessary to create an additional job in the near 
future. We stated last year that it will be necessary to re-classify the 
jobs of the present dental technicians in order to persuade them to stay 
in our program. They are constantly being offered job opportunities by 
outside concerns. It will be an impossible situation to replace these 
men at their present salary. It is my understanding that action is 
finally being taken in this matter. The auxiliary personnel of the 
dental staff is presently the most efficient we have yet had. It will 
probably, however, be necessary to obtain the services of one additional 
girl in the near future. 


Serial No. 


The Clinical Pathology Department, Clinical Center, I.JIK, le organ 
to provide laboratory coaaultation and services of research quality in the 
areas of hematology, clinical chemistry, microbiology end special diagr. 
procednres. One of the most Important contrlbutlone of our department 
the application of methods from the rapidly advancing fields of research in 
medicine and basic sciences to serve the purposes of daily patient a 
Growth of work in this phase mill continue to require core attention 
often t las-consuming meticulous operations. These efforts not only vidan 
the scope of signifies tie information, but frequently yield more 
economic, accurate and rapid technics, thereby increasing our capabilities. 

Work Load 

An average patient census of 392 In the seven Institutes resultec 
99,080 patient work days during 1958. Thsy required 319,048 test procedures 
en Increase of 14%. Concurrently the average number of equlvalc 
nologlsts on duty decreased from 41.7 in 1957 to 38.2 1 
rise in the average teot load for each technologic .7 to 33 per 
The increasing number of compllcsted and time-conuuming testo t&lcl. 
individual attention end meticulous operations add considerably to 
pressure of work, bet these cannot be proportionately reflected In product- 
ion etatlstlcs. Short supply of adequately qualified codlca). tec'.r vises 
prevented recruitment to full strength of our staff and we wore unable to 
adjust the individual load to 25 which is the maximum eompatib; 
sum efficiency, accuracy and research quality, even with automatic methods. 
Thus, the major problem was the increasing requirement for patient c 
laboratory work considerably exceeding the capacity of our staff end space. 
A vigorous campaign to reach 80Z bed occupancy during the winter and summer 
of 1958 exeggeratad this difficulty and unfortunately coincided with the 
msxlnw attrition of technical personnel. We wero o curtail several 
procedures temporarily until more staff could be recruited. Additional 
space and recruitment have Improved the situation and Implemented the 
department for a won': load of 20,000. However, the volume has 
steadily grows to 32,000 (October) end undoubtedly will remain above 30,000 
with continuing 30?. occupancy. Five additional modules were made available 
In October, modified and occupied in November and December. This corrected 
the overcrowded conditions and facilitated more efficient arrangement of 
the working spaces in Clinical Chemistry, Microbiology and Hematology. 
Smployomst of additional authorized personnel will enable the Clinical 
Pathology Deportment to process efficiently 30,000 tests each month and 
maintain the necessary high quality. 

of fche research patient requires thorougb evaluation ox nis basal c 
sad biological r ctition of teste 

detect early clianges and many mora meticulous and time* ires 

thaa would be needed In a general hospital. 


Growing preosuro from the American Society o L Tochnologir 
and governasnt pathologists and the American Society of Clinical Patholo- 
gists, or.peclally through their Joint Civil Service Armed Forces Committee, 
forced the Civil Service Commission to review, revise and reissue standardo 
for technicians. Strong representations from RIB supported the urgent and 
coordinated reccatendatlons of the Civil Service Armed Forces Committee to 
establish a new professional series of Medical Technologists similar to 
existing series for professional scientiots. The Civil Service Coir 
invited this committee, the Armed Forces, Veterans Administration and HIH 
to advisa and participate in writing the new standards and specifications. 
This departatsnt and Clinical Center Personnel otaff cooperated enthus^leetl* 
cully and submitted detailed specifications which were fully utilised h 
Commission. In late September the new series for Medical Teehnolog 
(professional grade) wae published. This recognition of the professional 
level of qualified Medical Technologists and their improved opportunities 
for advaaceweat in gcvernaent service should improve recruitment. Bet; 
tlon descriptions for this series have been written and subaitted for 
approval. Within the next few weeks the qualified technologists of our 
staff will be reassigned to the new positions* 


Automation and Specific Test Xaprovencntq 

During this year, further improvements , streamlining and shortening 
of test procedures were accomplished. In 1956 the average cost per tact 
itea was $1.61. This was reduced to $1.57 in 1957 (131 decrease). On. 
the first half of 1958 the coot dropped to $1.47, a further economy of 
(19% belo* 195$). The use of automation is largely responsible 
overall reduction in cost per test. The effect of mechanisation is more 
vivid in the comparison of former and current costs of individual to 

Costs of Individual Tests: Automatic vs. Manual Methods 



C : :Ic"iG3 

$ 0.33 

$ 0.06 





Co 32 


Blood Brea Bitrogea 



Red Blood Count 



vatic vs. 

Manual Method 


Automat i 



15 tain. 

5 min. 

24 hrs. 

8 hrs. 

3*4 tech. 

1 tech 

$ 500 

$ 3,850 

For specific items the former cost is reduced 601 to 901 by automation. 
Intangible, but no less important benefits Include the use of such smaller 
amounts of blood sample for each test, significant reduction of both tech- 
nical and human errors, and prevention of staff fatigue. Working vith 
these machines is much less frustrating than the older manual methods; the 
technologists are therefore, less apt to search for a change and resulting 
personnel turnover is less. Most Important, however, is the fact that these 
automatic Instruments greatly increase individual technologist capacity and 
permit more time for other more stimulating work. On the other hand, the 
test results obtained by these cut emetic methods are as accurate and fre- 
quently more accurate than by conventional methods. An example is the 
comparison of automatic and manual methods of red blood cell counting. 

Blood Cell Counting: Comparison of < 

Accuracy (Standard Error of Mean) 
Time per 1 determination 
Tims per 100 determinations 
Staff required per day 
Initial cost of equipment 
Coat of operation per year 

(751 capacity) $12,000 $ 4,000 

The advantages of automation in the clinical laboratory have stimu- 
lated much Interest among pathologists, hospital administrators, and the 
Instrument industry. Our experiences mere presented In two papers read 
at the Industrial Exhibition and Symposium on Automation in Kern York City 
in June and at the annual meeting of the American Society of Clinical 
Pathologists In Hovember 1958. Both vers received enthusiastically and 
have resulted In many inquiries concerning our methods. 

The Hematology Service has made further Improvements in electronic 
cell counting by the adaptation of the Coulter Counter for enumerating 
leukocytes. This new method has reduced the time needed for a single 
count from 90 seconds to 16 seconds. 100 white blood counts can be 
accomplished now without fatigue and more accurately by 1 technologist 
in 2 hours. Poratsrly this volume required full time of 2% technologists. 
Studies are In progress to modify the electronic counter for platelets. 

By the use of a semi-automatic drop method, a considerable savings 
In time has been achieved In preparing bottles vith anticoagulants for 
blood collection. A comparison study indicates that the method is more 
accurate and faster than individual pipetting. A similar Improvement in 
the time-consuming osmotic fragility tests is being explored. 

In relation to rapid diagnostic aids in hematology, a new microscope 
accessory developed by RCA engineers in their Lancaster Tube Plant was 
evaluated la clinical laboratory use. It is an ultraviolet image converter 
which makes possible direct visualisation of the ultraviolet absorption of 
blood and marrow cells on a phosphorescent screen without the interposition 
of elaborate television instrumentation. This small tube is attached to 
the microscope body tube in place of the ocular and on looking Into its 
eye piece the image may be examined. Ve found the resolution satisfactory, 

camera for Loedlate photography. This small Instrument obey Id greatly 
facilitate InmoHirm examination of fresh marrow apecimoas for W absorption 
Information to help differentiate early forms of hemoglobin containing cells, 
the high protein concentration in abnormal plasma cells, etc. Other velu- 
cble uses undoubtedly will be developed when this accessory become* general- 
ly available. 

The Clinical Chemistry Service has Introduced a serum tartrate-inhlbitabj 
(prostatic) acid phosphatase procedure* The serum salicylate procedure and 
the determination of cerebrospinal fluid protein have been simplified and 
shortened. A more rapid semi micro method for measuring serum CO2 content 
using 0.2 ml sample was developed and a spectrophotometry serum transaminase 
method was modified to permit testing groups of specimens. This can be done 
In about half of the time formerly required. 

Our Assistant Biochemist In Clinical Chemistry developed a direct readiu; 
clock on the electric chlorldlmeter by which we can now read results in mil 11 
equivalents of chloride, thoreby eliminating the calculation step. This 
saves time and avoids transcription and calculation errors. Be also has de- 
signed and tested mechanised pipettes which speed up several operations and 
Increase accuracy. 

The Chief of Control Chemistry and his assistant have modified and 
evaluated several procedures on the Autoanalyser which widens the scope of 
this automatic Instrument in our laboratory. 

In the Microbiology Service a number of minor improvements have been 
Instituted including elimination of most of the contamination of blood 
plates, utilisation of a new medium for TB cultures to enhance growth and 
virulence testing, and the time saving use of antibiotic disc dispensers. 
Emphasis has been placed on quality control measures end safety In handling 
pathogens. Mr. Zlerdt completed his studies on lyophllisatlon of stock 
cultures, particularly staphylococcal phages for minimising deterioration 
of titers. His contribution makes It possible for one bacteriologist to do 
In part time the routine phage typing which formerly required two full time 
staff. His excellent paper describing this unique method has been accepted 
for publication In the American Journal of Clinical Pathology. Survey work 
in hospital staphylococcal infections has continued. A new technic for the 
laboratory diagnosis of fungus diseases that promises to be more reliable 
end accurate than the previously available methods Is being evaluated. The 
senior resident in Microbiology has developed a simple and rapid presumptive 
test for antibiotic sensitivity which will produce results in two to four 
hours as compared to the two or three days needed at present. If evaluation 
substantiates preliminary results, this test promises to greatly enhance 
and speed up Institution of antibiotic treatment. 

The Clinical Center la one of the major centers of medical research 
in the world and because laboratory data furnished by our department con- 
stitutes an integral part of each research patient study, we maintain a 
Control Chemistry Service. Accuracy and reproducibility of test methods 
In Clinical Chemistry, Hematology and Special Procedures Services are 
constantly checked to maintain the high standards necessary for research 

instruments, or modifies existing ewthods or equipment to increase their 
efficiency or precision. ?or example, the available methods for the de- 
termination of serum magnesium ere inadequate because their accuracy and 
precision are poor. The growing demand for eerua magnesium analyses for 
the diagnosis and treatment of certain diseases has stimulated us to 
critically evaluate the existing methods and to develop an accurate and 
reliable method. It hao been noted that the concentration of serum 
inorganic phosphate determined by dialysis does not agrse with the results 
obtained by the usual deproteinlEatlon procedures. The reasons for this 
difference are being investigated. The dlalyaable phosphate of the blood 
may be an Important constituent. 


This department is accumulating a unique mess of detailed accurate 
scientific data on a large group of research patients whose diseases and 
response to therapy are veil documented. However, the customary methods 
of recording and storing this clinical laboratory information do not per- 
mit correlative or statistical study of this valuable material. This is a 
rare opportunity to develop e pretties! and efficient method for recording, 
storing and retrieving this mass of information for studies in diagnostic 
corrections, individual clinical pathologic patterns, etc. A specifically 
developed data processing system, particularly adapted for the purpose of 
utilizing this Information is urgent. The available IBM cord system has 
been tried and found unsuitable. Various other technics are being explored t 
and the most efficient and practical should be suitably modified and in- 
stituted. Such a data processing system would constitute en invaluable 
contribution to clinical pathology. 

Examples of Spoclal Contributions to Patient Care 

Prompt and experienced application of the medical science "knowhov" 
of this Department's services hes provided Important and sometimes life 
saving assistance to Clinical Investlgatora with chemotherapeutlc, radio- 
therapeutic and surgical problems. 

Following heart surgery a patient suffered sudden and serious elevation 
of blood potassium. Our Clinical Chemistry staff measured serum potassium 
every four hours for 24 hours to maintain close watch on response to ex- 
change res in therapy. 

The staff of Clinical Chemistry contributed significantly to the 
elucidation of abnormal carbohydrate metabolism in a patient with an obscure 
muscular defect. Determination of blood sugar by the specific glucose oxi- 
dase technic indicated that glucose levels in this patient ware normal and 
established the validity of the normal glucagon test. Special paper chroma- 
tographic differentiation of galactose in the urine of twin Infants contri- 
buted valuable information concerning the possibility of galactosemia and 
we have established semi micro methods for performing galactose tolerance 
tests on infants and children. 

I 1 

nutrii:.'.oa on acids la urine, Or. Frame established ths procedure and 
demonstrated significant differences between states of relapse and rescission 
of patients with nephrotic syndrome. A marked amino aciduria was found in a 
young boy suffering an undiagnosed illness. Its significance is uncertain, 
but further tests are being done. 

The medical profession has been alerted world wide to the gravity of 
hospital epidemics, particularly coagulase positive staphylococcal infect- 
ions. Our service of Microbiology has done some of the pioneering survey 
work in comprehensive phage typing of these Infections comparing the strains 
found in the Clinical Center with those from other hospitals of the middle 
Bestam United States. All coagulase positive staphylococcal isolations 
from Clinical Center patients are phage typed and their origin spot-mapped 
to detect early any pre -epidemic focus. This has permitted prompt and 
appropriate preventive measures, and a true hospital epidemic has been avoid- 
ed thus far in our hospital. 

Of particular importance to operating room sterile security is the 
development of a strain of aerobic spore forming bacilli for testing thermal 
sterility of autoclaved supplies. Ths organism Is continuously propogated 
and used to spot check autoclave efficiency, particularly after shut down 
periods for repairs. 

Our Hematology Service Chief and his staff provide frequent clinical 
consultation concerning hematological problems at the request of many 
patient care physicians of several Institutes. Their unique method of 
sequential recording of hematologic data on Clinical Center patients provides 
the opportunity to spot minor or incidental abnormalities of no Immediate 
significance to the patient and of no Interest to the attending physician, 
but of Immediate concern to research interests of othere in the Clinical 
Center. For example, we called attention to three cases of e particular 
syndrome of refractory enemla and they now are being studied Intensively. 
Hew knowledge of basic regulation of red cell production is gained •» a 
direct result of cur "clearing house" function in coordinating the overall 
laboratory patterns of Clinical Center patients. 


The entire staff of this department, and more particularly the chief 
technologists and their assistant supervisory technologists of the four 
services have consistently performed their functions well beyond the call 
of duty and under discouraging circumstances of personnel shortages and 
consistent overload. They have persisted in maintaining a high quality of 
work despite long hours of necessary overtime. Many have displayed devotion 
to our mission of service to the research patient and manifested individual 
effort and initiative In Improving procedures, efficiency and developing 
better methods. Group awards have been recommended for outstanding and 
superior performance end individuals have been nominated for awards for 
specific contributions in methods and instrumentation from which the depart- 
ment is deriving considerable tangible benefits as well as significant 
though Intangible advantages. 



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as as 

as as 

Calendar Year 1958 




Serial No, 

The Rehabilitation Department, during calendar 
year 195#, continued as in previous years to carry out 
its primary function, viz., the treatment of patients 
referred by Institute physicians, usin<" r , recognized 
measures and techniques of physical, occupational,, 
and speech therapy. 

As previously reported, the Department's 
Physical Therapy Service provides the following: 

lo Tests and measurements 

a. Manual muscle evaluation. 

b. Joint ran^e of motion measurements., 

c. Girth, length and expansion measurements 
(linear, volumetric ) . 

d. Electrodia^nostic test with constant current 
impulse stimulator to determine chronaxia 
and strength-duration curve „ 

e. Faradic-^alvanic test for nerve or muscle 

fo Self-care evaluation (Activities of Daily 

Living) . 
'^. Skin temperature and oscillometric tests c 
h. Vital capacity determination with Collins 

Spirometer or portable McKesson. 
2. Heat, in various forms including 

a Q Superficial - hot packs (Hydrocollator) 9 

infrared, paraffin bath, whirlpool. 

b. Deep - short wave diathermy, microwave 8 

Co General body heat - Hubbard tank. Moistaire 

cabinet o 
3o Therapeutic exercises 

a. General exercise ~ passive, active assistive 9 

active and resistiveo 
bo Muscle re-education, usin<2 Warm Springs 

techniques, mass movement patterns and 

proprioceptive facilitation techniques o 

c. Ambulation training, including elevation 9 
with and without parallel bars 8 braces 8 
crutches, canes 

d. Posture exercises 

Rehabilitation Department 

e. Underwater exercises (Hubbard tank, 
therapeutic pool). 

f. Progressive relaxation. 

go Pre- and postoperative thoracic surgery 

program of breathing and coughing and range 
of motion exercises. 

h. Pre-prosthetic and prosthetic training. 

i. Training in self-care activities 

j. Exercise with apparatus such as weights and 
pulleys, shoulder wheel, finger ladder, 
quadriceps boot, stationary bicycle, 
restorator, and Elgin exercise table o 

k. Breathing exercises and postural drainage 
4o Miscellaneous 

a. General and local application of ultra-violet 

bo Contrast baths. 

c, Scientific massage* 

d. Neck traction (3ayre) e 
e Q Compression bandaging, 

f. Application of splints and casts to maintain 

joint in good anatomical and functional 

go Instruction to patients and family in home 

care program (application of heat, exerci 

use of self-care aids), 
h. Fitting and dispensing canes and crutches., 
i. Prescription -and procurement of corrective 

shoes, braces, corsets, splints 8 


Services in Occupational Therapy include: 

1. Maintaining or increasing joint range of motion 
and muscle strength; improving coordination; and 
developing endurance and work tolerance, 
through utilization of appropriate technics, 

2. Testing and training patients to aid them to 
become self-sufficient in eating, dressing^ 
horaemakin^ and the other significant activities 
of daily living. 

3. Exploring patient's skills, aptitudes and work 
habits in order to provide basic data for 
vocational planning. 

4. Observing and recording patient behavior patterns 
for use as diagnostic aids or in evaluating the 
patient's reactions in specific research studies 


Rehabilitation Department 

5o Substituting constructive interests for the 

insecurity ana anxiety which may develop during 
the research study 

6 n Providing normal developmental learning experiences 
for children. 

The Speech Therapy Service provides: 

lo Speech lan^ua^e and audiometric evaluation. 
2„ Stimulation and education or re-education of 

lan^ua-^e and lan*ua T ,e concepts , 
3» Exercises for improved control and coordination 

of the speech musculature. 
4° Breathing exercises. 
5o auditory training. 

6. Articulation exercises. 

7. Voice exerciseso 

80 Teaching of post-laryn^ectorny speech. 

9o Observing and recording patient behavior patterns 

for use as diagnostic aids in specific research 

10o Counselling patients' families with regard to 

speech problems and home procedures following 

discharge, and providing information re^ardin? 

availability of speech therapy services in the 

home vicinity 

Despite the annual turn over of clinical associates, 
we were happy to note a continuance of the generally 
increased awareness on the part of Institute physicians 
with respect to the functions of the Rehabilitation 
Department and the assistance which we can render inpatient 
care. Some of this may be derived through our participation 
in in-service training courses for members of the nursing 
staff and social service workers o 

The acquisition of new equipment has 9 as mi?;ht be 
anticipated „ slowed down. An additional mat table has been 
purchased for the Physical Therapy area 8 and an additional 
improved loom was acquired by the Occupational Therapy 
Service. The Physical Therapy Service also has on loan 
the new Circ-O-lectric Bed. This bed functions as a tilt 
table ;, with many variations of reclining, sitting „ and 
vertical postures and includes the advantages of the 
Stryker frame 9 enabling patient to be turned from prone 
to supine position and vice versa e 


Rehabilitation Department 

A tape recorder which has ^ood fidelity and 
is easily portable is essential to the work of the speech 
therapist. Such recorder v/ill aid in initial evaluation, 
provide periodic record of speech for comparative 
purposes, and also enable patient to hear and evaluate 
his own speech more objectively and to follow his progress o 

An audiometer is also essential for an adequate 
and complete evaluation of speech problems. Not infrequent! 
hearing problems exist in these patients, either concomitantly 
with speech problems or as the primary or contributing 
cause. Presently, a hearing loss may remain undetected in 
general observation, because of a patient's ability to 
compensate (through lip-reading, and so on), but with 
remaining inability to monitor his own speech, hence 
either contributing to or causing the speech problem,, 
(In particular, these "subtle" hearing losses frequently 
exist in patients with aphasia, cleft palate, or 
laryngectomies. ) 

The program of application of plaster casts for 
rheumatoid arthritic patients, described in last year's 
annual report , has fulfilled all expectations, proving 
of substantial value in the treatment of these patients 
'.Ve had not anticipated, however, its increasing use 
and value in a wide variety of other disease categories 
where limitations in joint ranf^e seriously impair normal 
function of patients » Thus, we have applied casts to 
patients with hemophilia, Friedreich's ataxia, infantile 
myopathy, salmonella infection, osteoarthritis, Guillain- 
Barre syndrome, periarteritis nodosa., Parkinsonism;, 
dystonia musculorum deformans and muscular dystrophy. 
Curing 195#, a total of 167 casts or splints was applied 
to 77 patients o 

The relationship of recreational activities to 
the Rehabilitation Department has to date not been 
clearly defined a One important recreational area p 
viz 08 Arts and Crafts, has been returned to the 
Rehabilitation Department Because of the nature of 
this activity, it has been assi<5ned to the Occupational 
Therapy Service with space in the O.T. area set aside 
and services rendered by the Chief 8 Occupational Therapy 
Service,, with assistance in the conduct of the program 
from the Gray Ladies We believe, however, that a 
program of this size and importance requires full-time 
professional services, and we have recommended that 
an occupational therapist whose skills and interests 


Rehabilitation Department 

lie in this direction be placed in charge of the 

program. Such a person has been employed and is 

expected to come on duty during the month of January, 

Other recreational activities (movies, gymnasium, 
patient library, etc.) are in the charge of and 
supervised entirely by the Patient Activity Section. 
The latter is not a part of the Rehabilitation Depart- 
ment. A Children's Program is provided by the 
Occupational Therapy Service in its own area. However^ 
the Patient Activity Section is also engaged in a 
variety of activities for children. Relationships 
here have not been clarifiedo 

The reduction in staff of the Occupational 
Therapy Service (from 11 to #) has resulted in a 
difficult situation in respect to covering patients 
on the units who are unable to come to the Occupational 
Therapy area but require occupational therapy services.. 
In an attempt to provide coverage, the Chief of the 
Occupational Therapy Service has been treating patients 
on the units and a therapist assigned to the 
occupational therapy mental health program has also 
been giving part-time services to unit patients* 
The latter will no longer be possible because of 
increased demands in the mental health program,. 
The Chief of the Service cannot possibly take more 
time from her primary and essential supervisory and 
administrative duties to see individual unit patients 
It is 3 therefore^ recommended that an additional 
occupational therapist be brought on duty to devote 
her time and services for the benefit of patients v/ho 
are confined to the units-. 

Another physiatrist was added to the staff of 
the Rehabilitation Department on June 30, 195# 
Dr. Tillye Cornman was at one time the Acting Chief 
of the Rehabilitation Department and more recently a 
clinical associate in the NINDB C Dr. Cornman' s 
transfer to the Rehabilitation Department as a 
full-time medical officer has made it possible for 
the Chief of the Department to devote more time to 
administrative problems and also assures continuous 
coverage for the Department in the absence of the 
Chief during meetings „ vacations 8 etc Q 


Rehabilitation Department 

We again urge, as we did in last year's annual 
report ? that five beds be provided for Clinical Center 
patients ready for discharge by the Institutes by 
reason of the completion of the research study, but 
who are still in need of continued rehabilitation 

We regret to report the loss durin? the past year 
of three modules which were turned over to the 
Pathology Department. We now have a serious problem 
in respect to the need for space . The speech therapist 
has no treatment space whatever and has been required 
to borrow such space from day to day and from hour to 
hour as it :aay be available. For the past few months^ 
she has had part-time use of the office of Dr. Cornraan, 
Assistant Chief of the Department. Since the office 
of Dr. Cornman is in reality our room for testing 
and training patients in activities of daily living, 
the entire situation is manifestly a decidedly 
unsatisfactory one» We have in the past proposed that 
the locker room now being used by our female physical 
therapists be converted into a speech therapy area 
It will still be useful, with some simple screening, 
as a locker room before 8:30 A.M. and after 5:00 P'.M 
A small expenditure of money can thereby provide 
additional and essential treatment and office space Q 

During calendar year 1957, Miss Helen Applebaum 9 
staff occupational therapist s attended Miss Margaret Rood v s 
course on "Neurophysiology in the Treatment of Neuro- 
muscular Dysfunction" This year Mr. Otho Bell 9 staff 
physical therapist, took the same course a They are now 
using thse new technics with what we consider to be 
favorable results We believe that we should continue 
to provide training for our staff members in the newer 
technics of rehabilitation medicine as they are 
developed for the benefit of Clinical Center patients „ 

Annexed hereto are statistical reports for 
Physical Therapy Service, Occupational Therapy Service^ 
and Speech Therapy Service c 

Calendar Year 195© 





Serial No. 

The servicee ccaprlsing the Ifcanaacy Department, Pharmacy Service, 
Central Sterile Supply Service, Phamaeeutical Development Service and 
an unofficial Radiopharmaceutical Service, nave continued to expand end 
provide the highest quality pharmaceuticals available within the 
limitations of facilities available. 

She Pharmacy Sarrice has continued to improve the routine 
pharmaceutical service available in several -nays. Sterile ophthalmic 
solutions are being dispensed routinely in $ ml. squeeze dropper bottles. 
Ibis mas introduced after approximately eight to nine months' packaging 
research by the Department. Improved packaging, following additional 
packaging research in the Department, has provided for control numbers 
on all Inpatient medications and outpatient prescriptions. 

Drugs in ointment form are being dispensed in tubes, thereby 
replacing the glass Jars. Personnel time Involved is unchanged, 
however, it is felt that the end user benefits greatly. Improved 
packaging is being provided to the Employee Health Service. As the 
demand for this service increases from the Employee Health Service, 
adjustments in the packaging service may have %o be made. 

The microfilming of prescriptions is continuing and, thereby, ve 
are reducing the storage space required for older prescriptions to 
approxitsately a l/2?th. 

There continues to be an increase in demand for service in 
prescriptions and total units dispensed. Some of this has been absorbed 
by purchasing suitable commercial packages vfeen the comparative costs 
are Justified. Other products are being switched to oompounding in the 
Clinical Center i&tore coats Justified savings. Many products "Hhich ■were 
previously prepared by the Phenaaceutical Development Service are new 
being supplied from the Fbaraacy Service in order to free professional 
men for true developmental service. 

Changes have again been made in the use of non-professional 
personnel in order to further extend their usefulness and increase the 
output of supplies. 

Sees of the major problems in the Pharmacy Service are: 

1. A 25# increase in prescriptions resulting in a real need for 
restyling the outpatient prescription area. 

- 1 

2. Insufficient information Is received in a great number of 
proscriptions requiring daisy for the patients and unnecessary 
telephone calls, thereby delaying the physicians and the 
pharmacists in completing their assigned duties. Size of 
prescriptions, both in cost and total supply, is still 

3. Prior plans or arrangements are rarely node with the 
Pharmacy vhen as unusual amount of a drug is required. The 
Department requests that serious consideration be given to 
Including the name of the aedication on all outpatient 
prescriptions, the exception being where the physician requests 
that it not be included. This leaves the perogative vith each 
individual prescrlber and would reduce a dangerous situation 
wherein the patient reeelvee nuserous pre s cr i ptions from the 
Pharmacy, vith only "Instructions for use" and prescription 

h, The need for sore experienced pharmacy officers has become 
critical during the past nine to twelve months. Every 
attempt will be made to replace vacancies vith older officers 
rather than vith the young, inexperienced pharmacists. 

The areas of iiiproveaant to be stressed in the Pharmacy Service 
are to commence by rearrangement of the physical facilities to improve 
working efficiency, and to continue to carry on service research. 


The Central Sterile Supply Service has continued to offer service 
and supplies within the limitation of facilities and personnel. It is 
to be noted that there was a considerable Increase in units issued during 
the 1958 Fiscal Year and an even greater Increase during the 1953 
Calendar Tear. 

The Service is continuing to evaluate new types of packaging 
techniques and materials in order to increase the amount of supplies 
which nay he processed. With improved disposable materials becoming 
available a shift is being made to purchased materials thereby releasing 
personnel for other services, which were previously not available to the 
requisitlonsrs. This does furnish the users with more units of materials, 
but it also increases the overall supply costs. When more automation 
t^pe of equipment is available, the number of units can be increased and 
eooa of the presently purchased materials nay again be packaged in the 
Departmsnt to partly offsot the cost of this increased service. 

The co<splc:cIty of some trays and items issued continues to be a 
problem. A real nesd exists for simplification and standardisation of 

- 2 - 

many troys. A means of accomplishing this has been very difficult and 
it is hoped that procedures nay he forthcoming toward this end. 

ProbleiES exist in packaging and sterilizing of eoa» of the newer 
supplies used,, thereby necessitating the purchasing of same units or 
furnishing materials \&Ach are not ideal. 

Personnel of the CSSS hare cooperated actively in aany highly 
specialized projects conducted by clinical investigators by asking 
available "special handling" supplies on an almost routine basis. The 
aoccmpliehaanta and problems listed in the 1957 report continue. A stare 
acute need nov exists for aore automation in packaging and for some type 
of gas sterilisation service. 

The Department has been offering Radlopharaaceutlcal service to 
various investigators. Since this service has not been officially 
recognised, there la no definitive policy concerning the role of the 
Departaent in this area. This has caused sens problems In producing 
radioactive dosage forms. A definite need exists for a policy in this 
area, along vita eaple, suitable space for use, depending upon the 
extent to shlcb service Is to be offered. Confusion exists as to the 
extent the Deportment any offer service and to idiat extent the Radiation 
Safety Office or NCI personnel are responsible for sons of these services. 

The Radiopharmaceutical Service has provided a superior service, 
i&ich voa not previously available at the CC or HIE - and in a few cases 
service Vhich vas not available elae*toere. Our personnel have provided 
consultation for the clinical and laboratory staffs concerning proper 
radioisotope preparation for use in the patient areas. In several 
instances, treaendous cost savings for the NXH have been exhibited by 
the purification and preparation of radioisotope products for injectable 


The Pharmaceutical Development Service appears to have Issued a 
lesser curiber of items than In the previous year. Early in the year 
requests for service idiich required less than one hour of professional 
tlae to prepare vers shifted to the Pharmacy Service. Projects 
requiriEg between one to six hours tsare encouraged to be shifted, 
ifterever possible, so as to free the PDS personnel for developmental verk 
rather than routine service. This has somewhat reduced the total 
auaber of units issued, but has resulted in aore service to the 
invest! •Ts.'ior?; . 

In September, 1958 the Investigational drug policy was Issued by 
Dr. Masur. The responsibility for registration, proper labeling, and 
packaging -were assigned to the PDS. After being properly registered, 

- 3 

packaged cxsd labeled these drags are Issued by the Pharmacy Service 
through their usual distribution channels* 

With chances in the Division of Biologies Standards personnel, 
there was seme difficulty in obtaining pyrogen end sterility tests for 
our injectable products. For acme weeks, a pharmacist from the PDS 
assisted in tests in order to alleviate the situation caused by a 
backlog of tests. 

Routine sterility testing of Central Sterile Supply Service 
products and the Pharmacy Service sterile ophthalsdc solutions have been 
carried on by the PD3. CDs of the pharaaeists from PD3 has been attending 
the weekly Drug Steering Committee meetings cf BCI. Ibis has resulted 
In increased co op e ra tion, understanding cf autual problems, a pboraaclst 
assisting in better planning by furnishing inferential before the final 
protocols are established. It is felt that by attending these committee 
noetlngs the PD and BCI will co o per at e better in all phases of service. 

Several examples follow of products which were furnished to the 
investigator by industry in a ebeodeal fom rather than in an injectable 

a. D. Searle and Company was unable to supply a satisfactory 
injectable product of SC-8109. Personnel of the PDS reerystolllsed the 
notorial into a mlcroc r y stall ine chemical, prepared the product satis- 
factorily for injection and lypholised the prepared vials for stability. 
She product prepared was fry satisfactory and after seme weeks of use, 
the representatives from Ssarls asked if the formulation Information 
could be given to them. 

The identical situation existed with Testolsctoae Suspension for 
Injection and on anti -carcinogen product, known as A-139* 

Several Investigational products prepared by this service are being 
shipped to c oo p erat ing hospitals in Cancer Research. Products ere 
prepared, bottled and packaged according to a randomisation schedule end 
shipped to cooperating hospitals. The coop e rating hospitals also ship 
materials to the Clinical Canter for use in the patient area. 

Recruitment and retention of trained personnel In this area is 
wry difficult. A person with a proper background, training and 
experience is soon offered a higher salary in the some type of activity 
in another area. Trained personnel can receive a such higher salary 
in industry and pharmacy colleges. Consequently, we can retain only 
one officer by raeaaa of a graded position and must rely upon untrained 
personnel to assist in the staff positions. This lessens the potential 
usefulness of this service. 

Suitable equipment for certain types of pr epa ra tions is lacking 
and, in addition, the space in which to place this equipment is lacking. 

„ k „ 

ami art 

¥te area of greatest need is a separate roan or cubicle is ubich to 
prepare pharaoceutical products such as anti-fcHc acid products, 
estrogens, endroge&s end some Bustards. There is a pressing need for 
a cubicle type of space Vhlch is Tented under negative pressure so as 
not to spread these products over the entire area. 

In cooperation with the Cancer Chemotherapy national Service 
Center a -valuable service has been offered to the Clin ical Center and 
many other hospitals cooperating in research. A pharmacist employed by 
CCHSC works In our area under the supervision of the Chief of PDS to 
prepare products for the CCBSC. There is continuing pressure and 
ilmanil to increase this service, but we are Halted by personnel, 
equipment and space in vbich to place such equipment, or allow personnel 
to function. 

The p ro cu rem e nt of supplies has becone a acre acute problea than 
vas reported in the 1957 Annual Report. The availability of suitable 
supplies when necessary for our requisltioners and users has now reached 
a critical stage due to proniTewant, policies and storage facilities. 

The Departaent has had a problea with trainees froa the Division 
of International Health, which was reported to the Inforsation Officer 
of the CC. Iap r op o x advance planning for trainees has resulted in 
great inconvenience to the Departaent, end in soaswhat lass than an 
ideal training situation for the individuals concerned. It is hoped 
that this problea can be elladnated. If not, it will bo necessary to 
refuse trainees the opportunity due to inability of the Departaent to 
deny service to the patient area in order to acooaplish the desires of 
the Division of International Health. 

The Departaent baa great need for relocating soae of the physical 
facilities, thereby iaprovlng efficiency, adding new services, and 
bringing certain services up to the level at which we should be operating. 
However, all of these have had to be postponed pending the outocae of the 
Iharaacy Departaent survey, which is now overdue for ocapletion. It is 
hoped that this nay be ~"tp*«*'»* so that the Departaent say proceed with 
i apr ov e nents, which we feel are necessary. 

- 5 - 

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i« gg IM 3 

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8 $ 










to *r* 






SI ! SM 



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• 2* 









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s 1* 































































Milton Wo Skolaut 

"Hov the Pharmacist Can Help Solve Problems in Purchasing Medical 
Supplies", HOSPITALS, Vol. 32, Hov. 1, 1958. 

Speech at Maryland -D-C -Delaware Hospital Association, Pharmacy Section, 
"Labeling and Its Responsibilities", Nov., 1953- 

Speech at AAAS Meeting, Pharmacy Section, "Impact on the Hospital 
Pharmacy Program" (of the CCHSC Program), Dec., 1958. 

Chairman, Research and Development Committee, American Society of Hospital 
Pharmacists, completed three year term. 

John A. Sclgllano 

"Studies on a Bacitracin Peridental Dressing, " Co-authored vith 
Paul R. Baer, D.D.S. and Henry M. Goldman, D.H.D., ORAL SURGERY, 
11:712-720, July, 1958. 

William H. Brlner 

"Certain Aspects of Radiological Health, " THE AMERICAN JOURNAL OP HOSPITAL 
PHARMACY, Vol. 15, No. 1, Jan., 1958. 

Speech at Maryland State Nurses Association Meeting, Rockrille, Maryland, 
"The Nurses' Role in the Use of Radioisotopes." 

Speech to Surgical Nursing Service, Clinical Center, "Certain Aspects of 
Radioisotopes " . 

Training at Oak Ridge Institute of Nuclear Studies in Radioisotope 
Techniques, May-June, 1958. 

Samuel Merrill 

Training by Taft Engineering Center Staff at B.I.H., "Occupational 
Radiation Protection", Dec., 1958. 

Numerous visiting pharmacists vere oriented in certain phases of 
the Department. These were hospital pharmacists, hospital pharmacy 
internes and foreign hospital pharmacists. 

. 8 - 

i* OP >. .JBitM 


SailMtf &AAA 

■•■ ' 

LJMff * ft 

•vxutl otf?" 

Various staff taenbers have continued to assist the Nursing 
Department In the orientation of new nursing personnel. 






The Med delegated the major 

onsibillty o ag the completeness, accuracy, safe- 

1 records by maintaining the 
• -nal controls; tl 1 of 

medical I . at the . Center 

;r and accrediting agencii squired 

discharged patients and the 
diseases and operation) Standard Nomenclatur 

nnd Operatlom:) \rlth a view toward providing adequ 
data to physicians participating esearch; 

maintenance of a clinic a; b projects index for i 
etion end. die administrative , 

■enance, compilation and 
distribution of statistical data, concerning patient care, for 
zation by administrative, clinical and research directors; 
maintenance at of medical records to 

,te accessibility 3es of patient care 
ch; (6) of a system whereby 

ty and quality of dictated mi :?.ord reports are 
7) maintenance of an edequate control system with 
o.'d to the release of medical repc: nutation and 

;al records to authorized personnel; (8; maintenance of a 
letual patients' index (Soundex) and patient registry. 


The foregoing basic internal controls are adequate and 
d the administration of a medical record program; 
because of the peculiar organization of this institution., 
controls require constant evaluation and revision and a variety 
linimun standards in order to meet the needs of the respective 
Institutes. To properly execute basic controls, S"^andard operative 
lures were instituted and developed; namely, (l) for the 
; purpose of reducing the increasing number of incomplete 

ohe sped jf screening medicel 

joj-ds to 
e of same to authorized i for the 

of dictated 
:.sion in the medical 

In order to meet the minimum requirements of the hospital and 

L research, t! i ant 
development and supervision of record end report eyotems io essential; 
progress or improvements in accomplishing asld responsibility w^re 
achieved by: 

1. Transfer of the addressograph equipment and the 
rasponsibility for the verification of admission 
forms to the Admissions Unit. 

2. Development of new format for recording of information 
to the patient index card (Soundex - perpetual file). 

3- Establishment and maintenance of "house file" for 
current inpatients (incorporated with the perpetual 
patient index file) . 

k. Revision of the Patient Registry (permanent file) and 
procedure for issuing unit numbers and delivery of 
medical records to nursing units for readmiflslons. 

5. Completion of the installation of Terminal Digit 
System of filing medical records. 

6. Purchase of multigraph machine and establishment of 
procedure for the preparation of medicel reports 
(history and physical examination, narrative suranary, 
operation) and Daily Census, Admission and Discharge 
Report on multilith masters and duplication of same 

on raultigr&ph machine. The avex-ege daily word production, 
resulting from use of multilith mastere, increased from 
35CO to 56OO. All backlog dictation received through 
December 31> 1958 has been completed. 

7- Development of a system for maintaining and filing 
multilith masters and duplicate copies of medical 

sal Record Department Page 3 

8. Revision of the Discharge Analysis Registry. 

9. Development of the index of diseases and operations; 
posting and coding to the disease and operation index 
cases up to August 1955 • 

10. Revision of the Death Registry. 

11. Elimination of the backlog of required statistical 

12. Completion of an index consisting of diagnoses, 
diagnostic and operative procedures and their respective 
codes for National Heart Institute, Surgery Branch. 

13. Preparation, in draft form, of the Daily Census, Admission 
and Discharge Record by the Administrative Officer of 

the Day. 


Intradepartmental problems which are major and critical to 
Hie fulfillment of the functions and responsibilities of the 
.^dical Record Department are: 

A. Personnel 

1. Difficulty encountered in recruiting qualified 
medical record librarians. 

2. Lack of sufficient number of qualified medical 
record librarians. 

3. Loss of personnel in clerical positions due to low 
grade classification. 

U. Lack of action and cooperation, on part of personnel 
section to alleviate and correct personnel problems. 

Bo Backlog of unindexed medical records. 

C. Lack of adequately trained and experienced multigraph 
operators (two messengers detailed to operate multigraph 

Medical Record Department Page h 

D. Incomplete medical records (approximately 500 as of 
December 31, 1958). 

Extra-departmental problems have existed previous to this 
year and will continue to plague the department in the future. 

A. Generally, there is a failure on the part of the Clinical 
Center personnel participating in patient care to 

accept their respective responsibilities for the completion 
and accuracy of the following medical reports contained 
in the medical record: 

1. Consultation Report 

2. Operation Report 
3- Outpatient Notes 
h. Progress Notes 

5 . Doctors ' Orders 

6. Nurses' Notes 

7. Sunmary Sheet 

8. Authorization for Operation 

9- Release of Responsibility for Discharge of a 

10. Death Certificate, Report of Death, pronunciation 
of death 

11. Discharge Note 

B. Care/Disposition of the medical record - as of this date 
there are eight (8) lost medical records. 

1. Failure on the part of the nursing department 
personnel to forward all component parts of the 
medical record to the Medical Record Department 
at time of discharge. 

:al Record Department Page 5 

2. Failure, on -the part of the original recipient of 
said record, to notify the Medical Record Department 
at the time said record is transferred to another 
person and/or area. 

3. Removal of medical record from Building 10; locking 
medical records in files, desks, etc.; burying 
medical records in boxes, papers, etc., particularly 
in the laboratory areas. 

C. Necessity of verifying the name and address of referring 
physician, at the time the narrative summary and/or medical 
reports are released, due to failure on part of supervising 
institute to notify appropriate departments of changes 
and/or corrections. 

D. Add-Ons - In comparison to scheduled visits, the add-on 
requests represent a small percentage of the patients seen 
in Admissions and Follovup Department, and the Medical 
Record Department must give special consideration to 
said requests in order to locate, process and immediately 
deliver the medical record. 

•artment Page 6 


Recruitment of three medical record librarians to fill the 
following needs: Supervising Medical Record Transcribing Unit; 
Supervising Medical Record Files Unit and the third to devote 
full time to the coding and indexing of diseases and operations 
in order to eliminate the existing backlog of unindexed cases. 

Review and audit, in cooperation with Personnel Section, all 
positions in the department. 

Transferring information now contained in three separate 
patient registration books into one revised permanent Patient 

Initiation of a new order of filing medical record contents in 
order to facilitate location and filing of medical reports. 

Purchase and installation of a mechanical rotary file in which 
the patient index cards will be maintained. 

Purchase and installation of unitized office furniture in all 
sections in order to efficiently utilize the limited amount 
of space available. 

Establishment of a skeleton medical record staff on weekends 
in order to provide a continuing service on a limited basis. 

cord Department 


.oration, Calendar Year 

Page 7 





First Registration 





patient (First Registration) 












5. Employee Registration 



f »nt Care Statistics, Calendar Year 

. ^r of Admissions 

>er of Discharges (includes deaths) 

2nt Days 

Patient Days 

Bed Days 
rage Number of Patients 

cigth of Stay 
;e of Bed Occupancy 

ar of Deaths 
bar of Autopsies 
























































: Department Fage 8 
harged Inpatients by Institute and Georgraphic Origin, Calendar Year 1958 
























l Central 


























1 Central 

























1. S. 
















fliington, D.C. 146 


rginia 171 

La are included in reported figures under South. Atlantic Region 





















>rd Deparv 

Page 9 

^patient visits, Calendar Year 1958 







Janu2ry - Mexch 






•• June 







•- September 

262 " 






jer - Decembe 













21J-5 1 *! 

Calendar Year 1958 



Serial No. 

A. Program Goals 

The goals of the Nursing Department are wide in scope and include 
the provision of a high quality nursing care to patients; providing 
expert assistance to the clinical investigative staffs; orientation of 
personnel to the requirements in a research environment; an inservice 
education program which will help keep personnel alert to the ever 
changing demands of research projects and their responsibilities in 
this regard; a supervisory program which will insure the high quality 
of nursing care required in clinical research; and a nursing research 
program which will assist in improving nursing practice. 

To meet these responsibilities has been a difficult task during 
the past year as we have been operating with fewer personnel and the 
patient load has been somewhat increased over calendar year 1957- 

Miss Jane Wilcox has completed the academic requirements for 
the Doctor of Science degree and has returned to full time duty a 3 
Special Assistant for Nursing Research. She will be responsible for 
giving consultation to the Chiefs, Nursing Services relative to 
nursing research they may wish to undertake, for carrying on projects 

- 2 - 

as chief investigator and coordinating all projects toward attainment 
of our general goal of improvement of nursing services at the Clinical 
Center. The three specific goals in our nursing Research program are: 

1. lb study the particular requirements for nursing services in 
an institution where medical research is the primary goal. 

2. To study administrative practices which facilitate the pro- 
vision of nursing services in such an institution. 

3. To study specific elements of nursing practice In terns of 
their contribution to effective nursing services and nursing 
care to patients. 

B. Develop ment and Trends 

During the past year certain trends have been manifested which 
necessitate soma careful consideration in program planning for 1959/ 

1. The number of infants end children admitted to all services 
has increased and continues to grow. 

2. As more patients are released from in-bed status to follow- 
up status the demands for service in the Admissions and 
Followup Department increase proportionately. In addition, 
an increasing number of research projects are concerned with 
the patient only on an outpatient basis, which contributes 
to the growth of this departaant. 

3. The number of surgical cases continues to increase as does 
the complexity of surgical procedure. This increase has been 
x-efleeted especially on the Heart Cursing Service where 237 

- 3 - 

major operations were performed and where the hours of con- 
tinuous nursing care provided for critically 111 patients or 
patients requiring constant observations have increased 
67 percent* 
it. dare has been a decided emphasis in the clinical investi- 
gation of certain eye conditions with resulting need for 
intensive orientation of nurses to this particular specialty- 
5. As the information related to the Nursing Department of the 
Clinical Center becomes more widespread, the flannndB for 
information and consultative service grow. There is every 
evidence that this kind of demand will continue. 
C. Pro gress Made 

Although soma progress has been made in carrying out the goals 
we have set, we have not been able to accomplish as much as we would 
lilae because of the pressure of our dally work. 

Recruitment of professional nurses has continued; however, we 
still have a serious shortage and the outlook at present is not 
promising. A full time Nurse Recruiter is in the field and all 
written inquiries from nurses ere answered promptly. On December 1, 
1957 ve had ^93 personnel on duty, of which ^jg were professional 
nurses. On December 1, 1958 we have p&J personnel, of which 3J-U are 
professional nurses. *Bte number of unfilled professional nurse posi- 
tions as of Dacemter 1, 1958 is 8j, with only 10 committed. In an 
attempt to meet the needs for professional nurses we have continued 
to employ part time nurses and at present have 38 on the rolls. 

Because c lility to sec nuatoer c 
needed, we increased our jorsr -oe pcaiticns frost 60 -to 76. 
Silo Increase la positions was aade on the Cancer, Heart and Kteurology 
Suraing Services *.jfcere it ens felt acre nonprofessional personnel 
could he safely utilised* 

By tic end of this year all eligible practical nurses OS-3 '---iLl 
have had the addlSioool 5-0 hours of instruction which is required Y. 
they can be considered for promotion to gsade GS-t. 

The position) cf the Assistant (Deputy) Chief, Hursing Departr 
Special. Assistant Per Horsing aesoareh, and Chief, Sur.g»vy Hursing 
Service hove been re-vrltten and re-#radad and the incuribento proa*: 
to the aov grade* 

'iSw Chief , i\ : : aisslons end Follavup Horsing Service has been 
relieved of sons of the administrative functions related to opar&t I 
of the Admissions r.ud Follomip Departasnt through a reaeeign»»r : 
functions to the .Amissions Unit vithln the Adsdnistrative Branch of 
Office of the Director, Clinical Center. She changsover la this re- 
assignaant was a&scqpllshed without too such difficult 

She evslvatltm of tha utilisation of nursing sersottael beguo 
1956 was euagdeted during tills year. Probably the aoat al®alfle»,nt 
outcosaacf this c^lmtioi. wore (l) eoaa assurance that east personnel 
were wor&leg In accordance with their job dssorlptiom as writteaj 
(2) corwbor&tloa of what we suspected that there vm little differ.- 
batmen the functions of the nursing assistant (sale) GS-3 and the 
praetlee& nurse GS;»3j (3) saaa^ for sOarifica&ioa tw& ftvthor In 

m of the functions of tfcft - 

- 5 - 

Based on this evaluation, Increased time and effort has been 
given (l) to developing a supervisory program on the nursing units 
during the evening and night tours of duty; (2) to pointing up train- 
ing needs of nonprofessional personnel and planning programs to meet 
these needs; and (3) to directing staff education programs toward the 
particular needs of the staff nurse OS-7 and the head nurses. 

An intensive care unit of four beds was planned and constructed 
on 10 East and 8 Bast* This has served to make It possible to utilize 
the professional nurses u»re effectively on these two nursing units. 
D. Ma jor Problems 

Some of tho problems were reported last year and are reported 
again this year as they are becoming more acute. Others are directly 
related to changes in program and Increased demands for nursing 

As indicated previously, we have fewer professional nurses on duty 
than at this same time last year. Although our turnover of professional 
nurses does not appear to be excessive, it must be pointed out that at 
no time since the Clinical Center opened have we had all professional 
nurse positions filled. Some dissatisfaction has been expressed by 
professional nurses over the amount of time they spend on evening and 
night duty because of the number of separations and accessions to the 
staff and the amount of orientation needed by new staff members before 
they can assume the responsibilities of these assignments. 

With these continuing staffing shortages, the increasing demands 
for services create cany problems which must be solved. The increasing 

flmwTYiP for oervlces are directly related to (l) the increases In the 
number of surgical procedures both major and minor; (2) the increasing 
pediatric census with the dispersal of children on many nursing units; 
(3) changes in emphasis in the research programs of the seven Insti- 
tutes without adequate planning for how the ueeds for additional 
services will be net; (U) the overall Increase in the number of patients 
who require more nursing care because of the acuity of their Illnesses. 

Another problem is related to lack of space. More playroom space 
for children is needed on the Arthritis and Metabolic Diseases, 
Heurology and Heart Services. Although plans have been cade for a 
parents* waiting room on 2 Bast, the delay In oarrying then out has 
made it difficult for both the nursing staff and the parents who spend 
so much time on this unit. On the Psychiatric Service the utilization 
of patlont and nursing care space for laboratory use and offices for 
the research staff has created problems In the care and management of 
patients on the cursing units. With the reassignment of space in the 
Admissions and Followup Department, there are periods during the day 
when the waiting room in the South wing is congested and problems 
arise in providing space and nursing services on the South wing when 
three Institute groups hold their clinics at the seme time. The 
wrfn-<TMrt amount of office space for the Chief, Nursing Service and her 
staff on each of the six categorical. Services continues to be a 

The number of employees of the Eational Institutes of Health seen 
on weekends and holidays again points out the need for the Employees 

- 7 - 

Health Service to be In operation every day of the year. Although 
much tine io frequently spent by the staff of the Surging Department 
in providing care to employees after 3:00 p. a. each day, this could 
be provided durlrg thin casing year unless there is a Barbed increase 
in the number of employes* needing care after 5:00 p.m. 
E. Accomplishments 

1. Tovard Qoala 

Despite the problorcs associated vith the shortage of professional 
nurse personnel and the lack of space, the quality of nursing care 
has remained at a high level. The supervisory program has demon- 
strated improvement, and the in-service education program continues 
to grow in quality and in line vith the needs of personnel to meet 
their on-the-job obligations. 

2. Publications 

a. "She Role of The Burse in Clinical Research, " Ruth L. Johnson. 

The Yearbook of Madera Hurslng, 1957-58. 

b. "Collecting Continuous Urine Specimens from Infants, " 

Virginia Sllvis. (To be published in Hg). 

c. "An Accurate Method for Measuring Urinary Drainage," 

Isabella DsBella. (To be published in the American Journal 
of Huraing ). 

d. Members of the nursing staff of the Arthritis and Metabolic 
Diseases nursing Service assisted in the compilation of "A 
Manual for Metabolic Balance Studies" to be published in 
December 1958, by the U. S. Departoant of Health, Muoation, 
and Welfare, Public Health Service, Sational Institutes of 
Health, Clinical Center. 

Calendar Year 1958 




Serial No. 


In -a total research hospital such as the Clinical Center, the Nutrition 
:\)artment ' s role is * unique one for it must not only assume the usual responsi- 
oi.ities of a hospital food service organization, but must take an active part in 
ol. research programs in a supporting manner. Because of the exacting requirements 
v its program, the Nutrition Department can delegate no part of the patient food 
3crvice to another department as is done in other hospitals, but assumes full re- 
3pnsibility for the entire operation. 

Some of the unique responsibilities assumed by the Nutrition Department and 
samples of the kind of work involved are these: 

1. Controlling a single nutrient intake in varying amounts for an 
individual patient. For example: 

(a) Controlled sodium intakes used by the National Heart 
Institute in evaluating new drugs in treatment and con- 
trol of hypertension- -these diets require controlled 
levels of sodium beginning with a level of 200 milligrams 
sodium and increasing the salt intake to normal levels of 
10 grams of sodium chloride. This is accomplished by 
weighing all food and calculating its sodium content. 
Higher levels of sodium intake are provided by the addition 
of weighed amounts of sodium chloride to the previously cal- 
culated low sodium diets . 

(b) In the National Institute of Arthritis and Metabolic Diseases, 
the request for purine free diets, controlled protein intake, 
and maintenance calories is routine in connection with their 
studies of uric acid metabolism. A controlled purine free diet 
is a means of reducing the amount of exogenous purine and al- 
lowing for a constant intake of protein and calories daily. 
Maintenance calories is important because it is essential that 
the patient neither gain nor lose weight during the course of 
the study. All food refused is replaced in protein and calories. 
In many instances these patients are fluid restricted. Re- 
placement of protein and calories to patients on these rather 
monotonous, meat free, restricted fluid type diets becomes a 
real problem when the diet continues over a period of months 

as is often the case. VJith the use of these diets several ad- 

urition Department Annual Report, 1958 Page 2 

vances in the understanding of gcut cave "been repoi'ted. 
Outstanding was the demonstration with, specific tracer 
techniques that over-production of uric acid is the 
metabolic defect of primary gcut. 

The National Cancer Institute has used the same type 
of diet in investigation of leukemia. Elevation of 
serum and urinary concentration of uric acid have been 
described in leukemia and patients with solid turners. In 
fact, it has been known since IC70 that the excretion of 
uric acid may be increased in seme patients with leukmla. 
However, reported findings are not consistent and the;y? 
studies are to further the investigation. 

2. Controlling more than one nutrient intake for an individual patient. 

An example here is the diet controlling calcium and phos- 
phorous, ueed by the National Institute of Allergy and 
Infectious Diseases and investigators from other institutes 
such as National Heart Institute, in the study of sarcoid. 
The diet order is to maintain each individual patient on a 
constant intake daily of 200 milligrams calcium and 500 
milligrams phosphorous. The protocol in these studies in- 
cludes an increase in calcium to 3bQ milligrams with phos- 
phorous levels constant at 50u milligrams depending on the 
daily results from the laboratory. 

3- Controlling nutrient and/ or calorie intakes for individual patients 
to he iso-nutrient or iso-caloric, or both, daily. This means that 
a diet of very near the same composition is calculated daily and 
food refused is replaced so that the nutrient and calorie intakes of 
the patient stay essentially the same. For example: 

(a) A study by National Institute of Arthritis-and-'Metabelic 
Diseases involving two groups of investigators working in 
three different areas in a study of energy and riboflavin 
metabolism — the order was for a liquid diet of known compo- 
sition requiring an ad lib feeding program and yet a constant 
intake of riboflavin. The procedure used in administering 
the diet was as follows : 

A liquid formula of known composition was calcu- 
lated and prepared. The calories and total amount 
of the feeding for each patient were calculated to 
more than the patient would actually eat. A level 
of riboflavin to be given in the diet daily was de- 
cided upon by the investigator and the dietitian. 

f.trition Department .Annual Report, 195S Vage 3 

The subjects were then placed on an ad lib program 

requesting the feedings "as they felt a need for 

them. Feedings requested were- measured" by- the- 

dietitian and recorded". Subjects were required to 

take the-- amount- asked for. Daily/ 'at 3: GO p.m.. -the 

ad- lib feeding'" schedule stopped and the : dietitian- 1 

planned with 'each subject-the^ambunt" <6f feeding^he 

would take -for the : remainder- of- the day, calculated 

the- total amount-: taken for- the dayV and. calculated - 

the amount of riboflavin in that amount of feeding. 

The riboflavin was then adjusted to the constant 

level by pipetting the calculated amount of vitamin 

concentrate into the last feeding. The fluid intake 

of total formula and other clear liquids was reported 

to the nursing station. Intakes of protein, fat, 

carbohydrate, calories, and riboflavin were calculated 

and charted on separate intake records for the investigators. 

(b) In the National Institute of Arthritis "and Metabolic "Diseases, there 
has been a need for diets administered frcm two food service units 
to provide the diets used in connection with a study of energy 
metabolism in a metabolic chamber. The specific dynamic effect 
of food on energy expended was a part of this project. The study- 
required a 2600 calorie diet with 23$ of the calories from protein, 
314 from fat, and kjf, frcm carbohydrate, with one meal of the diet 
in liquid form containing 63$ of the total day's protein, 23$ of 
the total day's fat, and 30$ of the total day's carbohydrate. 
The liquid meal was calculated by the 9th Floor Metabolic Kitchen 
dietitian and served under her supervision. The breakfast and 
dinner meals were calculated by the 9th Floor Kitchen unit dieti- 
tian and prepared and served under her supervision. After discussing 
the requirements of the diet with the investigators, the dietitians 
from each of the food service units calculated the diet- -subtracting 
the amount of protein, fat, and carbohydrate to be included in the 
liquid meal from the total day's intake (composition of the liquid 
meal was determined by the problem under investigation), and allotted. 
the remainder of the total diet to the other two meals. The patient 
was consulted regarding food he would accept and one constant 
breakfast and once constant dinner meal were planned. The usual 
meal pattern of the patient was breakfast, the liquid metabolic 
meal at noon, and dinner. However, on the days he was put in 
the metabolic chamber, he entered the chamber in a fasting state 
in the morning, received the liquid meals while in the chamber, 
and was served his constant breakfast after leaving the chamber 
at 2: CO p.m. His dinner was served as usual. The patient was 

,'urition Department Annual Report, 1958 Page k 

observed in the chamber two or three times a week. He was on 
the controlled diet for the entire study period. Nutrition 
Department routine in administering this diet required continu- 
ous weighing, checking, and recording of intake. The investi- 
gator planned the days of observation in the chamber which 
altered the diet administration routine of the two kitchens 
and required vigilance and cooperation of personnel administer- 
ing the diet. 

(c) For the National Institute of Neurological Diseases and Blindness, 
a constant diet is being used in the study of periodic familial 
paralysis. The daily intake of carbohydrate, protein, fat, 
sodium, and potassium is Kept constant. The dietary requirements 
for this study began in February, 1958, and are still in effect. 
Cn a constant diet, calories are controlled for the purpose of 
making the diet iso-caloric. The sodium content is 3-° grams 

and potassium 5»5 grams. Two constant diets of comparable compo- 
sition were calculated and served to the patient on alternate days. 
This was done for the patient's benefit because he tired quickly 
of his constant diets. New constant diets of the same nutrient 
composition were calculated every two weeks to help the subject 
stay satisfied on his controlled regime. Food refused is re- 
placed for calories and potassium. Potassium level of 5-5 grams 
not met by food intake is made up with a 25/? potassium chloride 
solution which is administered by nursing personnel. The amount 
of 25^ potassium chloride needed to meet the level of 5-5 grams 
is calculated by the dietitian and reported to the nursing station 
in writing. Since December 11, 1958, this patient has been per- 
mitted to be on a selected diet allowing a variation of foods, 
but the diet order requires that the levels of sodium and potassium 
be kept the same as on the constant diet. Now the patient is 
visited daily and permitted to select what food he wishes. The 
amount of sodium and potassium in the food selected is calculated 
and the sodium is made up to the required $.0 grams by weighed 
salt af.rved to the patient with his meals . The remainder of the 
routine remains the same. 

(d) In the National Heart Institute fat studies of atherosclerosis, 
the patient may be placed on a five-gram fat iso-caloric diet. 
The diet is used to stabilize the patient on a minimal fat intake 
in preparation for testing and evaluating effect of various types 
of fats on blood lipids . 

.itition Department Annual Report, 1958 ?age 5 

h. Controlling group intakes qualitatively --as control groups and therapy- 
groups. For example: 

(a) For the National Institute of Mental Health it was necessary 
to maintain a normal control group of patients (4 West) on a 
control diet for studies of patients on 2 West. This means 
that the exact foods were served to both groups without vari- 
ation. These studies were not quantitatively controlled. 
This study was developed by the investigators studying the 
biochemical aspects of schizophrenia. 

(b) Another study by the same investigators placed a group of 
normal control patients on an ascorbic acid free diet. The 
study was an investigation of a probable disturbed metabolism 
in schizophrenia, specifically, the rate of oxidation of 
adrenaline which is affected by low levels of ascorbic acid. 
The patients acted as a control group for a group of patients 
with a diagnosis of schizophrenia in another nursing unit who 
were served the same ascorbic acid free menu. 

5. Providing diets for short term studies or tests involving numerous controls. 
Some examples are: 

(a) The use of a controlled diet by National Heart In6titue with 
hourly feedings of 30 grams of carbohydrate for 48 consecutive 
hours . This study was designed to evaluate the effect of oral, 
hourly, 30-gram carbohydrate feedings on lipid levels in the 
blood and was undertaken on four patients at a time. One of 
the four was on a special formula with 60$ of the calories from 
fat. This type of study involved a continuous feeding procedure 
which included re -weighing and replacing feedings not accepted. 

(b) The "2 Deoxy-D-Glucose" study of National Cancer Institute re- 
quires a constant diet for a patient on a regular diet. The 
patient may eat anything so long as he eats the same amount of 
the same specific type of food at the same time each mealtime 
for a 2| day period. The patient must be served at exactly 9 
a.m., 4:30 p.m., (a doughnut), 5 p.m., and 8 p. m. (milkshake). 
All food must be eaten--no substitutions allowed. The investi- 
gator must be furnished a detailed calculation of nutrient 

(c) For the National Institute of Arthritis and Metabolic Diseases 
in their hematology study, it has teen necessary to decrease the 
amount of dietary fat to very low levels in hemophilia patients. 
This is to aid in the study of the factors involved in blood 

furition Department Annual Report, 1958 Page 6 

C. Preparing semi -synthetic liquid diets for the purpose of supplying pro- 
tein, fat, carbohydrate, and calories of specific amounts. Since the 
only sources of these nutrients are cornstarch, oil, vitamin-free casein, 
sucrose, and distilled water, the problem of developing a product ac- 
ceptable to the patient makes this one of the most difficult and time 
consuming diets to prepare. Vitamins and minerals are regulated by 
the investigator in known quantitative preparations of supplements de- 
pending on the type of study under investigation. For example: 

(a) The National Institute of Dental Research uses the experimental 
semi -synthetic liquid diet to investigate possible allergic 
reactions to foods in the disease entity aphthous stomatitis. 
The patient remains on the diet until ulcerations disappear, 
then natural foods are added periodically, are recorded, and 
included in his diet as long as he remains symptom free. This 
in-patient study i6 done in connection with an out-patient 
study which requires the dietitian to collect and evaluate 

30 -day diet records for nutrient content. 

(b) The experimental semi -synthetic liquid diet is also used by 
the National Cancer Institute to make a diet free of folic 
acid in conjunction with drug studies (antagonist of folic 
acid) with choriocarcinoma (food composition tables are not 
adequate to allow for the preparation of a diet containing 
natural foods devoid of folic acid.) 

(c) In other studies of the National Cancer Institute, this type 
of diet is used to make a diet free of pantothenic acid and 
pyridoxine. These diets are used in conjunction with virus 

(d) National Cancer Institute's study of a chemically defined 
water soluble diet had previously been tested on animals and 
is now being applied to studies in man to determine its use- 
fulness in future studies requiring purified diets. The diet 
is composed of vitamins, minerals, amino acids, carbohydrate, 
and fat which are prepared and ordered by the doctor and then 
mixed in the Nutrition Department formula room. One study was 
for a 20- day period during which time the patient had only the 
diet and distilled water. The study was planned as follows: 

(l) Control period of Ik- days: the patient received 
the same number of calories each day (kC calories 
per kilogram of body weight), intake was charted 
daily, and patient's weight observed for gain or 

.'- / 

- ■- n 

Iijrltion Department Annual Report, 1958 Page 7 

(2) Experimental period of 20 days --patient re- 
received same number of calories as during con- 
trol period. 

(3) Control period for 14 days after experimental 
period- -patient received same diet as in pre- 
vious control period. 

This study will continue, using variations in fat and carbohydrate con- 
tent of diet to try to solve problems of patient intolerance to fat and 
carbohydrate . 

7. Assuming the responsibility for measuring the daily nutrient intakes of 
individual patients. 

For instance, in the National Cancer Institute units, it 
has been necessary on occasion to measure and calculate 
daily intakes of 27 patients at a time frcm one floor 
(2 nursing units). In the studies of neoplastic diseases 
in the area of chemotherapy, the total number of calories 
a patient consumes may have seme bearing on the interpre- 
tation of the effectiveness of the specific drug used. 
Lack of sufficient calories may change the interpretation 
of the effectiveness of the drug under investigation. The 
investigator is keenly interested in total calorie intakes 
in this particular type of study. A regular diet is usually 
ordered for these patients but a great deal of professional 
time must be spent with them to see that the needs of the 
investigator are met. 

The foregoing gives only a few examples of Nutrition Department responsibili 
tes, but demonstrates the exacting details of its work, the extreme importance of 
cose supervision of non -professional personnel, and consequently its need for 
jofessional know how. To explain also the complexity and variation of the depart- 
nnt's daily accomplishments, it should be pointed out that most of the examples 
sown here apply to work involved with just one patient. The Nutrition Department 
cily renders this same exacting and detailed service to man y patients simultane- 

Aside frcm the twelve floor kitchens, a kitchen in the residential treat- 
t:nt facility, 1-h, a formula room, and the main kitchen, all of which are important 
"> the services of the type described above, three metabolic kitchen units have beer 
nintained. These metabolic kitchen units have serviced the research programs for 

ie National Cancer Institute, National Institute of Arthritis and Metabolic 
Iseases, and the National Heart Institute. As an outgrowth of continuous need 

)r these units this year, two of them, the one for the National Cancer Institute 

iurition Department Annual Report, 1958 Page 8 

■the National Institute of Arthritis and Metabolic Diseases were maintained 

Ive months of the year. This facilitated the continuation of studies and 

-er usage of the beds for these areas. This too vas unique in that as far 

-is know, the Clinical Center is the only facility which operates more than 

i metabolic kitchen and as far as is known, such units in other institutions 

t closed for one month during the year, thus limiting the term of studies as 

1 as the time in vhich certain studies could be undertaken. 

Examples of the types of diets served from the Metabolic Kitchens have 
boa omitted from this report since they have appeared in previous annual reports. 

In addition to the patient area food service, the Nutrition Department 
\-rated a public cafeteria serving National Institutes of Health Personnel and 

The Nutrition Department makes a definite contribution to the Clinical 
losarch progrsm by providing precise and accurate information concerning intake 
t assist research investigators in such activities as: 

1. Interpreting the mode of action of new drugs which they may 
be investigating. 

2. The possible discovery of additional knowledge concerning a 
disease, which could lead to the development of a new medica- 
tion for use in the treatment of a specific disease or 
totally unrelated disease. 

3- Evaluating the course of a disease or the status of a patient. 

In accomplishing its objectives during the period December 1, 1957, through 
ivember 30, 1958, the Nutrition Department provided 25,222 recorded daily dietary 
:itakes (an average of more than 69 a day), 76,666 patient meals, and weighed food 
a 226,998 individual trays. In addition, during this period 175 diet aliquots 
';re weighed and provided for laboratory analysis, 9^36 days of fluid intakes were 
nasured, and 312 individual patients were given discharge or follow-up diet in- 
fractions each of which required a complete nutrition history. 


Nutrition services ^.n a total research hospital such as this must be 
Lexible in order to meet the changing research program need. This fluctuation 
n workload within a unit with a stationary staffing pattern such as ours pre- 

ents considerable problems . 

For example, in the national Institute of Mental Health the project of 
valuating psychotherapeutic methods of treatment of schizophrenia was discon- 
inued and a project study in the bio-chemical aspects of the disease was initiated. 

litrition Department Annual Report, 1958 Page 9 

.Jder the former study, nutrition services included only family style meal service 

th arrangement of periodic cook -outs, parties, and cooking classes, requiring 
^taall staff. As the biochemical study has developed, the Institute is working 
rward controlled diets for severely regressed schizophrenia patients and the 
athorized staff will be inadequate to handle the anticipated workload. 

In areas servicing patients of the National Institute of Neurological 
Cseases and Blindness, ordinarily a relatively small number of rather routine 
terapeutic diets is anticipated thus requiring limited professional coverage. 
le dietary workload has at times, however, reached unusually high levels at 
te onset of studies evaluating electrolyte metabolism before and after brain 

Studies of amino acid metabolism in patients with an inability to raetab- 
cism phenylalanine have necessitated careful calculation of severely restricted 
cets and their administration to feeble minded patients. It has been necessary 
t> add a dietitian to the staff to cover this workload. 

These fluctuations in workload are experienced in all areas as research 
rojects are started and discontinued and, together with fluctuating usage of 
]itient areas, make it increasingly difficult tc administer the type of food 
sjrvice required in the Clinical Center, within the allocated budget. 

Food costs of the Nutrition Department have continued to increase, 
litient raw food cost has shown a gradual increase over the last two years. 
fe are now faced with one of two alternatives --either to make no alterations 
:i the food service of the Clinical Center, which will leave us at the end of 
lis year above the budgeted raw food cost, or to make some drastic changes 

1 the food service, which would more than likely produce the serious impact 
F patient dissatisfaction, which in turn leads to the same dissatisfaction 
aong the medical staff. 

In anticipation of this problem, during the period August 1, 1957 > to 
jgust 1, 1958» w e developed fifty-two sets of menus with the idea that as of 
jgust 1, 1958, having an approximately equivalent patient occupancy, we would 

2 in a position to begin repeating the identical menus. The ration costs 
ould then be compared on a day to day basis with those of the preceding year 
s well as on a monthly basis. A study of the cost figures for the period 
ugust, September, and October 1957? and the same months for 1958 (menus identi- 
al and approximately the same patient occupancy), indicates an increased focd 
ost. This is true of a representative group of food items --particularly the 
sat items (entrees). 

In order to evaluate this increased raw food cost, a detailed analysis 
as made using as a guage the consumer price index of the Bureau of labor 
tatistics. This study indicates that raw food cost increases are indicative 
f the conditions of the food market rather than inadequate controls of the 
utrition Department. 

: v Jurition Department Annual Report, 1958 

Page 10 

Below is a chart giving a ccmparison of Bureau of Labor Statistics Price 
;r.ex figures for the years 1957 and 195$ as well as the Clinical Center patient 
rj.ion food costs for those years. (The 1952 cost of a certain group of food 
»ms is considered equal to ICO. The index for the later date measures in per- 
tage terns how much the cost of the same foods have changed. An index of 110 
cjis a 10$ increase over the 1952 price.) 

Comparison of Price Index Figures Published by Bureau of Labor Statistics 
and Clinical Center Patient Ration Cost Figures for 
1957 and 1958 



J auary 






C tober 



-rerage 115-4 

Reflects inventory adjustment. 

Clinical Center 

Clinical Cente 

Price Index 

Ration Cost 

Price Index 

Ration Cost 

112. 8# 















Ilk. 6 








117. h 






















It will be noted that there was an increase in price index costs each month 
0? I958 over the same month in 1957 and Clinical Center ration food costs show the 
ame increase. 

Bureau of Labor Statistics figures are available through the month of 
sptember 1953. Assuming that the first nine months are indicative of the year 
958, the average price index shows an increase of 5-2f>. By the same token, the 
verage Clinical Center ration food cost shows an increase of 5-1$- 

Ntrition Department Annual Report, 1958 Page 11 

The increases in food costs have been reflected also in operation of 
te Cafeteria. 

Another problem which is increasing in the Cafeteria operation is that 
o over-loaded facilities. Remodeling and expansion need to be undertaken in 
srving, dining, and dishwashJng areas. 

Major problems continue to be presented by inadequacies of some equip- 
■ nt and layout --namely, the lack of cooling air in the Main Kitchen which 
rt only contributes to excessive heat, but to high humidity, causing danger- 
csly slippery floors; lack of adequate food storage facilities; and lack of 
cnference rocm and classroom space. 

The lack of an overall preventive maintenance program for equipment 
cntinues to hamper operations. 

The turnover in clerical staff was a serious handicap in the operation 
c' the Department this past calendar year. 

The problem of insufficient parking facilities became so acute that it 
\is necessary, in the Nutrition Department, to set up a policy to schedule time 
:*f during working hours for employees of the late shift to re-park their cars. 
Uen these employees report for duty, parking areas are so crowded they must park 
■leir cars at great distances from the building. By the time they go off duty, 
:; is dark and unsafe for employees to walk the distance to their original 
]irking spaces. 

Insect infestation became acute in the Main Kitchen areas in June 1958- 
'ie staff has worked continuously with the Sanitary Engineering Branch since 
lat time to eliminate the problem and progress has been shown. Numerous 
langes in the physical layout of the Main Kitchen have been recommended to 
■ffect complete extermination. 


Technique of application is one of the most important functions of the 
atrition Department. The department must be ever alert in evaluating its prac- 
ices, and streamlining, shortening, and developing new procedures in order to 
sep up with and meet the new and ever changing demands which result from the 
piraling developments of modern science. 

Training and staff development have been found to be valuable tools in 
aintaining an efficient and smoothly running operation. Up to date equipment 
nd sufficient space in which to operate are basic requirements. 

;:trition Department Annual Report, 1958 Page 12 

Seme of the more important Nutrition Department achievements made along 
+ese lines during the past year are: 

1. Completed three manuals, (l) Operating Procedure for Each Patient 

Dietetic Service Unit 

(2) Food Portions Manual 

(3) Manual for Training Food Production 
Area Dietitians 

2. Organized plans and procedures for keeping two Metabolic Kitchen 
Units open 12 months a year. 

3. Reorganized Food Production Service professional and non -professional 

h. Revised and reissued policies for food service to Mental Health units 

5. Developed a complete set of specification for all perishable food items 
used by the Nutrition Department and revised general requirements for 
each cf the perishable food controls. 

6. Put in effect a preventive maintenance program in the dishwashing 

7- Initiated a monthly review of operations with Purchasing Department. 

8. Formed a committee to study and develop criteria of achievement 
levels for each non-professional position in the Department. 

9- The entire non-professional staff completed a one week course in 

Food Sanitation and a set of group meetings dealing with the subject, 
"How Can We Eetter Function as a Group?" 

10. The non-professional supervisors classes were extended to teach more 
about other departments of the Clinical Center. 

11. Assisted, to some extent, in the Nursing Department staff training 
program . 

12. Added the following equipment and space: in the Main Kitchen- -a 
larger safe, a pie dough roller, garbage disposal, separate space 
unit for sandwich making; in the patient area- -an additional office, 
rinse injector, and new motors for 12 floor kitchen refrigerators. 

Utrition Department Annual Report, 1958 Page 13 


Definite plans in the Nutrition Department for 1959 include: 

1. Completion of the revision of the manual of current diet procedures. 

2. A comprehensive review of the Department ' s staffing structure. 

3- Continued extension of in-service training for professional and 
clerical staff. 

k. Continued emphasis on developing more efficient operating pro- 
cedures in practical dietary application to research programs 
and reporting seme. 

5- The opening of a fourth Metabolic Kitchen unit. 

6. Completion of a study on, "The Factors Affecting Calorie Intakes 
of Undernourished Patients." 

7- Revision of the supplement of the Food Composition Books used in 
each Patient unit. 

8. Extension of the preventive maintenance program to other areas of 
equipment maintenance. 

Along with these there will be, in the Nutrition Department, the usual 
ontinuation of evaluating present practices, and streamlining and shortening 
j;w procedures in order to meet the ever-changing needs of our research 
avirorjnent . 

Research Projects 
Clinical Pathology Department 

Cell Volume and Distribution Pattern in Ascites Tumor Populations 

The Coulter Electronic Counter, originally designed for red blood 
cells , has been modified to permit accurate enumeration of larger cells, 
such as mouse ascites tumor cells and cells growing in tissue culture. 
This determination is based on the principle that cells displace their 
volume of electrolyte, therby causing proportionate changes in the re- 
sistance of an electrolyte column. Adjustment of the circuit charac- 
teristics permits enumeration of only those cells larger than a selected 
size. A serieB of counts at different adjustments permits the construc- 
tion of a size frequency distribution curve. As many as 5^000 cells may 
be measured in a few minutes. 

That this method measures cell volume was experimentally verified 
by the close agreement of the instrumental results with mean cell vol- 
umes determined by the hematocrit of different sized erythrocytes from 
various species . This was further corroborated by calculation of cell 
volumes from direct microscopic measurements of cell diameter. 

Ascitic tumors studies and the median volumes obtained were; 
Krebs K-2 lSOOn 3 , Lymphoma #1 800^3, lymphoma #1210 550u3. These 
results are in good agreement with published values. In addition the 
Krebs K-2 tumor was studied 3* 5, 7 and 12 o^ye after transplantation, 
during which time no significant changes were noted, tela tissue cells 
were readily counted and sized; median volume 2800|j3. 

If the logarithm of the volume is used, a normal distribution is 
obtained in almost all cases. 

G. Z. Williams 

A. C. Peacock (NCI) 

H, F. Mengoli 

Cytochemical Studies of Human Marrow by Ultraviolet Television Micro- 

The investigation of the ultraviolet absorption patterns produced 
by proteins and nucleo -proteins of cells in leukemia, anemia, myeloma, 
and normal marrows is developing an accumulated experience in the recog- 
nition of these changes. We are particularly interested in the changes 
which may occur following chemotherapy in leukemias. 

G. Z. Williams 
G. Brecher 
P. A. Fox 

- 2 - 

CP 58-03 Properties of Acridine -Orange ae a Supravital Stain for Bone Marrow 
aiV. rumor Cells 

Acridine -orange may prove to be a useful supravital stain and 
.;heBe experiments were designed to determine its properties related 
to examination of tumor cells and human marrov cells. Excellent dif- 
ferentiation of certain types of cells, particularly those containing 
hemoglobin and the young cells and more maturing cells are found in 
the bone marrow. The Acridine -orange was found to transfer very 
rapidly from stained cells to non-stained living cells when two cell 
populations are mixed either in the test tube or in vivo- pH and 
concentration are extremely critical factors in the differential 
staining of nucleo -proteins and ribo nucleo -proteins. The fluores- 
cence is greenish-yellow for the former and orange -red for the latter . 
Comparison of the fluorescence and ultraviolet absorption patterns is 
being studied. 

0. Z. Williams 

A. C. Peacock (NCI) 

CP 58-O'f Cytochemical Determination of Phosphatases in Living Cells 

Dr. Peacock has synthesized a formazan phosphate which appears 
to be readily split by phosphatases in living cells. This causes im- 
mediate precipitation of the insoluble formazan presumably at the point 
where the enzyme attacked the soluble formazan phosphate. The formazan 
intensely absorbs ultraviolet and therefore, ultraviolet television 
microscopic pictures with time-lapse motion studies indicate the pro- 
gression of this enzyme activity in living cells. 

G. Z. Williams 

A. C. Peacock (NCI) 

CP 58-05 Biological Reactivity of Substituted 2, 3, 5 Triphenyl Tetrazoliums 

A number of variously substituted 2, 3, 5 Triphenyl tetrazoliums 
were studied as hydrogen acceptors in the following types of systems: 
washing mouse liver mitochondria, fortified with DPN, TPH and lactate, 
succinate or malate; with purified diaphorase and DPNHj with sulfhy- 
dryl compounds (cysteine, glutathione, lipoic acid). In all systems 
studied tetrazolium salts containing a nitro group were reduced much 
more readily than otherwise similar salts not containing a nitro group. 
The nitro tetrazoliums are further divisible into groups depending on 
the rate of reaction in the liver mitochondrial system. Some react 
best with succinate, some best with malate, and other equally well with 
succinate, lactate or malate. Those reacting best with succinate were 
characterized by one or two nitro groups and a para -met hoxyphenyl group 
on C5. With sulfhydryl compounds tetrazoliums containing two nitro 
groups were reduced most rapidly, those with one nitro group more 
slowly, and those without a nitro group, not at all. 

G. Z. Williams 

A. C. Peacock (NCI) 

Improvements in the Ultraviolet Television Microscopy for Time-Lapse 
Cinemat ography 

The use of high quality broadcast type television components has 
markedly improved the accuracy of the ultraviolet absorption informa- 
tion obtainable by ultraviolet television microscopy. Interrupted 
ultraviolet light cycles and increase of the sensitivity of the system 
by pulsed grid bias blanking of the vidicon makes feasible time -lapse 
ultraviolet cinematography of living cells for protracted periods with 
minimal a»i delayed damage. An electron trapping phenomena may be 
responsible for the loss of vidicon sensitivity and response when low 
intensity light is interrupted. A method for maintaining point light 
activation or sensitization of the vidicon target plate during the 
dark interval between ultraviolet illumination cycles has been employed. 
These improvements have made it possible to take time-lapse motion picture* 
of cell changes resulting from the damage by ultraviolet light , of enzyme 
changes indicated by tetrazolium and forma zan production in living cells. 

G. Z. Williams 

R. Beuhauser (RCA) 

0. Johnston (Instrument Section) 

J. Vurek (Instrument Section) 

Ultraviolet Microscopy of Tritiated Thymidine Tagged Human Blood and 
Marrow Cells 

In experiments designed to study the incorporation of tritiated 
thymidine into human marrow cells in vitro, the difficulty of poor 
visualization of the stained cells after radio -autography was encounter- 
ed. This is due to the poor staining of the cells when they are 
covered by a gelatin film which is necessary for the radio-autographic 
procedure. A method has been devised for inverting the coverslip film 
preparation so that the ultraviolet light traverses the film first, 
then the cells and finally the quartz coverslip. This permits a clear 
and detailed ultraviolet absorption image to be observed and photo- 
graphed for purposes of identification of the cell types. 

G. Z. Williams 

T. M. meaner (Brookhaven 

National Lab) 

Cooperative research with other Institutes and individuals. The 
following numbers designate the cooperative research projects detailed 
above with the Office of the Assistant Director of Research, NCI and the 
Field Investigations Branch. 

CP 58-01 
cp 58-03 
CP 58-01+ 
cp 58-05 

Pathol; t, Medical Division, Brookhaven Ka vonal 

Laboratory (Dr. Fliedner) 

cp 58-07 

The following projects have been supported by the Office of 
the Associate Director for Research. 

CP 58-04 

cp 58-05 
CP 58-06 
cp 58-07 

P 58-O8 The Effect of Light on BCG 

Studies from the World Health Organization Tuberculosis 
Research Office revealed the killing effect of sunlight and sky- 
light and the production of higher allergy by light-treated vac- 
cine than an admixture of live and heat-killed vaccine containing 
similar numbers of live and dead BCG units. Effort was made to 
develop a light-resistant strain and to test itc immunizing and 
allargy-produc Lng qualities. Skylight and electric light do not 
kill BCG even under direct exposure conditions . Direct exposure 
to ultraviolet produces lethal effects, but no truly resistant 
variant has been countered. 

J. J. Huang 

P 58-09 Use of HeLa Cell Culture as Tool for Studies on Jicid-fast Bacilli 

Attempt is being made to use Shepard's technic of infect- 
ing HeLa cells with acid-fast bacilli as a possible tcol in the 
evaluation of immunity status produced by BCG vaccines in animals 
and in the study of the effects of adjuvants on acid-fast infec- 
tions . 

C. Zierdt 
J. J. Huang 

3» 58-10 Immediate Antibiotic Sensitivity Tests on Urine Specimens 

The present procedure of doing a colony count, isolating 
and identifying organise and antibiotic sensitivity testing on 
urine specimens of patients suspected of having urinary tract 
disease is helpful to the physician. However, there in too much 
time lost in this procedure in patients with clear-cut clinical 
urinary tract disease. 

The present project concerns adding the urine directly to a 
liquid nutritive medium containing an antibiotic and determining its 
optical density immediately and again after two hours intubation. 

H. E. Harsh 

- 5 - 

Development of a Biological Thermal Sterility Check for Autoclaves 
Utilizing an Aerobic Spore Forming Soil Bacillus 

A carefully standardized and well characterized strain has 
been developed and preserved. Multiple tests of the O.R. auto- 
claves disclosed satisfactory operation. The organism is now used 
as an occasional spot check and whenever an autoclave has had 
repairs . 

H. H. Marsh 

Survey of Staphylococcal Epidemics in Hospital 

The results of a year and one half study by this department 
on organisms received from twenty hospitals on the eastern seaboard 
were presented at the Baltimore Pathology Society. The study has 
been limited to the Clinical Center since the above presentation 
and represents an excellent monthly check on isolation technic in 
this hospital. 

C. Zierdt 
H. H. Marsh 

Preservation of Staphylococcal Bacteriophage 

Due to the laborious procedure involved in propagating bac- 
teriophage, the phage typing of Staphylococci has not been practical 
for most laboratories. This laboratory has been able to lyophilize 
the phage and maintain them for one year with only insignificant 
loss of titre. There is great saving of time and materials and the 
method has been accepted for publication in the American Journal of 
Clinical Pathology. 

C Zierdt 

Adaptation of Agar Diffusion Antigen -Antibody Reactions to the 
Laboratory Diagnosis of Fungal Diseases 

Present Eethods of skin testing and complement fixation 
reactions for antibodies indicative of systemic fungus diseases 
are bereft with false positive and negative reactions. Early 
results using a modification of Ouchterlony ' s agar diffusion 
method for separating antigen -antibody reactions appear promis- 
ing with regard to sensitivity and reliability in the diagnosis 
of disseminated fungal disease. 

R. L. Bell 

- 6 - 

CP 58-15 Chemical Induction of lysis in Members of Pseudomonas Species 

A new chemical agent has been four*! that induces lysis in 
a variety of Pseudomonas species cultures. This phenomenon is 
being investigated with respect to the presence or absence of bac- 
teriophage in this lytic system. 

C. ;lLerdt 

Cooperative Research 

In addition, the members of the K :robiology Service, par- 
ticularly Dr. Marsh, Mr. Zierdt and Miss .5alta have collaborated 
closely with Dr. Leon Smith of NIAID in t-a.luating the Greiss test 
for screening urinary infections and in r;ucying the effect of 
antibiotics on throat end intestinal flo;u i.\ patients with fever 
of undetermined origin. 

The Measurement of the Concentration and '.telati\* Distribution of 
Human Serum Proteins by Electrophoretic t .nd Chem val Methods on 
Normal Subjects in Active and Resting States 

Diagnosis end treatment of certain diseases \rc partly 
based on serum protein analyses of patients compared ;■;> protein 
analyses on normal active subjects. Because most patients are 
bed resting, it is necessary to determine values on normu^ cub- 
jects who are also in the resting itate, if comparative du-U. j.s 
to be properly assessed. It has irsen observed that out-patiei; ', 
show a significant difference in elative distribution and con- 
centration of Bc-urn proteins corr/ared to in-patients, and seeming!., 
independent of the disease statr . 

This study is being con 4 ,.nued for several years. Very few 
normal persons are put to bed 'or several days and the accumulation 
of this data will be slow. 

A. Ness 

H- Dickerson 

A Study of the Factors which Affect the Electrophoretic Mobility 
and Dye Absorption of Proteins in Paper Electrophoresis. 

Paper electrophoresis is rapidly establishing itself as 
a routine procedure for determining the distribution of serum pro- 
teins. It is important to evaluate critically the experimental 
factors involved in this technic, since the voluminous literature 
of recent years has reported confusing and conflicting results. 

A. Ness 

H. Dickerson 

58-I8 An Investigation of the Various Factors and Conditi-.ns Required for 
the Specific Determination of Blood Glucose 

Accurate blood glucose analyses are necessary i-.r the diagno- 
sis and treatment of certain metabolic diseases. Our r\?ent compara- 
tive evaluation of various me+hods of determining blood -^lucose lias 
revealed that the currently used methods too frequently j'.eld erratic 
results. Our study has resulted in some degree of improvt_vnt, but 
further investigation is being continued. 

A. Ness 

H. Dickerson 

58-19 A Study of the Factors which Influence the Binding of the Dye IJyd . 
benezeneazobenzoic Acid with Protein, with Particular Reference to 
Human Serum Albumin 

This is the dye used for the determination of serum album n 
by a procedure developed in this laboratory. Rarely a serum is en- 
countered in which the albumin does not quantitatively bind the eye. 
It seems that the albumin of these subjects is either qualitati 
different from normal albumin or that some of the dye -binding si.\~ 
are occupied by other molecules which cannot be displaced by the eye. 
The exact cause of this interference has not been ascertained, and 
the investigation is being continued. 

A. Begs 

H. Dickers on 

58-20 Determination of Copper 

Certain neurological diseases are accompanied by defects in 
the metabolism of copper. Diagnosis and treatment is followed by 
the analysis of copper in blood and urine. A simple method was 
developed for the specific and sensitive determination of copper in 
biological materials and its evaluation was completed this year. A 
manuscript for publication is being prepared. 

A. Ness 

58-21 The Effect of a High Protein Heal on the Level of Individual Plasma 
Amino Acids in Humans at Various Intervals after Feeding 

Because of the lack until recently of suitable analytical 
technics, little is known about the absorption from the intestine 
of the products of protein feeding. With the new technic of chro- 
matography on columns of ion-exchange resins for the quantitative 
determination of all the common amino acids, a fresh approach is 
being made to this study. Normal persons are given a high-protein 

meal, and the plasma drawn at varying intervals after the meal is 
analyzed for its individual amino acid content . Changes from the 
fasting values are compared with the composition of the protein 
fed. Preliminary results indicated that the individual amino acids 
are absorbed into, or removed from, the plasma at different rates. 
Studies on normal subjects and a few patients were continued and the 
results were published in Journal of Clinical Investigations 37:1710, 

CP 58-22 The Effect of the Administration of Bromsulfalein on Serum Protein- 
bound Iodine Levels 

It has been suggested that the administration of Bromsulfal- 
ein may cause falsely elevated values of serum protein-bound iodine. 
It is important to have definite information on this point for the 
interpretation of results in certain patients . Preliminary results 
indicate that Bromsulfalein administration is without significant 
effect on the level of ?erum protein-bound iodine in patients with 
normal B3P retention. Continuing studies have shown the protein- 
bound iodine level to bo elevated in those patients with abnormal 
BSP retention. 

J. Paatewka 
E. Frame 

CP 58- "'. A Study of the Electrolyte Levels of Serum from Blood Drawn Under 
Oil and Without Oil 

It is known that when blood is exposed to air, carbon diox- 
ide escapes from the plasma and there are exchanges of sodium, pot- 
assium and chloride between cells and plasma. In order to minimize 
such changes in the determination of serum electrolytes, our present 
practice is to analyze serum from blood drawn under oil. Informa- 
tion is not available as to the magnitude or direction of changes 
which ore encountered when this precaution is not taken. It is 
important to obtain this knowledge because an occasional specimen 
is received in our laboratory which has not been drawn under oil, 
and also for purposes of comparison of results with those from 
other laboratories where this precautionary measure is taken. This 
project is being continued and additional data collected. 

E. Frame 

Although important to the quality of work in our laboratories, 
note it was necessary to suspend work on items 21 and 22 during the 
latter three quarters of 1958 because the pressure of service work 
load left no time or energy-available for this developmental program. 

- 9 - 

p 58 2k The labelling of leukocytes with tritiated thymidine (Sponsored by 

Thymidine labelled with tritium has been shown elsewhere to 
be an excellent radioautographic marker for potentially dividing 
nuclei. The technic has now been extended to in vitro use on 
leukemic blood and bone marrow. Direct evidence for growth and 
turnover rates of leukemic cells is being measured in these studies 
(Cooperative project with Chemotherapy Service, NCI Dr. Schroeder). 
Preliminary results indicate that turnover rates of both acute and 
chronic leukemias are less than for normal marrow. 

G. Brecher 

P 58 25 Abnormalities of the blood coagulation mechanisms of patients under- 
going perfusion with the mechanical pump- oxgenator during cardiac surgery 

It has been shown in this laboratory, that patients maintained 
on extracorporeal circulation during heart surgery frequently have 
post - operative drops in their blood platelets. The etiology of this 
change is being studied by means of bone marrow aspiration and sur- 
vival studies of platelets. The search for other possible abnormal 
ities of blood coagulation which would explain the post- operative 
hemorrhagic diathesis in these patientB is continuing. 

It has been shown in this laboratory that the adequacy of post 
operative neutralization of heparin can best be determined by use of 
the plasma thrombin time. This work is to be published in Blood and 
has been submitted for presentation to the Society for Vascular Sur- 
gery in May. 

J. Peden 

G . Brecher 

J. McParland (NHI) 

A. Morrow (NHl) 

P 58 26 The role of cephalin and normal plasma lipids in blood coagulation, 
tested with the aid of Russell's viper venom 

This work continues, with the major effort being directed at 
isolatinn of single, pure coagulat on factors. A preparation of 
purified prothrombin has been obtained by an original procedure which 
is now being evaluated. A portion of this work was reported in the 
Journal of Laboratory and Clinical Medicine. , July, 1958. 

"G. Br e cEer 

A. Peacock (NCl) 

- 10 - 

p 58 27 The normal turnover of leukocytes as determined by experimental 
transfusion of wb.te blood cells 

Turnover o:? white cell had been studied elsewhere by means of 
an apparently st-.ble atabrine tag. Experiments in this laboratory, 
carried out in conjunction with other studies on white cell turnover, 
established thf.; the marker is not stable and therefore unsuitable 
for turnover oxudies . This work was reported at the Conference on 
Kinetics of CeJJ.ular Proliferation in Salt Lake City in January 1959. 
Animal studies uiing tritiated thymidine as a white cell tag have 
been begun. 

G. Brecher 
J. Peden 

P 55 _d Studies of the requirements for clot retraction in a purified system 

Prior work in this laboratory indicated that a plasma or serum 
factor is needed to permit platelets to accomplish clot retraction. 
It has not been possible to identify this factor beyond establishing 
its heat stability. The existence of this factor has been questioned 
by a group at Johns Hopkins University, so studies on this problem 
are being renewed. 

J. Peden 
G. Brecher 



,rt oi; AraxKisiaariC/ri 


Project Title 


S. J. Kreshover 

Prenatal Influences on Fetal 

NIDR No . Investigator Project Title 

2 D. B. Scott Studies of the Structural, Phys? 

& Chemical Lee of Calcified 
Tissues by Electron Microscopy j 
Diffraction & X-B.ay Microscopy. 

3 M. U. Hylen Studies of the Structure of Deve 

ing Calcified Tissuas by Blec 
Microscopy & X-Ray Microscopy . 

4 S. Takuma Electron Microscopic Studies of 

Ultrastructure of Developing and 
Mature Bone. 

5 M. S. Burstone Diagnostic Surgical Pathology Utilise 

Ing Histochesolcal Techniques. 

6 M. S. Burstone Development & Application of His to- 

chemical Methods in the Study of 
Hard and Soft Tissues. 

7 H. M. Fullmer Histochemical Studies of Connective 


8 P. H. Keyes Experimental Animal Carles. 


Individual Project Reports 

Calendar Year 1958 



1UM SS» Investigator 

9 R. J. Fitzgerald & 
H. V. Jordan 

10 S. E. Mergenhagen 

11 H. W. ocherp & 

S. E. Mergenhagen 



W. Scherp 






Howell, Jr. 



V. Jordan & 


J. Fltzgera: 









R. Onata 



6. Haopp 



A. Kevin & 


Go Hastpp 

Project Title 

The Use of Germ- free Animals in the 
Study of Problems Relating to Oral 

Mucopolysaccharase Production by 
Oral Bacteria in Relation to Perio- 
dontal Disease. 

Digestion of Native & Altered Col- 
lagens by Oral Microorganisms in 
Relation tc Periodontal Disease. 

Manifest & Inapparent Viral Infec- 
tions of Oral Tissues. 

The Selationship of Pathogenic Fungi 
to Diseases of the Oral Cavity. I„ 
Incidence and Identity. 

A Study of the Genera Actinomyces & 
Leptotrichia . 

Inhibitors and Dental Caries. 

Microbiology of Saliva. 

Hutrition & Metabolism of Lactic Acid 

Studies on the Microorganisms As- 
sociated with Periodontal Diseases, 

Investigations In the Field of the 

Studies of Interactions Between Oral 


Individual Project Reports 

Calendar Year 1958 


Hir& Ho . Investigator Prolect Title 
21 W. D. McBride 



M. Rogosa 

A. A. Rizzo 

Studies on the Host Cell-Virus Re- 
lationship with Specie! Reference 
to Latent Infections. 

Immunological, Biochemical, & Patho- 
logical Studies of Oral Bacteria, 

The Rat as an Experimental Animal 
for Research on Periodontal Disease. 






K. A. Pier 

K. A. Piez 
F. J. McClure 
J. E. Folk 

H. G. McCann 

Prolect Title 

Biochemistry of Amino Acids & Proteins 
in Mammalian Cell Cultures, 

Analytical & Structural Aspects of 
Collagen in Relation to Function and 

Nutrition & Dental Caries with Emphasis 
on Froteln & Cereal Foods, Lysine, & 
Phosphate Supplements. 

Protein & Amino Acid Chemistry. I. 
Proteolytic Enzymes; Chemistry, 
Kinetics, Isolation & Purification. 
H# Blood Clotting; Enzymology and 
Amino Acid Sequence Studies. 

Study of the Inorganic Chemistry of 
Mineralized Dental Tissues, Surface 
Enamel Chemistry as Affected by Diet 
& Oral Environment & their Relation 
to Dental Carles. 


Individual Project Reports 

Calendar Year 1958 



NIDR No . 

R. H. Larson 

Proiect Title 

A Study of EDTA & DHA as Caries Po- 
tentiating Agents & Prenatal Factors 
Affecting Carles Susceptibility in 
White Rats. 









S. Levis 





R. C. Likins 

S. A. Narrod 

Biochemistry & Physiology of the 
Salivary Secretions. 

Non-dental Physiological Effects of 

Biochemistry of Salivary Proteins. 

A Study of the Biochemistry of the 
Submaxillary Glands of the Rat with 
Particular Reference to Polysac- 
charide Synthesis & Intermediary 
Carbohydrate Metabolism. 

Biochemical & Biophysical Studies of 

Enzymological Studies in Relation to 
Oral Tissue & Periodontal Disease. 


NIDR No . 


A. L. Russell 

A. L. Russell 

Jo C Greene & 
A. L„ Russell 

Proiect Title 

Epidemiology of Oral Diseases Among 
Alaskan Natives. 

The Epidemiology of Dental Caries, 
Including the Fluorine-Caries Relation- 

Study of the Epidemiology of Perio- 
dontal Disease in India. 


Individual Project Reports 

Calendar Year 1958 



nidr No. 



C J. Donnelly 


N. W. Littletoi 


A. L. Russell 


C. L. White 

Project Title 

Study of Periodontal Disease and 
Carles in Adventlsts Families. 

Epidemiology of Oral Diseases Among 
Ethiopian Natives. 

The Epidemiology of Periodontal 

Relation Between Dental Caries and 
Periodontal Disease. 

NIDR N o, 



49 -C 


C. J, Witkop 

C, J, Witkop 

C. J. Witlcop 

C. J. Witkop 

C. J. Witkop 

D. L. Rucknngel 

J. D. Biswander 

R. M, Stephan 

R. K, Stephan 

Project Title 

Trl-racial Isolates In Eastern 
United States. 

Hereditary Defects of Enamel & Dentin 
(Michigan Study). 

Genetic Study in Southern Maryland, 

Familial Osteoarthritis. 

Hereditary Eye Defects. 

Sickle Cell Anemias & Other Inherited 
Hematological Disorders. 

Oral Aspects of the Results of Con- 
sanguineous Marriage in Hiroshima & 
Nagasaki, Japan. 

Studies of the Oral Microbiota & Oral 
Environment in Relation to Dental & 
Systemic Diseases. 

Interrelationships Between Local & 
Systemic Factors in the Etiology of 
Dental Diseases in Rats. 


Individual Project Reports 

Calendar Year 1958 


NIDk No . Investigator 
A. D. Merrit-: 




58 -C 


P. J. Coccaro 

54-C B. J. Drieco.l 

55-C I. I. Ship 

I. I. Ship 
P. N. Baer 

P. H. Baer 

59-C H. R. Stanley, Jr. 

60-C H. R. Stanley, Jr, 

61-C H. Sverdlow 

62-C R. S. Lloyd 

63-C R. S, Lloyd 

» 64-C H. Sverdlow 

D„ R. Poultoa 

BRANCH (Continued) 

Project Title 

Studies on Enzymatic Reactions of 
Cyclic Alcohols and Steroids. 

Serial Cephalometric Study of the 
Longitudinal Growth of the Soft 
Palate & Nasopharynx in Cleft Palate 

General Anesthesia (Dental) Baseline 

Investigations into the Etiology of 
Recurrent Aphthous Ulcerations and 
Other Oral Hon -malignant Soft Tissue 

Stress in the Dental Patient. 

Esterase Activity Associated with the 
Formation of Deposits on. Teeth. 

Some Genetic Aspects of the Periodontium 
& Periodontal Disease in Mice. 

Histopathologic Study of Periodontal 
Disease in BALB/Cann Mice. 

Hletopathology of the Human Pulp. 

Pulp Response to High Speed Cutting & 
Grinding of Teeth. 

Cephalometrlc Evaluation in Full 
Denture Prosthesis. 

Maxillofacial Prosthetics. 

Malocclusion as Related to Traumatic 

Twelve-Month Changes in Class II Maloc- 
clusions with & without Occipital 
Headgear Therapy. 

Sumcf.ry - Research 

Estii.« t«d Obi l- '.aLione for ?.Y. 1959 









January-December 1958 

aural Activitie s 

One of die more conspicuous ireas of operational emphasis 
during 1953 was the graduate training program, to which was im- 

■:d further stability through special programming effort. Such 
stability was achieved by bringing together, for the first time, 
all institutional training program directors for the free exchange 

deas with selected members of the National Advisory Dental Re- 
search Council, thur. enabling all participants to learn of the fie 

ty possible; in use of these fi rrding to the particular 
needs of eac tion. In addition, the comprehensive for 
reports prepared by program directors were compiled and distributed 
to serve 33 valuable reference data for formulating improved opera- 
tional standards. Although initial.".} no increase in appropriated 
training grant funds was made for FY 1959, it was possible to adjust 
a portion of the FY 1959 funds lecording to the critical needs of 
various research training centers. The increase of $200,000 in the 
FY 1959 appropriation for training grants has resulted in a total of 
S650.000 avai this important program. Judicious use of 
total available funds allowed the activation of two new training 
centers during the year, bringing the total number of active centers 
in 12 States at the end of the year to 17, in comparison to the 15 
of 1957. It was also possible to pay supplemental requests to those 
schools requiring program expansion. A conspicuous increase in the 
number of der, iring postgraduate training as investigators 
or teachers was evident during the latter part of the year, and 
served as a source of encouragement, as well as justification for 
establishment of a permanent Training Grant Committee which met for 
the first time during October. Between February and October, all 
research training centers were visited by Council members and NIDP 
staff members to evaluate each program in terms of progress and futu:a 
development. Complete reports were prepared on each program. 

In keeping with Council's recommendation to intensify research 
and training as a means of attacking the periodontal disease problem, 
a special Ad Hcc Committee on Periodontal Research was organized and 
held meetings during June and September. Specific recommendations 
by the Committee were made to the Council at the fall meeting and 
are summarized as follows: (1) establishment of periodontal research 
training centers, (2) set up program- type research centers, (3) compile 
a review of pertinent literature, and (4) hold conferences for exchange 

j ted by 

he Council requested the Branch to integr 
variev or a concentrate n effort 

connection w :nt 
Cental X-ray procedure. Discussion of 'cl- 
an be tw L school si, Courci: members and numer 

cf a coordinated 
e . The amout 
11 meeting of >rt 

As roblems of national oral health, 

■ -.-■■■ i t . era Lacuast tout the 
ced to r c analysis and ( 

pro: :ct of the general problem is consideration of 

in of speech impc I 
y, and f ed research grant funds 

rt of the year enabled the establishment of broad;: 

al of the leading academic institutions con- 
vas also made available 

;.or inclusion of sociological and psycho- 
cal aspects throughout th* entire field cf oral research, and es- 

: attitudes, both as regards 
! lay populaticr. The latter part of 
forth several suggestions uncil as to methods 
for forming : ing relationship between professional dental 

..el . 

By way of correlating dental research and training at an inter- 
national level, approval was given during the year to a proposal from 

\merican Eental Association for a representative group of: professional 
:1 person vlall Leal research and training centers 

\va. the Soviet Unio:i during 1959. 

The research project grants program continued during the year 
principal source ::or obtaining essential date, from both the 
: research find clinical areas of the oral health field. Funda- 
vb\ research categories, such as biochemistry, microbiology, pathology, 
and physiology continued to remain prominent: as constituting the majority 
"c&l research projects, while notable stimulation was given to 

Udemiology, now 
■: in the oral hec 
or the support of pre. 
a 3ound means for bi^ 
leal departments, thereby for 
on oral Hess of 

became available at the beginning of the 
t only approximately half of the new 
he caler- ided for 

tinned integrity of all reviewing boc" 
of all additional funds sei 
it the pre ; 1 is not yet being 

Lr. \t the year's end the nv 
:ts totalled to near 350 in 32 states and 
red to '.' 57 and i 

lal clini 
;ers , the 
survey to determine 
I Journal publication for mutual satisfaction 
ivation of the survey during the 
1 of Dental .: 
es . 

>n by the Branch during the year r 
-ore efficient liaison bet.-ean medical and den' 
M extramural prograir laid the founda 
L health aspects in several lr 
led for the study of chronic diseases 
and / th problems related to aging. 

nleo mar! litiation of support for conference 
as, designed in each instance to 
foreaost research personnel according to research 
disease srea. Because of the unquestioned value of 

L lowing nge of informa- 

- progtess reports, it seems certain that the con- 
ferer 3ft palate research, microbiology, and related dental 
fie'.< j centered within the Dental Study Section, DRG, will 

rda accelerating the rate of resei- 

fellowships continued to demonstrate their great worth 
mans for introducing both dental students and dental 
graduates in; -2 arch field, thereby maintaining a continual 

;ormulited to 
£ regular fell jere dv 

and toto 

jnts, pax cellowship a* 

len:nl schools, and requf::= 
inue to king that the pre. 

sed to 8 uni 

ral progr Dental Institute during 1958 covered 
de range roject receiving support proportional 

: productivity mce to the c 

Whi - ne a major responsibility for the conduct 

. caries related particularly to 
ii, the !IIDR program duri.- 
gavf. increas "'ogy of periodontal diseuse; 

ni facial nn I formations; morpho- factors in oral dices: 
case oriented studies in the biophysical and t riel 

In th of electron py and related physical s: 
ies continued to be directed toward the development of new 
information on the formation and calcification of teeth and bones, and on 

of fully mineralized tissues. It should be indie 
chat interest in the work on tooth formation extends beyond the dentil field 
because tl .-ation derived bears on the function and structure >f 

i detailed study of dentinogenesis, which has just 
been published in monograph form, and the collection of data for a sim^ 
major publication on enamel formation will probably lead to changes in 

ctural concepts that may clarify some of the basic histological tenets 
that have long had to be accepted and taught due to limitations in the 
avciLifole information. With reference to the difficult and complex problem 
i g and demonstrating the crystal structure of fully mal ora~ 
lized tissues ; it iB becoming increasingly evident that maximal utiliza- 
tion of all of the modern physical tools, such as electron microsc: 
end diffraction, and X~ray nricroscopy, will be required to compile the 
visual picture. A further aspect of the work in electron microscopy 

if fraction is the investigation of the mechanism by which fluoride 
compounds exert a protective effect on tooth enamel. Inasmuch as the 
tin compounds have received considerable attention as possible agents 
for the control of dental caries, studies on stannous fluoride will 
continued and extended to include other metallic salts. 

^r.ion during 1958 was 
techniques to the investigation 
.? dev<<_ lor cent of r?v method.? for pro- 
id nnrpbt changes 
in i Leads to a bettsr understanding of 

as well a6 providing promts i 
. Other 

the nature of normal 
>.ges th^ disease. 

ta pertinent to tha 
lich the e tissues s •;.- 
:il disease. 

in both rats and 
hasten :o Rome degzee and under <- 

;gests a new cencepfc in regard to 

microbiology during 1958 continued in the 

lal physiology, mechanisms 
the more significant 
solation, characterization, end class 

ood oral bacterid 
I Veil lone I la; 2) a better under z tan ling 
:s and metabolic activities of the 
3) obser langes brought About 

lonstration of essential 
y certain oral bacteria; 5) the observa- 
is organisms predominjte at certain early 
js of dentil caries thereby xxj. i fresh appraisal of 

the causatior and 6) the demonstration th; 

cocci, K>st numerous :nd in the smith 

cause car - ise gem- free rats. 

ty in microbiology were related to an 
increased enp srimental infections with the oral flora 

and the rr. their development; and studies of inhibition 

of caries in rats by ir corporation of antioxidants in the diet. Also, 
investigations in the field of periodontal disease were expanded by 
initiation of studies of the role of oral filamentous bacteria in 
dental calculus formation and by extension of studies of the digestion 
of essential polysaccharide and protein compo7ients of the gingivae by 
oral bacterial enaymes. Another significant accomplishment during 1958 
was that, for the first time, means were found to produce regularly 
an experimental infection with oral spirochetes which are commonly 
associated with periodontal disease. It was of further Interest to 
note the eiihance^ent of such infections by either the enzymes men- 
tioned, or by concomitant syngeristlc infection with the bacteria 
producing Che infection. 

The 1958 pro&rcm of i • tory of Blochea: .: 

continued to eiaphr.size pro relation to calcifi- 

cation nn< le. A.s e major protein of skin, tendon, bones 

and teeth, collagen is Implicated as a vital factor in calcification, 
as well as ir. ntal disease. With respect to the amino acid 
analysis of c Urates (.: oon, bone and den 

of the rat and of the steer), it has been tat, except for 
lysine and hydroxylysina contents, these col, i gens are identical. 
Col lagens. from invertebr' , however, vr.ried widely in proline, 
hydroxy pro line and hydroxy lysine. 

To elucidate the relation of collagen to mineral metabolism, 
the calcifying tendons of the young turkey were the object of parti- 
cular experimental study. Age-associated changes in the amino acid 
composition of calcifying tendons hove been indicated by ar.alyt 
data, an have differences in calcified bone r.r.d tendon with respect 
to carbonate and citrate consent. Concomitant changes in the histology 
and physical properties of the turkey tendon undergoing the process 
of calcificaticn have also b*en under study. 

In other studies, observations on the skeletal metabolism of 
calcium as compared with strontium hr.v ad calcium to be the 
more stable in the hydroxyipitite structure of bone. Thu3, excretion 
of radios trontium by the kidney was approximately four times that of 

The pre, 'experimental carles research moved forward in 
1958 mainly in areas pertinent to the cariostttic effect of phosphates. 
A significant finding was that whereas insoluble phosphate (Ca3(PG c 
and CaHPO^ alone, had no i tic effect, the addition of 
NaCl to CaHPC>4 rendered it significantly car ic static. A very signi- 
ficant inhibition of experimental caries also was produced by 
Ca(H2K)4)2 'H2O. This continuing evidence of a pronounced anticaries 
effect of the more soluble phosphates lends further support to the 
proposed study of the effect of phosphated fleur on human dental caries. 

Continuing studies during 1958 on basic enzyme and amino acid 
chemistry gave the following significant results: 1) More extensive 
purification of She Folk enzyme, carboxypepticose B, whereby specific 
activity values were increased approximately 10 fold over previous 
preparations; b) Respective increases of 100 end 400 percent in 
hydrolytic activity of carboxypeptidase A and B when preincubated with 
Co++, Ni++ or Fe ++ ; and c) Advancement of new theories of fibrin 
polymerization and thrombin action relative tc blood clotting; these 
being derived from a study ox the amino acid sequence of the peptides 
A and B of cofibrin. 

Calendar year 1958 also marked the completion of analytical 
data pertinent to the relation of fluoride content of human bone 
and its chemistry findings demonstrated that increases in 
fluoride in bone n 0.09% to 0.73% were accompanied by a sig. 
f leant increase in the ash and magnesium content, whereas carbonate 
and citrate decreased. On the other hand, calcium, phosphorus, 
sodium and potassium concentrations in the skeleton were not affected 
by bone fluoride changes. 

In the field of epidemiology and biometry the broad goals of 
1958 included an amplification of the descriptive and the determina- 
tive epidemiology of oral diseases. Particular emphasis was placed 
on patterns of prevalence and severity of periodontal disease as re- 
lated to age, sex, race, and social status; and an evaluation of 
etiologic role of variour chronic systemic illnesses. The following 
studies may be cited as particularly productive: 

1. Preliminary survey of Alaskan Sskiac (in cooperation 
with an Alaskan Survey Medical Team of the Interdepartmental Com- 
mittee on Nutrition for National Defense) . A still Incomplete analysis 
of findings in 700 Eskimo men reverled significant data regarding 

the oral health of this group. For example, comparing the prevalence 
of oral disease in individuals living under relatively civilized 
conditions with that of men from primitive villages, it was found 
that dental carles, as well as periodontal disease, in very rare 
in ! :he latter group. On the other hand, when Eskimo c have lived 
for some time under relatively civilized conditions, the prevalence 
of oral disease increases to a point quite comparable to that seen in 
average male population groups in the United States. 

2. Studies of periodontal disease in India. This study, con- 
ducted in collaboration with the University of Michigan, showed that 
a considerably mere advanced gingivitis exists at earlier ages in 
India than in the United States. An important sequel to this pre 
liminary stud) has been the encouragement and education of Indian 
dentists to continue the program. As further data becomes available 
it is transmitted to the Epidemiology Branch of the Dental Institute, 
for analysis. 

3. Study of Ethiopian natives (conducted in collaboration 
with the Interdepartmental Committee on Nutrition for National Defense) 
This recently initiated study is providing material for comparison 

of the patterns of disease in primitive Ethiopian groups with the 
patterns already observed in equally primitive Eskimo. Since these 
populations differ ethnically, in ecology, and in dietary and other 
habits, it should be possible to eliminate some accidental associa- 
tions on the basis of this comparison and to re-enter the Alaskan 
area in the next year with a plan of study that promises even greater 
rewards . 

./•.omen* ii 
cy .are still 
:al oral hygiene 

Branch have 
Lucidatir L biologr- 

■ral cavity, with the 
aim jj sntion and tr 

ment. our understanding of 

disc M Because the development of most c 

r:.ety of complex factors vhich involve 
the -he range of 

thes » :ope. Also, as has been 

illi ry study of the oral 

s for detecting, sur- 
veyi i hich may not 

• tion groups . 

vith increased momen t 

end ry encourag:.rg. Outpatients and 

indj Jor portion of mater 

6tudi Htecl on an inpatient 

basis, ore has beer little difficulty in 

keepL NIDR occupied within 

the nrxatf space anc timing. As the clinical 

res: ch; t more beds can be effectively 

util <-£ ; he pre j ;tr. . 

Th; majority of scribed in the following 

sect. or. ar i cc oi: si.vdies begun in previous 

year.;. D. irch pi-cjects in the Clinical 

ve now been completed or dis- 
continued. Dur^ ht other projects have been added. Sir 

the subjects of these :ta have varied widely, an effort has bf 
made to further define their scope tnd to arrenge them into relatively 

,e pre »ran a::eae *, i.e . , (1) Periodontal Disease. (2) Oral and Facial 
Prosthetic h and Development, (4) Genetics, (5) Oral Surgery, 

i etiology, (8) Intermediary Metabolism, 
and [9) Diseases of the Teeth. Several examples of specific projects 
with:'.n the bread program areas mr.y be cited: 

Hun a n Genetics Study . This rrogram started as a study of 
hereditary defects in the teeth of school children in Michigan, and 
lad to the discovery in the Eastern United States of several inbred 
population groups having an unusually high rate of genetic defects. 

ry dental 

abnormal i-ies, such >is al 

iciancy, Albers-Schonberg 
Extensive investigati :ns of the phys 
1 anemia found in these inbred groups 
Heretofore j.irecoynized. A new 

been an found ir %noth'r inbred group 

he succes 
demo for the prediction and detection of 


^ is survey were ( 
cribed as generalised hereditary 
OSS:. rited as a r s trait, and 

tion of , closely associated 


! t . 

- :al Institute program 
of Michigan 
by the National Research Council 
and the Atcrc?.:' Initiated in A\ B, this 

1 and cral conditions of approxi- 
mate nusin mating s in Ji.pan Whereas the 

iiited States ie concerned with the effects 
of continuous .nese survej has the unique adv ntage 

one type of mating pattern. 

Periodontal disease is a major cause of 
die-aged end older pe id m?.ny local and systemic 
factors amy affect its devi Irpnent. Research directed at the etiology 
as inclu: es on the rubbers and different types 
of microorgai his topathc logical and histochemical 

changes; and etjeies on calculus formation in periodontal lesions com- 

periodo .ues. As noted in the individual 
project reports ;or '953, each of these areas of study has contri- 
buted new :.nfctn the disease process. Periodontal 
dis: experimentally in laboratory animals 
so that interact ?japortance cf several different 
factors in the aiialogy of the disease can be determined. 

al Caries . Dental caries is the major c=use of tooth loss 
ildren ^r idults. The general program of study of this 
common drl- s been directed so as to secure more precise informa- 
tion concern!, the bacteriology, physiology and bio- 
chemical ;he ore! cavity, which in soms cases lead to 

pant ct ries and in other cases none. Thus, 3uch 
:3 ha : e bean axplored as variations in eating habits, salivary 
flow, intraoral pH and microbial flora. 

o <• 

Ac ■i ronic Stomatitis . Recurrent aphthous stomatitis 
painful disease of the oral mucous, the etiology and 
treitmonl h is poorly understood at i:he present time. Rest; 
in ^bis d tsetse category seeks to determine whether causative agents, 
can be demonstrated either by using virus cultures, allergic, hema- 
tological and o':her diagnostic tests, or by therapeutic agents, 
as corticosteroids, which may relieve the condition. Findings, to 
date, serve to ecphasize the complex nature of aphthous lesions. 

Oral and F acial Grew-h end Developmen t. Research in this 
important field has been conducted with the objective of (1) better 
understanding of the normal and abnormal growth patterns of the facial 
structure:; end cranium, and (2) evaluating orthodontic methods for 

;nent of developmental abnormalities. Particularly emphasized 
have been a laoinographic study of the gTowth of the soft palate in 

t palate patients, and the refining of those technics of there 
that hold promise of relatively widespread explication. 

De ntal Anesthesia . General anesthesia for the more conmon 
dental procedures on ambulai ory patients poses certain special problems 
and considerations. Among the more important are that the field of 
operation in and around the mouth and throat where blood and secre- 
tions are present poses a constant threat to a patent airway, and 
that complete relaxation is not essential since work can be performed 
in rather light planes of anesthesia. Unfortunately, dental anesthesia 
in non-horpitalizad patients has been subjected to considerable criti- 
cism due tek of physiological data. A necessity, therefore, 
exists for denial surgeons to know more about the ?.gents they use if 
patient srfety snd smooth anesthesia are to be assured. Although 
many studies have been carried out on the physiological effects of 
general anesthetic agents, the results are not genarally applicable 
in the care of dental anesthesia administered in the office. The 
Dental Institute study was vrdertaken to obtain physiological data 
on ambulatory patients receiving various general anesthetic agents 
for standard types of dental surgical treatment, bindings to date, 
as noted in the following section of individual project reports, 
indicate the importance of this study to the gener-l irield of anes- 
thesiology and the likelihood of long-range applicability. 

Maxillofacial Prosthetics . Different types of prosthetic 
appliances have been studied and constructed for individuals who 
have had surgical destruction of tissues of the head and neck, and 
for patients afflicted with cleft palate and other defects. By 
utilizing cephalonetric measurements and other tests in their design, 
there has been greater assurance of maximum functioning for eating, 
breathing, speech, and esthetics. 

"ued need foi nd re-evalu- 
of ligations program to the 

~cjii planning for the future, good lia 
has beer hed with the Institute Board of Sciertific Couns? 
By such .8 well as better utilization of consultants' servi 
there should evolve a program that will bring both basic and ap- 
areas to on purpose of solution of the major oral health pr 
of the 



January-December 1958 

NIDR recognises that its laboratories and branches can function 
most effectively if they are relieved of as many of the details of 
administration as possible. The administrative office therefore 
endeavors to provide as many of these services as are best performed 
centrally and continues to process all personnel papers from the 
office of the Administrative Officer. However, because ol the delivery 
and redistribution problems inherent in a central requisitioning 
activity, and because the laboratories end branches now maintain a 
simplified memorandum accounting record of expenditures, orders for 
equipoient and supplies are prepared by the laboratory and branch secretaries 

NIDR endorses additional training for its employees and lias 
sponsored Civil Service employees for off-the-Job courses and Com- 
missioned Officers for training. Four Commissioned Officers continued 
their training outside the Servlce--two finished special training by 
the erd of June, one has been in training status for the whole calendar 
year, and one pursued evening courses while on full-time duty. A 
Civil Service employee at the professional level completed require- 
ments for his Master's degree; one Clerk was endorsed for a Department 
of Agriculture scholarship, the scholarship was subsequently awarded 
and the individual enrolled in an evening class entitled 'Adminis- 
trative Procedures"; and an Administrative Assistant, after successfully 
passing a required qualifying examination, was permitted to attend the 
Civil Service Commission's first Financial Management Institute for 
one week. 

Recognising the advantages of tht periodic efficiency ratings 
on Commissioned Officers, NIDR has amended the rating form for use 
with Civil Service employees. The rating does not become part of the 
persocnel file, however, it is extremely useful In evaluating the 
employee, particularly in respect to prcootions and career planning, 
and serves as a basis for completing the HEW Performance Rating Report. 

Throughout the year the various administrative functions were 
re-evaluated for the purpose of determining how effectively they pro- 
vided service to the operating personnel . The following changes were 
made as a result of this program analysis: 

1 . rhe revised property accountability procedure outlined 
In last year's annual report was put into effect in 1958. Each princi= 
pal investigator Is charged with property in his own area. Although 
this aeens that there are some 35 custodial areas, the system has proven 
itself in that inventory is much easier to check out in the event an 

leaves detailed avay from the reaervati< 

I proper :y checked out to 

ta Institute was established the Administrative 
e has had to rely on the serviced of a part-time Budget ExamJ • 
Thii a assistance laiposed a great deal of hardship on the 

small admin ir.trative which wai endeavoring to keep pace v 
an eqwndirif program. With che beginning of fiscal year 1959 the 
Chief of the ?lnanclal Management Branch agreed to Dental's request 
for .i Eull-tiaa Examiner. The ExamlneT is now physically located 

ilr.istraclve Office and has greatly contributed to the 
efficiency of budget preparation and execution, particularly the taak 
of administering the memorandum accounting system for each labo 
and branch. 

Kb the research expanded, a problem developed 
relative to providing a central reference poin: for reprints from 
both tflDR investigators and those received from outside sources. A 
Byei.ii was devised whereby such reprints are classified and coded by 

. author, and aubject on McBae cards. This system will pr 
a ready reference for i variety of dental research material and will 
save count] ens hours in searching out reprints concerning specific 

eta. This system was initiated near the and of 1958 end should 
be completed for ell reprints on hand in the early part of next year. 

An analysis wee made of the various types of letters 
directed to dentists end physicians by our clinical staff In coi 
with, p tieut raftrrels. It was found chat the major portion of these 
replies could be answered by adopting three carefully prepared form 

tra. Thia procedure has effected considerable savings in manpower 
formerly spent io dictating and tl net riblng individual letters. 

In order tc place requests for reservations within 
traveler's immediate area, due to the necessity of consulting on 
schedules, cases, etc., this function was decentralised to the labora- 
tory and branch secretaries. This arrangement has improved the pro- 
re for obtaining tickets for crewel; made it easier to make changes 
in itineraries end cancellations; and, in addition, the change has 
enabled the secretaries to become more familiar with seme of the various 
travel regulations, thereby improving the completion of the expense 
account fo 

In March of 1953 cur request was approved by the Surgeon Geno 
to change the titles of two laboratories In the Institute. The Labora- 
tory of Orel and Biological Chemistry became the Laboratory of Bio- 
chemistry, and the Laboratory of Oral bacteriology became the Laboratory 
of Microbiology. The new tittles more adequately describe the type of 

- 3 - 

rest; - g conducted, and ore in I: le with our philosophy that 

dent* h is not corfineci to the ore I 

ifl. r.taff of the Office of th Director, Associate Director, 
Chiefs devotee considerable tins assisting in de- 
veloping various plans for the construction of the new Dental Building 
whlcl i ic a uled for completion in November 196C . Numeraus xeetings 
were he d t roughout the year with key oersonn i: In the Research Facili- 
ties :?L.nni::g Bratch, DRS, to establ .sh certain types of requirements; 
i.e., interior construction. Installation of special types of equip- 
ment, and office and laboratory layouts. In addition, plans were 
developed for staffing the various components that will occupy the 
new building, plus all movable equipment needs, t.s it is anticipated 
that construction will begin la Marc'' or April of 1959, It is reasonable 
to believe that the meetings will be continued throughout most of 
next ;'Ci.r. 

The pay raise for both Comir. stoned Of ?:.cers and Civil Service 
personnel during the calendar year resulted in additional demands on 
KIDS' iy liialted funds. It han been necessary to block a number 
of pc nit ion: in an effort to accrue sufficient i'unds to pay salaries, 
and thin action has placed an undesirable restriction on research 
activities. The financial problem was further complicated by the 
change-over from advance formula reimbursement to a fee-for-senrice 
basis for ar.iaitls supplied from the laboratory Aids Branch. Aa a 
resul':, it bei been found than the cum returned tc IflDR for this 
purpes e is not adequate to pay for animals used during the last six 
month i. < f the calendar year; and the Institute hat had to divert other 
funds to purchase animals under the . e<' for- ue .-vie 2 arrangement. 
These tvo problems, in face of a static budget, haa placed a real 
econonlc hardoip on the Institute which has a Personal Services»Ot 
Objects ratio of approximately S2: 18 at the prauant time. 

Space liml cations have continued to prohibit related research 
studies fron occupying space within the same general area, thus making 
administration and coordination more difficult. However, this problem 
will he solved when tha n«» Dental Building is completed in the Fall 
of 19<<0. 



Individual Project Reports 
Calendar Year 1958 


1. S. J. Kreshover: Prenatal Influences on Fetal Development 

Ottiem •£ A— odf r.i?Gc(;ar 

Estimated Obllftat.'onc for F.Y. 1959 




Project Kuabers; 


Serial So. HIDR-1 

1. Office of Associate Director 
3. Bethesda, Md. 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Prenatal Influences on Fetal Development. 

Principal Investigator: Dr. S. J. Kreshover 

Other Investigators: Miss Lee E. Feild 

Cooperating Units: None. 

Man Years (calendar year 1958) : 
Total. 1 1/4 
Professional: 1/4 
Other: 1 

Project Description 

Objectives : 

To study such factors as anoxia and external pressure 
on the growing fetus in the causation of congenital 

Method s Employed : 

1. In order to study the effects of amniotic sac com- 
pression, 79 gravid rats (ranging in gestational de- 
velopment from 13 to 16 days) had their uteri exposed 
through a midline abdominal incision, and a fine needle 
puncture made through the amniotic sac of each embryo 
contained in the right or left uterine horn. The opposite, 
non-punctured horns served as controls. Cesarean sections 
on the twentieth gestational day permitted examination 

of the uterine contents. These totaled 461 experimental 
and 289 control fetuses. 

2. In order to evaluate disturbances in maternal- fetal 
vascular supply, 124 gravid rats (6 to 15 days in gestation) 
had their right or left uterine and uteric arteries 
occluded for periods ranging from 10 to 120 minutes. 
Circulation to the opposite uterine horns was not disturbed 
and thus provided a source of control fetuses. Cesarean^ 
delivered young numbered 726 experimental and 440 control. 

Part A (Continued) Serial No. NIDR-1 

- 2 - 
Major Findings ; 

1. The incidence of fetal resorptions following 
amniotic puncture was 7 percent in rats operated at 
16 days gestation, 28 percent at 15 days, 60 percent 
at 14 days, and 36 percent at 13 days. 

2. Examination for incidence of congenital defects 
after amniotic sac puncture showed palatal clefts and 
limb deformities (malrotatlon, agenesis, and syndactyly) 
in 0, 5, 33 and 66 percent, respectively, of the offspring 
of rats operated at 13, 14, 15, and 16 days of gestation. 

3. In contrast to amniotic sac puncture, the study to 
evaluate the effects of anoxia by circulatory arrest 
showed a 31 to 66 percent incidence of fetal resorptions 
in animals operated on the 14th to 15th days of gestation; 
50 to 80 percent in 12 to 13 day gestational animals; 

and 13 to 46 percent in rats treated during the first 

4. A two and one-half to 5 percent incidence of cleft 
palate and limb deformities in the anoxia experiment was 
without apparent relationship to either duration of 
circulatory arrest or period of gestation when occlusion 

Significance to Dental Research : 

Although congenital abnormalities of the oral structures, 
including the palate, teeth and their supporting tissues, 
have been the subject of considerable attention, much 
remains to be learned of the various etiologic and con- 
tributing factors involved. 

Proposed Course of Project : 

Further exploration of maternal factors influencing 
fetal development. 

Part B included: No 


Individual Project Reports 
Calendar Year 1958 


2 D. B. Scott: Studies on the Structural, Physical, & Chemical 
Properties of Calcified Tissues by Electron Microscopy, 
Electron Diffraction and X-ray Microscopy* 

3. M. U. Nylen : Studies of the Structure of Developing Calcified 
Tissues by Electron Microscopy and X-Ray Microscopy. 

4. S. Talcuma ; Electron Microscopic Studies of the Ultrastructure 
of Developing and Mature Bone. 

5 - M. S. Burstone : Diagnostic Surgical Pathology Utilizing Histo- 
chemical Techniques. 

6 - M. S. Burstone : Development and Application of Histochemical 
Methods in the Study of Hard and Soft Tissues. 

7 H. M. Fullmer : Histochemical Studies of Connective Tissues,, 


P. H. Keyes . Experimental Animal Caries 

laboratory of Histology & 

E stimated Obligations for 





Project Numbers: 




:< - 7 

HIDE -8 

lo Histology & Pathology 
2c Bethesda, Kd„ 

Indivi J ual Pr r?crt 

sndar Year 1958 

Stud'.ss of the Structural, Physical, and Ches 
cal properties cf Calcified Tissues by Electron 
Micrr3Copy, Electron Eifiivrction r.nd X~ray Mi- 
cros r:jpy., 

Principal Investigator ; Dr. 0. Be Scott 

Other Investigators: Dr» M. U. Hylen 

Cooperating Units: Kyckcff , NIAMD, laboratory of 

ysical Biology, 

Man Yearr (calendar : sar 195i 
Total; 3 1 

Profestiimal: 1 
Other: 2 1 

Project Description: 

Objective s 

a. Determination of the crystal structure of enamel by 
electron microscopy and electron diffraction,, 

b Investigation of the reaction of various chemical agentB, 
such as fluoride compounds, <?\ too :h surfaces, with em 
phasis on th< mechanisms by which icid-solubility of 
enamel can be altered „ 

Co Davelopmen: of techniques for the application of contact 
and projeccicn X-ray microscopy la studies of hard and 
soft tissues r 

H Bthods L g loy a 1 : 

For electron m c oscopic studies of crystal structure, siiad- 
owed positive and negative replicas and pseudorepllcao are 
made from the 3u- faces of fully calcified enamel and dentine 
To reveal crystal outlines* samples are treated, prior to 
replication, with various acids for light etching, or with 
solvents for removal of organic components o ^aeudorepllcaa 

Part A. (Continue Serial No. H] 

containing actual crystallites, dried suspensions of com- 
minuted tissue, -tid specially prepared surfaces of' solid 
enamel are atudi.d by various electron diffraction tat 
niqusso Similar methods are used in investigating the 
changes in enamel structure and composition which result 
from various chemical treatments » In X-ray microscopy 
work primarily concerns technical aspects of soft and hard 
tlnsue preparation, effectiveness of types and conditions 
of radiation, and possible applications of both contact and 
projection equipment. These latter projects are carried 
out in close collaboration with Dr. Wyckoff's group (KIAHD> , 

Major Findings ; 

A. Further information has been gained about the morpho- 
logical characteristics and orientation of the crystallites 
in fully calcified enamel . The most interesting findings 
concern enamel pre- treated with ethylene diamine and thio- 
glycol late solutions. When organic matter was thus removed 
the remaining inorganic apatite, which could be identified 
by diffraction, ften appeared in the form of chains of 
very small particles. The observations suggest that organic 
matter may actually be incorporated within the long ribbon™ 
like crystals ordinarily found in intact mature enamel. By 
actual direct and indirect measurement, the average particle 
sise Is on the order of 300A. Further work with electron 
diffraction has indicated that the crystallites are oriented 
in a regular pattern, with major exes parallel to the prism 

3. Studies of tee effects of fluorides on the acid-solu- 
bility of the enamel surface have indicated that a much 
greater protection is afforded by the stannous than by the 
sodium compound. Experiments, aimed at ascertaining the 
mechanism of the protective reaction, have been continued, 
the most interesting being those in which it was found that 
stannous compounds not containing fluoride also reduce enamel 
solubility very effectively. These results have led to in° 
vestigations of 'he reaction of tin itself with enamel a It 
has been found that treatment with aqueous solutions of all 
tin compounds tested results in formation of a surface layer 
of hydrous hydrated stannous oxide. This layer is highly 
acid and water resistant, accounting for the previously 
observed protection, but it can be damaged or removed in 
varying degrees by mechanical means, such as brushing,, It 

Part A. (Continued) Serial Ho. nwz 

- 3 - 

has been found further that brushing for short periods with 
a very fine abrasive removes the protective layer complete 
lye Tests with fluoride-containing dentifrices have indi- 
cated that «hen enamel is treated by immersion in prepare 
tiona containing either the sodium or tin salts a pronounced 
protection against acid-etching is afforded,, However, no 
effect has been noted when the dentifrices were applied by 
brushing. This suggests a paradox in formation and simul- 
taneous removal of the surface layer = 

Co The principal results of work with X-ray microscopes 
concern technical development, The main effort has been 
applied in the direction of devising methods for specimen 
preparations and in determining the applicability of ad- 
vanced radiographic methods (developed by HIAMD) to studies 
of developing and mature calcified tissues. 

Significance to Dental Resear ch; 

The exact structure of the mineral component of calcified 
tissues is still relatively unknown. Although both the in= 
organic and organic structures are submicroscoplc in their 
dimensions, more has been learned about the latter through 
electron microscopy than about the former. The present work 
should help fill in some of the facts needed for an under- 
standing of the total composition of the enamel. Further 
studies should provide similar information about dentin and 

Since fluoride compounds are of inmediate interest In the 
clinical control of dental caries, the value of information 
on basic reactions with enamel on tooth surfaces is self- 
evident o X-ray microscopes are still in the early stages 
of technical development, and It is of Importance to investi- 
gate their applicability in problems of interest in dental 
histology and pathology. Information of general use should 
be forthcoming from this project and others being carried on 
in this laboratory. 

Proposed Course of Project ; 

Continuation of these studies along similar lines „ 

Part B. included; Yes 


r-d Tissue 


jponcnt of Dental Enamel. 

Pre / (accepted 

chlo projGc; : 

Serial No„ NIDR-3 

1. Histology & Pathology 

2, Bechesda, Md. 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Studies of the Structure of Developing Calcified 
Tissues by Electron Microscopy and X-ray Micros 

Principal Investigator: Dr c M. D. Nylen 

Other Investigators: Dr. D. Bo Scott 

Cooperating Units: Nona 

Man Years (calendar year 1958): 
Totals 3 
Professional: 1 
Other: 2 

Project Description: 

Objectives : 

a. Study of the suhmicroscopic morphology of the develop- 
ing enamel region including the enamel- forming cells 
and the organic and inorganic components of enamel. 

b. Investigation into the structural changes which take 
place in turkey leg tendons during initial calcific 
cation e 

Methods Employed : 

Specimens fixed, dehydrated and embedded by special methods. 
Thin sections are cut on a modified Spencer microtome or a 
Porter microtome and studied in the electron microscope. 
Thicker sections for X-ray microscopy are cut from identi- 
cal blocks on a standard Spencer microtome and are mounted 
directly on the photographic emulsion. 

Amelogenesis. Prior to enamel formation, the ameloblasts 
inner enamel epithelium) change from cuboidal cells to 

Part A. (Continued) Serial Ho. NJEDR-3 

- 2 - 

tall columnar cells Which are lined tip along the first 
formed dentin- With the beginning of enamel matrix elabo* 
ration there is a further differentiation as evidenced by 
an increase in amount and organization of the cytological 
components o Thus the functional ameloblast can be divided 
into 6 different areas on the basis of cytological charac 
teristics.; 1. the supranuclear or basal area where all 
the mitochondria are located, 2. the nucleus , 3. the 
subnuclear concentration of endoplasmic reticulum, 4. the 
Golgi area, 5. the distal concentration of endoplasmic 
reticulum and 6. Tomes 1 process where only secretory gran- 
ules are present . Of these various areas the Golgi zone 
occupies the major part of the cell and is perhaps the one 
of particular interest because of possible connection be- 
tween the organelle and cellular secretory activity. In 
the present work there is some evidence that secretory 
granules may form within the Golgi vacuoles o Observations 
so far seem to indicate that enamel matrix formation takes 
place in two distinctly different steps. The first step 
is the elaboration of an Intercellular or interred sub" 
stance which is separated from the cells by distinct cell 
membranes - The part of the cell which is surrounded by 
the interred matrix is the Tomes' process, and in the second 
step this part of the cell is gradually filled up with ma" 
trix substance while new interred substance is laid down 
on top of the old. Selected areas of this newly formed 
matrix were subjected to electron diffraction, and it ap- 
pears that small amounts of crystalline apatite are present 
in the interrod matrix from when it is first laid down while 
the initial rod matrix is devoid of crystalline material. 

b. A study of the calcifying turkey leg tendon was under- 
taken in the hope that it would increase our understanding 
of the morphological collagen-mineral relationship, as well 
as provide a clue to why some collagens as in bone and den- 
tin calcify while others remain uncalcified. In contrast 
to' the Intricate network found in dentin and bone, these 
tendons are characterized by a simple parallel arrangement 
of the collagen fibrils. Furthermore, the mineral incorpo- 
rated in this matrix is the same (hydroxyapatite) as in 
the other more complex calcified tissues. An initial part 
of the study was an Investigation into the effect on the 
fibril structure of various fixatives. Thus in osmic acid 

Part A. (Continued) Serial No s WIDR-3 

- 3 - 

fixed material it was found that the collagen fibrils 
appeared as individual entities while they remained in 
close contact to form sheet like structures in material 
fixed in chilled acetone followed by chilled ethylene 
chloride. It is possible that the sheets represent a 
truer picture of the actual state of the tendon collagen 
than the array of individual fibrils -*ich may have been 
torn apart during fixation. The changes accompanying 
calcification were best observed in specimens which con- 
tained both calcified and uncalcified areas Prior to 
calcification the flattened fibroblasts change into 
cuboldal eel Is o The collagen fibers, in some instances, 
appear to thicken while in other instances there is an 
actual breakdown and reorganization of the f iber- struc- 
ture c A sharp transition between calcified and uncalci- 
fied areas is particularly evident under the X-ray micro- 
scope, whereas under the electron microscope the calcified 
areas consist of a mixture of both calcified and uncalci- 
fied tendon. Crystallisation appears to begin in certain 
localized regions and extends laterally with the first 
crystals being positioned on the collagen cross bands. 
Using selected area electron diffraction it is possible 
to identify the crystals as apatite oriented with their 
c-axis parallel to the fibril direction. 

Significance to Dental Research : 

An increased knowledge of the origin and ultrastructure 
of the mineralized tissues is of importance for the under- 
standing of changes in the dental hard tissues which may 
take place as the result of systemic disorders during de- 
velopment, or thru post-eruptive pathologic involvement. 

Proposed Course of Project ; 

Continuation of current program and initiation of additional 
studies of developing dental tissues from animals subjected 
to systemic disorders of either nutritional, toxic, or in- 
fectious origin o 

Part Bo included; Yes 

- 4 » Serial Ho. KIDR°3 


Individual Project Report 

Calendar Year 1958 

Part B : Honors, Awards, and Publications 

Publications other than abstracts from this project . 

Hylen, Mo U« and Scott, D. B., An Electron Microscopic Study 
of the Early Stages of Dentinogenesis (Public Health Service 
Publication Ho, 613), Washington, UoS. Government Printing 
Office, 1958. 

Scott, D. Bo, Hylen, Mo Uo, and Takuma, S., Electron Micros- 
copy of Developing and Mature Calcified Tissues , Revue Beige 
de Science dentalre (accepted for publication). 

Honors and Awards relating to this project: 
Hot • 

Serial No. H1DR-4 
1. Histology & Pathology 
2o Bethesda. Md. 


Individual Project Report 

Calendar Tear 1958 

Part A. 

Project Title; Electron Microscopic Studies of the Ultra- 
structure of Developing and Mature Bone., 

Principal Investigator: Dr. S. Takuma 

Other Investigators: None 

Cooperating Units: None 

Man Years (calendar year 1958): 
Total: 11/3 
Professional: 1 
Other: 1/3 

Project Description: 

Objectives : 

a. Observation of normal osteogenesis of endochondral 

and periosteal bone, 
bo Observation of the subnicroscoplc structure of mature 


Methods Employed ; 

The specimens used were developing phalangeal bones from 
mice ranging in age from 1 to 8 days<> The bonee were fix- 
ed with a mixture of osmium tetroxide and potassium dichro- 
mate, and were embedded in a mefchacrylate , Microtomy was 
carried out with either a Porter-Blum microtome or a modi- 
fied Spencer microtome <> In certain instances the methacry- 
late was dissolved and the sections were shadowed with 
tungsten oxide . Some sections, following removal of the 
methacrylate, were decalcified in place on the specimen 
screens by immersion for 30 minutes in 2% hydrochloric 

Part A. ((Continued Serial No=> NIDR-4 

Major Findings; 

I. Cartilage 

Chondrc r.ytes: The stain organelles in the cytoplasm of 
the chondrocyte are endoplasmic reticulum, mitochondria, 
and microsomal granules^. Localized dilatations of the 
endoplasmic reticulum are observed which resemble thr 
globular bodies described in the odontoblasts by Nylen 
and Scor Co 

In the < y topi asm of the chondrocyte in the prolifera- 
tive zoi4, there can be seen numerous vacuoles of various 
sites which contain matter of low density. The remainder 
of the c |' topi asm is taken up by closely packed endoplas- 
mic retl :ulua, mitochondria, and microsomal granules. 
The first', ltdlcation of retrograde changes in the. chondro- 
cytes sevtas to occur as a dispersion of the cytoplasmic 
organelle tio This change is followed by destruction of 
the organ il .e* Deterioration of the mitochondria appears 
to begin / cer the endoplasmic reticulum has started to 
change, lie limiting membrane seems to be the last por- 
tion of thi mitochondria to be lost. 

Cartilage liatrix: The fibrils in the cartilage matrix 
are arrant d at random in the form of a fine networks 
It is dif cult to observe a well defined periodicity 
in the fill lis although some fibrils with a 220A stri- 
atlon are • cc asternal ly found* 

In the da: ( nerative cone there is a remarkable change 
in the ch : setter of the matrix, which takes on a solidi- 
fied appe : ance. This solidified matrix can be subd 
vlded int two different morphological types. In one 
type the teratlon seems to have been brought about by 
investment of the matrix fibrils with an amorphous 01 
granular > i o stance. There is no indication of the pre- 
sence of n I aeral in the altered regions which can be 
detected t; electron diffraction. 

The other I ype of dense cartilage seems to develop as 
the resul of accumulation of thread* like filaments, 
each of w! . ch is outlined by a thin electron dense 
border. I i such changed areas the presence of crystal- 
line apat I e can be detected through electron diffraction. 

Part A. (Continued) Serial No. HXD&-4 

- 3 - 

When this mineral- containing type of solidified matrix is 
detainer alised on the speciaen screen, it becomes similar 
in structure to the undecalcifled type of solidified ma- 

On the basis of these findings it may be that maturat ion 
of the cartilage matrix proceeds through at least three 
stages, which may be tentatively called fibrillar, solidi- 
fication, and mineralisation pbaseso If this should prove 
to be the case, the sequence parallels that which hat) been 
shown in the development of dentin by Hylen and Scott:. 

II. Bone 

The first indication of bone formation is an accumulation 
of collagen fibrils in the area between the cartilage ma* 
trix and the osteoblasts. In the next stage the collagen 
fibrils become covered with an unidentified amorphoun 
substance. Through the use of selected area electron 
diffraction, it is sometimes possible to detect the pre- 
sence of crystallites in such amorphous structures. At 
higher magnification thread-like filaments outlined by a 
thin dense border, as in the calcified cartilage matrix, 
are revealed In resorption the bone matrix loses it* 
high density and changes into a somewhat porous structure. 
It appears as if the Investing substance is removed first, 
leaving a network of fine matrix fibrils behind. 

Significance to Dental Research ; 

This study is essential to a further understanding of the 
mechanism of calcification, aa well as to clarification 
of the process of bone development Itself. 

Proposed Course of Project : 

a, Continuation of the present study. 

b, Application of replica methods to mature bone. 

c, Further extension into the study of abnormal osteo» 

Part B. included: Yes 

- 4 - Serial No, H IPR-4 


Individual Project Report 
Calendar Year 1958 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts frosi this project. 

Scott, D. Bo, Hvlen, M. D. ( and Takuna, S. Electron Micros- 
copy of Developing and Mature Calcified Tissues. Revue Beige 
de Science dentaire (accepted for publication). 

Honors and Awards relating to this project: 

Serial No. KjgR°5 

1. Histology & Pathc? 

2. Bethesda, Hdc 


Individual Project Report 

Calendar Year 1938 

Part A. 

Project Title: Diagnostic Surgical Pathology Utilising Histo- 
chemical Techniques . 

Principal Investigator: Dr. Mo S. Buretone 

Other Investigators: Hone 

Cooperating Units: Kone 

Han Tears (calendar year 1958): 
Total; 11/3 

Professional: 2/3 
Other: 2/3 

Project Description: 

Objectives : 

The object of this project is to apply existing and newly 
developed histochenical techniques to the study of fr< 
removed huuan surgical specimens, with an emphasis upon 
cancers. This type of study reveals tissue changes «l 
are not visible with routine stains and may contribute to 
the diagnosis of tumors as well an to the elucidation of 
the pathogenesis of neoplasia. 

Methods Employed : 

For the last four years a systematic histochemical study 
haa been made in this laboratory of practically every 
human neoplasm removed in the Clinical Center. To date, 
specimens from over 300 surgical cases have been processed. 
These include patients scheduled £or general and dental 
surgery and also selected cases to be admitted to the 
Clinical Center specifically for this 3tudy. Normal 
tissues such as salivary glands and oral epithelia have 
also been studied. 

Part a. (Continued) Serial No, HIDR-5 

- 2 - 

Major Finding s: 

lc The histochemical finding of aminopeptidase activity 
in the connective tiocue stroma adjacent to human ncc 
plasms has been confirmed in continued studies. This 
finding has also been confirmed in several foreign labor" 
stories; (BraucFalco, Germany) .Sylvan, Sweden). 

2. High acid phosphatase activity has been correlated 
with dyskeratotlc cells in human epidermoid carcinoma, 
This finding may have diagnostic significance. The new 
re '.table histochomlcal technique used was developed in 
this laboratory. It employs new chemical substrates 
synthesized by the principal investigator. The older 
ac'.d phosphatase techniques, by contrast, have been con- 
sidered Co be highly unreliable. 

3. New techniques for the demonstration of cytochrome 
oxidase have been developed. These methods should have 
wide application in the study of oxidative systems in 
pathological oral tissues. Salivary gland ducts, for 
example, show high cytochrome oxidase activity. This is 
a new finding and should contribute to our understanding 
of salivary gland function in health and disease. 

4. Application of alkaline phosphatase techniques de~ 
veloped in this laboratory have proved of va'.ue in study- 
ing the phosphatase activity of leukemic cells. This is 
an important clinical tool. 

Significance to Dental Research ; 

The application of histochemical techniques to investi- 
gations of neoplastic tissues will provide considerable 
new information about the pathological alterations and 
processes involved, which until recently were only studied 
on a morphologic basis. The development of standardised 
methods enables histochemical techniques to be utilized 
in various disciplines. 

Pgoposed Course of Project ; 

It would be desirable to expand this program at the pro- 
fessional level so as to increase the scope of the histo- 
chemical approach, and to utilize more fully, at the 

Part A. (Continued) Serial No. KB)R"5 

technical level, special procedures for diagnosis «. Enzyme 
techniques designed for use with specially fixed frozen 
sections are being planned and developed in order to facili- 
tate rapid routine diagnosis of surgical specimens without 
the use of coo? lea equipment for freezing and drying B 

Part B. included: Yes 

- 4 <* Serial Ho. KXDR-5 


Individual Project Report 

Calendar Year 1958 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts frou this project. 

Burstone, Ho So, Histocheaical Comparison of Haphthol AS-phos- 
phates £o? the Demonstration of Phosphatases Jo Hato Cancer 
Inst. 20;601°615, 1958. 

Burstone, M. So, Histocheaical Demonstration of Acid Phospha- 
tase Using Haphthol AS -phosphates, J. Hato Cancer Inst. 21; 
523-539, 1958. 

Honors and Awards relating to this project: 

Serial No, MXDR-6 
lo Histology & Pathology 
2. Bethesda, Hd 


Individual Project Report 

Calendar Year 1938 

Part A. 

Project Title: Development and Application of Histochemical 

Methods In the Study of Hard and Soft Tissues., 

Principle Investigator: Dr<> Mo So Bur stone 

Other Investigators: Dr. G. Glenner 

Cooperating Unito: Dr. G. Glenner, N1AMD, Laboratory of Histo- 
chemistry and Pathology . 

Man Years < calendar year 1958): 
Total: 2 3 
Professional: 1/3 

Project Description: 

Objectives : 

The object of this project is to develop new histochemical 
procedures and modify existing ones for the microscopic 
characterization of hard and soft tissues . 

Methods Em ployed : 

Histochemical methods have been employed for demonstration 
of enzymes o The latter Include esterases , peptidases t 
phosphatases, sulfatases, and glucuronidases . For the 
most part azo-dye procedures have been utilized. The cub- 
strates consist of esters which release potentially chro» 
oogenic naphthols upon enzymatic hydrolysis. In addition 
to applying existing techniques, new substrates have been, 
and are being, synthesized in this laboratory . A series of 
new methods for the demonstration of oxidases are also being 
utilized. The oxidase methods employ new naphthol and 
methylene compound substrates which form highly substantive 
(dye fastness) indbamline and azamel trine dyes, and thus 
permit more accurate microscopic localizations. 

Part A. (Continued) Serial No. HH)3i6 

- 2 - 

The use of frozen dried tissue makes it possible In many 
instances to utilize raany poorly hydrolysed substrates 
which would be unsatisfactory with conventionally fixed 

A. A new finding, namely the presence of high acid phos- 
phatase activity of osteoclasts associated with physio- 
logical bone resorption, has been studied during the lost 
year. This enzyme appears to be the only one which ban 
ever been correctly assigned to osteoclasts. 

B. A hitherto undescribed property of calcifying bone and 
dentin matrices, namely, their ability to hydrolyze certain 
fatty acid esters, has been observed in several species. 
The enzyme was found to be inhibited "in vivo" by toxic 
doses of fluoride. 

C. Comparative histoenzymo logical studies in necturus 
maculo8us (with Dr. G< Glenner) have shown a striking dis- 
similarity of enzyme patterns in organs of this animal as 
compared with those of other species. This suggests that 
considerable caution be esployed in conclusions based 
upon comparative physiological studies. 

Significance to Dental Research ; 

Since normal and abnormal hard and soft tissues are probably 
characterized by basic protein and enzyme differences, newer 
methods and modifications for demonstrating these differ- 
ences are of use in relating morphology of various tissues 
to their chemical composition. 

Proposed Course of Project ; 

Present investigations will be continued and extended. In- 
cluded are studies on the effect of various hormone ele- 
ments upon the protein and enzyme characteristics of develop- 
ing bone. 

Part B. Included; Yes 

- 3 - Serial No N1DR-6 


Individual Project Report 

Calendar Year 1956 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts from this project. 

Bur a t one r Mo So and Keyes, P, H. The Effect of Inhibition of 
Enzyme Activity on Developing Bone and Dentin. Am. J. of Path. 
33:1229-1235, 1957, 

Glenner, G. G. and Burstone, M. S. Esterase and Phosphatase 
Activity in Necturus Haculosus. A. Study in Comparative 
Hlstoenzymology. Anat. Rec, 130:243-252, February, 1958. 

Burstone, M. S. The Relationship Between Fixation and Tech 
niques for the Hlstochemlcal Localization of Hydrolytic En- 
zymes. J, Hlstochem. and Cytochem 6:322-339, September, 1958. 

Honors and Awards relating to this project: 

Serial No. N3DR-7 
lo Histology & Pathology 
2. Bethesda, Md. 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Histochealcal Studies of Connective Tissues » 

Principal Investigator: Dr» Ho Mo Fullmer 

Other Investigators: Hone 

Cooperating Units: None 

Man Years (calendar year 1958): 
Total: 3 
Professional: 1 
Other: 2 

Project Description: 

Objectives : 

To determine the chemical composition and the reactive 
groups on normal and diseased connective tissues through 
the use of hlstochemical methods • 

Methods Employed : 

Hunan and animal tissues are obtained from autopsy and 
surgical material o Specimens to be sectioned are embedded 
either after freeze drying or after chemical fixation and 
dehydration o Under controlled conditions, the sections 
are exposed to various reagents designed to determine the 
site and quantity of a reactive group or substance o 

Major Findings : 

1< A new stain, the per acetic-aldehyde fuchsin°Halml 
stain, was developed which led to the discovery of a new 
connective tissue fiber = This has been designated the 
oxytalan fiber and is located principally in areas of 
stress such as the periodontal membranes, ligaments, 
tendons, the adventitia of blood vessels, eplneurium and 

Part A. (Centime Serial No„ HH>3 

- 2 - 

perineurium, and in the connective tissue sheaths surround- 
ing the hair follicles and sebaceous glands of the skin„ 
To date, the fibers have been found in man and in sev 
animals o The fibers have protein and mucopolysaccharide 
components o During the course of studies of the embryology 
cal development of the fibers, it was found that the! 
velopment was associated with mucopolysaccharide production 
and maturation* The histc chemical reactions of oxy talon 
fibers appears to indicate a relation to elastic fibers o 

In addition, this stain has for the first time permitted 
a distinction to be made between Reed* Sternberg cells In 
volved in Hodgkln's disease, and megakaryocytes o Consider- 
able value may thus be related to its use in establishing 
a diagnosis of Hodgkin's disease, Since oxytalan fibers 
are found in the epineurium and perineurium, it is possible 
that the stain would be of value in differentiating neuro 
fibromas from fibroma* =. To date, we have examined several 
flbromas and one neurofibroma, and found a considerable 
difference in staining characteristics of these two types 
of lesions The stain has also been found to be useful in 
the study of pigment in aging cells of the brain, 

2o The development of another new stain, the per ace tic 
orcein -Halmi stain, has resulted in the demonstration of 
an elastic like component of mucous connective tissues 
It is believed that the substance reactive with the pera 
ceric=orcein-Halmi stain is sometimes the same as that 
reactive with the peracetlc~aldehyde fuchsln-Halml stain, 
However, the peracetic°orcein~Halmi stain is more restricted 
in Its reactivity; i,e it reacts with fewer types of muco- 
polysaccharides, and, aa a result, demonstrates a previously 
undescrlbed fibril in mucous connective tissues (in the 
pulps of teeth, in umbilical cord and in the rooster combK 
The development and use of the two stains described above 
has aided considerably in the development of a unified con- 
cept of connective tissues This has permitted the obser- 
vation that tendon and mucous connective tissue cells, 
just as connective tissue cells generally B produce an 
elastic-like component, and that the nature of this product 
varies with the anatomic site and function. 

A relation during development between one connective tissue 
product (mucopolysaccharide) and another connective tissue 
product ([elastic-like fibers) was also demonstrated* 

Part Ac (Continued) Serial No NIDR-7 

- 3 - 

3c Histochemical reactions for proteins were carried out 
for the first time on the jaws of human embryos and chil- 
dren at various ages and development Principle findings 
were (a) the histochemical demonstration of protein in 
pre- enamel and enamel; and (b) the demonstration that 
changes in the reactivity in proteins and mucopolysaccha- 
rides takes place in ameloblasts and odontoblasts coinci- 
dent with cytodifferentiation and functional activity* 

Significance to Dental Research ; 

Periodontitis and periodontosis are diseases of connec- 
tive tissueso It is believed that information gained 
about the nature of normal connective tissues and their 
alterations in the diseased states will contribute to a 
better basic understanding of the pathological processes 
involved in periodontal disease „ The ultimate Importance 
of the discovery of a new connective tissue fiber in the 
periodontal tissues, and in other tissues subjected to 
stress, is still unknown. However, there is some indi- 
cation (not yet fully demonstrated) that oxytalan fibers 
are more resistant than collagen to destruction by the 
lesion of periodontitis. 

The finding of changes in protein and mucopolysaccharide 
reactivity in cells coincident with cytodifferentiation 
and function may give some insight into the complicated 
mechanisms of cellular function? 

Proposed Course of Project ; 

Continued attention will be given to the histochemical 
study of normal and diseased connective tissues and their 
alterations during development and with age. Time will 
also be devoted to the further exploration of the chemical 
composition, function and distribution of the newly found 

Part Bo included; Yes 

Serial No. NIDR-7 


Individual Project Report 

Calendar Year 1938 

Part_B: Honor s„ Awards, and Publications 

Publications other than abstracts from this project., 

Pullmer, Harold Ho Histochetaical Protein Reactions in Hunan 
Developing Teeth. Labo Inves 7:48-51, 1958c 

Fullner, Harold M» and Alpher, Norman. Histocheaical Poly- 
saccharide Reactions in Human Developing Teeth. Lab, Inves „ 
71163-170, 1958c 

Pulliaer, Harold M. Differential Staining of Connective Tissue 
Fibers in Areas of Stress. Science 127:1240, 1958. 

Fullmer, Harold M. and Lillie, R» D. The Oxytalan Fiber: A 
Previously Undescrlbed Connective Tissue Fiber. J. Hlstochenu 
& Cytochea. 6:425-430, 1958, 


Fullmer, Harold M, The Peracetic-Orcein-Halmi Stain: A Stain 
for Connective Tissues. Stain Technol. (in press). 

Honors and Awards related to this project: 

Serial No. NIDR-8 
lc Histology & Pathology 
2o Bethesda, Md, 

Individual Project Report 
Calendar Year 1958 

Part Ac 

Project Title: Experimental Animal caries. 

Principal Investigator: Dr. P« W. «e/es 

Other Investigators: None 

Cooperating Units; None 

Man Years (calendar year 1958): 
Total: 3 1/3 
Professional; 1 
Other: 2 1/3 

Project Description: 

Objectives : 

a. To study the transmloGable nature of experimental 
dental caries and procedures for influencing the 

b, To develop technics for inoculating a "carlogenlc" 
flora into animals whose caries- inducing flora in 
absent or artificially depressed* 

Methods Sap loved ; 

A. Dental caries activity has been induced in hamsters 
by feeding a high-carbohydrate low-fat diet and by inocu- 
lation of fecal material from animals which have active 
caries o When this inoculum has been added to the water 
supply or smeared into the mouths of smimale which do 
not carry the "carlogenlc" flora, active carles has follow- 
ed * 

Bo It has also been possible to transmit the flora from 
one animal to another by placing active and inactive anl~ 
nkils in the same cage > This continues to be an easy and 
effective method of contamination, as previously reported. 

Part A, (Continued) Serial No„ NH>R°8 

Hamsters have been fed a pre-experlmental diet containing 
erythromycin to assess possible methods for depressing 
the "cariogenic" flora prior to introduction of various 
bacterial inoculums » 

In rats, penicillin has been fed to the highly susceptible 
Osborne -Mendel strain during the suckling period to de- 
termine the subsequent effect on caries activity* Rats 
raised on this antibiotic diet have been exposed to non° 
treated animals to determine if their caries activity 
would be al tared » 

Inasmuch as black rats have been leas susceptible to caries 
than Osborne -Mendel rats when fed the same diet for equal 
periods of time, breeding experiments are in progress to 
determine if this difference may be related to genetic 
factors o 

A. In newly wemed Syrian hamsters, fed a high- carbohydrate 
low-fat diet, dental caries can be induced by inoculation 
of fecal material added to the drinking water or smeared 
into the mouth 

Caries-inactive hamsters sent from the University of Ala- 
bama have developed carious lesions after 14 days of ex- 
posure to active animals of our colony „ These animals 
usually require de salivation in order to become caries 
active. In fact llttermates kept at Alabama and fed a 
diet similar to that used in this laboratory remained caries 
fir— . 

Inactive hamsters made active by inoculation will produce 
susceptible litters <. Also a small group of hamsters made 
inactive by means of penicillin have produced four genera- 
tions of Inactive animals without additional use of the 
drugo Difficulty has been experienced with hamsters fed 
erythromycin prior to commencement of the experimental dieto 
Animals so treated have developed serious and generally 
fatal diarrhea The reasons for this complication are not 
known but may be related to a parasitic flora among other 
factor So Efforts are being made to overcome this diffi- 

Pert A. (Continued) Serial Bo. NXDR-8 

3 - 

Bo In Osborne-Mendel race the pattern and degree of carle. 

S£«?« 8 < UerCd *° d rCduCed by feedin * ™<*li»S animals 
penicillin prior to commencement of the experimental diet. 
The pattern of caries and the degree of activity can be 
almost restored to usual levels by exposing the penicillin 
tre.ted animals to untreated animals, an observation indi- 
cating a measure of transmissabillty between animals of 
the same "strain" . By mating female Osborne-Mendel rats 
simultaneously to CM ("susceptible") and Black ("resistant") 
rats it is possible to tag the fathers of the offspring 
and to Jceep the maternal and suckling environments identi- 
cal c With back-crosses to this semi-hybrid generation it 
nay be possible to determine if the difference in caries 
activity between the two strains is influenced by genetic 

Significance to Dental Resea rch; 

The observation in hamsters that dental caries can be inocu 
lated under certain conditions, makes possible studies with 
specific microorganisms. Findings in Osborne -Mendel rats 
suggest that within a given strain it is possible to trans- 
mit the flora from an animal which has it to one tbat does 
not . 

These observations open for reconsideration many Interpre- 
tations of results in the field of experimental animal caries 
and may possibly explain a number of seemingly inconsistent 
bindings reported over the past 35 years . 

Proposed Course of Project ; 

A. Additional work is needed to establish the validity of 
the above theory. This is expected to entail cooperation 
with investigators in the Laboratory of Microbiology,, 

B. Additional work is in progress with regard to genetic 
factors which may influence the caries pattern in the 
Osborne- Mendel and Black rats. 

Pert B. included: Yes 

- 4 - Serial Ho, HH« 8 


Individual Project Report 

Calendar Year 1958 

Part B : Honors , Awards, and Publications 

Publications other than abstracts from this project, 

Keyes, Paul Ho Dental Carles in the Molar Teeth of Rats> 
I. Distribution of lesions induced by high-carbohydrate 
lov-fat diets. J, Dent. Res., 37:1077, 1958. 

Keyes, Paul H. Dental Carles in the Molar Teeth of Rats, 
lie A method for diagnosing and scoring several types of 
lesions simultaneously, Jo Dento Res<> 37:1088, 1958 „ 

Reyes. Paul R. and White, Carl L. Dental Carles in the 
Molar Teeth of Rats. III. A bio-assay of sodium fluoride 
and sodium lauroyl sarcosinate as caries inhibitory agents. 
(Accepted for publication in J. Am. Dent? Assoc.) 

Keyes, Paul H. Dental Caries in the Syrian Hamster. 
VIII. The induction of rampant caries activity in albino 
and golden animals- (Accepted for publication in J. D. 

Honors and Awards relating to this project: 


Individual Project Reports 
Calendar Year 1958 


<i " R - J- Fitzgerald and H. V. Jordan : The Use of Germ-free 
Animals In the Study of Problems Relating to Oral Disease. 

1°- S. E. Mergenhogen: Mucopolysaccharase Production by Oral 
Bacteria in Relation to Periodontal Disease. 

U« H. W. Sc herp and S. E. Mergenhagen : Digestion of Native and 
Altered Collagens by Oral Microorganisms in Relation to 
Periodontal Disease. 

12. H. W. Scherp: Manifest and Inapparent Viral Infections of 
Oral Tissues. 

13 - A. Howell, Jr. : The Relationship of Pathogenic Fungi to 
Diseases of the Oral Cavity. I. Incidence and Identity. 

1^« A. Howell. Jr. : A Study of the Genera Actinomyces and 


H. V. Jor dan and R. J. Fitzgerald : Inhibitors and Dental 

16. T. Shlota : Microbiology of Saliva. 

17. T. Shiota: Nutrition and Metabolism of Lactic Aeid Bacteria. 

18. R. R. Qmata: Studies on the Microorganisms Associated with 
Periodontal Diseases. 

19. B. G. Hampp: Investigations in the Field of the Spirochetes. 

20 • T. A. Ne vin and E. G. Hampp ; Studies of Interactions between 
Oral Microorganisms. 

21. W. D. McBrlde: Studies on the Host Cell-Virus Relationship 
with Special Reference to Latent Infections. 

22. M. Rogosa: Immunological, Biochemical, and Pathological 
Studies of Oral Bacteria. 

23. A. A. Rizzo; The Rat as an Experimental Animal for Research 
on Periodontal Disease. 

La ' of yictpbtolc 
Estimated Obligations for P.Y 1959 



Rain bur 5 assents 

Project Numbers 
SIDE -9 
HIDR 11 
HIDR 14 
HIDR- 17 
HIDR -20 
HIDR -22 
NIDR 23 

$ 93,700 

Serial No, NIDR-9 
1. Microbiology 
3, Bethesda, Md 

Individual Project Report 
Calendar Year 19 58 

Fart A 

Project Title: The Use of Germ-Free Animals in the Study of 
Problems Relating to Oral Disease. 

Principal Investigators: Dr. Ro J. Fitzgerald 

Dr. H. V. Jordan 

Other Investigators: Dr. H. R, Stanley, Jr. 

Cooperating Units: Germ-Free Unit, Sanitary Engineering 
Branch, Division of Research Services 
Lobund Institute, University of Notre 

Man Years (calendar year 1958): 
Total: 2 
Professional: 1 
Other: 1 

Project Description 

Objectives : 

To use germ- free animals in the study of the etiology of 
dental carles, calculus formation and periodontal disease. 

Methods Employed : 

All three of the commercially available germ-free systems 
(Reyniers, Gustafsson and Trexler) have been employed in 
our studies One group of germ- free rats is provided with 
a sterilised cariogenic diet (i.e., cariogenlc for conven- 
tional animals) and remains germ- free. A second group is 
reared under the same conditions except that pure strains 
of selected oral microorganisms are introduced into the 
oral cavity of the animals to determine if it is possible 
to induce some oral pathological condition. 

- 2 - Serial No. HIDR-9 

Part A (continued) 

Major Findingt : 

1. It has been possible to confirm the original observations 
of Orland that caries does not occur in germ- free rats even 
when they are maintained on a cariogenic diet. The diet used 
was a modification of NIDR coarse-corn, high-sugar diet 585 
and differed in composition from that employed by Orlando 

2 We have been able to induce extensive tooth decay in 
germ- free rats on this diet when they were mono infected 
orally with a tingle streptococcus of the mltis type 
This organism was isolated from the oral cavity of conven- 
tional rats maintained on cariogenic diet 3"S 

Orland previously had reported the production of caries in 
rats aonolnfected with enterococcl Our findings have, there- 
fore, added a second microorganism which is able to induce 
carles in the rat in the absence of other organisms, 

3. We have suggestive evidence that rat oral diphtheroid 
organism* may Initiate lesions of the enamel,, but these 
did not progress into the dentin within the normal experi- 
mental period. 

4. Histopathologic examination of the tissues of the germ- 
free animals by Dr. Stanley has revealed that a leucocyte 
response, epithelial hyperplasia, foreign body reaction, 
and occasionally ulceration may occur in the gingivae in 
conjunction with Impacted hairs. This indicates that at 
least some of the initial processes leading to periodontal 
disease may occur in the absence of a microbial factor. 

5o We have been able to select dietary and environmental 
conditions which have permitted us to breed and rear three 
generations of Lund (Gustafsson) rats germ- free at NIB. 
This means that we could become independent of an external 
supply of germ- free rats should the necessity arise. 

Significance to Dental Research : 

The use of germ- free animal technic i» potentially of great 
usefulness in the study of a number of oral diseases because 
by this means one can first dissociate the vast and complex 
oral flora from the tissue under study, and then reintroduce 

- 3 - Serial No. NIDR-9 

Part A (continued) 

known types of Rlcroorganlsma at will. Thus it should 
be possible to work out the precise etiology of conditions 
such as caries, calculus deposition, and periodontal 

Proposed Course of Project ; 

Future studies will attenpt to confirm and extend the 
observations reported herein. The principal deterrents 
to the progress of the project had been the scarcity of 
gem- free animals and the lack of apparatus for housing 
them. The former is no longer a factor; however, since 
NIDR still has only two germ-free tanks of its own, our 
full potential haa not yet been realized. 

Part B included Yes 

4 - Serial No NIDR-9 

Individual Project Report 
Calendar Year 1958 

Part B °. Honors, Awards, and Publications 

Publications otber than abstracts from this project: 


Honors and Awards relating to this project: 

1. Invited participant as a Specialist in Germ-Free 
Animal Symposium, VII International Congress of 
Microbiology, Stockholm, Sweden, August 1958. 

2, Awarded a National Science Foundation Travel Grant 

to attend the VII International Congress of Microbiology. 

Serial No. NIDR-10 

lo Microbiology 
3. Bethesda, Md. 


Individual project Report 

Calendar Year 1958 

Part A 

Project Title: Mucopolysaccharase Production by Oral Bacteria 
in Relation to Periodontal Disease 

principal Investigator: or. S. E. Mergenhagen 

Other inveatl gators: None 

Cooperating units: None 

Man Years (calendar year 1958): 
Total 1 1/6 
Professional: 1/2 
Other 2/3 

project Description: 

Objectives ; 

To determine the conditions governing the production of 
mucopolysaccharases and other enzymes, which probably 
operate in the pathogenesis of periodontal disease, by 
bacteria indigenous to the human oral cavity* and to analyze 
their action on host tissue. 

Methods Eaiploved: 

Present investigations are concerned mainly with the produc- 
tion by oral bacteria In vitro of extracellular and Intra- 
cellular enzymes capable of breaking down the essential 
mucopolysaccharides of the ground substance of connective 
tissue* special emphasis has been given to a study of the 
cultural conditions necessary to suppress or Increase 
selectively the synthesis of chondral tinase, hyaluronldase, 
or both enzymes by a mlcroaerophi 1 ic diphtheroid bacillus 
of oral origin, in addition, the effects of salt concentra- 
tion, pK, metallic ions, and reducing agents have been 
studied in relation to the activity of these polysaccharases 
in vitro . The action of these enzymes in vivo is being in- 
vestigated by studying the role they may play in synergistic 

- 2 - Serial no„ NIDR-10 

Part A (continued) 

intracutaneous infections with oral bacteria, by observations on 
their spreading factor activity in connective tissue, by their 

effecta on capillary permeability, and by their ability to release 

histamine like substances.. 

Both intracellular and extracellular chondroltinaae and hyaluronidase 
have been found to be produced by oral diphtheroid bacilli., By the 
addition of certain substances to the culture medium (glucose, 
reducing agents), extracellular chondroltinaae production can be 
suppressed, while hyaluronidase and intracellular chondroitinase 
production are unaffected,, In vitro action of hyaluronidase can 
be Increased 5-fold by the addition of Mg**» Mn or Ca** Ions to 
the enzyme substrste mixture, whereas chondroltinaae activity is not 
significantly altered by these additives, certain aulfhydryl- 
containing compounds (glutathione, hioglycollate) enhance chon- 
droltinaae activity JU\ vitro ; cysteine, however, had no effects 
Present results indicate that these enzymes potentiate the enhance 
ment and extension of experimental intracutaneous Infections produced 
by oral anaerobic streptococci and spirochetes, when injected Intra 
cutaneous ly in the vicinity of the lesions; and moreover, that a 
mixture of the diphtheroid bacilli with oral spirochetes acts 
synerglstlcally on intracutaneous Injection to produce a spreading 
necrotic infection. 

Significance to Dental Research ; 

Depolymerization of the mucopolysaccharide tissue cement substances, 
hyaluronic acid and chondroitin sulfuric acid, Is a prominent feature 
of periodontal disease, as seen in the disappearance of the intra- 
cellular substance from the crevicular epithelium and the vacuoliza- 
tion of the ground substance of the subjacent connective tissue of the 
gingivae. Therefore, investigations of the oral microbial sources 
of hyaluronidase and chondroltinaae and of the factors influencing 
their production and activity contribute directly to our understand- 
ing of this disease. 

Proposed course of Project ; 

To continue and expand the studies outlined above. (Project initiated 
1 September 1958). 

Part B included No 

Serial no. NIDR-11 

1. Microbiology 
3. Bethesda, Md. 

Individual project Report 
Calendar Year 1958 

Part A 

Project Title: Digestion of Native and Altered Collagens by oral 
Microorganism* in Relation to Periodontal Disease, 

Principal Investigators: Dr. H. W. scherp 

Dr. So E. Mergenhagen 

Other investigators: None 

Cooperating unit*: None 

Man years (calendar year 1958): 
Total: 1 2/3 
Professional; 1 
Other 2/3 

Project Description; 

Objectives ; 

1. To determine whether members of the oral mlcrobiota, singly 
or in combinations, can digest native collagen. 

2. To determine whether collagen altered in vivo , as by in- 
flammation, becomes more susceptible to digestion by the oral 

Methods Employed ; 

Animal skin was selected as a collagenous substrate resembling 
gingival tissue in essential features. Areas of inflammation 
are induced by four methods: a sterile (turpentine) abscess; 
infection, as by staphylococci and mixed infection with oral 
microorganisms; cutaneous application of irritants such as 
xylene and croton oil; and allergic reaction of sensitized 
animals to cutaneous application of a simple allergen, such as 
picryl chloride. Areas of normal and inflamed skin from each 
animal are defatted, dehydrated, and sterilized by immersion 
in acetone, samples are exposed to cultures in a special 
medium, inoculated with accumulations from human periodontal 

- 2 - Serial No. NIDR-11 

Part A (continued) 

pockets or organisms Isolated from such cultures. Residual 
tissue and microbial growth is removed by centrifugation. 
Collagenolysis is estimated by determination of hydroxypro line- 
containing peptides in the supernatants, compared to the 
hydroxypro line content of the undigested residues. 

Major Findings : 

Control tests of normal skin confirm previous findings that the 
readily cultivable oral bacteria do not produce true collagenases, 
though they attack promptly collagen that has been only slightly 
altered _i_n vitro by simple physical and chemical procedures 
discovered in our previous investigations. Similarly, the 
collagen of skin inflamed by xylene resisted digestion by 
mixed cultures of the oral flora. On the other hand, the 
collagen of samples of inflamed skin from the periphery of 
staphylococcal and turpentine abscesses had become definitely 
digestible by such cultures. The collagen of skin treated 
in vitro with testicular hyaluronidase, as a source of 
"spreading fsctor," also became digestible by the orsl flora. 
Recently we have discovered that various of the oral bacteria 
tested decompose hydroxypro line rapidly and hydroxyproline- 
contalning peptides slowly. This observation necessitates 
repetition of the previously negative experlmente and the 
development of an alternative test for collagenolysis. 

Significance to pe ats! Research ; 

Degenerative changes of collagen fibers in the marginal 
gingivae are a prominent feature of periodontitis. Disruption 
of the collagenous fiber attachment of the root of the tooth 
to the alveolar bone with eventual loss of the tooth is the 
most important consequence of periodontal disease. It is 
essential, therefore, to determine whether such collagenolysis 
results from the inflammatory reaction per se, from proteolysis 
by the oral microbiote, or from a combination of these processes . 

Proposed Course of Project ; 

To continue ar.d extend the studies outlined above. (Project 
initiated 1 September 1958). 

Part B included No 

Serial No. NIDR-12 

1* Microbiology 
3. Betheada, Md. 

Individual project Report 
Calendar Year 1958 

Part A 

Project Title: Manifest and inapparent viral infectlona of 
Oral "issues 

Principal investigator: Dr. H. W. Scherp 

Other investigators: Dra. R. Mo Stephen, R. j. Fitzgerald, 
I. I. Ship 

Cooperating units: None 

Man Years (calendar year 1958): 

Total: 3 1/2 

Professional; 1/2 

Other: 3 

Project Description; 

Objectives ; 

1. To determine whether, as often claimed, herpes simplex 
virus is the etiological agent in recurrent aphthous leslona 
of the oral mucosae 

2. To Isolate, identify, and determine the etiological 
significance of viruses from various other lesions of the 
oral mucosa of undetermined etiology. 

3. To test for the presence of inapparent viruses In the 
tlsaueB of "germ-free" animals. 

Methods Employed ; 

Samples (from lesions) containing tissue cells are frozen and 
thawed to liberate intracellular vlruees. To test for viruses, 
the samples are inoculated into standard tissue cultures of 
rabbit kidney cells, monkey kidney cells, Hela cells, chicken 
embryo cells, and human skin cells. After incubation, cultures 
are frozen, thawed, and transferred to a fresh tissue culture; 

" 2 - Serial No, NIDR 12 

Part A (continued) 

frequently, a second serial transfer is made. In addition,, 
specimens of biopsy tissue are maintained in tissue culture 
medium for protracted periods to allow development of latent 
viruses. Evidence for a virus is sought by microscopic 
examination for cytopathologic changes in the cultured cells 
and by hemagglutination tests of the culture fluids with 
guinea pig and chicken erythrocytes. Isolates of herpes 
virus are identified by neutralization tests with specific 
ant 1 sera. 

Major Findings ; 

Herpes virus was recovered from only 1 of 47 samples taken 
from recurrent oral aphthous ulcers (Dr. ship); in contrast, 
this virus was recovered from clinically diagnosed herpetic 
lesions of the 3 patients tested to date. This control 
series will be enlarged, No evidence of a virus was found 
in 200 specimens from various other oral lesions, mostly 
perlodontsl pockets (Dr. stephan) . A number of specimens of 
gingival tissue have been maintained for varying periods under 
the conditions of tissue culture without exhibiting obvious 
cytopathologic changes Tests with the cellular outgrowths 
from these tissues indicate that it should be possible to 
establish pure fibroblastic and epithelial cell lines of 
gingival origin, which might be more suitable than available 
cell lines for the present type of investigation. 

To date, 206 samples from germ- free animals (throat and rectal 
swabs, feces, urine, brain, kidney, pancreas, liver, lung, 
spleen, and salivary gland) have been tested (Dr. Fitzgerald). 
A transmissible cytopathogenic agent has been isolated from 
11 of the 18 salivary glands tested. In comparison, such an 
agent waa isolated from the salivary glands of 6 of 7 ordinary 
laboratory rats. These observations support the suspected 
possibility that gnotoblotic animals may harbor viruses. 
Optimal conditions are still being sought for the maintenance 
of these agents. They are too rapidly lethal for rabbit 
kidney cells and do not affect monkey kidney cells or Hela 
cells but seem to establish a usable infection in human skin 

Recently, satisfactory conditions have been worked out for 
producing plaques with herpes virus in monolayers of tissue 
cells In culture, as a preliminary to a serological comparison 

- 3 - Serial No„ NIDR-12 

Part A (continued) 

of strains of this virus by measurement of the rate constants 
of their neutralization by homologous and heterologous 
antisera, using s procedure previously applied with success 
to polloviruses by Dr. W. Do MCBride in this Laboratory., 

Significance to Dental Research ; 

Orel soft tissue lesions of unestablished but possibly viral 
etiology are frequently encountered by the dentisto The still 
incompletely understood phenomena of latency and activation 
of herpes virus are accordingly of direct dental interest; 
they also provide prototypes of fundamental viral relation 
ships, Demonstration of an lnapparent viral infection in 
otherwise germ-free animals would open another approach to 
the study of the consequences of such infections on "normal" 

Propose d Course of Project ; 

Studies of the rat salivary gland agent and of the serological 
comparison of strains of herpes virus will be continued pend 
ing the return of Dr« w, D- McBride from training project at 
California Institute of Technology, 

Part B included No 

Serial No- NIDR-13 
1„ Microbiology 

3 J Betheade, Md,. 

individual Project Report 
Calendar Year 1958 

Part A 

Protect Title- The Relationship of Pathogenic Fungi to Diseases 
of the Oral cavity. I. Incidence and identity, 

principal investigator: DTo A. Howell, jr. 

Other investigators: Dr. R. M. Stephen 

Cooperating units: None 

Man Years (calendar year 1958): 
Total: 1 2/3 
professional: 2/3 
Other: 1 

project Description: 

Objectives : 

1 to study the incidence and identity of pathogenic or 
potentially pathogenic fungi in the "normal" mouthc 

2 To study the possible relationship of these organisms to 
dental caries and to other diseases of the oral cavity 
or related areas. 

Methods employed : 

As in 1957, considerable emphasis has again been placed on 
the incidence and identity of both filamentous and yeast-like 
organisms in the orel cavity. For this purpose standard 
bacteriological methods for the isolation and identification 
of these organisms, modified for the organisms under study, 
have been employed. 

patient Material : 

Selected oral samples are obtained from various typee of 
individuals, including patientu with caries, periodontal 
disease, and stomatitis. To date over 250 oral samples 
(plaque material, calculus, etc.) and 63 saliva samples 
from over 50 patients have been studied,, 

Serial no. NIDR-13 

Part A (continued) 

M»jor Finding* ; 

An analysis of Ch« findings on 231 oral samples (other than 
saliva) and 56 sallvaa has revealed probably significant 
differences in the incidence of filamentous form in different 
areae of the oral cavity, For example, Actinomyces wse 
isolated from over 70% of plaques from early stages of carles, 
45 to 60 par cent of samples from periodontal lesions, but 
from only 23 per cent of saliva samples* Large numbers of 
these orga/.lama were Isolated from 20 25 per cent of plaques 
from early carles and periodontal disease, but no saliva 
gave high counts. Similar results were obtained with 
Leptotrichla buccalla , a previously little studied organism,, 
which has been isolated with some regularity from specimens 
of this series. Yeasts, on the other hand, while prevalent 
in plaques from early caries, were seldom found in material 
from periodontal lesions and were quite prevalent in saliva. 

As pointed out previously (Annual Report, 1957), the absence 
of a selective medium for filamentous oral organisms and the 
variation in colonial morphology make the above reeults 
qualitative rather than quantitative. 

Significance to Dental Research ; 

The finding that the flora varies in different locations in 
the oral cavity of the same individual at a given time, or 
perhaps in different types of oral disease, strongly indicates 
that a more careful and detailed study of the flora in different 
sites must be undertaken in order to provide knowledge of the 
true incidence of these forms and the exact etiology of disease^ 
It further emphasizes that bacteriological study of saliva 
need not necessarily be a reliable indicator of the predominant 
organisms of the oral flora. Added emphasis must be placed 
on the care with which oral samples are taken for study, 

Proposed Course of project ; 

1. To continue the study outlined above for approximately 
one year to increase the number of samples studied, 

2o To study the bacteriology of calculus formation in collabora- 
tion with Dr„ A. Rizzo, Laboratory of Microbiology, NIDRo 

Part B included No 

Serial No NIDR-14 

L. Microbiology 
3 Bethesda, Md. 

Individual Project Report 
Calendar Year 1958 

Part A 

Project Title: A Study of the Genera Actinomyces and 
Leptotrlchla . 

Principal Investigator: Dr. A. Howell, Jr, 

Other Invest lgatore: Mr. W. C. Murphy, III 

Cooperating Unite: None 

Man Years (calendar year 1958): 
Total: 1 1/3 
Professional: 1 1/3 
Other: None 

Project Description. 

Objectives : 

1 To study strains of Actinomyces isolated from the 
oral cavity and to compare such organisms vith strains 
isolated from clinical cases of actinomycosis 

2. To study the incidence, identity, and characteristics 
of the organism previously described in the literature as 
Leptotrlchla buccal is . 

Methods Employed : 

Standard bacteriological procedures, modified as 
necessary foir the organisms under study, are being 
used. Organisms are being studied on the basis of 
(1) fermentation of 16 carbohydrates; (2) oxygen toler- 
ance; (3) other biochemical tests; and (4) animal 
pathogenicity « 

Major Findings: 

Approximately 200 oral strains of Actinomyces , 

isolated from the oral specimens studied In Project No. 13, 

- 2 - Serial No, NIDR-14 

Part A (continued) 

have been compared with 11 strains isolated from 
clinical actinomycosis. Bases for comparison have been 
their microscopic and colonial morphology, ability to 
grov and produce acid on each of 16 carbohydrates, 
oxygen tolerance, catalase reaction, ability to digest 
casein, hydrolyze gelatin, and reduce nitrates, and, in 
some instances, ability to produce diseases in experi- 
mental animals, particularly hamsters and mice. One 
group of the oral isolates corresponded closely to the 
previously reported, but not officially recognized, 
A, naeslundl , which is accordingly recommended as a 
valid species. The remaining oral isolates fit the 
description of A. israelii , the cause of human actino- 
mycosis; 88 per cent of these strains could be sub- 
classified into three major subgroups on the bases 
listed above. The results of this study have been 
incorporated into a manuscript for publication. 

Some estimate of the incidence of Leptotrlchla buccal is 
in the oral cavity has been obtained (Project No 13). 
The morphology and physiological characteristics of 20 
Isolates have been determined. A short note describing 
the morphology has been published (see Part B attached). 
A report of the physiological properties of this organism 
will be prepared In the near future and submitted for 

Significance to Dental Research : 

A more complete description of human strains of Actinomyces 
and of Leptotrlchla buccalis will lead to a clearer under- 
standing of both of these groups of organisms. This will 
facilitate future work on the possible role of ttoese 
organisms in oral disease, 

Proposed Course of Project : 

1. To study bovine isolates of Actinomyces , in collabora- 
tion with Dr Leo Pine, Duke University, School of 
Medicine and compare them with human strains. 

2 To study, in more detail, the human diphtheroids and 
comoare them with species of Actinomyces ^. 

Part B included Yes 

Serial No, NIDR-14 

Individual Project Report 
Calendar Year 1958 

P«rt_B: Honors, Awards, and Publicationa 

Publication, other than abstracts from this project: 

Howell A,, and Rogosa, M. Isolation of Leptotrichia 
buccjU., J. Bact., 26(3): 330-331. 1958 . ° CrlCllla 

Honors and Awards relating to this project: 

Serial no. NIDR=15 

lo Microbiology 
3 Bethesda, Mdc 

individual Project Report 
Calendar Year 1958 

Part A 

Project Title: inhibitors and Dental caries 

Principal investigator; Dr. H. V. Jordan 

DTo R. J. Fitzgerald 

Other investigators: None 

Cooperating units; None 

Man Years (calendar year 1958): 

Total: 2 1/3 

Professional; 1 

Other 1 1/3 

Project Description; 

Objectives ; 

To study agents which can be reasonably expected to exert 
some effect on experimental carles by inhibition of growth 
or metabolic activities of the oral microflora. It is 
expected that such agents, if found to be active, would 
have potential use in the study and control of oral disease 
and other conditions in humans. 

Methods Employed ; 

A number of antioxidants or compounds which have potential 
antioxidant activity are being tested for their effect on 
experimental carles. To date testing has been largely 
confined to compounds which have been used in the food 
processing Industry or In other commercial applications. 
These compounds are incorporated in a carlogenic rat diet 
at fairly high levels in order to assess their in vivo 
effect on experimental caries. The _in. vitro effects of 
these antioxidants are also being observed on the growth and 
acid production of representative oral microorganisms isolated 
from the rat and humans. 

- 2 - Serial no. NIDR-15 

Part A (continued) 

Major Findings: 

Previous tests have shown that sodium metabisulfite and also 
the synergistic antioxidant combination of a-tocopherol »nd 
ascorbic acid inhibited occlusal fissure caries in the rat. 
These tests are being repeated, inhibitory end points for 
selected oral bacteria have been already determined for soma 
antioxidants, and this work is continuing. 

Significance to pentsl Research ; 

This work and studies mentioned in previous reports form part 
of a program of testing of agents which may find some use in 
the control of certain oral conditions. In addition, the 
study of compounds which undoubtedly have differential effects 
on the metabolism of the various bacteria of oral flora pro- 
vides a possible approach to the study of the etiology of 
these conditions from s bscteriological point of view. 

Proposed Course of Project ; 

Future studies will be influenced to a large extent by the 
results of current studies. Compounds showing promising 
in vivo effects will be studied further as regards effective 
dosagSp toxicity, and possible synergistic combinations,, 
Attempts will be made to correlate in vitro effects of 
antioxidants on oral bacteria with in vivo effects on 
experiments! csrles in rsts. It will also be necessary to 
determine the precise mode of action of the antioxidants. 

Part B included Yes 

Serial no. NIDR-15 

Individual project Reports 
Calender Year 1958 

Pert B : Honors, Awards, and Publications 

Publications other than abstracts from this project; 

Fitsgerald, R. j., parramore, m. L., and Macintosh, m. Ec 
Antibiotic sensitivity o£ Oral strains of Veillonella . 
Accepted for publication in Antibiotics and Chemotherapy. 

Honors and Award* Relating to this Project- 


Serial No. NIDR°16 

lo Microbiology 
3. Bethesda, Mdo 

Individual project eport 
Calendar Year 1958 

Part A 

Project Title: Microbiology of saliva. 

Principal investigator: Dr. To Shiota 

Other Investigators: None 

Cooperating units : American Dental Association 

Man years (calendar year 1958) 

Total: 12/3 
Professional; 2/3 

Other : 1 

Project Description: 

Objectives : 

1. To study certain environmental effects in the in vitro 
bacterial and chemical changes in saliva. 

2> To study the origin and content of bacterial growth- 
promoting factors in saliva. 

3. To relate certain chemical changes to the action or 
interaction of oral microorganisms. 

Methods Employed : 

Bacterial enumeration and isolation were made by using specific 
isolation media; determination of vitamins by microbiological 
assays; and identification of vitamins by chromatography. 

Major Findings; 

It was shown previously that certain chemical and bacterial 
changes occurred In Incubated saliva, under similar conditions ( 
levels of certain B- vitamins were found to increase, chroma- 
tography of nonincubated saliva revealed traces of free folic 
acid and a compound similar to coenzyme A. Chroma tograms of 

- 2 - serial no. NIDR-16 
P»rt A (continued) 

incubated .alive showed compounds having mobilities similar 
to folic acid, citrovorum factor, an unidentified compound 
intermediate between folic acid and citrovorum factor 
pyridoxine. pantothenate, and coenzyme A . The result, of 
the study showing the concomitant bacterial changes 
indicated that the lactic acid bacteria could be^ctive in 
increasing the levels of pyridoxine end folic acid- like com- 
pounds; and that fusobacteria and veillonellae could increase 
the levels of pantothenic acid-like compounds. This con 
elusion does not exclude the possibility thst other organisms 
present in saliva function similarly. 

Significance to pental Research ; 

The occurrence of e variety of microorganisms in great numbers 
in sslivs indicstes that saliva is fsvorable for the growth 
of orel microorganisms, which as the result of their growth 
slter their environment. The study of chemical alterations 
by microorganisms, and conditions favoring their growth, may 
be of great importance to the understanding of the health of 
teeth and oral mucosa. 

Proposed Course of project ; 

It is proposed that the project continue along the following 
lines; (1) to study the bacterial and chemical composition 
of whole saliva and of pure salivary gland secretions; 
(2) to follow certain chemlcel changes of saliva due to 
bacterial action; (3) to study the effects by varying the 
environments! conditions on bacterial growth in saliva. 

Part B included Yes 

- 3 - 

Serial No. NIDR-16 

Individual Project Report 
Calandar Year 1958 

P*rt_B: Honors, Awards, and Publication* 

Publication* other than abstract* from this project: 

5£Zl.V^? KU ? k,l ' I M - »•• *■ P» Vitro Chemical .nd 
Bacterial Changs. i„ ^liva. j. D.nt7 rST~37: 780. 1958. 

Honor* and Awrdi relating to this project: 


Serial no. NIDR-17 

1. Microbiology 
3. Bethesda, Md. 

individual project Report 
Calendar year 1958 

Part A 

Project Title: Nutrition and Metabolism of Lactic Acid Bacteria 

Principal Investigator: Dr. To shiota 

Other investigators: None 

Cooperating units: American Dental Association 

Man years (calendar year 1958) 
Total: 2/3 

Professional: 1/3 
Other: 1/3 

Project Description: 

Objectives : 

To study the enzymatic synthesis of folic acid-like compounds 
by extracts of Lactobacillus arablnosus . 

Methods Employed ; 

The determination of folic acid was carried out using micro- 
biological assay, chromatography and spectrophotometry. The 
bacterial extracts were prepared by methods commonly employed 
in protein purification. 

Major Findings : 

Previously it was reported that extracts of Lo arabinosus 
synthesized folic acid-like compounds in the presence of a 
reduced pteridine, p°aminobenzoic acid or p-aminobenzoylglutamic 
acid, adenosine triphosphate and magnesium. - we now have evidence 
which indicates that this reduced pteridine is a tetrahydro- 
pteridine and that the product of the reaction is tetrahydro= 
folic acid. 

- 2 - serial No. NIDR°17 
Part A (continued) 

Significance to pental Research ; 

The presence of the lactic acid bacteria in saliva and 
their great capacity to produce acid rapidly have in- 
criminated them as being etiologically associated with 
caries. The study of their biochemical activities is 
essential to an understanding of their part in the 
oral microbiota. 

Proposed Course of project ; 

The work started on the purification of the cell extract 
to obtain a purified enzyme preparation is to be -on 
tinued. The preparation and storage of the reduced 
pterldlne has not been altogether satisfactory since 
this pteridine is very labile. It is hoped that a 
suitable preparation can be obtained. 

Part B included Yes 

- 3 - serial no. NIDR-17 

Individual project Report 
Calendar Year 1958 

Part_B: Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Shiota, T.. Folk, J. e. . and Tietse, F. Inhibition of 
Lysine Utilisation in Bacteria by s- (3 -Arainoethyl ) 
cysteine and its Reversal by Lysine Peptides. 
Arch. Biochem. Biophys. 72: 372 (1958). 

Shiota, t. Enxymic Synthesis of Folic Acid-like 
Compounds by Cell Free Extracts of Lactobacillus 
Arablnosus . Arch. Biochem. Biophys., in press. 

Honors and Awards Relating to this project: 


Serial No. NIDR-18 
lo Microbiology 
3. Bethesde, Md, 

Individual project Report 
Calendar Year 1958 

Part A 

Project Title: studies on the Microorganisms Associated with 
Periodontal Diseases 

Principal Investigator: Dr. a. R. onata 

Other investigators: Dr. s. A. Narrod, Laboratory of Biochemistry 

Cooperating units; None 

Man Years (calendar year 1958) 
Total; 2 1/3 
Professional: 1 1/3 
Other: 1 

Project Description; 

Objectives ; 

To investigate various microbial flora and the nature of 
their products, specifically the proteolytic enzymes, 
as factors in periodontal disease. 

Methods Employed ; 

1. Methodology for the fractionation of collagenase 
elaborated by Clostridium hlstolytlcum is being extended. 
Although partial purification of this extracellular proteolytic 
enzyme has been achieved by salt and alcohol precipitation 
and sponge electrophoresis, further work is being extended 
with the use of column chromatography with some of the 
newer cellulose ion-exchange adsorbents. Newer methods 
are being investigated to measure enzyme activity using 
collagen and derived gelatins from various animal 

- 2 = Serial No. NIDR~18 
Part a (continued) 

2. studies are being conducted on the isolation and 
classification of the oral microflora and also on the 
physiology of these organisms, namely the nutritional 
requirements and the elaboration of proteolytic enzymes. 

Major Findings ; 

1. Partial purification of bacterial collagenase has 
been accomplished with the use of sponge electrophoresis; 
however, the presence of small amounts of protease has 
been an obstacle in determining the enzymatic degradation 
of collagen. Preliminary experiments with the cellulose 
ion-exchange resins have been more effective in the 
purification procedure and are still in progress. 

2. The nutritional requirements of Fusobacterium 
nucleetum, isolated from the human mouth, were investigated 
employing a simplified culture medium composed of vitamin- 
free casein hydrolyzate, synthetic B-vitamins, and deriva- 
tives of nucleic acids, it was found that fusobacteria 
required pantothenate, pantetheine, pantothenol or 
coenzyme A as vitamins for growth, m addition they 
required tryptophan, adenine, hypoxanthine and 
4-amlno-5-imidozole carboxaraide for optimum growth. 

Significance to pental Research ; 

The present investigation is based on the concept of the 
role of different groups of microorganisms and their 
products as contributory factors in the initiation of 
periodontal disease. There are implications that various 
bacteria and their enzymes may have an integral relation- 
ship in the etiology of oral diseases, and basic under- 
standing of these factors may be a fruitful approach. 

Proposed Course of project ; 

Investigations will be conducted on the action and degrada- 
tion of various types of native and purified substrates by 
different types of oral bacteria and on the effect of 
proteolytic bacteria on collagenous substances. 

Part B included yes 

Serial No, NIDR-18 

Individual Project Report 
Calendar Year 1958 

part B : Honors, Awards, and Publications 

Publications other than abstracts from this project; 

R. R. omata: studies on the Nutritional Requirements of the 
Fuaobactsrla. II. Requirements for Pantothenate and Purines 
journal of Bacteriology (in press). 

Honors and Awards relating to this project t 


Serial No, N1DR-19 

1 „ Microbiology 
3. Bethesda, Md 

Individual Project Report 
Calendar Year 1958 

Part A 

Project Title: Investigations in the Pield of the Spirochetes 

Principal Investigator: Dr E, Go Rampp 

Other Investigators: Dr. S, B, Mergenhageo 

Cooperating Units: American Dental Association 

Man Years (calendar year 1958) : 
Total : 1 
Professional: 1/3 
Other: 2/3 

Project Description: 

Objectives : 

1. To investigate the infectlvlty of certain pure strains of 
spirochetes for guinea pigs and rabbits by the intracutaneous 

2. To investigate the relationship of hyaluronldase and chon- 
droltinase to the spread of spirochetal infections in rabbits . 

3. To study the relationship of the combination of a micro- 
aerophllic diphtheroid organism and a small oral treponeme in 
the Initiation of lesions in rabbits. 

4. To determine the relationship of oral spirochetes to diseases 
of dental origin. 

Methods Employed : 

1. Inoculation of rabbits intracutaneously with pure strains of 
representative cultures of the small oral treponeme Borrelia 
vincentli and Borrelia buccal is, 

* 2 " Serial No. NLDR-19 
P«rt A (continued) 

2. Culture of the Inoculation sites for spirochetes at 
various time interval, for the duration of ^such legions. 

Li^'foi !!^ 1 / 6 ", 10 " 8 ""P 10 * 1 ** hematoxylin and eosin 
stains for studying the progress of lesions. 

L.™!™^^ 1 " 108 ° £ • nlMl tl68ue8 to demonstrate and 
determine the location of ,h e spirochete, in tissue sections, 

Major Findings : 

It was found that pure cultures of the small oral trenonemeB 
f n . Vincent" «d B. buccall, •hen introduced i^acu^Zsly 

yp. 8 . oTsn'msir h"*"" Pr .° dUCed 1 " i0nB lD ^ both 
types of animal. However, the lesions produced in rabbit. 

-ere more fulminating than in the guinea pigs! and wrt wis 

lesion h t0 T " bbU - ThU8 f "» 16 ' "> 1*<Z«9?IL™ 
lesions have been studied In rabbits. Wet mounts of material 

^J^iU^T: '^ b " n 8CUdied by d.r^-"f ie'iriicr: copy 
ana ajotlle organism, demonstrated In the case of the small or*) 
treponeme and B. vlncentil. but not for B. buccal s Te sin 

up to e 4 o d r . 8 :: 18n L> have been recovered *&&&*£ i.: s 

w?^ t y l v Abscesses •« produced within 4 days in rabbits 
aXc^es" In"™ T "»"°«**^< "rains of or"' 
1«^- h " 8t * lned CU8Ue eect *°ns of various 

virgin' fr^ n i 8 6 r * ted ^T 1 ^" ln Sundance in lesion. 
d?7i f ^ hOUr " t0 4 d * ys of du «tion. The organisms 

tat ««L r :; T iD : hC aube P^ h *^«l connective tfssue 
.««< ! COn ! ln « d to the inoculation sites. Hematoxylin and 
Jnf L^r l ° ed 8eCti ° n8 exhibited vnrlou. gradations^ .cute 
«.!Jr^ y u re8P0n " includi *S abscess formation. Preliminary 
studies are being conducted in rabbits employing various »rm2 

ijfh ;he°ILn d * *«—«**»» diphth^oiS iS co^ination 
with the small oral treponeme. Large spreading lesions are 

or'th^se •!*£" °ir 1 ^ 8tUdlC8 - "»- -"ions 
r cnxs pb ** e of tQ e problem have not been completed. 

Significant to Dental Research : 

le.i^i 8 i8 5 he / ir8t P°»itive evidence that reproducible 
.Piroc 8 J an be A1 lni£i * ted b v pure cultures of the various oral 

ge'nlcity of the e eTT aVail * ble in *• ^ ° D the ^ 
semcicy ot these organisms was presumptive. 

3 = Serial No, KIDR-19 

Part A (continued) 

2. A new approach has been developed which will enable I 
investigators to study the etlologic significance of the oral 
spirochetes in certain types of ulcerative conditions in- 
volving the oral car: 

Proposed Course of Project ; 

1. To continue the investigation of the lesions prodi> 
in animals to determine the mechanism of action of the 

2. To investigate the effect of spreading factors, such as and chondroitinase, on the progress of experi- 
mental lesions 

3 To 8'uqv the effect of heated vs, nonheated culture medium 
free of organisms on the initiation of experimental lesions. 

4. To investigate the effects of living cells, nonliving 
cells, and cellular fractions on the initiation of lesions 
in animals, 

5, To study fh i effects of the spirochetes on the various fixed 
tissue elements of the rabbit sk; 

6 To Investigate oral organisms that may act as aymfolo 1 
organisms in the production of experimental lesions in rabbits 

Part 8 included Yes 

- 4 Serial No„ NIDR-19 

individual project Report 
Calendar year 1958 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Hatnpp, E° Go Research Division: American Dental Association, 
National Institutes of Health, J„AoD.Ao, 57: 150, July 1958 

Honors and Awards Relating to this project: 

Serial No. MIPR-20 
1. Microbiology 
3. Bethee<3a, Hd, 

Individual Project Report 
Calendar Year 1938 

Part A 

Project Title: Studies of Interactions Between Oral 
Microorganl — 

Principal Investigators: Dr. T. A. Kevin 

Dr. E. Go Eatapp 

Other Investigators: None 

Cooperating Units: American Dental Association 

Man Years (calendar year 1958) : 
Total: 3 
Professional: 12/3 
Other: 11/3 

Project Description: 

Objectives : 

1. To study the biocheaical bases of the dynamic equilibria 
which determine the complex nature of the flora of the couth. 

2. To investigate the occurrence of, and the nutritional 
basis of. symbiosis among the more common microorganisms 
in the mouth. 

3. To employ more fastidious organisms, such as spirilla c 
various filamentous forms and spirochetes, in order to 
discover less fastidious microbes whose products are required 
by, or enhance, the growth of the more exacting types. 

Methods Employed ; 

1. The principal method is a modified ausanographic 
technique. It employs suitably formulated culture media, 
seeded with a selected bacterium which serves as an indicator 
of reaction. Standard amounts of test culture suspensions 
are pipetted to sterile filter paper disks which are then 
placed on the surface of the seeded medium. Controls are 

, of! 


efJectec rlate reference solution.; 

place of th< Thus, quantitative as well 

qualitative data are obtainable 

2. Standard or modified chemical, physical or micro- 
biological methods are used to identify common metabolite 

Major Findings 

1 In the annual report for 1957, it was pointed out that 
Borrelia vincen tii was stimulated markedly when cultivated 
in the presence of an anaerobic diphtheroid. Further, it 
wan pointed out that in order to understand the mechanism 
of the Interaction, it would be necessary to determine 
some of the spirochetes* nutritional requirements- The 
present report is concerned, primarily, with the nutriu 
of 8. vincentii . 

The basal medium consisted of casamino acids (Difco), g'iu. 
K RPO, , Fe , Mn , Mg , and a supplement of purines and 
pyrimldlnes. Results were determined on a growth yj no 
basis when a required factor was included in, or omitted from, 
the culture medium, When quantitation was necessary, dlr 
cell counts in a Petrof f-Hauser chamber were performed 
When the basal medium was enriched with ascitic fluid, oleic 
acid, folic acid, biotin, riboflavin, and cocarboxylase were 
required for growths Dibydrofolic acid and folic acid are 
both active, whereas citrovorum factor, tetrahydro folic acid 
and p-aminobenzoic acid are not. Thiamin cannot be substi- 
tuted for cocarboxylase . 

The ascitic fluid enrichment has been reduced from 10% to 
0,3% by volume by substituting adenosine triphosphate, 
coenzyme A :1 glucose- 1- phosphate, L-asparaglne and NaHCO- 
(as a source of CO.) There is no appreciable loss in 
eel 1 yields by sucS sub»6itu£i««s Pantetheine or calcium 
pantothenate can be utilized by the spirochete, but yield 
only about 1/2 the number of cells that can be obtained 
with a molar equivalent of coenzyme A, 

2c In a parallel study, the ascitic fluid requirement of 
Bo vincentii and several other Tre ponemat aceae has been 
completely satisfied by cocarboxylase, coenzyme A, adenosine 
triphosphate, diphosphopyridine nucleotide, pyridoxal phosphate, 

- 2 - Serial No. HIDR-20 

Part A (continued) 

effected by the use of appropriate reference solutions in 
place of the test culture. Thus, quantitative as well as 
qualitative data ore obtainable. 

2. Standard or modified chemical, physical or micro- 
biological methods are used to identify common metabolites. 

Major Findings : 

1. In the annual report for 1957, it was pointed out that 
Sorrel la viacantil was stimulated markedly when cultivated 
in the presence of an anaerobic diphtheroid. Further, it 
was pointed out that in order to understand the mechanism 
of the interaction, it would be necessary to determine 
some of the spirochetes 4 nutritional requirements. The 
present report is concerned, primarily, with the nutrition 
of B. vincentii. 

The aaeal medium consisted of casamino acids (Difco) , glucose, 
K HPO, , Fe , Mn , Kg , and a supplement of purines and 
pyrimldincs. Results were determined on a growth vs. no growth 
basis when a required factor was included in, or omitted from, 
the culture medium. When quantitation was necessary, direct 
cell counts in a Petrof f-Hauser chamber were performed,. 
When the basal medium was enriched with ascitic fluid, oleic 
acid, folic acid, biotin, riboflavin, and cocerboxylase were 
required for growth. Dihydrofollc acid and folic acid are 
both active, whereas citrovorum factor, tetrahydro folic acid 
and p-emlnobenxolc acid are not. Thiamin cannot be substi- 
tuted for cocerboxylase. 

The ascitic fluid enrichment has been reduced from 10% to 
0.3X by volume by substituting adenosine triphosphate, 
coenzyme A, glucoee-1- phosphate, L-asparagine and KaHCC, 
(as a source of CO.) There is no appreciable loss in 
cell yields by sucl subsfclfcut&fiaso Pantetheine or calcium 
pantothenate can be utilized by the spirochete, but yield 
only about 1/2 the number of cells that can be obtained 
with a molar equivalent of coenzyme A, 

2. In a parallel study, the ascitic fluid requirement of 
Bo vincentii and several other Ireponem ataceae has been 
completely satisfied by cocarbosylase, coenzyme A 3 adenosine 
triphosphate, diphosphopyridine nucleotide, pyridoxal phosphate, 

- 4 - Serial No. NIDR-20 

Part A (continued) 

may be stated that B, vlncentit deamlnates glutamine, 
but does not attack asparagine, lysine or arginine to 
a measurable degree. NH_ liberation was determined in 
spot plates by Neaslerization, Similarly, Cu reduction 
tests suggest that glucose is degraded. Further explora- 
tions In this area are planned. 

Part B included No 

Serial No, N r.DR-21 
1. Microbiology 

3. Bevhesca, Md» 

Ine ividual Project Report 
Calendar year 1958 

Part A 

project Title: stv-dies on the Host Cell-Virus Relationship with 
Special Reference to Latent infections 

principal investigator: Dr. w, D. McBride 

Other investigators i None 

Cooperating units: California institute of Technology 

Man years (calendar year 1958): 
Total : 1 
professional; 1 
Other : None 

Project Description; 
Objectives : 

To study the host cell-virus relationship with special 
reference to those conditions which might permit a moderate 
virus infect icr., equivalent to the lysogenic infection of 
bacteria and bacteriophages. During the past year, this 
investigation has been carried on as part of i training 
program under the supervision of or, Renato Dulbecco, 
at the California institute of Technology, 

Methods pnployed ; 

The work for the past year has centered primarily around 
attempts to analyse genetic recombination with a ribonucleic 
acid (RKA) viruso Polic virus has been chosen for this work 
because of the following considerations., The only successes 
claimed for recombination of viral genetic material have been 
with several ceoxyribonucleic acid (DNA) viruses and influensa, 
and RNA virus. The systemo with which the DKA. studies were 
done are clean and straightforward, The results are un- 
equivocal. With influenza, the system is not clean., A 
careful scrutiny of the results raises many objections to 

- 2 - s«ri«l No. NIDR-21 
Part A (continued) 

the claims for demonstrating genetic recombination. Even 
granting that it has been shown with influenza virus, then 
why has it been so difficult with other RNA viruses? Many 
attempts have been made with polio, Newcastle disease, and 
tobacco mosaic viruses, in each case results are either 
completely negative or equivocal at best. 

Major Findings : 

Specifically, our first aim baa been to develop and isolste 
strains of virus which have genetic markers appropriate for 
these studies, we now have four with which we can work com- 
paratively easily* These are: 

1. Temperature resistance, under standard conditions used 
the survival rate varies from 10" 1 with the resistant strains 
to 10'* with the most sensitive. 

2. Cystine effects. The wild type requires systine in the 
medium and gives clear plaques in its presence. The most 
divergent mutsnt from this grows best in the absence of 
cystine and gives turbid plaques in its presence due to a 
delay in virus growth in the presence of cystine. 

3. "Delayed" (d) character, d-yirus has low efficiency of 
plating (EOP) under acid agar as compared with alkaline agar. 
The wild type, d fc , has equal EOP on acid and alkaline agar. 

4. Resistance to ox- serum inhibitor. The wild type Is 
sensitive to this inhibitor whereas the mutsnt is quite 

Crosses have been made with strains carrying various 
combinations of these markers. Each cross is made by mixedly 
infecting uela cells at high multiplicity, isolating the 
cells in microdrops and harvesting individual cell yields. 
These ere then plated on a nonselective medium and plaques 
are isolated. Bach plaque isolation is grown into a stock 
and then each stock is tested for the characters used in 
the cross. To increase the probability of positive results, 
some parental stocks and/or the cells are treated with 
mutagenic agents such as ultraviolet light, x=rays, and 

- 3 - serial No. NIDR-21 

Part A (continued) 

To date, we cannot state unequivocally that we have 
demonstrated recombination,. The impression we get is that 
we do have it. However, there are many tests and retests 
involved before we can state with any finality just what 
the results mean. 

in addition to our primary work as outlined above, 
we have been conducting various exploratory experiments 
with herpes virus, in this case the primary object is to 
find a truly moderate animal virus infection in tissue 
culture. These experiments have included studies of the 
exclusion of herpes virus from monkey kidney cells by other 
viruses and, conversely, the exclusion of other viruses 
from monkey k-'dney cells by herpes virus. 

Significance to Dental Research ; 

It would be of great biological significance to find 
recombination of an RNA virus in a system that can be 
accurately studied, not for the importance of RNA re- 
combination per se (although it would be easy to make a. 
case for the utility of this for developing good vaccine 
strains of many viruses), but rather for analyzing how 
the RNA is replicated and handled with cells. Many of the 
most pressing problems in biology will surely find their 
solution when the mechanisms controlling RNA and DNA 
synthesis, recombination and mutation are understood. 
These are of infinitely greater importance than polio 
or any other virus infection. They certainly apply to 
neoplastic and degenerative diseases, both of which are 
within the range of interest of stomatology. 

Proposed course of project ; 

The studies outlined above will be continued until completion 
of the training program in 1959. 

Part B included yes 

Serial NOo NIDR=21 

Individual project Report 
Calender Year 1958 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts from this project; 

McBrlde, Wo D. Antigenic Analyses of Polioviruses by 
Kinetic Studies of Serum Neutralisation, virology, 
in press. 

Honors and Awards Relating to this project; 

Serial no. NIDR-22 
lo Microbiology 
3. Bethesda, Md, 

Individual project Report 
Calendar year 1958 

Part a 

Project Title; immunological, Biochemical, and Pathological 
Studies of oral Bacteria. 

Principal investigator: Mr. m. Rogoss 

Other investigator*: Dr. R. J. pltsgerald 
Dr. E° G. Hampp 

Cooperating unite: None 

Man Years (calendar year 1953): 
Total: 2 
Professional: 1 
Other: 1 

Project Description; 

Object- Ives : 

To determine the immunological, biochemical, and pathological 
characteristics of the ye 11 lone 11a and other orel organisms. 

Methods Employed ; 

1. Antisere, prepared in rabbits against bacterial cells, 
are tested against the organisms or antigens extracted from 
them by the techniques of agglutination, precipitation, and 
selective entibody absorption, 

2. Biochemical characteristics are determined by standard 

3. Pathogenicity studies are conducted in appropriate animal 
hosts, inoculated by various routes. 

- 2 - Serial No. NIDR-22 

P«rt a (continued) 

Major Findings: 

1* serology: Agglutination, cross-agglutination and Agglutinin 
absorption tests were carried out with high-titer antisera 
prepared against a number of representative strains of 
Veil lone lis Isolated from the human moutho These were tested 
against 33 human strains, 19 rabbit strains, 20 hamster 
strains, and 21 rat strains of yell lone 11a , The human 
strains could be divided into 5 serotypes, 2 of which were 
closely related serologically to a number of the rabbit 
strains, present evidence lndicstes the existence of 
other, sharply distinct, serotypes among the animal strains. 
The various serotypes are characterized also by differences 
in biochemical and nutritional properties. 

2. pathogenicity: Representative strains of human oral 
Vet Hone 11a were found to be uniformly nonpathogenic when 
suspended in various media and Injected intraperitoneal ly 
into mice. Certain strains, however, were sometimes lethal 
for rabbits when injected intravenously. This variable 
pathogenicity it not understood and requires further study. 

3. Since August, 1958, studies have been under way in 
cooperation with or. M. Elizabeth sharpe of the National 
institute for Research in Dairying, shinfield, Reading, 
England, where Mr. Rogosa is presently Visiting scholar, 
on the Immunological character of the lactobacilli as it 
relates to the biochemical properties and cellular composi- 
tion of these organisms. The latter two phases are being 
studied by the English group by means of paper chromato- 
graphy and related techniques. It appears that species can 
be differentiated according to their contents of amino acids,, 
peptides, hexosamine, and other cellular constituents. 

Significance to Dental Research : 

Since systematic immunological and pathogenicity studies are 
lacking, these studies are important in differentiating 
organisms; in increasing knowledge of the structure of the 
cell; and in determining the pathogenic potential of the 
mouth microbiota. 

- 3 - Serial no., NIDR-22 

Pwt A (continued) 

Proposed Coune o£ project ; 

The immediate future will be devoted to the immunology 
end chemical composition of the lactobacllli and their 
taxonomlc relationship to the oral diphtheroids, particularly 
the catalase-rtegative diphtheroids; and to the serological 
characteristics of streptococci Isolated previously from the 
rat. Further antigenic and pathogenic snalysls of the 
Velllonelle is required, particularly of the animal strains,, 
Present techniques will be adapted for precipitin studies 
and the use of. conjugated labeled antibodies for the 
demonstration of the organisms in situ in tissues. 

Part B included yes 

- u 

Serial no„ NIDR-22 

Individual Project Report 
Calendar Year 1958 

Pert B ; Honors, Awards, and Publications 

Publications other than abatracta from this project: 

Rogosa, M,, Fitzgerald, R. j , Macintosh, m. Elizabeth, 
and Beaman, Alfred J. improved Medium for Selective 
isolation of Vei llonella ,, j, Bacteriol. 76 : 4, 455-456. 
Oct., 1958„ — 

Honors and Awards relating to this project; 

Resident Visiting scholar, university of Reading, England 

Serial No, NIDR-23 

lo Microbiology 
3. Bethesda, Md. 

individual Project Report 
Calendar year 1958 

Part A 

Project Title; The Rat as An Experimental Animal for Research 
on periodontal Disease Q 

Principal investigator: Dr. A. A. Rlszo 

Other investigators: None 

Cooperating Units: None 

Man Years (calendar year 1958): 
Total 1 1/3 
Professional: 1 
Other: 1/3 

project Description: 

Objectives : 

1. To discover the nature and prevalence of periodontal 
disease in young and adult laboratory rats on various 
diets and to note particularly the tendency to form cal- 
careous deposits. 

2. To develop methods for the clinical inspection of rats 
so that observation of their periodontal health can be made 
throughout an experimental period. 

3c To study the effects upon gingiva and upon the accumula* 
tion of dental deposits, of altering the diet and salivary 

4. To determine whether or not filamentous microorganisms 
comprise part of the resident oral flora of rats. 

Methods Employed ; 

1. Examination of available material ,, A number of fixed „ 
of def Jeshed, and of microscopic specimens of rat and mouse 

- 2 - serial no NIDR-23 

Part A (continued) 

jaws collected from other investigators were examined 
In addition, fresh animals, including germ-free rats, 
were obtained and studied^ The periodontal findings 
observed in this material were correlated with age and 

2. oral examinati on of liv e ra ta., The otoscope, cystoscope, 
sigmoidoscope, nasal speculum, vaginal speculum, and the 
johannaen hamster device were tested for their effectiveness 
aa alda to the clinical oral inspection of live rats. 
Finally, a completely intraoral rat mouth opener was de- 
veloped and tested. 

3. Dietary and salivary alteration . Experimental diets 
were fed to groups of rats to determine whether these diets 
cause gingival accumulations, calculus deposits, and 
periodontal breakdown. The dietary components tested 
individually, and in combinations, included whole milk 
powder „ mucin powder, dibasic calcium phospha&»„ and a 
standard salt mixture. The Influence of the physical con- 
sistency of the diet was tested by maintaining different 
groups on Purina laboratory chow In the form of pellets, 

of coarse particles and of powder. 

The effect upon periodontal health of altering the salivary 
flow was studied by applying conventional desalivation 
techniques to rats, and by performing clinical oral examina- 
tion periodically over a period of several months. First- 
hand observations of the extent of xerostomia were made. 

4 Bacteriological studies . Specimens of gingival accumula- 
tion were taken directly from the teeth of live rats and 
cultured for filamentous organisms using the Isolation and 
identification methods of Dr. A. Howell, Jr. 

Major Findings : 

1. Heavy, but friable, calcareous deposits were found on 
the molar teeth of rats maintained for 40 days on a diet 
consisting mainly of sucrose and dried milk (Stephen #580). 
Similar deposits were observed In rats maintained for 56 
days on a diet consisting of unenrlched white flour, 
vitamins and salts (Hundley, #4256C). Heavy, firm, tenacious 

- 3 - Serial No, NIDR-23 
Part A (continued) 

deposits were observed in rats maintained for 90-120 days 
on a diet, containing corn starch, whole milk powder and 
yeast (Fitzgerald, #550). Finn deposits gave X-ray 
diffraction patterns resembling that of hydroxyapatite, 
whereas the friable precipitates did not, 

Generalized alveolar bone loss was minimal in rats fed 
adequate diets up to 12-18 months of age. The gross and 
microscopic anatomy of the oral tissues of germ- free rats 
up to 6 months of age did not appear to differ from that 
of conventionsl rats. 

2. A suitable method of clinically examining the complete 
dentition of live rats was made possible by the fabrication 
of a new type of Intraoral mouth opener from orthodontic 
wire. In addition to holding the jaws apart, this device 
spreads the cheeks apart so that all of (he oral structures 
are accessible for direct inspection. Since it takes up 
very little space and offers no obstruction to the entrance 
of light or to manipulation, the opener makes operative 
procedures and photography simple tasks. Since its develop- 
ment, this device has been tested successfully in more than 
100 rats, weighing from 70-600 grams, in hamsters of 100 
grams, and in mice as small as 20 grams. 

3. Soft gelatinous gingival accumulations were observed in 
adult rats maintained for 4 months on powdered laboratory 
chow diets to which were added individually, and in com- 
bination, the following powdered components: whole milk,, 
mucin, dibasic calcium phosphate, and a standard salt 
mixture, A smaller amount of similar accumulations were 
seen in rats fed ground laboratory chow alone, and little 
or none in rats fed pellets of laboratory chow. 

Adult rats desalivated by removing or defunctionalizing all 
major salivary glands, and fed moist diet for several months 
thereafter, seem to suffer no loss in weight or periodontal 
health. However, frequent clinical examination shows that 
the mouth is not dry as a result of conventional desallva- 
tion techniques, and indicates that remaining secretions 
arise mainly from minor sublingual glands „ Therefore, in an 
effort to create a more nearly complete xerostomia,, a saethod 
of removing the minor sublingual glands was developed. With 
the aid of the previously mentioned rat mouth opener, an 
intraoral approach was successfully employed. 

- 4 - Serial Noo NIDR-23 
Part A (continued) 

4. Prom accumulations around the teeth of several rats, 
a filamentous organism producing true mycella has been 
Isolated in pure culture, 

Significance to Dental Research : 

The formation and retention on teeth of precipitates of 
calcium salts in an organic matrix containing many fila- 
mentous organisms are phenomena of paramount Importance to 
the initiation of periodontal disease, since the presence 
of these deposits seams to be responsible for the subsequent 
pocket formation and alveolar bone loss. However, fundfl 
mental animal research on periodontal disease has been 
hampered so far by the apparent lack of a suitable small 
animal subject. The rat has been considered unsuitable 
primarily because it develops neither calculus deposits 
nor periodontal disease under average laboratory conditions.. 
Since it is now known that firm, tenacious calcareous pre- 
cipitates can be induced to deposit on the teeth of the 
rat, and since filamentous organisms have been isolated 
from gingival accumulations on the teeth of rats, the use 
of this animal in the experimental investigation of perio- 
dontal disease is promising. 

Proposed Course of Project : 

Studies of calculus deposits in adult rats are to be con- 
tinued with observations to be made throughout extended 
experimental periods. The relation of these deposits 
to filamentous organisms and to periodontal breakdown 
are to be Investigated. 

The nature and occurrence of the filamentous organism 
Isolated from rats is to be determined by further study.. 
(Cultures of this organism are to be furnished to 
Dr. R. J. Fitzgerald for studies with germ-free rats). 

In collaboration with Or. Arden Howell, a study of the 
relationship of oral filamentous organism to the develop- 
ment of dental calculus deposits in humans is to be 

Part B included No 


Individual Project Reports 
Calendar Tear 1958 


24. K. A. Piez : Biochemistry of Amino Acids and Proteins in 

Mammalian Cell Cultures. 

25 • *• A - piez : Analytical and Structural Aspects of Collagen 
in Relation to Function and Disease. 

26. P.J. McClure : Nutrition and Dental Carles with Emphasis on 
Protein and Cereal Foods, Lysine, and Phosphate Supplements. 

27 « J- E. Folk ; Protein and Amino Acid Chemistry. I. Pro- 
teolytic Enzymes; Chemistry, Kinetics, Isolation and Purifi- 
cation. II. Blood Clotting; Enzymology and Amino Acid 
Sequence Studies. 

28. H. C. McCann : Study of the Inorganic Chemistry of Mineralized 
Dental Tissues, Surface Enamel Chemistry as Affected by Diet 
and Oral Environment and their Relation to Dental Caries. 

29. R. H. Larson : A Study of EDTA and DHA as Caries Potentiating 
Agents and Prenatal Factors Affecting Caries Susceptibility in 
White Rats. 

30. I. Zipkln : Biochemistry and Physiology of the Salivary Secre- 

31. I. Zlpkin ; Non-dental Physiological Effects of Fluoride. 

32. M. S. Levis : Biochemistry of Salivary Proteins. 

33. H. B lumen t ha 1 : A Study of the Biochemistry of the Submaxil- 
lary Glands of the Rat with Particular Reference to Poly- 
saccharide Synthesis and Intermediary Carbohydrate Metabolism. 

34. R. C. Liking : Biochemical & Biophysical Studies of Calcifi- 

35. S. A. Narrod : Enzysao logical Studies in Relation to Oral Tissue 
and Periodontal Disease. 

005 '16 $ 
00J C V6T$ 








1^3 OX 

6S6£ A'S ^oj eacrj3BTiftqo pa^Bsrjrjsa 
Tj iBfBaqoo-jg jo Ooaeaoqe'f 

1, Biochemist 
3. Bethesda. 

^ject Report 
Calendar Year 1958 

Biocheraintry of Amine Acids and Pre 
Mammalian Cell Cultures. 

A. Piez 

sstigators: Dr. EllLnor Weiss, Visiting Scientist 

Dr. H. EagU (NIAID No. 50) 

dar year 1958 ) : 



is the broed purpose of this project to obtain basic 

. concerned with protein and amino ecid metabolism 
mammal inn cells in cu/.ture. The current studies are 

turnover, intracellular free amino acids, 
cystine biosynthesis. 

Met hoc 

I laboratory procedures. 

dinfi s: 

as been found that protein turnover in cell cultures 
proceeds at a rapid rate, about 1 percent per hour. This is 
about one fourth of the riaximal net synthetic rate. 

aover process is largely, if not entirely, intra- 
cellular. That is cell death and replacement are not 

lved. It proceeds undiminished whether the celXs are 
growing or are in a steady state; a very large proportion of 
the proteins of the cell are involved. 

The composition of the free amino acid pool of several cell 
lines has been examined. Both quantitatively and qualitatively. 

Part A, Cont'd. - 2 - Serial No. MIBR.-24 

the pool resembles that found in tissues in vivo . Most 
amino acids are concentrated by the cell with distribution 
ratios of about eight for the essential amino acids under 
the usual conditions of growth, but much higher for the 
nonessential amino acids. The distribution ratios increase 
with decreasing extracellular concentration. 

The biosynthesis of cystine has been studied. Preliminary 
results indicate that cystine is not an essential amino 
acid if a source of sulfur is available. This can be 
inorganic sulfur or a variety of organic compounds. The 
carbon skeleton of cystine is derived in part from glucose 
and In part from unidentified sources. 

Significance to Dental Research ; 

Cell culture is a relatively new tool suitable for the. study 
of many metabolic processes. These basic studies are 
important to all branches of the medical sciences. Topics 
more immediately related to dental research, such as 
collagen biosynthesis, are amenable to study with these 

Proposed Course of Project : 

It is planned to continue these studies along the lines 
already started. Preliminary work is underway to examine 
the possibility of studying collagen biosynthesis in pure 
cell cultures. 

Part B included Yes 

- 3 - Serial No. NIDR-24 

Individual Project Report 
Calendar Year 1958 

Part B . Honor 8, Awards and Publications 

Publications other than abstracts from this project: 

Pies, K. A. and Hagle, H. The Free Amino Acid Pool of Cultured 
Human Cells. J. Biol. Chem. 231:533-545, March 1958. 

Eagle, H. , Piez, K. A., Fleischman, R. and Oyama, V. I. Protein 
Turnover in Mammalian Cell Cultures. J. Biol. Chem. In Press. 

Individual Project Report 
Calendar Year 1958 

Serial No. HIDK 25 
1. Biochemistry 
3. Bethesda, Md. 

Part A 

Project Title: Analytical and Structural Aspects of Collagen 
in Relation to Function and Disease. 

Principal Investigator: Dr. R. A. Plex 

Other Investigators: Dr. R. C. Liklns 

Cooperating Units: Dr. B. Witkop (NIAMD , Lab. of Chemistry) 

Man Years (calendar year 1958): 
Total i 1% 
Professional : % 
Other: 1 

Project Description: 

Objectives : 

It is the long-range purpose of this project to examine 
co Hag ens from various sources vith respect to their chemical 
and physical nature (such as amino acid composition, amino 
acid sequence, crosslinking and solubility) in an attempt 
to correlate these factors with function of the tissue or 
with disease states. The immediate objectives are (1) to 
compare the amino acid composition of collagens from 
different tissues of the same animal, and (2) to compare 
the amino acid composition of collagens from different 
invertebrate phyla. 

Methods Employed: 

Usual laboratory procedures. 

Major Findings : 

The amino acid composition of collagen from akin, tendon, 
bone, and dentin of the rat and of the steer have been 
determined. It vas found that these proteins are identical 
except for the lysine and hydroxy lysine contents. The sum 
of these two amino acids was constant but the ratio differed 

Part A . Cont'd. ~ 

" c " Serial No. H.IDR-25 

from about six (rat skin) to nearly one (steer dentin) The 
variation between tissues was not the same for the So 
%£?*'* , SiaCe the hydr ° Xyl 8rou P of hydroxylysine 2, be 

Pojifera has been determined. The amounts of proline" 
hydroxyproline. and hydroxyzine, amino acids characteristic 
of collagen, varied widely in these proteins. Howler? the 
glycine content was constant at a value approaching one third 
of the total amino acid residues. This is in agreement with 
recent structure models, based primarily on pSsTcTdata 
which require that every third residue be glycine It 
££. "Eft therefo "« that ""I. is a nectary and 
oTaTrot'^ras'Tco"! 1 ^ 00 '" ^ ^^ l *&**»«- 
Significance to Dental Research : 

a^dMeth^denti^^rr ' 610 ^' "^ (dermU >» tendon » b ^s, 
thf« ?<« <*•»"»>• It may play a role in the function of 

in! r%5 T 8 a ? d lQ P«»»»lo«ic«l ".tes which involve them 
studL^f T<* Carl68 ^ P erio ^tal disease. Basic ' 
llttjt L ^° teln may « therefore, contribute to a 

better understanding of function and disease. 

Proposed Course of Project : 

These studies are to be expanded in the direction of both 
additional animals (and tissues) and further chemical and 

cor 8 i?*J, fltU K le8 - S P ecifi ""y ^ is planned to look Tor a 
correlation between hydroxyzine content and crosslinkiL 
Another study will be concerned with collagens bearing ^' 
evidence <?>f genetic defects. S 

Part B included Yes 

- 3 - Serial No. NIDR-25 


Individual Project Report 

Calendar Year 1958 

Part B . Honor 8, Awards and Publications 

Publications other than abstracts from this project: 

Piez, K. A. and Gross, J. The Amino Acid Composition and 
Structure of Some Invertebrate and Vertebrate Collagens. 
Blochlm. et Blophys. Acta. In Press. 

Serial Ho. NIDR-26 
1. Biochemistry 
3. Bethesda, Md„ 

Individual Project Report 
Calendar Year 1958 

Part A. 

Project Title: Nutrition and Dental CarleB with Smphasis on 

Protein and Cereal Foods. Lysine, and Phosphate 

Principal Investigator: Dr. F. J. McClure 

Other Invest igatorn: None 

Cooperating Units: None 

Man Years (calendar year 1958): 
Total: 5 
Professional : 1 
Other: 4 

Project Description: 

Objectives : 

To obtain information on (a) the rei.ation of dietary protein 
to caries etiology, (b) the effect of heat processing and 
cereal foods on the development of experimental rat caries, 
and (c) the effect of phosphate minerals on dental caries. 

Methods Employed : 

The quantity nnd quality of protein in the diet are varied 
to determine their relation to the development of experimental 
rat caries. Special diets contain wheat flours, bread, 
wheat biscuit,, and millet as the cariogenic agents. Dietary 
protein and mineral phosphates ere evaluated ae factors 
in the development of experimental caries in white rats. 

The major interest of this year's research has been the 
effect of phonphate minerals on experimental rat carles. 
It now appears that the insoluble phosphates Ca-(PO > 2 and 

P art A , Cont'd. - 2 - Serial No,, HxDR-26 

CaHPO, alone have no carlostatlc effect. The presence of 
NaCl end CaHPO, combined, in the die:, proved significantly 
ci.-ies inhibitory, thus confirming previous evidence that 
CiHPO, in bread containing fcaCl was <:ario8tatic. A very 
pronounced inhibition of caries was produced by Cad^PO^^. 
particularly in. the presence of KaCl in the diet. 

In addition to the results with phosphates, a significant 
production of caries was brought about by diets containing 
millat. Millet is an imbalanced protein food widely used 
tl.iougiout the Middle East and Asia. It holds particular 
it tere^t for experimental purposes because of lysine and 
other amino acid imbalance. 

S ignifi c ance to Dental Research : 

The pronounced effect of pho3phates en experimental caries 
supports the proposals to study the effect of phosphated 
flour on human caries. The results vith Ca(E2PO^)2 are 
ptrticularly provocative because of the presence of this 
phosphate at a level of 1.75% in self rising flours, These 
flours and other prepared mixes containing CaC^PCv^ are 
used extensively in human diets. 

The extensive consumption of millet--"the poor man's cereal"- 
in many foreign lands adds interest to its study as a factor 
in humrn caries etiology. 

P ropose d Course of P roject ; 

Continued observations will be made oi experimental rat 
caries as affec:ed by phosphate minerals, the wheat flours, 
mi'. let and variable protein factors o:: the diet. A 
conprehensive field study to elucidatu the caries 
inhibitory effect of a phosphated bread is now in the stage. 

Part B included Yes 

- 3 - Serial No. NIDR-26 


Individual Project Report 

Calendar Year 1958 

Part B . Honors, Awards and Publications 

Publications other than abstracts from this project: 

McClure, F. J. Wheat Cereal Diets, Rat Caries, Lysine and 
Minerals. J. Nutrition 65:619-632, Aug. 1958. 

McClure, P. J. and A. Muller, Jr. The Caries-inhibiting Effect 
of Dibasic Sodium Phosphate and Dibasic Calcium Phosphate Added 
to Wheat Flour and Bread Diets. J. Amer. Dent. Assoc. 58:36-41, 
January 1959. 

Serial No. KIDR-27 
1. Biochemistry 
3. Bethesda, 

Individual Project Report 
Calendar Year 1958 

Part A. 

Project Title: Protein and Amino Acid Chemistry. I. Proteolytic 
Enzymes; Chemistry, Kinetics, Isolation and 
Purification. II. Blood Clotting; Ensyraology 
and Amino Acid Sequence Studies. 

Principal Investigator: Dr. J. E. Polk 

Other Investigators: None 

Cooperating Units: American Dental Association 

Drs. K. Laki, J. A. Gladner, Y. Levin, 
W. Carroll, and Miss I. Knoller (NIAMO, tab. of 
Physical Biolo»j). 
Man Years (calendar year 1958): 
Total: 2 
Professional : 1 
Other: 1 

Project Dnscription: 

Objectives : 

I. a. Purification, physical chemical characterization, 
studies on activation and specificity of carboxypeptidase B 
and its application to end group analysis. 

b. A study of the activity increasing effects of certain 
metal ions on the carboxypeptidases. 

c. Development of simplified procedures for the purification 
of leucine aminopeptidase. 

II. A study of these aspects of the fibrinogen to fibrin 
conversion by thrombin: 

a. Mode of action and specificity of thrombin. 

b. Physical end chemical changes in fibrinogen. 

c. The structure and physiological properties of peptides formed. 

Part A. Cont'd. - 2 - Serial No. NIDR-27 

Methods Employed ; 

I. a. During this year some time has been devoted to the 
further purification of carboxypeptidase B as measured by 
physical chemical methods. The possibility of this enzyme 
being a metal activated one has been realized by employing 
metal chelators to block activity. Ester substrates for B have been prepared and employed in 
preliminary kinetic studies. 

b. Studies on the activation of carboxypeptidasee A and 
B by preincubation with certain metal ions has been 
investigated- Comparison on amidase and esterase activities 
following met.l treatment is underway. 

c. A method for the cellulose column chromatography of 
leucine aminopeptidase employing a rapid elution with a 
straight line gradient of salt has been studied. 

II. a. Amino acid sequence studies of the peptides A and 

B of cofibrin as described in II. b and c have been used to 
formulate a possible hypothesis of mechanism of the action 
of thrombin on fibrinogen. 

b and c. Cle.'.vage of peptides A and B with trypsin and 
chymocrypsln 'ollowed by amino acid and end group analysis 
of the result ag fractions has led to a partial structural 
elucidation o:: these peptides. 

Major Findings : 

I. a. Procarboxypeptidase B has been more extensively 
purified by means of cellulose column chromatography. How- 
ever physical chemical methods indicate that the material 
is still heterogeneous even though specific activity values 
have been increased approximately 10 fold over previously 
reported preparations. The action of carboxypeptidase B Is 
complately inhibited by certain metal chelators strongly 
indicating the. presence of an essential metal. It has been 
demonstrated that this enzyme has a pronounced esterase 
activity toward hippuryl-L-arglninic acid as well as 
chloroacetyl-I -argininic acid. 

b. The hydrolytic activity of carboxypeptidase A and B is 
increased 100 6 400X respectively by preincubation with Co"* 4 ", 

Fart A. Cont'd, - 3 - Serial No. HIDR-27 

Ni and Pe^"*" ions. Certain hypotheses concerning the role 
of these metals in enhancing activity have been formulated. 

c. The cellulose chromatography of leucine amino peptidase 
offers a very simple method of purifying this enzyme in 
preparation for its use in end group analysis. 

II. a. It hcs been observed that the peptides A and 3 of 
cofibrin have two outstanding structural similarltieo; (1) 
C-terminal arginine preceded by a group of mono-amino, 
mono-carboxyl amino acids and (2) a group of negative 
charges exerted by glutamic and aspartic acids toward an 
R-terminus. Theories of fibrin polymerization and thrombin 
action have teen derived from these findings. 

b and c. See II. a. 

Significance to Dent al Research : 

1. An understanding of the mechanism of protein digestion 
is fundament el to any biological research program. A clear 
evaluation of the specificity and mode of action of 
carboxypept lease fl contributes knowledge to the ovevall 
protein metabolism. 

Carboxypept lease B has proven to be a valuable tool in 
both natural and chemically modified protein-structure 
detei-Tninat ior. . It also complements the classical 
carboxypept tease In this respect. 

A specific vrlue of carboxypeptidaee to dental research may 

lie in some under standing of lysine metabolism as it 

relates to the etiology of experimental smooth surface 
dental cariee. 

2. A thorough understanding of the structures and 
mechanisms involved in the fibrinogen to fibrin reaction 
should open tie vey for a more systematic pharmacological 
and clinical approach to the correction of certain abnormal 
blood clotting reactions. 

Proposed Course of Project : 

I. a. Attempts at complete purification of procarboxypeptidase 
B will be continued, followed by studies of mechanism of 
activation to the active enzyme and role of metal in the 

Part A. Cont'd. - A - serial No. NIDR-27 

b. The met a.', activation of carboxypeptidasas presents 

a complicated problem requiring a good deal more study of 
mechanism and significance. 

c. Repatition of work and preparation of manuscript. 

II. a, b, anl c. Future \*>rk will involve the study of 
peptides forced during the clottinj of human fibrinogen 
and bovine fibrinogen by thrombin :rom the above sources, 
in s hope to gain a more complete inderstanding of the 
specificity and mode of action of :hrombin. 

Part B Included Yes 

- J - Serial No. NIDR-27 


Individual Project Report 

Calendar Year 1958 

Part B . Honors, Awards and Publications 

Publications other than abstracts from this project: 

Polk ., J. E. The Properties of a New Proteolytic Enzyme — 
Carboxypeptidase B. Med. Science 3:749-751, 1958. 

Polk .. J._E. and Gladner, J. A. Carboxypeptidase B. I. 
Purification of the Zymogen and Specificity of the Enzyme. 
J. Biol. Chem. 231:379-391, March 1958. 

Gladner, J. A. and Folk , J. E. Carboxypeptidase B. II. Mode 
of Action on Protean Substrates and its Application to Carboxyl 
Terminal Group Analysis. J. Biol. Chem. 23^:393-401, March 1958. 

Shiota, T., Folk, J. E. and Tietze, F. Inhibition of Lysine 
Utilization in Bacteria by S-( »3 -Aminoethyl) -cysteine and its 
Reversal by Lysine Peptides. Arch. Biochem. and Biophys. 
77:372-377, Oct. 1958. 

Laki : K., Gladner, J. A., Folk , J. E. and Komitz, D. The Mode 
of Action of Thrombin. Thromboeis et Diathesis Haemorrhagica 
2:205-217, 1958. 

Gladner, J. A., Folk , J. E . , Laki, K. and Carroll, W. R. The 
Thrombin Induced Formation of Co-fibrin. I. Isolation, 
Purification and Characterization of Co-fibrin. J. 3iol. Chem. 
In Press. 

Folk , J. E. , Gladner, J. A. and Laki, K. The Thrombin Induced 
Formation of Co-fibrin. II. Preliminary Amino Acid Sequence 
Studies on Peptides A and B. J. Biol, Chem. In Press. 

Folk , J. E. and Gladner, J. A. Carboxypeptidase B. III. The 
Specific Esterase Activity. Biochim. et Biophys. Acta. In Press. 


Individual Project Report 

Calendar Year 1958 

Serial No. NIDR-28 
1. Biochemistry 
3. Bethesda, Md. 

Part A. 

Project Title: Study of the Inorganic Chemistry of 

Mineralized Dental Tissues, Surface Enamel 
Chemistry as Affected by Diet and Oral 
Environment and their Relation to Dental Caries. 

Principal Investigator: H. G. McCann 

Other Investigators: None 

Man Years (calendar year 1958): 
Total: 2 
Professional : 1 
Other : 1 

Project Description: 

Objectives : 

1. To find what reactions take place between mineralized 
tissues and certain other substances, such as fluoride, 
phosphate, or calcium, which may furnish some protection 
to the teeth against caries. 

2. To obtain information on the properties and structure 
of the mineralized tissues from these reactions. 

3. To determine what effect the inorganic composition of 
the teeth, as regulated by diet or other factors, may have 
on caries susceptibility. 

4. To determine how the mechanism of calcification is 
affected by diet. 

5. To evplvc methods of analysis suitable for the Inorganic 
constituents of mineralized tissues. 

Methods Employed : 

The teeth and bones of rats receiving diets having variable 

Part A . Cont'd. - 2 - Serial No. HIDR-28 

calcium-phosphorus ratios, and various magnesium-phosphorus- 
calcium ratios are analyzed. A micro-method was adapted 
for the direct determination of magnesium in mineralized 
tissues. A comparison of dietary vs. injected Ca^5 end p32 
vas also made. 

(2) An in vitro study of the composition, solubility and 
radioactive exchange of Ca^5 and P^2 is being carried out 
on the surface enamel of the teeth of rats which have been 
fed various mineral-supplemented diets. For this purpose 
whole crowns, or powder obtained by grinding off the 
surface of the teeth. are employed. 

(3) The effect of various methods of treatment of rat femurs 
on the exchange with radioactive phosphorus is being 
investigated to obtain a better understanding of the role 

of the inorganic crystals vs. the organic matrix. 

Major Findings : 

Further experiments with a skim milk powder diet with added 
mineral supplements have confirmed previous results. Thus, 
rats fed this diet (with Xa^HPO, added so that the Ca:P 
ratio was decreased to 1:3 as compared with the normal 1:1 
ratio) have & significantly reduced caries incidence and 
number of teeth affected. In the teeth and bones of these 
rets there was a decided tendency toward a alight decrease 
in the Ca:P ratio, an increase in magnesium content and 
n decrease in carbon dioxide. However, it seems unlikely 
that these are factors associated with the decrease in 
caries, as one group of rats had a significant decrease in 
caries when fed a diet with a high Ca:P ratio. This was 
accomplished by the addition of CaCO, to the diet which had 
the opposite effect on the inorganic chemistry of the 
mineralized tissues, i.e. an increased Ca:P ratio, lowered 
magnesium, and increased carbon dioxide. 

In vivo studies, employing radioactive tracers in rats, have 
uhown that enamel has an uptake differing from other 
mineralized tissues (dentin and bone). The uptake of soluble 
calcium and particularly soluble phosphate was greater in the 
enamel than in the other mineralized tissues as compared with 
the uptake from the relatively insoluble CaHPO, . This is 
apparently due to a direct exchange of Ca45 and P-*2 i n the oral 
surface enamel. Other tissues which must obtain all or nearly 

Part A. Cont'd. - 3 - Serial Wo. NIDR-28 

all of their minerals systemically vill utilize equally 
any source of Ca and P providing it is completely 
solubilized in the intestinal tract. This mineral 
exchange on oral surface enamel, the apparent lack of 
effect on caries of changes in inorganic tooth chemistry, 
and the apparent requirement that a phosphate be soluble 
in order to bring about a reduction in caries, all suggest 
that caries reduction may be caused by a suppression of 
enamel solubility through a common ion effect. 

Various treatment of rat bone, such as autoclaving, 
extracting cold or hot, removal of organic matter, etc., 
which would chiefly affect the relationship of the 
inorganic mineral vs. the organic matrix, were compared 
by measurement of the exchange of P^2 ± a a phosphate buffer 
solution. The coadition of the organic matrix was found 
to have an important effect on the rate of exchange. 

Significance to Dental Research ; 

As more is learned about the effect of dietary factors 
on oral tooth surfaces and their concommitant Influence 
on dental caries, less empirical methods may be employed 
in the prevention of this disease. 

Pre po tied Course of Project : 

(1) An in vitro study of the surface chemistry and reactions 
of the enamel crowns of rats fed various diets supplemented 
vith various minerals will be continued with the aid of 
chemical analyses, radiochemical tracers, and solubility 

(2) Studies have been initiated to determine the effect of 
magnesium supplementation on cariogenic diets and the 
effect of various magnesium-calcium-phosphorus ratios on 
carles and tooth and bone composition. 

(3) It is proposed to study the kinetics of and the 
composition of calcium phosphates formed from salt 
colutions similar in composition to those present during 
calcification, and to study the effect of various changes 
in the composition, pH, etc. of this solution. 

(4) Additional work on the micro-analysis of magnesium will be 
carried out to overcome the interference present in rat enamel 
but not found in human enamel. 

Part B included Yes 

- 4 - Serial No. NIDR-28 


Individual Project Report 

Calendar Tear 1958 

Part B: Honors, Awards and Publications 

Publications other than abstracts from this report: 

McCann, H. G. and Path, E. H. Phosphate Exchange in Hydroxyl- 
apatite, Enamel, Dentin, and Bone. I. Mechanism of Exchange, 
J. Biol. Chem. 231:863-868, 1958. (April). 

McCann, H. G. and Fath, E. H. Phosphate Exchange in Hydroxyl- 
apatite, Enamel, Dentin, and Bone. II. Effect of Pluoride on 
the Exchange. J. Biol. Chem. 231:869-872, 1958, (April), 

1 . Bioche - 
3. Bethesda, 




:b: IT I. Zlpfc 

Han • 8): 

ilciura, m 

F.OTA wa3 added to die 
aad fed for 56 days ac.d to di< : a 

ayp. After sacrifice, molar 
tef caries in the usual manner. 

Hajor Pindi." 

It was shown that the dioodium and the magnet' 

EDTA ctxeed e significant increase in cai The cl 

salt was less cariogenic than the other two sa 

58f> the calcium chelate was the least toxic as she 

md weight gair.g. On diet 632. 
caused siitilar reductions in food con sump: 

lias, Incisor pigmentation and hemogl 

.'ental Resea rch: 

The data suggest that calcium chelat .on may play a role in 
carles potentiation by EDTA. The elucidation of the 
etiology of dental caries may be promoted through the 
study of potentiators such as EDTA compounds. 

jabpr o ject B: 

Obje ctives; 

To sti dy prenatal toxicity of EDTA and DHA as a factor 
affecting dental caries development. 

Methods Employed : 

or 5.0Z) end DHA (0.1, 0.3 or 0„5%) were 
added to an adequate basal diet which was fed for varying 
periods in order to produce a highly toxic reaction in 

iung adult female rtt.s Toxicity was evidenced by 
anorexia, weight losses jp to 30%, and reduced hemoglobin. 
The animals were then fed the diet without EDTA or DHA 
supplementation, and allowed to return to normal weight 
before being bred. Following conception, as determined 
by vaginal smear, these experimental rats and an equal 
number of pregnant normal rats were fed diets containing 
either 1.0% EDTA or LA for periods ranging from 2 
to 15 days during gestation. 

Chronic toxicity was produced by feeding breeding animals 
the basal diet containing 0„4% EDTA from prior to conception 
through the first two weeks of lactation. Pellets were fed 
the last week before weaning. 

21 days of age all the rats in these experiments were 
weaned and then fed a ceriogenic diet for two months They 
were sacrificed and the teeth examined in the usual manner. 
Forty-one litters comprising 297 offspring completed the study. 

Major Findings ; 

Toxicity produced by EDTA or DHA, and from which the rats 
recovered prior to conception, gave no evidence of interfer- 
ence with pregnancy. On the other hand, 18 of 21 animals 

Part A . Cont'd. - 3 - Serial No. NIOR-29 

receiving 1.0% EDTA in their diets for a period of 7 or more 
days after conception either resorbed their fetuses or 
delivered non-viable young. The three exceptions were rats 
that delivered normal, viable litters after receiving EDTA for 
10, 12 and 14 days. Eight rats fed 0.3% DHA in the diet for 
a period of 7 or more days resorbed their young, and 3 others 
delivered normal, viable litters after receiving DHA for 8, 9 
and 12 days respectively. 

The mothers on the diet containing 0.4% EDTA had a lover birth 
rate and higher neonatal death rate than the controls. However, 
only 5 litters were reared on this regimen. 

In spite of having been born of mothers subjected to near 
maximum EDTA or DHA toxicity prior to or during pregnancy 
(compatible with delivery of live offspring) there was no 
apparent prenatal influence on dental caries susceptibility. 

Significance to Dental R esearch : 

Since one of the major symptoms of EDTA and DHA toxicity is 
anorexia and consequent malnutrition, it is of interest that 
dental caries was not related to these factors in these 

Proposed Course of Project : 

It appears that although pre-natally administered EDTA and DHA 
are ineffective in the development of dental caries in the 
offspring, administration of EDTA to the weanling rat promotes 
dental caries. It is proposed now to determine whether EDTA 
toxicity induced immediately after weaning will predispose 
the rat to the development of caries when subsequently placed 
on a cariogenic diet. 

Part B included Yes 

- A - Serial No. HIDR-29 

Individual Project Report 
Calendar Year 1958 

Part B . Honors, Awards and Publications 

Publications other than abstracts from this project: 

Zipkin, I. and McClure, F. J. Potentiation of Smooth Surface Caries 
by Dehydroacetate Variously Administered to the White Rat. Proc. 
Soc. Exptl. Biol. & Med. 9_7:318-320, February 1958. 

Serial No. NIDR-30 

3. Bethesda, Md. 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Biochemistry and Physiology of the Salivary 

Principal Investigator: Dr. I. Zipkin 

Other Investigators: None 

Cooperating Units: Dr. ?. Pronove, Dr. P. Chen (NHT, General 
Med. & Exptl. Therapeutics Branch) 

Man Years (calendar year 1958): 
Total: 2 
Professional : 1 /3 
Other : 2 

Project Description: 

Objectives : 

To study the basic chemistry and phys5.ology of human 
parotid, submaxillary, and whole salivary secretions and 
their relation to oral and systemic disease. 

Subproject A. 

The tyrosine and tryptophan content of human parotid saliva. 

Methods Employed : 

Por tyrosine analysis, a modification of the Ddenfriend and 
Cooper procedure; for tryptophan analysis, a modification 
of the Steers and Sevag procedure. These constituents are 
being determined in the whole and hydrolysed parotid saliva, 
la the ultrafiltrate and in the TCA precipitate. 

Major Findings : 

At present, the methods have been developed for application 
to salivary analysis. 

Part A. Cont'd. - 2 - Serial No. NIDR-30 

The concentration of total tyrosine in normal salivas 
is as follows: whole parotid saliva 8.3 mg %, TCA 
precipitate 6.9 mg I, TCA supernatant 1.8 mg X, 
ultrafiltrate 1.9 mg Z. 

The concentration of tryptophan averaged 2.8 mg I. The 
values for unhydrolyzed and hydrolyzed saliva were 
essentially similar indicating that total tryptophan 
was being determined by the procedure. Only negligible 
quantities of tryptophan were found in the ultrafiltrate 
and in the TCA supernatant. 

Proposed Course of Project : 

The relation of salivary tyrosine and tryptophan to caries 
and periodontal disease as well as various systemic 
diseases will be studied. The contribution of the 
tyrosine and tryptophan content of saliva to its ultra- 
violet absorption spectrum will h« studied. 

Subproject B. 

The "salivary clearance" of phosphate and lnulin, 

Methoda Employed : 

Patients undergoing urinary clearance tests for phosphate 
and lnulin were studied. Whenever urine was obtained from 
t.'xe catheterized patient, parotid saliva was collected by 
means of the Lashley cup. Samples of blood were also 
collected. The urine and blood specimens have been 
analyzed for phosphate and lnulin by standard procedures. 
Analytical data on the saliva are not completed. 

Significance to Dental Research : 

Since saliva is in Intimate contact with the teeth and other 
oral structures, it is important to study its chemistry in 
both normal and pathological states. Little basic bio- 
chemistry of the saliva secretion is known and such informa- 
tion is necessary to study the relation of saliva to oral and 
perhaps systemic diseases. 

Proposed Course of Project : 

Study will be continued to include additional normal and 
pathological cases for the determination of salivary 
"clearance" of these and other constituents. 

Part B Included No 

Serial No. NT.DR- 31 
1. Biochemistry 
3. 3etrescla, Md. 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Non-dental Physiological Effects of Fluoride, 

Principal Invest igator: Dr. I. Zipkin 

Other Invastigators: Dr. F. J. McClure 

Coopi rating Units: None 

Man Vears (calendar year 1958): 
Tot si: 1 2/3 
Professional: 2/3 
Otter: 1 

Project Description: 

Obj ect lvej : 

To st:udy the relation of tie fluoride content of luraSn 
hones to their mineral composition. 

Met he d 8 Employed : 

S^ecjjnens of iliac crest, rib and vertebra from individuals 
exposed to various levels of fluoride in their drinking 
water were analysed for F, Ca, Mg, P, Na, K, CO and citrate. 

Major ?iofllngs : 

la the dry, fat -free bone t:here was a slight increase 
in ash, calcium and phosphorus with an increase in fluoride 
concentration. There was a definite increase in the 
n.ignesium concentration, a small decrease in the CCv,} and 

9 more pronounced decrease in the citrate concentration. 

iSo changes were observed in the sodium and potassium content 

of the bones. 

Signi f icance to Dental Research : 

'This is the first extensive study on the relation of the 
fluoride content of human bone to the content and composi- 
tion of the mineral phase. Although some changes were 
observed, they were relatively small and not related to any 
histologic observations. 
Part B included Yes 

- 2 - Serial No. NIDR-31 


Individual Project Report 

Calendar Year 1958 

Part B: Honors, Awards and Publications 

Publications other than abstracts from this report: 

Zlpkin, 1% Leone, N. C, and Lee, W. A. Fluoride Content of 
Urinary and Biliary Tract Calculi. Proc. Soc. Exptl. Biol, 
and Med. 9_7:650-653, 1958. 

Zlpkin, I., McClure, F. J., Leone, N. C. and Lee, W. A. Fluoride 
Deposition in Human Bones after Prolonged Ingestion of Fluoride 
in Drinking Water Pub. Hlth. Rep. 73:732-740, 1958. 

Zipkln, I., Llklns, R. C. and McClure, F. J. Deposition of 
Fluoride, Calcium and Phoaphorus in Experimental Low Phosphorus 
Rickets. J. Nutrition. la Press. 

McClure, F. J. and Zlpkin, I. Physiological Effects of Fluoride 
as Related to Water Fluoridation. Dental Clinics of North 
America, pub. by W. B. Saundere Company, July 1958. 

McClure, F. J., McCann, H. G. and Leone, N. C Excesnive 
Fluoride In Water and Bone Chemistry. Pub. Hlth. Rep, 73:741-746, 

Serial No. NIDR-3? . 
1. Biochemistry 
3. Bethesda, Md„ 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Biochemistry of Salivary Proteins. 

Principal Investigator: Dr. Marc S. Lewis 

Other Investigators: None 

Cooperating Units: Nona 

Man Years (calendar year 1958): 
Total : 1 
Professional : 1 
Other : none 

Project Description: 

1. To fractionate, isolate in pure form, and character i: 
physically and chemically the proteins of saliva. 

2. To attempt correlation of these properties to oral 
and systemic pathology generally, and to the normal and 
abnormal physiology of the salivary glands. 

Methods Employed : 

Samples of saliva are collected using suitable appliance;; 
for isolating the appropriate ducts if parotid or sub- 
maxillary secretions are specifically desired. The 
salivary samples are dialised against a Tris-phosphate 
buffer, pH 8.7, /i » .02, adsorbed on an anion exchange 
cellulose chromatography column (DEAE) and eluted with a 
buffer gradient of decreasing pH and increasing ionic 
strength. Appropriate volumes of effluent are collected 
with a fraction collector. The protein content of the 
various fractions is determined by measuring U.V. adsorp ion 
at 280 mi. Some of the fractions are rechromatographed n 
a cation exchange cellulose column (CM-W). The degree 
isolation of specific proteins in a chromatographic frac ion 

Part Ao Cont'd.! - 2 - Serial No. NIDR-32 

is evaluated by means of the analytical ultracentrifuge 
and by moving boundary electrophoresis. 

It has not been possible to fractionate saliva adsorbed 
on DBAS into individual components either by increment 
or by gradient elution. It has been found that one, 
and sometimes two discrete groups of components are 
eluted between pH 8 and pH 7, and another discrete group 
of components is eluted between pH 7 and pH 5. This hae 
been observed for both whole and parotid saliva. The group 
of components eluted above pH 7 have been rechromatographed 
on CM-W and have been separated into six relatively discreet 
components. The quantities obtained so far have been too 
small to permit evaluation of the purity of these components 
by electrophoresis or ultracentrifugation. 

Si gnificance to D ental Research : 

It is necessary to heve more information on the salivary 
proteins in order to make any study on the relation of 
salivary chemistry to normal and pathological oral 
conditions more meaningful. The qualitative and quantitative 
aspects of these problems may also reflect general systemic 

Proposed Course of Project : 

(1) Work is currently in progress on repeating proven 
isolation procedures on large enough quantities of material 
so that adequate quantities of the purified proteins are 
available for detailed physical and chemical studies. 

(2) The results of the preceding work should make possible 
more significant studies on discrete salivary secretions 
from individuals utilizing micro and semi-micro techniques. 

Part B included No 

Serial No. NIDR-33 
1. Biochemistry 
3. Bethesda, Md. 


Individual Project Report 

Calender Yeer 1958 

Par; A. 

Project Title: i> Study of the Biochemistry of the Submaxillary 
Glanda of the Rat with Particular Reference 
to Polysaccharide Synthesis and Intermediary 
Carbohydrate Metabolism. 

Principal Investigator: Dr. H. Blumenthal 

Other Investigators: None 

Cooperating Units: None 

Man Years (calendar year 1958): 

Other : 

Project Description: 

Objectives : 

a. To isolaea pure neutral polysaccharide(s) from rat 
submaxillary glands. Khen such a reproducible preparation 
is obtained and characterized a number of studies will be 
made. First, the metabolic pathways involved in the 
synthesis of polysaccharide substances will be investigated 
and an attempt nade to elucidate steps in this biosynthesis. 
A number of ouxilliar; studies will be made such as the. 
possible eff nets on submaxillary polysaccharide patterns of 
cariogenlc diets, die:s producing hyper- end hyposalvation, 
and diets causing hypertrophy or atrophy of the glands them- 

b. To study ;he intermediary carbohydrate and energy 
metabolism of rats' submaxillary glands. 

Methods Employed : 

a. General methods for extraction of tissues. 

b. Ion exchange and paper chromatography. 

Part A . Cont'd. - 2 - Serial No. NIDEl-33 

c. Standard eaalytical procedures for metabolites such as 
uronic acids, hexose amines, reducing sugars, proteins, etc. 

d. Spectrophctometric methods. 

Major Findings: 

«.. To date we have been able to Isolate a neutral poly- 
eaccheride fraction high in sugars (based on a general 
carbohydrate test like ancnrone) and in hexosamines, 
r.nd low in uronic acids. However, such preparations are 
contaminated with large amounts of protein (Lowry -positive 
nub stances). Thus far attempts to get rid of these proteins 
have not been very successful and are blocking further 
resolution by paper or sponge electrophoresis. 

b. The intermediary metabolism phase of this work has just 
utarted and -hue far we evidence that rat submaxillary 

glands have little or no galactokinase activity, but do 
ihow glucokiiase and ATPaee. 

Significance to D antal Research : 

The literature contains evidence that salivary glands may 
play a role La the reduction of caries susceptibility in 
various animal species. Vheri are also reports on the 
profound offsets of hormones such as thyroxin and steroids 
an the salivary glands. There is ample literature on the 
predominant place polysaccharides play in the make-up of 
whole saliv* On the other hand, there is almosc no 
literature on how the sal .vary glands might accomplish a 
role in caries prevention or what part they play in the 
synthesis of saliva polyBiccharides. Also, aside from some 
secreted enzymeo such as mylaae, lipase, and proteinase 
and some hietochemically iiscernable enzymes such as 
phosphatase*, there is li:tle or no knowledge of the 
enzyraology cf these glands. 

In view of the known effects of the salivary glands on 
oral health and on the secretion of so important a 
biological t ubstance as aaliva, any information on 
activity and function of these glands might prove of value. 

Proposed Course of Project : 

Work on both the isolation of polysaccharide material end 
the elucidation of metabolic patterns of carbohydrate 
metabolism cf rat salivary glands will continue. 

Pert B included No 

Individual Project Report 
Calendar Year 1958 

Serial No. NIDR-34 
7 , Biochemistry 
3. Bethesda, Hd. 

Part A 

Project Title: Biochemical and Biophysical Studies of 

Principal Investigator: Dr. R. C. Likins 

Other Investigators: Dr. K. A. Piez 
/br, A. S. Posnar, 
Cooperating Units:-* National Bureau of Standards and Armed 
Porces Institute of Pathology 

Man Tears (calendar year 1958): 
Total: 3 
Professional : 1 
Other: 2 

Project Description: 

Subproject A: (With A. S. Posner) 

Obj actives : 

To Investigate the skeletal metabolism of calcium and 

Methods Employed : 

(a) Synthetic hydroxyapatites labelled with Ca* 5 and Sr 89 
were prepared by precipitation, and by hydrolysis of 
CaHPO^, Exchange studies were carried out with these 
materials to investigate the comparative release of the 
isotopes in calcium solutions. 

(b) Radioactive calcium and strontium were administered 
to rats and the urinary excretion of these lone studied 
as a function of time. The relative amount of free and 
bound Ca^5 and Sr89 i n the plasma at different time 
intervals was investigated by ultrafiltration experiments. 

Part A . Cont'd. - 2 - Serial Ho. NIDR-34 

Major Findings : 

There was no evidence of any discrimination between Ca^ 
and Sr 89 in the formation of hydroxyapat ite from a solution 
of labelled elements. The ratio of Ce^.'Sr 89 in the 
CaHPO, on the other hand, was significantly higher than 
that of the solution from which it was prepared. It has 
been postulated that CaHPO. is the precursor of apatite 
in calcifying bone. In this light, the above finding 
would suggest a possible discrimination between Ca and Sr 
in the deposition of new bone mineral. The exchange 
experiments revealed that the synthetic apatites released 
relatively more SrS9 than Ca^5. xhis finding suggests 
that calcium is more stable in the hydroxyapat ite 
structure than is strontium, and may serve to explain 
the results with bone (see Subproject B — Calendar Year 

Analyses of the twenty-four hour urine revealed that the 
excretion of radiostrontlum by the kidney was approximately 
four times that of radlocalcium. Of interest was the finding 
that this renal discrimination Increased from the beginning 
of the time period to the end. It was considered possible 
that the latter finding was related to the relative amounts 
of free and bound Sr^ and Ca^5 of the plasma. The 
ultrafiltration experiment ruled out this possibility. 

Significance to Dental Research : 

This research will provide further information on the 
nature of the mineral phase of teeth as well as bones. 

Proposed Course of Project ; 

The possibility that the state of calcium balance in the 
animal is responsible for the changing Ce45;Sr89 ratio in 
the urine will be investigated. 

Subproject B; (With K. A. Pies) 

Objectives ; 

To investigate the nature and interrelation of the mineral 
phase and collagen matrix of bone as occurs in the calcifying 
tendons of the turkey. 

Part A . Cont'd. - 3 - Serial No. NlDR-34 

Methods Employed; 

The leg tendons of the turkey undergo regional 
calcification beginning at about 16 weeks of age. 
Possible age-associated changes In the amino acid 
composition of the collagen matrix are being investigated 
by ion exchange chromatography. In addition, a detailed 
chemical analysis of the mineral phase is in progress. 
This Includes assays for calcium, phosphorus, magnesium, 
sodium, potassium, carbonate, and citric acid. Radio- 
tracer techniques have been employed to study the 
deposition of Ca45 t n tendons and bone. 

Major Findings : 

Amino acid analyses of bone and tendon-collagen reveal 
a difference in the lysine and hydroxy lysine content. 
In bone, the mole ratio (lysine /hydroxy lysine) is 5.0 
while In early non-calcified tendon it is 1.9. The sum 
of these two amino acids is the same. At 29 weeks of 
age when tendon calcification was essentially complete 
the ratio in tendon was 2.4 indicating that the collagen 
was still primarily of the tendon type with, perhaps, 
beginning bone formation. While the inorganic analyses 
have not been completed, preliminary evidence suggests 
differences between bone and tendon with respect to 
their carbonate and citrate content. 

Significance to Dental Research ; 

Studies on the relationship of the organic matrix to 
mineralisation should further an understanding of the 
calcification process in teeth as well as bones. 

Proposed Course of Project : 

This must await completion of the study now in progress. 

Part B included Yes 

- 4 - Serial No. BIDR-34 


Individual Project Report 

Calendar Year 1958 

Part B: Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Likias, R. C, Posner, A. S. and Steere, A. C. Effect of 
Calcium Treatment on Solubility and Calcium Uptake of 
Synthetic Hydroxyapatite and Rat Molar Enamel. J. Am. 
Dent. Assoc. 57:335-339, Sept. 1958. 

Liking, R. C, Scow, R. 0., Zipkin, I. and Steere, A. C. 
The Deposition and Retention of Fluoride and Radiocalcium 
in the Growing Rat. Am. J. of Physiology. In Press. 

Serial No. NIDR-35 
1. Biochemistry 
3. Bethesda, Md. 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Enzymological Studies in Relation to Oral 
Tissue and periodontal Disease. 

Principal Investigator: Dr. S. A. Narrod 

Other Investigators: Dr. R. R. Omata 

Cooperating Doits: American Dental Association 

Dr. Wn. Jakoby (NIAMD, Lab. of Biochem. & 

Man Years (calendar year 1953): 
Total: 2/3 
Professional: 2/3 
Other: Kone 

Project Description: 
Ob 1 ect lves : 

To study enryme systems in relation to structural integrity 
and general metabolism of biological systems with emphasis on 
deatal supportive tissue. This includes (a) additional 
studies on the mechanism of collagen degradation and (b) 
a study of ethanolamine metabolism. 

Methods Employed : 

(a) Further fractionation of the growth medium of 
Clostridium histolytica* using the standard protein 
fractionation previously described. In addition, fractiona- 
tion on ceUulose columns is being developed. Activity 

of the enzymes has been measured by hydroxyprollne 
liberation" from collagen (collagenase) or by the disappearance 
of turbidity from rat tail collagen gel solution when 
ethenol is added. 

(b) Oxidation of ethanolamine by a pseudomonad type organism 

•irs to proce- metabolism by the convey 
tc le. The metabolic activity has been 

followed - glycoleldehyde tcnaation 

as the i -hydrazine derivative and by 

manometric techniques ot oxygen uptake. 

Ma jor F indings : 

(a) From the research done in this laboratory as well 

as that reported by ctb-r investigators, it appears that 
the "coliagenase" liberated by Clostridium histe lyt lcum 
is coaposed of several proteinases e.nd peptidases* 
At>.>>mpta to separate the enzyme, responsible for the 
I attack on collagen, rendering it capable of 
attack by ether enzymct has been unsuccessful, so far, 
although accumulation of peptides as shown by piper 

;matogri.phy indicate* the feasibility of this 
approach ii & study of t:ie nature of collagen and 
co.'.lagenast It i6 interesting to note that several 
pro snd peptidases can be identified as separate 
fr<nn tt i 1 collagen-attacking snsyme by variation 
of activity, as well «e oy different optiiaa pHs and 
«ct ivators such as calcijm and ferrous ions. Hove 
thi:ce hr.B t aen no dint inct ion between the initial 

tolutioc of collagtn or the action upon gelatin 
(gulatinast ) also obafcrvsd in these preparation* 

(b) Precl cellular protein of th«. pseuclomonad 

organism ising ei h-i iol amine as its eole source of 
carbon i ^gen hus show: t is organise cont . 

an enzyme capable of converting ethanolam .ne to glycol alde- 
hyde by oxidative deamination. Although the latter 

extremely slow, it has been shown that oxygen 
uptake, ao measured ixygen ele.troc-f., exactly 

i a molar baais with glycol aldehyde formation, 
as taeasurec by the 2,4 dinitrophenyl hydrazine 

Sign tjo Dental Resfcfi 

In keeping with a basic approach to the problems of 
periodontal diseases it is hoped that these studies will 
give some under a tending of the formation ao well as the 
destruction of supportive tissues. Thus collagen, the 
basic protean of bone and periodontal tissues, raighr. .be 
a key to understanding the pathological conditions 

PartA. Cont'd - 3 - Serial No, HIDR-35 

occurring in periodontal diseases. On the other hand, 
the production of complex proteins, such as collagen, 
from simple carbon compounds such as ethanolamine, 
mi^ht be functional in the healing states following 
collagen destruction. 

Proposed Course of Project ; 

Further studies on separation of the initial collagen 
attacking enzyme are being carried on by Dr» R. R. Omata, 
NICR, using the methodology described herein., 

No additional studies on ethanolamine metabolism are 

Part B included Yes 

Serial No, NIDR-35 

Individual Project Report 
Calendar Year 1958 

Part B: Honors, Awards, and Publ lections 

Publications other than abstracts from this project: 

Jakoby, W. B. and War rod , S. A. Aldehyde OxldBtlon IV. 

An Aldehyde Bufler for Growth Studies, J. of Bact, In Preus. 


Individual Project Reports 
Calendar Year 1958 


36. A. L. Russell : Epidemiology of Oral Diseases Among Alaskan 

37. A. L. Russell : The Epidemiology of Dental Caries, Including 
the Fluorine-Caries Relationship. 

38. J. C, Creene and A, L> Russell: Study of the Epidemiology of 
Periodontal Disease in India. 

39. C. J. Donnelly : Study of Periodontal Disease and Caries in 
Adventists Families. 

40. H. W. Littleton : Epidemiology of Oral Diseases Among 
Ethiopian Natives 

41. A. L. Russell : The Epidemiology of Periodontal Disease 

42. C. L. White Relation Between Dental Caries and Periodontal 

atology & B 

Esti m ated Obli^atlopa for FY. 1959 


R e iabur s eaen t s 

$ SI 

$ Ul 

ct Numbers: 








»1 No. S3 
>gy & Bis 

Individual Project R^ 
Calendar Year 1958 

Proje gy of Oral Di3ea: es Among Alaskan 


Principal Investigator: Dr. A. L. Russell 

Other Mr. C. L. WW 

Cooperating Onits: Interdepartmental Comm.'.ttee on Nutrition 
for National Defense. 

Hen Years (calendar year 1958): 
Tot 1 

Pr-: il: 


To determine the prevalence of dental caries, periodo 

and malocclusion among Alaskan natives and to in- 
:e the relationship of these disorders to cer.: 
:ionai factors and geograp :ion 

Me thods E nployed : 

examination of members of the /.laskan National Guard 
followed by statistical analysis of examination data to 
a nine relative risk, under varying conditions. 

Major Sinning s : 

Prel -minary examination of about 700 Eskimo men of the 
Mia tan Bational Guard has revealed significant data regard- 
ing oral health of the group. Compsr ng the prevalence of 

i disease in individuals living under relatively civilized 
conditions wi'ih that of men froa primitive villages, it was 
found that dental caries is virtually non-existent 

r group. Of further significance was the observation 

al No 

it uany of these primitive groups vere essentia) 
free of periodontal disease. Although other male 
viduals from similarly primitive areas exhibited 
fonaly prevalent gingivitis of sever- degree, the 
ase apparently did not progress to destruction 
er periodontal tissues even in the absence of 
caonal hygiene or professional dental care. However, 
3 significant to note that when Eskimos have 1 
some time under relatively civilized conditions, 
the prevalence of oral disease increases to a point 

sparable to that seen In average aale popula- 
groups in the United States. 

Significance to Dental Research : 

At present the Eskimo lives under conditions ranging 
from the most primitive to highly civilized, a spectrum 
which af unique opportunity to study oral die? 
uninfluenced by professional treatment. Thus, research 
efforts may be focused more directly on specific factors 
such as nutritional deficiencies, that might be respon- 
sible for the high prevalence of dental caries and perio- 
dontal disease in civilized populations. 

Propoaed Course of Fro |ect . 

Pollowing complete analysis of the National Guar J 
data, a study of carles and other oral conditions 
children, priirtrily from primitive village schools, wiil 
be Initiated. In extending our knowledge of the epidemio- 
logical statistics of oral disease, the information derived 
from these studies will undoubtedly contribute signifi- 
cantly to increasing the oral health statua of the Alaskan 

Part B included: No 

Serial Ho. HIIB^S? 
1. Epidemiology & Biosietry 
3. Be :hesda, Md. 

Individual Project Report 
Calendar Year 1958 

Per; A. 

Project: Title: The Epidemiology of Dental Caries. Including 
the Fluorine-Caries Relationship. 

Principal : nvostigaixr: Dr. A. L. Russell 

Othe ,^ a tors: Mr. C. L. White 

Coop Units: Board of Education. City of Grand RapldB, 

C .ty-County Health Department, Grand Rapids, 
M'.chi£,an: Michigan Department of Health-, 
Montgomery County and Prince Georges 
jnties, Maryland 

Man Years (calendar y^.ar 1958) 
Total: . 2 

Project Description: 


a) :?o observe the effects of the fluoridation of a com- 

.ty water supply on children whose Initial exposure 
fluoride occurred at different ages. 

b) To define the epidemiological characteristics of dental 

c) To elicit evidence as to the basic aature of denta . 
caries and the tissues it attacks so that present 
methods of control may be perfected and, possibly, new 
methods of control can be evolved. 

Methods Iteplo y ed ; 

Direct examination of population groups followed by statis- 
tical analysis of examination data to determine relative 
rlslc under varying conditions. 

Par * ,:l&l R°- lSSz2£ 

- 2 - 

a) The progres; ive diminution cf caries, previously 

«d la Grand Rapida c'r continued during 
whole population studied. Sodium 
which Is cheaper, is as effective as 
u* fluoride for fluoridation of public water 

b) After six years of fluoridation in Montgomery and 
Prince Georjeo Counties, Maryland, there has been 
significant reduction in caries in first solars wh 
ware newly -erupted at the ttae fluoridation began. 
Tie curve plotted for DMF retis of the 5, 6 7, and 3 
yaar olds if approximately the er:p ictancy curve. 

tlon s eea8 to b i a i ;ple function of the 
time available to a tooth for accumulation of fluoride 
prior to enption aid the co,i3eqi e -.t risk of carle 
a: tack. 

Si gnificanc e to Dan ie 1 Research ; 

Data f.xom direct observations of human populations pro- 
vide a bridge whereby findings fron animal studies way be 
appllad to humor diseasa. Present attention is concenti 
upon iieveral areas where ba3ic Information is scanty 
equi vocal; for example, analysis of the selective action of 

aoridated water used while taeth aire calcifying, w 
young calcified enamel is maturing, and after adult eti 
is pr«isent, is expected to add information as to the h 
mechanics and sequences In calcification and raaturatio 

Proposed Course of Project : 

Studies of the epidemiology of dental carles will continue 
with particular emphasis on lead.? that develop in experi- 
mental studies of this disease. Grand Rapids Junior and 
Senior High School students will continue to be observed 
until the full inhibitory effect becomes evident and ui 
the prevalence of fluorosis has reachec its maximum point. 

Part B included: Eo 

Serial Ko. SHEBc2& 
1. Epidemiology & Biometry 
3. Bethesda, He. 


Individual Project Report 
Calendar Year 1958 

Part A. 

Project Title: Study of the Epidemiology of Periodontal Disease 
in India. 

Principal Investigator: Dr. J. C. Greene 

Other Investigators: Dr. A. L Russell 

Coopemti.n-j Units: The World Health Organization, the Government 
of India (through the radian Council of 
Medical Research) ; the Epidemiology Branch of 
the Communicable Disease Center, Atlanta, 
Georgia; and the University of Michigan - 

Mar. Years (calendar j aar 1958) 
Total: 12/3 
Professional: 1 
Other: 2/3 

Project Descript* 

Objectives : 

a) To study the epidemiological and clinical characteristics 
of periodontal disease In a population reportedly having 
advanced f.lveolar bone destruction at relatively early 

b) To encourage and train dental investigators in India to 
di creditable research. It was hoped that these In- 
vestigators could be encouraged to carry on a long-term 
epidemiological study of dental diseases in their country. 

Methods Employed : 

Dr. Greene, accompanied by Dr. Sigurd Ramf jord of the Uni- 
versity of Michigan, visited India in July and August of 
1957 co set up a field study of periodontal disease. Two 
study technics were presented to and discussed with repre- 
sentatives of six Indian dental schools. Actual field 

Serial No. KIM -38 

both methods, and several native dentists 
i :ip&ted Cne procedure was based on clinical appear- 
of the i&ve.iting tissues of all the the teeth, while 
the o':hi;r involved measuring the depth: of the gingival 

ro ind selected teeth. Recor ings were also aade 

i i tatus of ear. it. Approximately 

.0 male school children between 11 and 17 years of cge, 

in and around Scabay, India, wer ; se e' ted for this study. data were collected on 577 male persons 11 - 17 

s of age In this country for comparison with the India 

Major Finding g : 

Tlie severity of ical disease was much greater 

the India group than in the comparison group 
er.amined in this country. 

b) Calculus were more numerous and larger In 

■up than :'.n the group studied in 

c) Periodontal disease was more severe in the rural 
ptrtr.or tndla grauo than in those from ur'; 

Lcaace to Dental Research : 

Tha iite show a -ery close association between the presence 
of orrl calculus and periodontal dijeaca. The difference 
in the severity of perl ir.& in the urban and 

rural groups was accompanied by a significant difference in 
calculus deposits. Further study of the characteristics of 
these and other -roups might ahed some light on the epidemi- 
ology of dental alculu: formation. 

The dental investigators in India are continuing to cc 

data on doniology of dental diseases in their country. 

Proposed Course of project: 

HIDR participation in the project has been completed except 
for publication of the findings. The cata have been 
analyzed and three reports have been prepared for the World 
Heal ± Organize t 

Part B included: Ho 

SSrial Ho. BIPR-39 
idemiology & 31: 
3„ Bs.thesda, Md. 


Indivi.d 1 al Project Repo::t: 

Calendar Year 1958 

Part A. 

Project Title: Study of Periodontal Disease and Caries in 
Adveniiats Families. 

Principal Inve3tiga-:or : Dr. C. J. Donnelly 

Other Xnvastigators: Hone 

Cooperating Units: Indiana Conference of Seventh Day Advent J.sts 
Horth Dakota " " " " " 
Wisconsin " " " 
Michigan " " " " " 

Kan Years (calendar year 1958): 
Total: 12 3 

Professional: 2/3 
Other : 1 

Project Description: 


a) To investigate the influence of familial factors and of 
geographic location on periodontal disease and dental 

les among Adventists families. 

b) To determine if the dental caries r.^te among Adventists 

<iren is lawer than among comparable non-Adventlsts 

c) To study the prevalence and severity of periodontal 
disease in children and adults from a related group of 
individuals . 

Met! ada 

Direct examination of parents and children attending Seventh 
Day Adventists Caap Conferences in different parts of 
United States The data will be treated statistically to de- 
ter .a:'.at; the influence of familial factors, geographic loca- 
tion and Adventists health reforms. 

A (Cor Serial Ho. KXPH-39 

- 2 - 

Major f^ndf .pya : 

.dren examine ; . at Adventists Camps .how a consistently 
Ties rate regardless of the part of the country in 
which they live. The severity of periodontal disease 
among Adventiats families shows a pattern different frost 
that of any othe population examined by the Epidemiology 
and Blotcetry Branch. 

Significanc e to Dental Research ; 

The lew-caries rate consistently found among Adventists 

Ldren may be elated to the dietary recommendations 
the t.ealth refo ~m" suggested by the Atventis'cs Church. 
Further study :ni ht add to our information on the influence 
of diet on carie raizes 

By examining Adventists in different pcrts of the country, 
sone Information will be obtained on the influence of geo- 
graphic location on carles and periodoctal disease. Since 
the severity of arlcdortal disease in a related popula- 
tion differs frcr that of the general populations, further 
study might yield Information as to the factors which effect 

Proposed Co urse of St; dj: 

A controlled study will be carried out to determine the 
magnitude of tbe difference in caries rates between com- 
parable groups o Adventists and non-Adventlsfca children. 
To determine the effects of geographic location on caries 
and periodontal <• Isesse, Adventists populations in parts 
of the country net previously included vill be examined. 

Part B included: No 

Serial Ho. MDR-AO 
1. Epidemiology & Biometry 
3. Bethesda, Md. 


Individual Project Report 

Caletdar Year 1958 

Part A. 

Projec . Title: Epidemiology of Oral Diseases Among Ethiopian 

Principal. Investigator: Dr. H. W. Littleton 

Other Vn^'estigatorc: None 

Cooperr.t;.nj 7nits: interdepartmental Committee on Nutrition 
Tor National Defense. 

Man Years (calendar year 1958) : 
Tota. : 1 
Professional: 1 
Other : None 

Project Description: 


::mine the prevalence of dental caries, periodontal 
disease and stale cclusion asaocg Ethiopian natives and to 

sstigate the relationship of these disorders to certain 
nutritional factors and geographic location. 

Methods Em ployed : 

Direct examination of Ethiopian natives followed by 
statistical analysis of examination data to determine 
relative risk under varying conditions. 

Major Findings: 

Field examinations are in progress. 

Significanc e to Dental Research : 

Studies among the Ethiopian natives allow an investigation 
of oral diseases in groups showing varying degrees of 

;i) Serial Ho. SK" 

- 2 - 
Proposed Course o£ Project; 

To raalyze the assenbled data over the course of the 
-iexf: year and correlate findings with thos? la Alaska India. 

Part B included: Bo 

al No, HIDR-41 
1« Epidemiology & Biometry 
3. Betheada, Md. 


Individual Project Report 
Calendar Year 1958 

Pa rt A . 

Project Title: The Epidemiology of Periodontal Disease. 
cipa] Investigator: Dr. A. L. RiiBsell 
er Investigators: Dr. C. J. Donnelly 

Unite: Jefferson County Health Department - 
Birmingham, Alabama. 

Man Years (calendar yoar 1958): 
al: 2 


ot;. l i 

Project ton: 

To compile a descriptive epidemiology of the group of 
ise entities commonly called "pyorrhea." 

Methods Em plo yed: 

Direct examination of population groups, particularly 
adul:s and older children, followed by statistical analysis 
of examination data to determine relative ri*k under 
ying conditio 

Major Findings : 

In previous studies it appeared that periodontal disease 
was nore prevalent and more severe in Hegro than in white 
populations. However, when data from examinations of 
white and Negro residents of Birmingham, Alabama were 
analyzed by equalizing the groups with respect to educa- 
tion and occupation, the differences in prevalence and 
severity of periodontal diseases between the two races were 
no;, significant. As in previous work the incidence pattern 
rad for white males and females. The relationship be- 

Lai lio, WID E 

- 2 - 

tocial factors and the severity of periodontal 
..sen was Investigated in sore dec, 11 in an attempt 
f.teraine the degree of influence of various sc 
tdltions. Ths true etiological factors in periodor 
di««aae may be independent of those factors which in- 
2 its course once initiated. 

Use of a fluoride domestic water has no relation to 

ity or pre alence of periodontal disease. 

S ig nific ance to Denta l Research : 

After dental carles, the periodontal diseases are the 
:nd most iaportant cause of tooth loss, and affect 

7 per ceut of all persons 50 years of age or 

jj . Epidemiological study of dental caries led to 

loridation of domestic, water supplies, which will re- 

ce the treat* int needs for caries by about two-thirds. 

Lmate aim of this study is mass preventive measures 

i he periodontal dise* 

Propos ad Course ojE Fr o \z < 

Field examinations of older children will be continued 

intended in an effort to determine the characteristics 
of t: d who 3how signs of the disease at the 

earlier age, and hence are presumably the more susceptible 

roups and of social factors will be con- 

Part B included: Ho 

1. Epidemiology & Biometry 
3o Be this da, Hd< 


Individual Project Report 

Calendar Tear 1953 

Part A. 

Project Title: Relation Between Dental Cables and Periodontal 

Prinelpii Investigator: Mr. C. L. White 

Other Investigators: Dr. A. L. Russell 

Cooperating Units: 3ooe 

Man Tears (calendar year 1958) 

Professional: 73 

Other: 2 3 

Project Description: 

Object ives: 

This study is designed to learn if a correlation exists 
between periodontal disease and dental caries (D,M,F). 

Ma thee 3 Bjployed: 

Tie procedure la to analyze the data frost periodontal 
studies conducted by the Epidemiology and Biometry Branch 
over the past several years. 

Major r eadings : 

Preliminary anaiyslo indicate that the per cent of sound 
teeth affected by periodontal disease is approximate;.? 
equal to the per cent of fcaeth with open carious lesions 
effected by periodontal disease. 

Signi fic ance to Dental Research : 

lis study increases the program of the Institute to in- 
clude statistical investigation of available data from 
other studies. 

Serial Ho. HIDR -^ 
- 2 - 
Fropcise d Course of Project : 

ontinue and expand the outlined study. 

Part B included: Ho 


Individual -.eporta 
Calendar Year L958 


43-C. C. J t W ltkop : Trl-raclal Isolates In Eastern Unit, 

44-C C. J, Wltkop : Hereditary Defects of Enamel & Dentin 
(Michigan Study). 

J, J. Wltkop ; Genetic Study in Southern Maryla 

J Wltkop ; Familial Osteoarthritis. 

47-C C . J. Wltkop : Hereditary Eye Deft 

D. L. Ru Anemias and Qthe 

Hematological Disorders. 

49-C J. D. llfwaad^r ; Orel Aapceta «f th« Reaolta ©t Co** 

Mar r lag* in llroahlau and Vagaaa 

50-C FL_M Step h" ' 

virooment in Relation to Dental t, 
Dli • - 

k. M. Stephan : Interrela'. Between 

in the Etiology of Denta; 

'dies on Enzymatic Reactioua o 
Alcohols and Steroids.. 

P. J„ Co . • Cephalometric Study of the I 

Growth of the Soft Palate and Ha: 
Cleft .ZMldren. 

54-C, E. J, Driscoli General Anesthesia (Dental) Basell 

I. I, Ship : Investigations into the T 

Aphthous Ulcerations and Other Oral H 

56-Co I, I. Ship ; Stress in the Dental Pati- 

V, S Baer ; Esterase Activity Associated with th? 
--posits on Teeth. 


Individual Project Reports 
Calendar Year 1958 


58-C. P» N. Baer ; Some Genetic Aspects of the Periodontium and 
Periodontal Disease in Mice, 

59 -C. H. R. Stanley, Jr , : Histopathologic Study of Periodontal 

Disease in BALB/Cann Mice. 

60-C. H. R. Stanley, Jr . : Hlstopathology of the Human Pulp. 

61 -C, H. Swerdlov ; Pulp Response to High Speed Cutting and 
Grinding of Teeth. 

62-C. R. S. Lloyd : Cephalometric Evaluation in Pull Denture 

63-C. R 3 S. Llo yd: Maxillofacial Prosthetics, 

64-C. H. Swerdlov : Malocclusion as Related to Traumatic Occlusion 

65-C D, R. Poulton : Twelve-Month Changes in Class II Malocc 

with & without Occipital Headgear Therapy 

Clinical Investigations Branch 

(Including Genetics, excluding Center Dental Department) 

Estimated Obi; Rations for ?.Y. 195? 

Total : §4-73,700 

Direct : $248,900 

Reimbursements: $159,800 

Project Numbers: 
NIDR-43 (C) 
NIDR-44 (C) 
NIDR-45 (C) 
HIDR-46 (C) 
NIDR-47 (C) 
NIDR-48 (C) 
NIDR-49 (C) 
N1DR-50 (C) 
NIDR-51 (C) 
N1DR-52 (C) 
NIDR-53 (C) 
HXDR-54 (C) 
HIDR-55 (C) 
HIDR-56 (C) 
NIDR-57 (C) 
NIDR-58 (C) 
NZDR-59 (C) 
NIDR-60 (C) 
NIDR-61 (C) 
NIDR-62 (C) 
HIDR-63 (C) 
NIDR-64 (C) 
NIDR-65 (C) 

Serial Ho. HIDR-43 (C) 

1. Clinical Investigations 

2. Human Genetics 

3. Bethesda, Md. 

Individual Project Report 
Calendar Tear 1958 

Part A 

Project Title: Tri-racial Isolates in Eastern United States. 

Principal Investigator: Dr. C. J. Witlcop, Jr. 

Other Investigators: Rone 

Cooperating Units: Calvin L. Beale, Agricultural Marketing 
Service, U.S. Department of Agriculture. 

Kan Tears (calendar year 1958): 
Total: 5/6 

Professional: 1/3 
Other: 12 

Project Description: 

Objectives : 

1. To determine the number, types, location and size of 
tri-racial Isolates in Eastern United States. 

2. To determine the degrees of inbreeding shown by various 

3. To determine the amount and types of hereditary illness 
shown by the groups. 

4. To verify or not the assumption that more individuals 
will show homozygous recessive hereditary illness in 
such populations. 

5. To investigate the social consequences of Isolate for- 
mation and their effects on the distribution and preva- 
lence of intrinsic disease. 

6. To investigate and compare the prevalence, types and 
inheritance patterns of Intrinsic Illness with in-group 
and out-group marriages, with degrees of consanguinity, 
with various other tri-racial groups, and with the 
general population. 

- 2 - Serial No. NIDR-43 (C) 

Part A (continued) 

7. In summary, the purpose of this investigation is to 
find, enumerate and describe all tri-raclal isolates 
residing in Eastern United States to investigate 
the degree of consanguinity in these groups; to look 
for unusual types of matings, rare in human material, 
that are of special value in determining modes of 
Inheritance, gene inter-action, and biochemical 
pathways; to investigate the hereditary diseases 
present in these groups; and to utilize the patient 
material found for further clinical and biochemical 
genetic lnveetigations. It is assumed that in isolate 
populations, consanguinity will be prevalent; and it 
is further assumed that an increased number of indi- 
viduals in such a population will show homozygous re- 
cessive illness. It is also assumed that all cases 
of a specific hereditary illness from any one group 
will nearly always be due to the same gene defect, 
thus providing an epidemiological method for select- 
ing a large sample of individuals with the same gene 
defect for biochemical investigations in other NIDR 
projects or projects in other Institutes. 

Methods Employed : 

This investigation requires three approaches: 

1. Analysis of census and other civil records. 

2. Survey of groups by letters to physicians and public 
health officials concerned. 

3. Field work by HIM teams. 

1. Analysis of census and other records: 
Because of the U.S. Census Bureau's wish to further class- 
ify people whose race entries were somewhat nebulous, the 
1950 Enumerators Reference Manual directed, "Report persons 
of mixed white, negro, and Indian ancestry living in certain 
communities in Eastern United States in terms of the name by 
which they are locally known." Such persons were then to be 
classified for publication purposes among "other nonwhlte 
races," that is, other than Negro, American Indian, Chinese, 
Japanese, or Filipino. Starting with these data, and tracing 
the surnames so recorded through previous U.S. and British 
Colonial Census records, it was determined that such surnames 
frequently residing in the same household or neighborhood, 

- 3 - Serial Bo, BIM-43 (G) 

Part A (continued) 

showed variable race entries from decade to decade, even 
for the sams individual (i.e. white, Hegro, Indian or 
other), and Cor the moot part had large families. She 
existence of suspected trl-racial isolates from these 
data ware then verified froa local civil, historical, 
and social records such as marriage records, historical 
accounts, and newspaper articles. A search of the 
anthropological and social literature frequently re- 
vealed studies on such groups. Visits to these com- 
munities, t» verify the existence and surname data, 
were made by collaborators and this investigator. 

Of the 116 counties checked, the population of trl- 
racial character was estiis&ted at 77,000 persons, of 
which 33,000 wire listed as Indian, 29,000 as white, 
14,000 as Bsgro and 1,000 under colloquial race names. 
Forty percent of the total reside in Bbrth Carolina. 

Local and census records were used to estimate coo- 
sangulnlty and fertility. For example, census data 
from one Borth Carolina group showed the highest 
fertility ratio for any known racial or ethnic group 
in the United States. The ratio of children under 5 
years old per 1,000 women, 15 to 49 years old, was 325 
as compared to 417, the U.S. average. Marriage records 
of one group showed that 171 required ecclesiastical 
dispensation for relationship within the first three 
degrees (1.3., 2nd cousins or closer). Investigation 
of the marriage records in Barbour and Taylor Counties, 
West Virginia, showed that 102 of the 112 marriages 
from 1856 to 1931 were to other trl-racial group members. 
School records of the same group in 1958 showed an un- 
usual proportion of students from the group in the higher 
ege levels for the grade in which they wer j in. 

2. Survey of groups by letters to physicians and 
public health officials concerned: After determining! the 
number and lurncmes of persons comprising these isolate 
populations, letters were sent to all county health of- 
ficials in the 116 counties where these people reside re- 
questing Information on specific listed hereditary illnesses 
among these groups and any illness that appeared unusually 
f r extent in such populations. Other local physicians and 
dentists were also contacted. These returns were tabulated. 
Literature by all writers was scanned for mentioned heredl- 

- 4 - Serial Bo. HIDR-43 (C) 

Part A (continued) 

tary illness in these groups, (ego, aicrophthalmla in the 
Moors of Delaware) . 

3. Field work by HIBH teams: It is planned to send 
investigators to these areas for 1-3 day preliminary re- 
connaissance work. If any Important leads develop or if 
the reconnaissance shows promise of further study, these 
studies will be planned around the genetic material 
found. Some of this work has been done. For example, a 
day's visit to Halifax County, Worth Carolina, revealed 
a very rare hereditary eye lesion that has not, to my 
knowledge, been reported in this country in this form. 
It has been reported from Pakistan in as many as 201 of 
some villages, but the definite hereditary character of 
this illness has not been previously described. A detailed 
clinical medical genetic study is being planned for the eye 
disease in this group. This study will entail a team of 
two ophthalmologists from VOBA, a hematologist, a nurse, 
and chief of the Genetics Section from BIDS., spending 
about two weeks in Halifax County, Borth Carolina. 

Patient Material : 

Patients found In this study of Isolates have been admitted, 
from time to time, for the study of particular diseases. 

Major Findings : 

1. The reported high Incidence of harelip and cleft 
palate in the Guinea group of West Virginia was not verified 
by our investigating team. 

2. A hereditary form of limbal vernal conjunctivitis 
was found to affect some 50 members of the Haliwar group in 
North Carolina. An unusual and undiagnosed hereditary neruo- 
loglcal disease was also present in these people. 

3. The reproductive rate of the Lumbee Indians of North 
Carolina Is the highest of any ethnic group in the United 

4. The size, location, distribution, emigration pattern, 
and a knowledge of the hereditary diseases present in many 
of these groups has been further described. 

Ser: i £1 

Several studies have been Initiated on these 
)s by or tigators tc whoa we furnished 
ita, as tc location, surname, 
Lation nix*. Thes« studies lsave already shown 
jnusuaL hereditary conditions present in these 
including hemoglobin disorders and sexual inter- 

£& Pgot<^ R esearc h , 

Thee*! sroups of £er unusual genetic material for the 
ioaactive tissue, eye, and metabolic 
de nt quantities so that the geneticist 

tical and biochemical investigator that 
the genes invcLved are ail of the aame type. rhiu offers 

:©r the selection of pa 
with hereditary die - rich the same metabolic de- 

To continue to ccumulat- Information on the hereditary 
diseases present in these Isolate populations. 

Part B ircluded: fes 

- 6 - Serial Wo. NIDK-43 (C) 


Individual Project Report 

Calendar Tear 1958 

Part B: Houors, Awards, and Publications 

Publications ctber than abstracts from this project: 

Bea'.e, CI, : Anerican Tri-raclal Isolates; Their Status 
and Pertinence Co Genetic Research. Eugenics Quarterly 
4:187-196, Decenber, 1957 (Published after 1957 project 

Honors aud Awards relating to this project: Done 

Serial Ho. Bg»-44 r .,,CC) 

1. Clinical Ifiiveatigatii 

2. Human Genet ice 

3. Bethesda, Maryland 


Individual Project Report 
Calendar Tear 1958 

Part A 

project Title: Hereditary Defects of Bnaael and Dentin 
(Michigan Study) 

Principal Investigator: Dr. C.J. Witkop, Jr. 

Other Investigators: Hone 

Cooperating Units: Dantai Section, Michigan State Haa~.fch Department. 

Man Tears (calendar year 1958) : 
Total: 1/3 
Professional: 1/3 
Other: Rone 

Project Description: 

Objectives : 

1. To define the various hereditary defects in enamel end 

2. To determine the prevalence and mutation rates in the 
general population. 

3. 7o determine the mode of inheritance. 

4. To investigate the histological processes involved in 
the defects. 

s. To describe any associated physical or chemical defect. 

6. To investigate possible linkage associations - 

I'o determine the various methods of restoration, pre- 
vention or treatment where possible*. 

Methods Employed : 

1. Field Study: A survey of 96,761 school children, be- 
tween the ages of 4 and 12, was conducted in 42 couatiee of 
the State of Michigan for hereditary defects and other for- 
mations. Subsequent pedigree studies on these Individuals 
were conducted to determine the mode of inheritance and pos- 
sible linkage with known genetic markers, such a3 blood type. 

■ - 2 - Serial Ho. HIDR- 44 CO 

Pare A (continued) 

secretor factor, and P.T.C. taste test. These de- 
fects vera classified by clinical descriptions, histo- 
logical findings, and inheritance patterns. Patients 
with representative difficulties were brought to HIE 
for clinical evaluation. 

2. Clinical Study: Complete physical and routine 
laboratory examinations were given to determine if 
any physical or metabolic defect was also associated 
with the tooth defect. Except in one particular 
defect, osteogenesis Imperfecta, no genetic defect 
was found associated with the tooth condition. 

3. Laboratory Study: Histochemical studies con- 
ducted on fresh ground and decalcified specimens using 
normal controls, showed that there was a possible bio- 
chemical defect in opalescent dentin. Linkage studies 
were analysed. 

Patient Material : 

Patient material was obtained from 1) individuals con- 
tacted during our field study in Michigan; 2) indivi- 
duals In our Genetic Study in Southern Maryland, Pro- 
ject Ho. D -4-002; and 3) in a few instances from indi- 
viduals referred by local practitioners. 

Major Findings : 

1. The mutation rate for opalescent dentin is ex- 
tremely low. Ho case that could be a new mutation was 
found among the 38 propositi in this study. A check 
of other cases reported in the literature and by other 
investigators failed to find a single instance of this 
disease that could possibly have arisen by mutation. 
This is the only autosomal dominant hereditary condi- 
tion in which a mutant has never been found. 

2. The homozygous state of opalescent dentin may be 
lethal causing intrauterine deaths. 

3. From a comparison of the Michigan population with 
the Maryland population, one of two possibilities eslsts. 
Either the defect known as opalescent dentin is the same 

- 3 - Serial Ho. NIDR-44 (C) 

Part A (continued) 

in both populations, with the gene manifesting more 
severe symptoms because it is present in a different 
gene complex in the Maryland population; or the gene 
is different in the Maryland population and, therefore, 
a different disease. If the genes are the same, the 
significance would indicate that defective genes 
adapt over many generations to the rest of the gene 
complex, and when put into a stock with different 
racial background, manifest themselves in a more 
severe form. 

4. Some evidence is present that a bone defect of a 
mild type is found in people with opalescent dentin. 

5. Found during this survey was a hereditary, tumor- 
like condition of the ;:e*th closely associated with 
dens-in-dente, which iii also probably hereditary, and 
with odontomas, which have bean described as dentin 
dysplasia- This is inherited as an autosomal domi- 
nant trait. 

6. We have found a new hereditary condition consist- 
ing of premature closures of the fontanels, ptoses of 
the eyelid, absence of tear ducts, and missing lateral 
incisors that is inherited as an autosomal condition. 
These individuals require prompt surgical intervention 
for the cranial defect. Many also need surgical cor- 
rection of an associated agenesis of the condyles of 
the mandible. To my knowledge, this condition has not 
been previously described in a hereditary form. 

Significance to Dental Research ; 

The etiology of several conditions may now be ascribed 
to hereditary causes. The clinical description of the 
oral defects has been given, and experience in surgical 
correction and restoration has been obtained. 

Proposed Course of Project: 

We recently have initiated, with Dr. Karl Piez, NIDR, 
an investigation to try to determine the biochemical 
defect present in opalescent dentin. Histochemical 
studies indicate that there are more chemically re- 
active groups in the matrix of the dentin in the af- 
fected teeth than in normal teeth. A basophilic P.A.S. 

- * - Serial So. 

Part ft (continued) 

positive substance is present in dentinogenesis 
Imperfecta similar to that found in the bone of 
osteo-genesls Imperfecta. We believe that this 
is an abnormal protein. The chemical methods are 
being perfected for analysis of normal bone, dentin, 
skin, and tendon, vhich will be utilized to inves- 
tigate a variety of hereditary defects. The first 
of these will be opalescent dentin. Continued ac- 
cumulation of pedigree material and linkage data is 

Part B Included: Yes 

- 5 - Serial Ho. HIDR-44 (C) 


Individual Project Report 
Calendar Tear 1958 

Part B : Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Wltkop, C.J. , Jr.: Genetics and Dentistry: Eugenics 
Quarterly, 5.:15-21, March, 1958. 

Honors and Awards relating to this project: None 

Serial No, 

lo Clinical Investigations 

2. Human Genetics 

3. Brandywine, Maryland 

Individual Project Report 
Calendar Tear 1958 

Payt A 

Project Title: Genetic Study in Southern Maryland. 

Principal Investigator: Dr. C.J,, Witkop, Jr„ 

Other Investigators: Dr, Do L. Rucknagel 
Dr. J. Do Nlswander 
Ho Ro Dyson, R.N. 

Cooperating Dnlts: Blood Bank DBS; NIAMD; NCI; NINDB; School 
of Dentistry, Howard University; Sociology 
Department, Catholic University* 

Patient Days: 319 

Man Tears (calendar year 1958): 
Total: 3 1/3 
Professional: 113 
Other : 2 

Project Description: 


1. To determine the total amount of hereditary Illness 
present in an isolate population of not over 5,000 
related individuals of white, American Indian, and 
Negro ancestry as revealed by dental and physical 
examinations and selected laboratory procedures. 

2. To determine the prevalence, mode of inheritance, 
effects on viability and longevity, clinical mani- 
festations and variations in expressivity of the 
hereditary illnesses found. 

3. To Investigate the relationships and possible linkages 
between these illnesses and blood types, secretor 
factor, P.T.C. taste test and other genetic markers. 

4. To investigate genetic factors in Illness not usually 
known to have a hereditary component. 

- 2 - Serial Mo. HIDR-45 (C) 

Part A (continued) 

5. To select patient and biological material from 
this population for clinical and biochemical in- 
vestigations of specific Illnesses and normal 
factors (these will constitute separate projects). 

6. To assess the effects of inbreeding as revealed 
by ln-group and out-group marriages and by degree 
of relationship (i.e., first cousins, first 
cousins once removed, second cousins, etc.) as 
shown by perinatal deaths, viability, longevity, 
and the presence of apparent homozygous recessive 

7. To try to trace the source and outflow of patho- 
logical genes in this population. 

8. To assess the social and genetic factors in mate 

9. To look for correlations between genetic illnesses 
and other normal and abnormal factors that may have 
etiological or diagnostic relationships. 

10. To try to reconstruct, on the basis of gene 
frequencies, the sources of the group's genetic 

11. To compare the prevalence of illness, gene 
frequencies, and vital statistics with other isolate 
populations and the general population. 

12. To detect carriers of recessive genes, if possible. 
Methods Employed ; 

This study is divided into three phases: 

1. Field Study: 

The field study consists of history, and dental, 
medical and laboratory examinations designed to 
detect as much hereditary illness as possible. 

1) History, genetic, and census data are obtained 
on a questionnaire covering 158 items which ar- 
ranges the data by family groups and by sibships. 

- 3 - Serial Ho. NIDR-45 (C> 

Part A (continued) 

Such items as birth order effects, parental 
age, consanguinity, age of onset, longevity, 
and perinatal mortality are included. A record 
of hospitalizations and institutionalizations is 
obtained in addition to a medical and genetic 
history. A master kindred chart showing the re- 
lationship of each individual is made. 

2) A search of existing social and church records 
for births and marriages is made by Fr. Thomas 
Harte. Sociological data on development in 
marriage patterns in this group are also ob- 

3) All subjects in this study are then given a com- 
plete dental and medical examination, designed 

to detect as much hereditary illness as possible. 
This Includes complete blood typing genetic 
markers; urine analysis; hematological work-up, 
especially for abnormal hemoglobins; and special 
procedures such as X-rays and ophthalmological 
examinations where indicated. These examinations 
are made during the summer months at mobile units. 

4) All data are then coded and tabulated for I.B.M. 
punch :ards. 

2. Clinical Study: 

Selected patients seen in the field study are brought 
to NIH for detailed clinical studies. These studies 
are designed to give a description of the hereditary 
diseases involved; to determine the limits of the signs 
and symptoms of the illness; and to describe any 
metabolic abnormalities detectable on routine labora- 
tory examinations, or other associated defects that 
might be found on X-rays or by special procedures. 

3. Biochemical Investigation: 

Selected genetic defects are being investigated bio- 
chemically to find the chemical error present. Cur- 
rently under study are sickle cell disease, dentino- 
genesis imperfecta, and diffuse goiter. The sickle 
cell disease is being investigated by tonometry, 

- 4 - Serial Ho. HIDR-45 (C) 

Part A (continued) 

electrophoresis, and the survival of this abnormal 
hemoglobin in transfused patients. Dentinogenesis 
imperfecta is being investigated histochemically 
and by chemical analysis for abnormal protein con- 
stituents. The goiter patients are being studied 
for the presence or absence of non-thyroglobulin 
thyroid proteins, the response to thyroid stimulat- 
ing hormone, serum electrophoresis, total protein 
bound and butanol ex tractable iodide, and incubation 
of thyroid biopsies with 1*31. 

Patient Material : 

Patient material is selected from the families seen 
in the field studies. Approximately 500 hospital 
days have been utilized by this and other institutes. 

Ma lor Findings : 

These data have not been analyzed statistically; how- 
ever, a preliminary examination indicated the following: 

1. There appears to be a hereditary block in 
thyroglobulln synthesis that accounts for the 
prevalence of goiter in about 8 percent of the 
women in this group. 

2. A new hereditary bone disease, best described 
as generalized hereditary osseous hyperplasia, 
has been found in this group to be inherited as 

a recessive trait, and is known to exist in three 
other families. This is a newly described bone 
condition in which all deaths, with one exception, 
of people showing this disease have been attributed 
to respiratory causes. 

3. Ankyloglos8ia appears to be inherited as a domi- 
nant traie. 

4. There appears to be a genetic basis for certain 
speech defects not necessarily associated with 
gross oral abnormalities. 

5. Four patients with a rare blood type, which has a 
frequency of .00008 in the general population, have 
been found. 

- 5 - Serial No, BIDR -45 (C) 

Par t A (continued) 

6. Genetic evidence has been obtained to show 
that either there is an alternate pathway 

in melanin synthesis (other than that presently 
proposed) or that a secondary pigoent system is 
present in humans. 

7. A familial form of gynecomastia exists that is 
not Klelnfelter 's syndrome. 

8. The frequencies of dentinogenesis imperfecta and 
albinism in the Brandywine group are the highest 
known in any population. 

Significance to Dental R esearch ; 

An isolate population has been found containing a vast 
number of hereditary diseases which may shed light on 
genetics and biochemical pathways of various hereditary 

Propose d Course of Project : 

To continue the field, clinical and biochemical studies 
for the uext years. 

Part B included: Yes 

- 6 - Serial Ho. NIDR-45 (C) 


Individual Project Report 

Calendar Year 1958 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts from this project: 

1. Dyson, Hazel R. , Witkop, C.J., Jr, , and Butters, Shirley 
S.: Genetics in Public Health Nursing. Eugenics 
Quarterly, 5:23-28, Karch, 1958. 

2. Career, J.F. : A description of some speech anomalies in 

a tri-racial Isolate group. A Thesis, presented in partial 
fulfillment of the requirements for a Master of Science 
Degree, May, 1958, University of Maryland. 

3. Harte, T.J.: The Use of Parish Records in Social Research. 
Am. Catholic Sociological Review, June, 1958. 

4. Harte, T. J. : Trends in Mate Selection in a Tri-racial 
Isolate. Social Forces, November, 1958. 

Honors and Awards relating to this project: None 


Individual Project Report 

Calendar Year 1958 

Serial No. HIDR-46 (C) 

1. Clinical Investigations 

2. Human Genetics 

3. Bethesda, Md, 

Part A. 

Project Title: Familial Osteoarthritis 

Principal Investigators: Dr. C. J. Witkop, Jr. 

Other Investigators: H. R. Dyson, R„ N. 

Cooperating Units: Dr< R. L. Black, BIAMD 

Man Tears (calendar year 1958): 
Total : 5/6 
Professional: 1/3 
Other: 1/2 

Project Description: 

Objective : 

A clinical and genetic investigation of the descendants of 
two short sisters (dated froa seven generations ago) is 
being undertaken to determine if osteoarthritis is inherited 
as suspected from our preliminary study. It was noted in 
this particular family that shortness of stature seemed to 
be inherited as a dominant trait and that the short indivi- 
duals shoved an early onset of osteoarthritis, degenerative 
disc disease, obesity, and gall bladder disease. The inheri- 
tance of these conditions in this family seems to be similar 
to the strain of rats found by L. Sokoloff of KIAlffi. 

Methods Employed : 

A kindred chart and census of the entire family was made by 
our genetic field worker. Following this, all individuals 
on the kindred chart ere asked to come to HIH for a physical 
screening examination. This examination includes skeletal 
X-rays, routine blood and urine analyses, and physical 
examinations with emphasis on the signs and symptoms of 
osteoarthritis; i e„ examination of joints, joint movements, 
spinal involvement, signs of degenerative disc disease, 
height, weight, and other physical measurements. The latter 

Part A . Cent's. - 2 - Serial No. HIDR-46 (C) 

findings are correlated with the X-ray findings and then 
compared with the observed inheritance of stature as 
shown by the kindred chart. The physician doing the 
examinations does not know if the individuals being ex- 
amined are in the affected line or not. 

Patient Material : 

Patient material is drawn from this kindred, all exami- 
nations being on an outpatient basis. 

1. In the affected line, the onset of osteoarthritis occurs 
at a much earlier age, about 30 years, than in the unaffected 

2. There appears to be a correlation between the age of 
onset and the stature of the individual, with the short 
individuals showing the earliest onset. 

3. The earliest onset of the osteoarthritis in this family 

was an individual 21 years of age. « 


4. There is no affected consanguinity. The condition u 
appears to be an autosomal dominant trait. m 

Significance to Dental Research : 

The finding of a large family with hereditary disease in [ 

a large number of individuals furnishes a mass of clinical <" 

material which is most likely to be of one gene defect and £', 

provides an epidemiological method for selecting a large * 

number of patients for biochemical studies. J 

Proposed Course of Project : 

To date, 105 individuals have been examined. It is pro- 
posed to continue examinations until as many individuals 
as possible have been seen. If the clinical findings show 
that osteoarthritis is inherited as a simple dominant trait, 
the patients will be admitted to look for a biochemical 
defect which may accompany this illness. 

Part B included: So 

Serial 8o„ HXPR-47 ftq 

1. Clinical Investigations 

2. Human Genetics 

3> Be thee da and Clinton» Md< 


Individual Project Report 
Calendar Year 1958 


Project Title: Hereditary Eye Defects 

Principal Investigator: Dr. C. J. Witkop, Jr. 

Other Investigators: H. R. Dyson, R.Ho 

Cooperating Units: Laboratory of Ophthalmology, HIEDB; 
University of North Carolina; and 
Ophthalmology Dept. , Washington 
Medical Center. 

Han Tears (calendar year 1958): Patient Days: 2A3 

Total: 5/6 PatienO admitted to MINDB* 

Professional: 1/3 
Other: 1/2 

Project Description: 

Objectives : 

To study the hereditary factors involved in eye diseases 
encountered in the Genetic Study in Southern Maryland aac> 
in the study of Trl-racial Isolates in Eastern United States c 

Three conditions have been encountered which have been 
included under this project: 

1. A study of chronic simple glaucoma in the Southern 
Maryland population. 

2. A study of hereditary limbal vernal conjunctivitis in 
the Haliwar population. 

3. A study of cranial dysostosis and ptosis occurring In 
a Southern Maryland faally, 

These families were discovered during a study of hereditary 
diseases in isolate populations. 

Part A . Cont'd. - 2 - Serial No. NIDR-47 (C) 

Methods Employed ; 

Field workers obtain history and kindred charting from the 
families concerned, make appointments for eye examinations, 
and record the data. Ail patients in the kindred are ex- 
amined by the usual ophthalmological methods. Blood and 
saliva examinations, and PTC taste test are given for 
evaluation of family relationships and for linkage purposes- 

Patient Material : 

Patient material is drawn from these families and admitted 
to the Ophthalmological Service of NINDB. 

Ma lor Findings : 

1. Mrs. Hazel R. Dyson, R.N. , assigned to the NIDR Human 
Genetics Section, has successfully demonstrated a genetic 
method for the prediction and detection of chronic simple 
glaucoma in individuals. She found that if an individual 
has an inherited disease, examination of predetermined 
members of the kindred may reveal additional examples of 
tha disease and thus increase the effectiveness of screening 
procedures. Further, if the mode of inheritance is dominant, 
as with glaucoma, one can accurately trace the affected 
individuals through family history and home visits to 
identify the sibships that should be examined. 

For example, during the early stages of a current dental 
examination program of an isolate population group in 
Maryland, two patients were found to have chronic glaucoma, 
A family history of eye disease was obtained for each, end 
sibships in which glaucoma was most likely to occur were 
established. Examination of the selected kindred revealed 
five cases of frank glaucoma and one glaucoma suspect. Two 
of the patients were discovered before they were aware of 
any eye trouble. Additional studies on other families have 
shown that relatives of affected patients frequently have 
the disease. 

The Washington Society for the Prevention of Blindness is 
new utilizing the described method of study for detecting 
new cases of glaucoma. The last report of the Society, 
following a one-month trial utilizing this method, showed 
that 21 new cases of glaucoma and 36 suspects were found in 
the Washington area. 

- 3 - Serial No. NIDR-47 (C) 

Pirt A . Cont'd. 

2. The llnbal form of vernal conjunctivitis had not 
previously been known to be hereditary. A study of a 
large family in Halifax County, North Carolina, has 
shown that the disease followed a hereditary autosomal 
dominant inheritance through five generations, affecting 
more than forty individuals in the family., This con- 
dition also has a strong allergic component, the eye 
lesion showing many eosinophiles and eventually leading 
to blindness. It was thought that this lesion was a 
carconoma in situ, and removal of the eye was recommended. 
Our study pointed out that many individuals lived to be 
over 90 with the aondition, and we recommended that 
enucleation not be performed. 

Significance to Dental Research : 

1. Genetic principles applied to such illnesses as chronic 
simple glaucoma provide a practical method for detecting 
this disease in an early stage when it is most susceptible 
to treatment. 


2. Hereditary defects may affect several structures, and n! 
a clue to their diagnosis may be provided by dental j|| 
practitioners. IH 

Proposed Course of Project " 1 

Examinations of individuals affected with limbal vernal 
conjunctivitis is being planned for December, 1959, to u< 

complete the family study. Patients will continue to be <j' 
admitted for biopsy and steroid treatment, and the data ^ 
will be analyzed for publicstion in the near future. <" 

Continued study of the eye defect occurring in these. 1 

groups will be made. The next condition to be studied 
will be microphthalmia occurring in a population in 

Part B included: No 


Individual Project Report 

Calendar Tear 1958 

Serial Hc KEDH- 

lo Clij&cai^nvestigf 

2 Human Genetics 

3o Betheeda 9 Maryland 

Part A 

Project Title: Sickle Cell Anemias and Other Inherited 
Hematological Disorders 

Principal Investigator: Dr e D* L« Rucknagel 

Other Investigators: Dr & C« Jo Witkop, Jr e 

Cooperating Units: Drub Jo E Q Seegmiller, Bo LaDu, L c Laster 9 

Man Years (calendar year 1958): Patient Bays: 730 
Total: 1 2/3 
Professional: 2/3 
Other: 1 

Project Descriptions 


To study in detail physiologic, biochemical, and genetic 
factors in various Inherited hematologic disorders in the 
Brandywine trifacial isolate., Laboratory and clinical 
data will be integrated with the pedigree and analyzed 
for the purpose of defining biochemical mechanisms and 
mode of inheritance of dieease 

Mgthpda Employed : 

Field surveys among the Wesort groups have detected indi- 
viduals with several hematologic disorderso Approximately 
fifteen percent of this group harbor the sickle cell gene 
Individuals with sickle cell anemia are hospitalized and 
evaluated by standard hematologic procedures,, In addition 
specialized investigations of intravascular sickling 
arterial oaqrhemoglobin saturation, and erythrocyte 
physiology using tonometric techniques are utilized. 

2 => Serial No c 

Part A (continued) 

Relatives of propositi are studied hematologically using 
standard hematologic t£chniques 9 paper and starch block 
electrophoresis and when indicated, oxygenation kino 

Non-sicklemic anemic individuals and their families a -e 
also being studied to elucidate ether genetic abnormalifcies„ 
One individual with pernicious aneaia was studied exten- 
sively (in cooperation with Drs e Seegailler» LaDu, and 
Laater) in an attempt to elucidate the effect of 
vitamin B- i2 deficiency upon purine biosyntheslso 

Patient Material ; 

This year another 1100 individuals were screened in the 
field examinationso Anemic individuals were selected 
fran this group for the mest parte A few non~group 
sickle cell anooia patients were studied as control 
subjects,, Fatal hemoglobin assays on blood of patients 
with choriocarcinoma ars continuing t and additional 
studies are under way on patients with an assort- 
mew; of other rualignancles being treated by the National 
Cancer Institute,, 

Major Findings : 

During the past year aerial clinical, studies and erythr©~ 
:yte investigations have been perform* < 
eighteen individuals with sickle .jell anemia These 
have indicated that the amount of intravascular sickling 
and the degree of arterial oxygen unsaturation are reasonably 
constant in a given individual over a long period of time 
Quantitative studies have documented several degrees of 
severity of these diseaae manifestatlonso Moreover, 
laboratory investigations have demonstrated that the 
"arlations in clinical manifestations and in arterial 
saturation can be correlated with abnormalities in 
erythrocyte physiology per se 9 and not with associated 
systemic disease 9 i e 08 pulmonary insufficiency^ 
cardiac anomalies? etc 

The patient with pernicious anemia was found to excrete 
large amounts of glutamic acid s a compound 

- 3 - Serial Noo 
Part A (contirn 

olved in folic acid metabolism, since it is al 
excreted by rata and humane receiving the folic acid 

aatabolite, amethopterin In addition^ carbe:i~l*f~ 
labelled formamidino aminoimidascle carboxamide was 
incorporated lesa efficiently into uric aoid than 
aminoldnidaiole carboxamide in our patiento Both of 
these obaervations implicate defective fcrmyl group 
metabolism in pernicious anemia 

To date 9 increased amounts of fetal hemoglobin have 
been found in six of twenty-eight patienta with chorion 
carcinoma studiedo Bloods from thirty individuals with 
other malignancies contained normal amounts of fe^al 
henog*.obino Since chorionic gonadotropin is considered 
the stimulus to fetal hemoglobin production in these 
patients, it may also increase the fetal hemoglobin 
content of oelis of patients with sickle cell anemias 
thereby allesriasing rhe clinical severity However 
preliminary tr'.als on a small numbe T * of sickle cell 
anemia patients have shown no significant results to 
date Q 

Si gnificance to Dental Research : 

e it is recognited that genetic factors are important 
in ohe study of dental diseases as well as other medical 
problems, this program is aimed at evaluating genetic 
mechanisms by e.o many approaches as are feasible. By 
studying specific entities as thoroughly as possible 
and correlating variations of disease and laboratory 
findings with pedigree data,, it is anticipated that 
conclusions may be forthcoming regarding genetic 

Proposed Course of Projects i 

Enlargement of the pedigree will continue but hematologic 
investigations are now being concentrated in areas of the 
pedigree deemed to be most promisingo These are designed 
to elucidate the presence and nature of modifying hereditary 
factors in sickle cell anemia, and to evaluate the presence 
of Thalassemic genes in the non~sieklemic anemia sibshipso 

- 4 ■ Serial No* jggjUjg, 
Part--' u*d) 

Other specific projects planned include: 

1) Extension of sickle cell erythrocyte physiology 

2) Formyl metabolism in pernicious anemia e 

3) Ery chrocyte en-syme anomalieso 

t) Haptoglobin genetics in the Brandywine Group<> 

Part B included Wo 

Serial No. NIDR-49 (C) 

1. Clinical Investigations 

2. atacan Genetics 

3. '.iethesda, &d. 

ladividual Project: Report 
Calender Year L958 

Part A. 

Project Title: Oral Aspects o.': the Results of Consanguineous 
Marriage in Bi.oshiaa and Nagasaki., Japan. 

Principal Investigator: Dr. J. D. Niswender 

Other Investigators: Dr. C. J, tfltkop, Jr. 

Cooperating Units: National Actdesy of Sciences, Washington, 
D. C. , under the direction of Jatses V. 
Seel, V7illi*» J. Schull, and Junes N. 

Kan Years (calendar year 1958) ; 
Tota;i: 2/3 
Professional: 2/3 
Other: Hone 

Project Description: 

Objectives : 

1. 'in co inaction with the genetic program of the Atcaaic 
Doe* Casualty Ccamissioa in Biroshlaa and Nagasaki, Japan , 
the details of which have been presented by Seel and Schull 
(Nee., J. 7. and Schull, tf.J.: The Effect of E:cposure to the 
Atomic Boabs on Pregnancy Termination in Eirouhima and 
Nagasaki. National Acadeeiy of Sciences, #461 Washington, 
1956), tie outcome of prejnancy was determined in 5,033 
instances in which the parents reported consanguinity of 
some degree. The products of these pregnancies would, if 
now alive, range in age between five and ten years at this 
tine. In essence, this proposal involves an attempt to 
obtain a detailed follow-u^ etudy on each of these 5,033 
pregnancy terminations, plus a suitable control Material. 

2. Of tha 5,033 pregnancy terminations, a ntr.ber are known 
to have resulted in stillbirths or deaths during the firs!; 
nine mentis of life. Starting from Che records of the Atomic 
Jiosab Casualty Commission, « trained utaff of "field workers' 
will atteapt to reestablisfc contacts with all of the regaining 

Part A . Cont'd. - 2 - 

Serial lio. NIDR-49 (C) 

children (i.e registered in the Genetics Program and 
not Known to have died during the period of timHovered 
by this Program). This attempt will result in a roster 
of livmg children, a roster of now-deceased children 
T ti _V T V °* children "*><> ^ve moved fro* the two study 
Ultl f 11 £ 5 f0rt WU1 be Mde to "tablish cause of * 
death for all deceased children. The background of the 
chilaren who have moved from the cities wil . be analyzed, 

ittLot*!iu r bl r ing factor8 - xt i8 p° 8aibie *•* « 

attempt will be made to contact theae latter children with 
reference to the question of present viability. The fact 
of death and, possibly, the cause of death will be aulvwd 
in relation to parental consanguinity. 

It „ A ? U ? t J , f? d ? Car f ied out ia 1P56 «*»" the direction 
of W. J. Schull has already revealed that the death rate 
"■ong the children of consanguineous marriages is approxi- 
mately twice the control death rate (cf. Appendix) . There 
is, then, already considerable assurance of significant 

?«*!! i ..« ^ in< J utr y- ***** e «« dat« can be analyzed 
in relation to the radiation history of the parents, thus 
yielding additional information concerning the genetic 
effecta of the atonic bombs. 

4. The dental portion of this study proposes to determine 
tne effects of consanguinity on DM? rates, periodontal 
disease indices, growth and development problems, and 
specific hereditary diseases affecting the oral cavity 

Methods Employed : 

Using ABCC facilities in Nagasaki aid Hiroshima, a team 
consisting of specialists in such fields as pediatrics, 
hematology, dentistry, and anthropology begaa examinations 
of offsprings of consanguineous marriage and appropriate 
control group in September, 1958. The following examinations 
are performed: 

1. Physical examination: This consists of a standard 
physical examination, with emphasis on characteristics 
which can ge graded in an objective manner lending itself 
to statistical analysis. 

a. Presence of gross physical defect; i.e. , abnor~ 
malities of skin, musculature, skeleton, etc. 

b. Defects of the auditory mechanism, with special 
reference to loss of hearing. 

c. Defects of the eye, including loss of visual acuity. 

d. Cardiac abnormality (apparent cardioaegaly disorders 
of rhythm, murmurs, etc.), 

PartA. Cont'd. - i = 

Serial gjo. Hlpg-49 (p) 

e. Puliaonary abnormality „ 

£. Apparent abnormality of abdominal viscera. 

g. Abnormality of external genitalia. 

h. Neurological evaluation. 

1. Bone age (single wrist film). 

J. Determination of leucocyte count and hemoglobin level, 

k. Standard urinalysis. 

2. The dental examination Includes pathology of the soft 
tissues, pathology of the teeth and index of malocclusion, 
index of tooth loss, developmental failures of oral struc 
tures, WS? and DBF rates, and the association of these 
conditions with and without generalized hereditary disease, 

3. Anthropometric examination: This will include such 
standard .teas as weight, sitting and standing height, span, 
measurements of head and face, subcutaneous fat index, etc. 

4. Psychometric examination: Discussion here centers on 
the use of the Koga intelligence test, one of the Japanese 
equivalents of the Stanford-Blnet. If possible, all tests 
will be individually, or seat -individually, administered. 

All results will be coded and transferred to IBM cardts for 
statistical analysis. As for the Genetics Program data, 
the statistical analysis will undoubtedly draw heavily on 
an approach which makes allowance for certain uncontrollable 
sources of bias. The first result will be a comparison of 
the different groups of children with respect to a wide 
variety of characteristics with particular reference to the 
regression of the findings of degree of Inbreeding, fieyonc'. 
this, howtVer, it is anticipated that the material will 
permit tentative conclusions concerning the number of 
"lethal-equivalents" carried by the average Japanese, 
utilising the mathematical approach suggested by Morton, 
Crow, and Killer (1956). 

Major Findings: Hone 

Significance to Dental Research : 

I. Properly executed studies on consanguinity effects will 
yield information on the genetic structure of human popu°> 
lations which at present can be obtained in no ether v>&y a 
Such information is vital to a realistic, quantitative 
appraisal of such problems as are posed by the increasing 
exposure of the human species to ionising radiation. 

Part A . Cont'd. - 4 - Serial Mo. HIM°49 (C) 

2. The Japanese Study ??ill offer an opportunity to study 
the generic effects of a specific type of int reeding, 
complementing our Brandywine Study. Whereas the latter 
program is assessing the effects of continuous inbreeding 
the Japanese Study has the unique advantage cf affording 
an evaluation of one type of suiting pattern. 

3. It would be expected that certain recessive character- 
istics determining normal growth sue. development, as well 
as pathological traits, would show i.p in the homozygous 
state vi. -Jh increased frequency. Maty of these conditions 
i».re probibly unknown, and the Japanese Study consequently 
offers a i opportunity to define both normal factors and 
pathological trsitu . 

4. The nrandywine population group is descended from 
relatively few ancestors. Consequently, unless an ancestor 
carried a hidden trait it could not appear ir the descent eats, 
that is, barring mutation. Therefore, the Brandywine group 
would praeeut only those inherited conditions carried by 
predecessor*. On the other hand, the first cousins in 

Japan ar; drat;o from a large unrelated population and, 
therefore, many characteristics and pathological traits 
should a>pear in their offspring. 

Propose d Cour m of Project : 

Examinations will continue until June, i960, following 
which Dr, Mlswander will return and spend the following 
year analyzing the data. 

Part B included: Jo 

Serial Mo. NIDR-50 (C) 

lo Clinical laves tigatloaa 

3o Bethesda, Md. 


Icdi/idual Project Report 
Calendar Year 1958 

Pare A 

Project Title: Studies of the Oral Microbiota and Oral 

Environment in Relation to Dental and Systemic 

Principal Investigator: Dr. R. M. Stephan 

Other Investigators: None 

Cooperating Units: Dental Clinic, Clinical Center, NIH 


Total : 

1 1/3 





Project Description: 
Objectives : 

1. To study tnacroscoplcally and microscopically the growth, 
localization, morphology, physical characteristics, and 
color reactions of microbic plaques on the teeth, ia 
gingival crevices, in periodontal pockets, and on the 
oral mucous membranes. 

2. To determine differences in microbic plaque formation, 
food retention, salivary flow, gingival exudates, and 
pH levels which are associated with caries, erosion, 
and periodontal disease in comparison with normal 
dental conditions. 

3. To investigate possible associations of specific types 
of oral microorganisms with dental and systemic diseases. 

Methods Employed : 

Direct sterecmicroscopic observations in the oral cavity 
are made at 6X-40X magnifications with the Zeieu ''Oto- 
scope." Direct pH measurements are made with micro 
antimony and glass electrodes and with micro-applicators 

- 2 - Serial No. HIDR-50 (C) 

Part A (continued) 

containing color ime trie indicators. Measurements of 
parotid salivary flow rates are made with the "Laahley 
Cup" apparatus. Samples of plaques and other surface 
material from the teeth, periodontal pockets, and 
mucous membranes are taken with dental explorers, 
scalers and swabs. Also, specially designed micro- 
sealers are used for taking quantitative samples 
which are weighed in a microbalance. These samples 
are used for phase microscopic examinations, stained 
smears, micro culture slides, and culture plates and 
also for related studies on certain types of organisms 
by Investigators in the Laboratory of Microbiology, 

Patient Material : 

Thus far 108 patients have been studied. Most of the 
cases have been outpatients of NIDR referred for study 
of rampant caries, dental erosion, severe periodontal 
disease, or stomatitis. Patients with various diseases 
who have been referred to the Dental Clinic from Che 
other Institutes at HIH also have been studied for pos- 
sible interrelationships between oral conditions and 
other diseases; 

Ma lor Findings ; 

Further information has been gained on several aspects 
of the complex host-parasite relationships in the oral 
cavity, particularly as they relate to carles, erosion, 
and periodontal disease. Quantitative estimates of the 
different forms of organisms in surface material removed 
from representative localized areas of the oral environ- 
ment have demonstrated wide variations in the miexobiota 
between different individuals, and between different 
areas in the same individual. At the same time, a more 
or less basic underlying pattern is emerging for the 
distribution of certain common types of organisms, such 
as spirochetes in gingival crevices and streptococci on 
tooth surfaces. The most interesting findings have been 
the unusually heavy growth of microbic plaques in rampant 
caries and the presence of many different forms of motile 
organisms in severe periodontal disease. Some of these 
motile forms have not yet been grown in pure culture and 

- 3 - Serial Ho. HXBR-50 (C) 

Tart A (continued) 

they have not been identified in histological material, 
so that their importance and general distribution is 
not yet known. 

Studies on intra-oral pH have continued to show normal 
pR levels between 6.0 and 8.0. In active caries, pH 
levels have been as low as 4.2. Recently, pH levels 
of 5.4 have been found in an unusual type of non- 
adherent "plaque" in active dental erosion areas. 
Patients who have received therapeutic X-ray irradiation 
of the head and neck have been of particular interest 
since, in some cases, they develop a lack of oral secre- 
tions and unusually rampant dental caries as a result of 
changes induced by the radiation. 

Significance to D ental Research ; 

This project has been planned to cover a neglected 
area in oral pathology and microbiology— namely, to 
furnish information on the total microbiota in the oral 
cavity as it grows "in situ 1 ' and as it is related to 
clinical disease processes. 

Progress has been made in developing a microculture 
slide method for carrying through observations on the 
oral microbiota from the time it is visualized micro- 
scopically in specific areas in the mouth to the time 
when the organisms can be identified by their selective 
growth characteristics. 

Proposed Course of Project: 

This project has explored several methods for study of 
the oral microbiota and oral environment and It is pro- 
posed to secure more extensive data and particularly to 
follow up observations on salivary flow and rampant 
carles following Z-ray therapy, on motile organises in 
periodontal disease, and on the pH changes la erosion 

Part B included: Ko 

Serial No. NIDR-51 (C) 
1. Clinical Investigations 
3. Bethesda, Hd. 
Individual Project Report 
Calendar Year 1958 

Part A„ 

Project Title: Interrelationships between Local and Systemic 
Factors in the Etiology of Dental Diseases in 

Principal Investigator: Dr. R. Mo Stephan 

Other Investigators: Hone 

Cooperating Units: None 

Man Tears (calendar year 1958): 
Total: 1 1/3 
Professional: 1/3 
Other: 1 

Project Description: 

Objectives : 

1. To deteraine the essential experimental conditions 
which control the development of dental caries and perio- 
dontal disease in rats. 

2. To study the relation between caries in rat incisors, 
and bacterial plaque, tooth formation, food retention, 
and intraoral pR. 

3. To develop the use of vital stains as indicators for 
the earliest stages of caries. 

4. To explore the caries-inhibiting action of some anti- 
bacterial dyes and the effects of dyes which stain tooth 

Methods Employed : 

Three basic diets have been used: (1) a nutritionally de- 
fined diet of sucrose, casein, salts and known 
vitamins which induces very extreme caries, (2) a nutrition- 
ally defined diet which induces little caries, and (3) a 
stock diet which does not induce caries. In aulti factorial 

Part A . Cont'd. - 2 - Serial No. NIDR-51 (C) 

experiments these basic control diets have been supple- 
mented with nutritional substances to determine systemic 
effects and the relation of growth rates to caries. Glu- 
cose and sucrose have been added to the drinking water to 
produce both local oral and systemic effects. Vital stains 
have been injected intraperitoneal ly, added to the diet, or 
added to the drinking water to serve as indicators for the 
early stages of carles. Stains have also been studied as 
indicators for pB and caries activity by direct application 
of the indicator to the teeth during examination of the 
oral cavity under the stereomlcroscope. 

Bacterial plaque formation has been studied microscopically 
on the incisor teeth in the live animal and the direct pfl 
measurements of plaques have been made with micro- antimony 
and glass electrodes and with pfl indicators. 

Mai or findings ; 

During the past year, a number of vital stains have been 
found to be good indicators for the initial stages of 
carles and over 60 stains were tested. One of the most 
interesting findings is that some dyes stain only the very 
earliest stages of caries in enamel whereas others stain 
only the advanced stages of caries. Although the 
hydroxyanthraquinone dyes continue to yield the best stain 
for bones and teeth, dyes from the oxazin and xanthene 
groups have been found useful for staining caries. It is 
becoming clear that there are several basic chemical re- 
actions which can be demonstrated in the caries process 
by improved vital staining methods. 

Toe effect of additions of sucrose and dextrose to the 
diet and drinking water has greatly increased caries in 
some experiments, and only moderately increased it in 
others. This has been shown to be due partly to the age 
of the animal and to the fact that when sugar is given 
early the effect on caries is greatest. 

These studies have continued to demonstrate that the 
development of caries in rats results from the interaction 
of many different factors including the strain of rat used, 
the physical properties of the diet, the nutritional value 
of the diet, the retention of diet in the oral cavity,, the 
growth of microorganisms on the teeth, and the relative 
growth and activity of the individual animal. 

p *rt A. Con'd. - 3 - Serial No„ NIDR-51 (C) 

Significance to Dental Research : 

The development of improved methods for studying the 
etiology of dental caries and periodontal disease in 
laboratory animals will permit more precise testing of 
the relative importance of many factors which may 
potentiate or control these diseases . The use of vital 
stains will aid in the early detection of dental caries 
and serve as an indicator for tooth formation in re- 
It tlon to caries susceptibility, 

Proposed Course o f Project 

It is clear that both dental caries and periodontal 
disease are very complex problems and that new methods 
are needed for determining their activity. It is 
planned to continue animal experiments on the vital 
staining of tooth substance and bone, and particularly 
to search foi more suitable indicators for demonstrating 
caries activity by color ime trie means. 

Part B included: No 

Serial No jSnibSLifil 
I. Clinical Investigations 
3. Betaesda, Maryland 
Individual Project Report 
Calendar Year 1958 

Part A 

Project Title: Studies on Enzymatic Reactions of Cyclic 
Alcohols and Steroids. 

Principal Investigator: Dr. A. D. Merritt 

Other Investigators: None 

Cooperating Units: Dr. G. M. Tonkins, RIAMD 
Dr. A. Rarmen, SHI 

Man Years (calendar year 1958): 
Total: 1 
Professional : 1 

Other : Hone 

Project Description: 

Objectives : 

(1) To purify a rat liver enzyme responsible for fcae 
reversible oxidation of cyclic secondary alcohols. 

(2) To further define the substrate specificity of horse 
liver alcohol dehydrogenase. 

(3) To specifically identify the stearic and conforma- 
tional characteristics of substrates for horse liver 
alcohol dehydrogenase. 

Methods Employed : 

The rat liver enzyme was prepared by standard procedures 
(hoaogenization, fractional ammonium sulfate precipita- 
tion , dialyzing) , followed by chromatography on 
diethylaminoethyl cellulose. Assays of enzymic activity 
were made by chemical, gas chromatographic and paper 
chromatographic techniques, as well as following the 
ultraviolet ebsorbtion of diphosphopyridine nucleotides. 

- 2 - Serial No. 

Part A (continued) 

Major Findings: 

Previous investigations by Dr G. M. Tomkins indicated 
that cyclic secondary alcohols were able to oxidize re- 
duced diphosphopyridine nucleotide in the presence of 
a rat liver homogenate. Studies were undertaken to 
purify the enzyme responsible for this activity. A 
partially purified enzyme was prepared and its kinetics, 
pE optimum, equilibrium constants, and substrate 
specificity were studied. Cyclohexanone was found to 
be stoichiometric4lly and reversibly reduced by 
diphosphopyridine' nucleotide. The products of the 
reaction were identified by means of paper chromatography. 
Since the substrate specificity paralleled that of horse 
liver alcohol dehydrogenase, the enzyme was thought to be 
a liver alcohol dehydrogenase. 

While Investigating the oxidation of cyclic secondary 
alcohols by a rat. liver preparation, it was found that 
horse liver alcohol dehydrogenase had activity with 
these substrates. Cyclohexanol was a somewhat better 
substrate than ethanol on a molar basis. The kinetics 
of this reaction were examined. 

Methyl substituted cyclic alcohols and ketones also were 
found to be active with liver alcohol dehydrogenase. The 
additional aubstituents allow the formation of cia- and 
trans- isomers which may be identified by means of gas 
chroma togr aphy . 

Significance to Dental Research ; 

These studies have broad biological import and tend to 
broaden the basic biochemical and medical aspects of 

Proposed Course of Project: 

Studies of cyclic ketone reduction products of liver 
alcohol dehydrogenase will be made to further define 
enzyme-substrate Interactions. 

In addition, some of the above techniques will be 
utilized in the study of »U reduction on ring A of 
steroid horsaones. 

- 3 - Serial Ho. 8XSR-52 (C) 

Part A (continued) 

Previous studies by Dr. 6. M. Tonkins on this enzyme 
indicated that there were a multiplicity of enzymes 
responsible for this activity. Attempts will be made 
to separate these enzymes into relatively pure com- 
ponents and then to prepare antibodies to these pro- 
teins in an effort to define similarities between the 
different enzymes. In addition, it is planned to 
study the antigenic relationships to a similar system 
of enzymes found in the microsomal fraction of rat 
liver hcoogenates . The two enzyme systems differ in 
that the soluble enzymes reduce & 4 bonds to 5? isomers 
while the microsomal enzymes form the 5a isomers. 

Part B included: Ho 

Serial Mo. MWBL-.S3 XC) 

1. Clinical Investigations 

3. Bethesda, Md. 


Individual Project Report 

Calendar Year 1958 

Part A 

Project Title: Serial Cephalometrlc Study of the Longi- 
tudinal Growth of the Soft Palate and 
Nasopharynx in Cleft Palate Children. 

Principal Investigator: Dr. P. J. Coccaro 

Other Investigators: Rone 

Cooperating Units: University of Illinois, Chicago Pro- 
fessional Colleges, Cleft Palate Center, 
Dr. Samuel Prut an sky, Associate Director. 

Man Tears (culendar year 1958) : 
Total: 1/2 

Professional: 1/2 

Other: Hone 'J|J 


Project Description: <«■ 

Objectives : uM 

1„ To study the growth pattern of velar length in cleft ""j 
palate subjects. X 


2. To determine incremental changes in nasopharyngeal , 
height and depth occurring as a result of growth., Jljjj 


3. To discern whether or not growth of the velum and Hj 
nasopharynx in cleft palate individuals differs 
significantly from that observed in the normal, 

Methods Employed : 

The method for this study was based on serial cephalo- 
metrlc roentgenology. The technique for taking the 
lateral headplates is well known and has been described 
by Broadbent, who helped develop the Broadbent- Bolton 
Cephalometer. A modification of the original cephalo- 
meter was designed to facilitate positioning the infant's 

- 2 - Serial No, NIDR-53 (C) 

Part A (continued) 

head into the specially constructed head holder which 
is in a fixed relation to the X-ray tube. The newly 
designed instrument was built around a table instead 
of a dental chair, and only one tube (the 37 lateral) 
was used. 

The cephalonetric records for the 37 subjects being 
analyzed case from the files of the Cleft Palate 
Center of the University of Illinois. The cleft 
cases were grouped according to type of involvement. 
The classification Is based upon general anatomic 

Type I - Posterior clefts. These Included clefts 

of the soft palate only, as well as clefts 
of the soft palate and the vault of the 
hard palate as far forward as the an- 
terior palatine foramen but not to Include 
the alveolar process or the lip. (Ten 
cases in this group.) 

Type II- Unilateral clefts. In this group the lip 
and palate are involved and either side 
of the face may be directly affected. In 
these cases the vomer is deviated from 
the midline at the line of attachment to 
the palatal process on the non-cleft side. 
(Fourteen cases.) 

Type Ill-Bilateral clefts. These involve the lip 
and palate also. The premaxillary seg- 
ment is projected considerably forward of 
the profile and there is a complete separa- 
tion of this segment from the lateral 
palatine processes of the maxilla as well 
as the horizontal processes of the palatine 
bone. There is no fusion of the palatine 
processes with the inferior border of the 
nasal septum. (Seven cases.) 

Type IV- Alveolar clefts. These may be defined as 
clefts of the alveolar process, varying in 
degree of severity and usually associated 
with a cleft lip. The palate is not in- 
volved in this group although the cleft 
may extend posteriorly as a hidden or sub- 
mucous cleft. (Six cases/) 

- 3 - Serial Ho. HIDR-53 (C) 

Part A (continued) 

Correctional scales cut from each film were utilized 
and the following linear measurements were determined: 

A. Velar Length 

1. The distance between the posterior border 
of the hard palate, as visualized on the 
lateral head plate, to the tip of the uvula 
of the resting soft palate, 

2 . The distance between a point where the In- 
ferior aspect of the pterygomaxlllary 
fissure approximates the palatal plane to 
the tip of the uvula of the resting soft 

B. nasopharyngeal Height 

The distance between a point, where the 
Inferior aspect of the pterygomaxlllary 
fissure approximates the palatal plane to 
the cranial base. This distance was 
measured along a line perpendicular to the 
Frankfort horizontal plane and projected to 
Intersect the cranial base line. The 
cranial base line extended from baslon to 

C. Hasopharyngeal Depth 

The distance between a point, where the In- 
ferior aspect of the pterygomaxlllary fissure 
approximates the palatal plane, to the soft 
tissue of the pharynx. 

Major Findings : 

1. Longitudinal growth of the soft palate appears to 
occur in both cleft palate and normal Individuals . 

2. The periods of active and less active growth seen 

In the cleft palate population do not parallel those 
observed for the normal ° 


- 4 - Serial So. BIDK-53 (C) 

Part A (continued) 

3. Shorter velar lengths were observed throughout 
the period studied for the cleft palate indivi- 
duals when compared with velar lengths recorded 
for the normal o 

4. Steady and consistent increments in the nasopharyn- 
geal height were recorded for the period evaluated. 

5. Shorter vertical measurements in the nasopharyn- 
geal area were observed for the cleft palate group 
when compared with the normal. 

6. Antero-posterior dimension of the nasopharynx in- 
creased noticeably up to the second year of ll£e° 
Periods of apparent increase and decrease in 
nasopharyngeal depth were seen to occur after two 
years of age. 

Significance to Dental Research : 

1. The cleft palate patient presents many complex oral' 
dental and facial deformities affecting mastication, 
speech, respiration and deglutition. 

2. The need for basic information on growth and de- 
velopment of structures Involved in these deformi- 
ties is quite apparent. 

3. Instituting therapeutic measures to produce adequate 
physiologic relationships of the anatomical parts la 
the veolpharyngeal area is essential for the produc- 
tion of satisfactory phonetic articulation. 

4. Adequate velar length along with the functional po- 
tential of velar tissue and its location to the 
posterior pharyngeal wall have beeu advanced as im- 
portant factors for the production of satisfactory 

Proposed Course of Project: 

1. Present study, consisting of 36 cases, will be ex- 
tended to obtain adequate numbers in each cleft 
group that would be significant. 

- 5 - Serial Ho„ 

Part A (continued) 

2c Future studies are anticipated from the 

analysis of this material relative to the growth 
of velar tissue before and after certain surgical 
procedures o 

Part B included: Ho 

Serial 80. WIER-54 (C) 
lo Clinical Investigatioea 
3. Bechesda, Kd„ 
Individual Project Report 
Calendar Tear 1958 

Part A 

Project Title: General Anesthesia (Dental) Baseline Data. 

Principal Investigator : Dr, E, J. Drlscoll 

Other Investigators: Rone 

Cooperating Units: Dr. Clarence L, Bebert and Dr. Gilbert 

R. Christenaon, Anesthesiology Department, 
Clinical Center. 

Man Tears (calendar year 1958): Patient Visits: 668 
Total: 1 1/3 
Professional: 2/3 
Other: 2/3 

Project Description: 

Objectives : 

Dental general anesthesia is different from ordinary 
hospital anesthesias In several important respects; 
a) The patients are generally ambulatory and not usually 
sedated; b) The operations are performed in as light a 
plane as possible; and c) The operations are performed in 
or about the airway. 

1. Because of these major differences, the back- 
ground of knowledge in general dental anesthesia is not 
for the most part complete, accurate or comparable. 
Thus, a great need exists for a background of fundamental 
data concerning alterations in the physiological mecha- 
nisms. Although body functions may be altered within 
certain limits, great hazards may be encountered when 
these limits are exceeded. It is Important, therefore, to 
define these limits. 

2. The accumulated data from this study will alao be 
used as a baseline of comparison for the many new anes- 
thetic drugs which are being, proposed for use in oral 

2 - Serial Ko. 

Part A (continued) 

Methods Employed : 

1. For the purposes of standardization of surgi- 
cal trauma, full south extractions have been utilised 
rather than random oral surgical cases. The teeth 
are extracted iu quadrants and the entire operation 

is systematized for procedure and traumatic experience 

2. During the current year, the following drug and 
drug combinations have been evaluated: Sodium Pentothal 
and the new barbiturate, Her aval, which is supposed to 
ensure a quicker recovery time than any previously u<jed 

The drugs Pentothal and Her aval have also been employed 
in a series of operations with the parasympathetic de- 
pressant, Banthine. The drugs are now being studied in 
another series of operations in which the anticholinergic 
drug, Tral (hexocycllum methyl sulphate) is used. This 
is an experimental drug supplied by Abbott. 

3. Supplemental Information is still being obtained 
by combining the barbiturates with nitrous Oxid-Oxygen. 

4a. Physiologic status and alterations being studied 
and recorded are pulse, blood pressure, circulatory 
changes, respiratory phenomena, and cortical (brain) 

b. In addition, information is being gathered by 
continuous recordings of E.K.G., E.E.G* , and a 
sphygmotonograph (continuous blood pressure) . 

c. Important technical data, such as amount of 
drug used in units of time, length of operation and dura- 
tion of sleep, are likewise recorded. 

d. Pertinent patient data such as age, race, sex, 
and emotional status in relation to the conduct of anes- 
thesia are being recorded, as well as post-anesthetic 
sequalae (headache, nausea and vomiting, depression and 
hiccoughs) . 


- 3 - Serial Bo. Mg^JliJLSl 

P art A (continued) 

Patient Material ; 

We now have over 300 detailed anesthesia records 
(E.E.G. , EKoGo, pulse, etc.) on almost 100 adult 
patients. Approximately half of the work has been 
performed this calendar year.. 

Ma )or P ladings : 

Sufficient data is now available to determine ob- 
jectively from brain wave activity the precise 
anesthetic levels at which various procedures are 
being carried out. 

1. Thus, it la definitely established that we 
are working In an extremely light plane. (Plane 1 
and II of the Kiersey Faulconer classification)) 
The E.E.G. -is also making it possible for us to state 
with certainty the precise anesthetic plane which is 
present at specific intervals when other physiologic 
measurements such as pulse and pressure are made<> 


2. Considerable disturbing arrythmias were noted 
in the study (when no parasympathetic depressant 

(Banthlne) was used). On the other hand, these „„. 

arrythmias were much less frequent where Banthlne was i(l) 

used. J 

3. The blood pressures have been found to con- 'L 
slstently rise during the operations; the mean rise p 
being approximately 30 mm, systolic regardless of ,,« 
whether or not Banthlne was used. This rise is inter- ''< 
preted as due to surgical stimulation under light 
anesthesia with a poor analgesic agent (Pentothal and 
Her aval). 

Significance t o Dental Research : 

1. The search is constantly being made in 
dentistry for safer general anesthetic agents, How- 
ever, without baseline data on physiologic responses, 
this search has no foundation. 

2. The possibility also exists for improvement 
and refinement of present methods of anesthesia as well 
as laying a sound foundation for further anesthesiology 


- 4 - Serial No<, 

Part A (continued) 

Proposed Course of Project : 

1. The study will continue on good risk out- 
patients. Additional baseline and comparative data 
will be accumulated, studied and evaluated, 

2= The experimental barbiturate, Sodium-Methyl - 
hexital (Lilly) and possibly the new anesthetic, 
Fluothane, will be studied. 

Part B included: Ho 


Individual froject Xaport 
Calendar Year 1958 

Serial Ho. MBBR-55 v. 

1. Clinical Investigation 

3. Bethesda, Maryland 

Fart A 

Project Title: Investigations into the Etiology of Recurrent 
Aphthous Dicerations and Other Oral Non- 
malignant Soft Tissue Diseases. 

Principal Investigator: Dr. I. I. Ship 

Other Investigators: Dr. E. J. ^riscoll 

Man Tears (calendar year 1958): Patient Visits: 400 
Total: I 1/3 400 (Outpatients) 

Professional: 2/3 
Other: 2/3 

Project Description: 

Ob jectives : 

1. To study and isolat? the various internal and 
environmental etiological factors involved in the patho- 
genesis of recurrent aphthous ulcerations and other non- 
malignant, chronic, oral, soft-tissue diseases* 

2., To determine the experience and incidence of re- 
current aphthous ulcerations, as well as related systemic 
conditions, in a graduate school population^ 

3- To investigate and elucidate the gross and micro- 
scopic pathology of the lesions. 

4. Investigations of new agents for the control and 
prevention of taese conditions. 

5„ To clarify the multiple diagnostic criteria for 
this group of diseases. 

6. Investigations of the relationship of adreno- 
cortical activity to the formation of aphthous ulcera- 

- 2 - Serial Ho. 

Fart A (continued/ 

Methods Employed : 

1. Comprehensive study of patients with these 
diseases comprising the following procedures: 

a. Viral cultures and serological anti- 
body testing. 

b. Allergic tests with utilization of 
synthetic diets. 

c. Hematological studies. 

d. Eosinophil counts and plasma cor- 
ticosteroid determinations. 

e. In vivo microscopy and micro-photog- 

f. Biopsy and pathological examination. 

g. Active clinical consultation. 

2a. Administration of questionnaire to approx- 
imately 2,000 graduate school students at 
the University of Pennsylvania to deter- 
mine the incidence of recurrent aphthous 
ulcerations and other oral soft tissue 

b. Active follow-up with examination of past 
history, review of the systems, and clinical 
examinations as well as monthly diaries of 
duration, location, and severity of indivi- 
dual lesions in each patient for the period 
of one year. 

3. Experimentation with new forms of topical 
anesthetics with low toxicity for amelioration of 

4. Double-blind clinical trial of agents showing 
promise for prophylactic administration to prevent re- 

Patient Material ; 

1; Patients suffering active disease referred from 
the local community (approximately 70). 

- 3 - Serial Ho, HXBR-55 (C). 

Part A (continued) 

2. Students attending the University of 
Pennsylvania Schools of Dentistry, Medicine, 
Veterinary medicine, and Graduate Nursing; 
Student Nurses at the Hospital of the University 
of Pennsylvania Hursing School, The Presbyterian 
Hospital Nursing School, and the Philadelphia 
General Hospital Nursing School; and students of 
the Dental Hygienist School of the University of 
Pennsylvania Dental School. Total of approxi- 
mately 2,000. 

Major Findings : 

1. Attempts to culture Herpes Simplex virus, or 
other viruses in tissue culture from 75 aphthous ulcera- 
tions in 53 patients, in various stages of development 
were unsuccessful. 

2. Acute exacerbations with the formation of 
multiple acute lesions appear to coincide with periods 
of emotional stress in many patients. No cause and 
effect relationship has been demonstrated. 

3. There has been a failure of regression of lesions 
when patients have been placed on synthetic, non-allergic 
diets with the elimination of all cosmetics, aromatic 
oils, and synthetic clothing. Dietary allergy did not 
have etiological significance in the progression of 
disease in the patients examined. 

4. Allergic etiology has been proven In one case. 
Formation of ulcerations has been successfully observed 
under select experimental conditions in this patient. 
Previously unrecognized mucosal changes have been ob- 
served during initiation, and photographic evidence is 

5. Hydrocortisone, taken systemlcally or applied 
locally to the affected tissues, has demonstrated con- 
siderable activity in the prevention of recurrences and 
the regression of aphthous ulceration. Standardization 
of vehicle, with Incorporation of various levels of 
active agent, is in progress; and the clinical testing 

in double-blind fashion is under way. Patient-maintained 
diaries of lesione,as well as repeated clinical examina- 
tion of patients, serve as criteria for beneficial re- 

- 4 - Serial Ho. MIBR-55 (C> 

Part A (continued) 

6. A solution containing 0.5£ Dyclonine Hydro- 
chloride and 0.5Z Diphenyihydramlne applied topically 
to acute aphthous ulcerations gives significant 
temporary relief from pain, appears to decrease the 
severity and duration of individual lesions, but has 
no effect on recurrences. 

7. In types other than aphthous ulcerations, no 
significant etiological patterns have yet been detected. 

Significance to Dental gesearch : 

Investigations into the many unknown factors in these 
diseases could lead to the eventual discovery of 
etiology and, subsequently, rational cure. 

Proposed Course of Project : 

To continue the experiment in progress. Examination 
of metabolic and enzymatic relationships within tissues 
prone to disease in affected patients will be Initiated. 

Part B included: Ho 


Individual Project Report 

Calendar Year 1958 

Serial No. NIDR-56 (C) 

1. Clinical Investigations 

3o Bethesda, Md. 

Part A. 

Project Title: Stress in the Dental Patient 

Principal Investigator: Dr. I. I. Ship 

Other Investigators: None 

Cooperating Units: None 

Man Years (calendar year 1958) Patient Visits: 200 
Total: 1 

Professional: 2/3 
Other: 1/3 

Project Description: 

Objectives : 

1. To determine and compare changes in adrenocortical '„l 
activity caused by various aspects of dental thereapy |" 
in healthy adult humans. L 

-•-••—- n« 

2. To investigate the changes in adrenocortical responses 

to dental stress induced by the use of local anesthesia ljj« 
and simple premedication. 

3. To establish a reliable index of physiological, response 
for future testing of dental technics and therapeutics. 

Methods Employed : 

1. Total eosinophile counts taken from peripheral capil- 
lary .blood by fijigeT-pudcture* prior to dentak -therapy are 
compared with counta taken four hours after treatment. 

2. . A1J. dental treatments arc standard and .divided into six 
categories: Interview Radiographic examinations, Oral 
examination, Prophylaxis, Operative dentistry, and Oral 

3. A 50 ago Pentobarbital Sodium capsule, and identical 
placebo capsule or no capsule, is dispensed by the pharmacy 
according to a regimen determined by a statistician and 
kept blind from patient and investigators. Capsules are 
taken one-half hour prior to dental appointments. 

Part A . Cont'd. - 2 - Serial No„ KIDR-56 (C) 

4 Two cc of 21 Lidocaine solution, 2 cc of sterile 
saline, or no solution, is injected at each appointment 
according to a schema established by the statistician,, 

5. Controls tor this study consist of: a) Ordinary 
dental appointments, randomly selected as regards 
sequence of treatments, at which time no procedures 
are attempted and patient rests for entire time in 
dental chair, b) Eosinophile counts, taken at four- 
hour intervals at unscheduled times, indicate the normal 
daily variation in counts. 

Patient Material : 

Twenty healthy adult employees of the SIH with extensive 
dental caries referred from the Employee Health Service, 

Major Findings: 

1. Preliminary results indicate that adrenocortical 
activity varies directly with the magnitude of stress 
involved in the dental procedure. 

2. The level of adrenocortical activity does not appear 
to be affected by the use of local anesthetics or barbitu- 
rate premedication in all categories except oral surgery. 
In this category, both local anesthetics and premedication 
make a profound change in the level of response, 

3. All dental procedures produced an adrenocortical re- 
sponse that was significantly greater than control dental 

4. Variations of procedures within each category produced 
no significant changes in adrenocortical responses. 

Significance to Dental Research : 

A physiological tool for the evaluation of technics and 
therapeutics in the dental field would be a valuable 
contribution to the profession. 

Proposed Course of Project J 

1, The conclusion of the experiment in progress. 

2. Further evaluation of the effects of general anes- 
thesia, extensive oral surgery, and prolonged periodontal 
therapy on adrenocortical activity. 

Partjfc. Cont'd . > 3 - Serial Ho, HIDR-56 (C) 

3. Investigation into the responses of individuals 
with compromised systemic resistance to stress and 
comparison with normal adults, 

4. Concomitant determinations of physiological and 
procedural variables to determine the mechanisms of 
the magnitude of activity demonstrated. These will 
Include : 

a) Blood pressure measurement before, during 

(at one minute intervals), and after procedures 

b) Measurement of the time and speed of the dental 
engine, with observation of Che effects of 
alteration of these on endogenously secreted 

Part B included No «J) 


Serial Ho. HIDR-57 (C) 
1. Clinical Investigations 
3. Bethesda, M, 
Individual Project Report 
Calendar Tear 19S8 

Part Ao 

Project Title: Esterase Activity Associated with the Formation 
of deposits on Teeth. 

Principal Investigator: Dr. P. H. Baer 

Other Investigators: Dr. M. S. Bur a tone 

Cooperating Units: Hone 

Man Tears (calendar year 1958): Patient Visits: 150 
Total: 1 
Professional: 1/2 
Other: 1/2 

Project Description: 

Objectives : 

To characterize and elucidate the role which esterase 
might play in the formation of dental "calculus." 

Methods Employed : 

Celluloid, Mylar and Hylon strips were secured to the 
teeth of patients for periods of 1-30 days. The deposits 
formed on these strips were then studied by histocheaicai 
techniques fox enzyme activity. 

1. Esterase activity is present in both cocci and fusi- 
form bacteria, with the former showing the highest 

2. Polymorphonuclear leukocytes reveal consistently 
high esterase activity. Degenerating leukocytes, how- 
ever, are inactive. 

3. Hot water (90°C.), Benzakonium chloride, and eseriae 
produce distlcct inhibition, 

4. Fluoride, diisopropyl f luorophosphate , and benaethoaium 
chloride prodtce minimal to no inhibition. 

Part_A„ Cont'd. - 2 - Serial No. NIDR-57 (C) 

5. The presence of high esterase activity in areas 
associated with calcified deposit material might 
possibly be correlated with a saponification process. 

Significance to Dental Research ; 

Since calculus is an etiologic factor in the develop- 
ment of periodontal disease, it is important to know 
the mechanism by which calculus is formed. It is 
hoped that this study will give some insight into 
this phase of the problem. 

Proposed Course of Project ; 

Continued studies of calculus formation, particularly 
as related to dietary factors in rats and mice. 

Part B included: No 

' Serial Ho. WIBR-58 (C) 
1. Clinical Investigations 
3. Bethesda, Maryland 


Individual Project Report 
Calendar Year 1958 

Part A 

Project Title: Sone Genetic Aspects of the Periodontium 
and Periodontal Disease in Mice. 

Principal Investigator: Dr. P. H. Baer 

Other Investigators: Rone 

Cooperating Units: Biometrics Branch, Division of Research 

Services: Mr. J. E. Lleberman; Laboratory 
of Physical Biology, National Institute of 
Arthritis and Metabolic Diseases: Dr. L. 

Man Years (calendar year 1958): 
Total: 1 
Professional: 1/2 
Other: 1/2 

Project Description: 

Subpreject A: The Effect of Diet 

Objectives : 

To study the role of diet on various components of th<s 
Java and teeth in 3 strains of inbred mice. 

Methods Employed : 

Male, inbred mice of strains C57L, BBA-2JU, end STR/H t?ere 
divided into 4 groups and fed the following diets: 

Group 1 - Purina lab. chow — hard physical consistency 
Group 2 - Diet 10,046 (high fat) — soft, greasy consistency 
Group 3 - Diet 10,046A « soft powdery consistency 
Group 4 - 10.005A — nutritious diet of watery consistency 

The animals were sacrificed at 16 months of age and the 
following structures studied: (a) mandibular weight, 

- 2 - Serial Ho. HIDB.-58 <\C} 

Pt rt A (cor.fciausd) 

(b) maxillary width (from first molar to first molar), 

(c) wear of molar teeth, (d) trabecular pattern of the 
alveolar bone, (e) presence or absence of periodontal 

1. Under the conditions of this experiment strain 
STR/N is susceptible to periodontal disease, whilt*. 
strains DBA/2JH and C57L are relatively resistant 
to periodontal disease. 

2. In strain DBA/2JH the maxillae of the animals on the 
high fat and high fat plus filler diets are wf.der 
than those on stock diet. 

3. Zn all strains the mandibles of the animals on the 
high fat and high fat plus filler diets weigh more 
than those on stock diet. 

4. Neither the physical consistency of the diet, width 
of maxilla, nor weight of mandible are important 
etlologlc factors in periodontal disease in these 
strains of mice. 

5. The trabecular pattern of the alveolar bone is distinct 
for each strain. Regardless of the nature of the diet, 
these patterns remained unchanged. 

Subpro lect B: Periodontal Disease in Six Strains of Inbred lice. 

Objectives ; 

To determine whether additional strains of mice, sus- 
ceptible to periodontal disease, can be found. 

Methods Employed ; 

Inbred mice, both males and females, of strains SS*R/H S 
Bl/Hen, 3RSUHT/H, C57X./6JH, A/Ben,, and A/LB were studied. 
The mice were fed a stock diet, and sacrificed at 12-24 
months of age. 

- 3 - Serial 

Fart A { continued) 
Major Findings : 

1. All strains, except strain SWR/H have higher 
mandibular periodontal scores (severer periodontal 
disease) than strain A/LN. 

2. All strains Including SWR/H have higher ©axillary 
scores (more severe periodontal disease) than the 
standard mandibular scores of strain A/LM. 

3. The females of all strains have greater perio- 
dontal involvement than the reference strain 

4. Strains A/LH and A/Ben have severe hair impaction on 
the palatal surface of the maxillary molars and severe 
loss of alveolar bone in the maxillary arch as compar- 
ed to the mandibular. 

5. Strain C57L/6JN shows the most marked sex difference 
in regards to alveolar bone loss, the females having 
the greater loss. 

6. Strain BRSUHT/H has the greatest loss of alveolar 
bone for both arches, in both sexes, and in addition 
develops a true periodontal pocket. This strain;, of 
all 6, would appear to be most valuable for use in 
periodontal research. 

Significance to Dental Research ; 

This study should contribute to our knowledge of the 
Interaction of diet and genetics as they affect the jaws, 
periodontium, and periodontal disease. 

Proposed Course o f Project : 

To continue the study of genetic influences on periodontal 

Part B included: Yes 

- 4 

Serial Ko. HZBR-58 (C) 


Individual Project Report 

Calendar Year 1958 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Baer, Paul N. , and Lieberman, J.E. : Observations on Seme 
Genetic Characteristics of the Periodontium and Periodontal 
Disease in Three Strains of Inbred Mice. Oral Surg. , Oral 
Med., and Oral Path. (In press). 

Honors and Awards relating to this project: None 

811$ - 


Part L 

Project Title: Hlstop i Periodof ease 

In BAI.T5/'Carin Mice. 

;pal Investigator: D:. H. R. Stanley, Jr. 

In vestige to.* c : "done 

ag Units: Bone 

aleadar 7231 
1: 1 1/3 


Objectives ; 

1. To det amine apical nigr&ticn of periodontal 
tissues occurs with age if annuals are kept under 


2. To assess the constancy of specific distances within 
the anatcaic ccatpGasntE' of thf; periodontal tissues 
in step-serial oectionB and how these distances 

fluctuate in relation to local pathologic factors. 

3. To determine how oany sections per ailliBeter of 
tissue need to be studied in order t© observe all 
obvious structural variations that sight occ 

Methods BBglo^ed: 

Step-seria:. rectioniae of the jaws of 3J&3 mice, ranging 
in age frosi 1 to 22 iscathe, is beiag carried out. 

cancel? are s^sft&ursd at a magnific 
X100. A .-.nroaeter la aaployed. Differences ia 
these distances changes in the aaatasBic coa-pon 
as successive sections are studied and these differences 
will be statistically analyzed in relation to local 
pathologic facte present*.. 

' 2 " Serial No. HIDR-59 (C) 

Part A (continued) 
Major Findings : 

1. Previous studies on huaan jaw sections indicate 
that essentially the same results are obtained by 
the study of every 20th section (6 microns thick) 
as every Sth section. However, the procedure re- 
quires that the number of sections examined be 
sufficiently large to guarantee a valid estimate 
of distribution; that is, provides a representa- 
tive sampling. In human material, distribution 
appears adequately described by 8 to 10 sections 
per millimeter of tissue. 

2. In mice, the distribution has not yet been 

Significance to Dental Research : 

With the emphasis now being placed on geriatrics, it 
is important to determine, under controlled conditions, 
just what part the ageing process can have in relation 
to the progress of periodontal disease. 

Proposed Course of Project : 

When the animal phase of study is completed, the investi- 
gation will return again to human material to evaluate 
the factor of age in relation to Che hlstopathology of 
periodontal lesions. 

Part B included: Yes 

- 3 - Serial fto. 


Individual Project Report 
Calendar Year 1958 

Part B : Honors, Awards, and Publications 

Publications other than abstracts frosa this project: 

Stanley, H.R. , Jr.: The Value of Step-Serial Sectioning 
in tha Morphologic Study of Human Periodontal Disease. 
The J. of Periodont. , 28:248-252, 1957. 

Honors and Awards relating to this project: Hone 


Individual Project Report 

Calendar Year 1958 

Serial No NIDR~60 (C) 

lo Clinical Investigations 

3o Betheada, Maryland 

Part A 

Project Title: Histopathology of the Human Pulp 

Principal Investigator: Dr H e R c Stanley, Jr 

Other Investigators: None 

Cooperating Units: Dental Department, Clinical Center 

Man Years (calendar year 1958) t 
Total: 2/3 
Professional: 2/3 
Other: None 

Project Description: 

Objectives : 

lo To determine the healing capacity of normal and diseased 
pulps following drilling procedures per se and the additional 
complications of filling materialso 

2 C To further evaluate the critioalness of the remaining 
dentin thickness beneath cavity preparations in order to 
compare pulp studies of other investigators 

3o To determine the mitotic frequency of odontoblasts and 
all other less differentiated cells of the human pulp 

k D To determine the range of pathologic findings to be 
expected in the pulp resulting from various degrees of 
carious involvement,, 

5c To determine the possible contributing effects of local 
infiltrating anesthesia versus block anesthesia on the total 
pathology of the pulp 

6 To determine what characteristics of pulp pathology might 
be the result of pressure changes per se when frictional 
heat is not presents 

- 2 - Serial No NIDIU60 {0} 
Part A (continued) 

7« To determine the amount of pathology that might 
be produced by super-cooling of teeth by air=water 
spray techniqueso 

8 a To determine the significance and pathogenesis of 
eosinophilic staining of the pulp» 

9o To determine whether or not certain morphologic 
alterations of the pulp are truly pathologic or merely 
the result of inadequate fixation,, 

10 o To gain information concerning the origin and 
formation of Korff*s fibers* 

Methods Employed * 

The principal method employed is the formalin fixation of 

extracted human teeth, followed by decalcification in 

formic acid, paraffin embedding, serial sectioning,, and 

staining with hematoxylin and eosin Masson'e trichromat. 

Wilder* s reticulum stain, Feulgen's reaction, periodic 

acidUSchiff , toluidine blue, and other special stains 

will be utilizedb For phase microscopy, freshly extracted 

intact teeth are fractured with surgical chisels and 

mallet and the pulp tissue scooped out with spoon ex~ 

cava tor So This procedure avoids formalin fixation and 

decalcification., The tissue is then fresh frozen, 

sectioned,, and examined unstained under the phase 

microscope., This technique is particularly valuable is>r ""| 

studying the fibrous structure of the predentin In the 

study of autolysis, the freshly extracted teeth are allowed 

to dry for specific periods of time,, After clipping the 

roots the teeth are placed in formalin., 

Major Findings t 

1, We have shown that the higher speed grinding techniques 9 
with adequate coolants, are not only biologically safe for 
the human pulp but actually are less damaging to the pulp 
than low speed techniques regardless of the type of accompany- 
ing scolantso 



- 3 - Serial No* N3DfU6o vC) 
Part A (continued) 

2« Odontoblasts have been considered incapable of 
mitotic division. Pre-odontoblastic cells in the cell 
rich layer are, however, believed capable of division 
into cells which mature into odontoblasts as neededo 
We have now accumulated a group of specimens revealing 
mitotic divisions approximating predentin Consequently, 
this subject of pulp regeneration must be re~evaluatecL 

3o Some carious teeth present little or no pulp pathology ( 
whereas others present severe changes,, A survey of ex- 
tracted carious teeth is needed to determine whether 
depth of carious penetration is the deciding factor,, 

4 Low speed grinding techniques that require 1^»24 os* 
of pressure or force seem to precipitate considerable 
pulpal response despite the presence of effective 
coolants that neutralize frictional heato High speed 
techniques that require very little pressure (1-3 o« ) ds 
not seem to produce as much pathology even without a 
coolant o 

5o Clinicians have reported that patients complain of $* 

pain from the sensation of coldness that results from the Z 

air-water sprays accompanying the use of the air turbines "" 

Conventional speeds only utilize about 25 ec/ndno as '*" 

compared to the turbines using close to 100 or more cc/mino ,„, 

6 Apparently certa:in characteristics of the pulp are \Z\ 

not due to pathology but autolysis, as, for example, the '» 

well-known characteristic of reticular atrophy Accepted J"| 

methods of fixation are evidently not sufficiently efficient 
to permit preservation of the pulp tissues before autolysis 
begins,, Thus, a combined picture of autolysis and fixation 
will often be presented for examinatlon 

Significance to Program of the Institute: 

lo Many times one finds in the literature statements contra= 
indicating or advocating the use of certain techniques or 
filling materials© Contradictory results from the study of 
similar procedures, but from different sources, are common,, 
We feel that much of this confusion results from the operator 
being unaware of pulpal response to operative trauma per se 
and of how the intensity of the response is related to the 


~ 4 = Serial No NIDR~60 (C) 

Part A (continued) 

thickness of the remaining dentin beneath the cavity 
preparation* It is anticipated that the described study 
will help establish some standardisation in pulp studies 
and decrease the confusion in future studies 

2 The current study should also contribute significantly 
to the better recognition of artifactual characteristics 
and thereby lessen existing confusion in interpretation of 
findings from pulpal studies 

3a Various investigators have suggested that dental caries 
does not necessarily provoke severe alteration of the pulp 
Therefore, to leave behind some decay in the cavity prepare^ 
tion is preferred tc exposing the pulp A survey of 
carious teeth is needed to determine the veracity of such 
a suggest ion 

Proposed Course of Project: 

lo Since we have shown that high speed techniques are 
biologically safe for the human pulp, emphasis will now 
be placed on the added effects of filling materials,, In 
view of the possible effects of pressure, per se, the use 
of hand amalgam pluggers versus pneumatic instruments 
will be evaluated. 

2 We have accumulated over 500 human teeth, all of which 
were prepared under local anesthesia All the maxillary 
teeth were prepared under local infiltration and all the 
mandibular teeth by block anesthesia By merely comparing 
the amount of pathology in the pulps from both jaws, this 
problem should be re solved* 

3o We will attempt to reproduce experimentally the eosino- 
philic staining characteristic recently named the "rebound 
phenomenon,, • Special staining procedures will also be 
carried out to help determine its source and significance,, 

*♦■«, Odontoblasts are not considered capable of producing 
Korff*8 fiberso However, in view of the fact that the fibers 
are found between the cell membranes of odontoblasts it is 
difficult to conceive of their being formed in the deeper 
tissues with eventual contouring into their characteristic 
spirals as the receding odontoblasts approach them*, With 
special stains on formalin fixed pulps, and the phase 
microscope on fresh pulps, the origin of Korffs fibers will 
be atudiedo 

Part B included Yes 

- 5 - Serial Ko NHJR-60 (C) 

Individual Project Report 
Calendar Tear 1958 

Part B s Honors, Awards, and Publications 

Publications other than abstracts from this projects 

Swerdlow, H,, and Stanley, H s9 Jr Reaction of the human 
dental pulp to cavity preparation, I, Effect of water 
spray at 20,000 rpm, J,A D A, j>6: 317=329 Mar, 1958o 

Stanley, H«, Jr , and Swerdlow, H c Aspiration of cells 
into dentinal tubules, J Q Oral Surge, Oral Hed 09 and 
Oral Path,, 11:1007-1017 Sept, 1958. 

Swerdlow, H e . and Stanley, H Reaction of the human dental 
pulp to cavity preparation, II, At 150,000 rpm with an 
air-water spray, J. Prosthetic Dentistry (in press) 
Jan, 1959o 

Stanley, H„, Jr, , and Swerdlow, H, Reaction of human pulp 
to cavity preparation (a comparative study based upon 
histopathological findings produced by eight different 
operative grinding techniques) (In manuscript form for 
the J0A0D0A0) 

Honors and Awards Relating to this Project: 

Serial Bo. 

1. Clinical Gentei? 

2. Dental Separtssat 

3. Betheada, Md. 


Individual Project Report 
Calendar Year 1958 

Part A 

Project Title: Pulp Response to High Speed Cutting and Grinding 
of Teeth. 

Principal Investigator: Dr. B. !5wsrdlow 

Other Investigator a: Hone 

Cooperating Units: Pathology service provided by Dr. H.R. Stanley s 
Jr. , HIDR 

Han Years (calendar year 1959) : 
Total 4/5 

Professional : 4/5 
Other Bone 

Project Description: 

Ob jectivee : 

To evaluate the effects on the dental pulp of standard 
operative procedures perforated at high r.p.ra. with and without 
a coolant and with various types of rotary instruments. 

Patient Material : 

Patients of either sex and any age over six years Here selected 
for study. Additional requiresaents were that they had vital, 
non-infected, caries-free teeth which were to fee extracted for 
reasons such as orthodontic therapy or a hopeless periodontal 

Methods Sep loved : 

Over 500 huaan teeth were utilised to cosapare pulpel lesions 
resulting free cavity preparations with iastrusente producing 
speeds of 6,000 and 20,000 rpa, with a conventional ball- 
bearing piece; 50,000 with the water turbine j 150, 0QC 
r.p.a. with the belt-drive; and 200,000 r.p.n. with the air- 
turbine, esQ>loying the turbine principle. The intensity and 

** 2 - Serial 

Part A (continued) 

size of the lesions were also evaluated on the basis of an 
accoaananying coolant, such as air -water spary, air spray, 
water, or the absence of any coolant. Inverted cone dla»sad 
stones (approximate size #37) were used for all speeds froa 
6,000 to 50,000 r.p.m., and a #35 carbide bar for all speeds 
in excess of 100,000 r.p.m. 

Typical Class V cavity preparations were cut in the gingival 
regions of the facial surfaces of all the teeth. All pre- 
parations were air-dried and filled with sine-oxide and 
eugenol. The teeth were res&oved at intervals of one hour : 
132 days. Multiple step-eerial sections of each speciaen 
were stained with hematoxylin and eosln. Histopathologic 
comparisons were made between speed categories by recording 
the. incidence of: burn-like lesions, (lesions not confined 
to cut dentinal tubules) , irregular secondary dentin forma- 
tion, cellular displacement, cellular exudate, intrapulpal 
abscess formation, delayed healing, etc. 

Major Finding s : 

Speeds at 50,000 r.p.m. and over, both of the belt driven 
or turbine principle cutting with the #36 diasaond stone and 
the #35 carbide bur, were found to be less traumatic to the 
human pulp than the 6,000 and 20,000 r.p.a. techniques using 
the convantional ball-bear in3 handpiece and the #37 diamond 
stone and the #35 carbide bur. The value of coolants becomes 
more significant at the higher speeds. In the absence of 
adequate coolants, intermittent grinding is of no appreciable 
benefit. The combination of high speed, controlled tempera- 
ture, and light load is conducive to minimal pulpal pathology. 
Excessive temperature rise was apparently controlled by air- 
water sprays and water at all speede except at 6,000 r.p.m., 
when used in combination with a #37 diamond stone and a load 
of 16-24 oz. 

Significance to Dental Research; 

Clinical Information concerning pulpal reactions to higher 
rotary speeds is greatly needed in order to determine the 
limits of safety and most efficient methods for cavity pre- 
paration. When the boundaries of response have been de- 
lineated., the practicing dentist may move with more assurance 
into an improved era of dental treatment. 

- 3 - KIM°6i fC) 

Pars & (continued) 

rropoaed Course of Project: 

To expand clinical Investigations of palpal response to 
high speeds of rotary instruments and to various other 
standard operative procedures. After establishing an 
accurate base line for each operative technique, attention 
will be focused on the superimposed reaction of the cavity 
liner and/or filling materials. The combination of the 
cavity preparation and the restoration which will produce 
the minimal reaction in the dental pulp would be toast ideal. 

Part B included: Yes 

- 4 - Serial Ho. HXD&-61 


Individual Project Report 
Calendar Year 1958 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Swerdlow, H. , and Stanley, H. , Jr.: Reaction of the huaan 
dental pulp to cavity preparation 2. Effect of water spray 
at 20,000 r.p.m. J.A.D.A. 56:317-329, March, 1958. 

Stanley, E. , Jr. and Swerdlow, H. : Aspiration of cells Into 
dentinal tubules. J. Oral Surg., Oral Med., and Oral Path., 
307-1017, September 1958. 

Swerdlow, H. , and Stanley, 3., Jr.: Reaction of the human 
dental pulp to cavity preparation. II At 150,000 r.p.a. with 
an air-water spray. J. Prosthetic Dentistry (in press) 
January 1959. 

Stanley, H. , Jr., and Swerdlow, H. : Seactlon of the husasa 
pulp to cavity preparation (a comparative study based upon 
histopachological findings produced by eight different operative 
grinding technique/). J.A.D.A. (in press) 

Eoaorn and Awards relating to this project: Btene 

Serial fflo, KipR-62 mi {p 
lo Clinical Center 
2o Dental Depsrtasafc 
3c Beehesda, m 
Individual Project Report 
Calendar Tear 1958 

Part A 

Project Titles Cephaloaetric Evaluation in Full Denture Prosthesis 

Principal Investigator : Dr Ro So Lloyd 

Other Investigators: Koue 

Cooperating Units : Orthodontia Activity, HIDR 

Man Tears ((calendar year 1958) ; Patient Visits; 200 

Total 1/4 

Professional: 1/4 

Other Hone 

Project Description; 


To utilise cephalonotrlc aeesureaeats to evaluate the pattern 
of bone resorption end the eventual positioning of prosthetic 
teeth in conparlsou to their natural predecessors* 

Patient Material : 

Individuals requiring Multiple tooth extractions. Referal3 
are by dental and nedical practitioners in the Washington 

Methods Ewp loved : 

Before the extraction of teeth, complete records are taken. 
These include Impressions, cephalcmetric x-rays and photo- 
graphs. At regular Intervals, these records are again ob- 
tained for longitudinal study. 

Ma lor Findings : 

It is thought that this project will be able to delineate 
the exact pattern of bone resorption and positioning of the 
succeeding prosthetic teeth. We also hope to utilize the 
findings in other ways such as to evaluate the relative 

Part A (continued) 

efficiency of standard procedures of obtaining the bite 
and the placement of teeth in the arch. The changes in 
the freeway space can also be observed. Seventy-one cases 
are now under study, of which soae are iraeediate denture 
insertion and soae have been allowed to heal a short tisae 
before Insertion of dentures. 

Significan ce to Dental Research : 

We hope to contribute soae knowledge which will be of 
assistance in the proper construction of prosthetic 
denture teeth replacements. 

Proposed Course of Project: 

It is anticipated that this project can have soae relative 
ianediate benefit. However s it is possible that the cases 
can be followed for aany years. 

Part B included No 

Serial Bo. 

1. Clinical" 

2. Dental Department 

3. Bethesda, 34d» 

Individual Project Report 
Calendar Year 1958 

Part A 

Project Title: Maxillofacial Prosthetics 

Principal Investigetor : Dr. R. S. Lloyd 

Other Investigators: 

Cooperating Units: Hone 

Man Years (calendar year 1958): Patient Visits: 120 

Total : 






Project Description: 

Object Ives : 

To evaluate methods of treatment by maxillofacial prosthetics; 
the design of appliances; and the response of patients to 
such treatment. 

Patient Material : 

1. Referals from private practitioners. 

2. Referals fron the Rational Cancer Institute. 

Methods Employed : 

1. Different types of appliances are being constructed for 

individuals who have had destruction of tissues of the head 
and neck with surgery, and patients afflicted with cleft 
palate. Improvement in esthetics, ability to ingest food 
(both soli-i and liquid), function of appliance, and improve- 
ment in speech are observed. Speech therapy is given where 
advisable. Cephalometric measurements also are utilised to 
help design appliances; g&. idteralne the amount of freeway 
space; and to properly place the bulb in cleft palate devices 

- 2 Serial Ho. 

Part A (continued) 

2. In connection with Cancer Institute patients, close 
cooperation is stressed with the Chief of Surgery, 
since the operative procedure is often influenced 
by plans of prosthetic replacement of tissues that 
are destroyed in the proper treatment of cancer. 
There is also close cooperation with the Radio- 
therapy Department of the National Cancer Institute 
in constructing and formulating new types of radium 
applicators and protective devices. 

Major Findings ; 

The well being of cancer patients receiving therapy sad 
patients with cleft palate is greatly influenced by a 
properly made prosthetic device. Obviously, the design 
of such a device is the basis of success. During the past 
year a total of 51 special prosthetic devices were con- 

Significance to Dental Research : 

This is an area where very little work has been done* 
and any contribution will be of great assistance in the 
treatment of these unfortunate patients. 

Proposed Course of Project: 

To further Investigate methods of construction and design 
of maxillofacial prosthetic appliances, and patient re- 
sponse to such treatment. 

Part B included: Ho 

Serial So JEBLJ&JSQ, 

1. Clinical Center 

2. Dental Department 

3. Bethesda, Md. 


Individual Project Report 

Calendar Year 1958 

Part A 

Project Title: Malocclusion as Related to Traumatic Occlusion. 

Principal Investigator: Dr. H. Swerdlow 

Other Investigators: Dr. A. W. Mann (NIDR Consultant) 

Cooperating Units: Photographic Unit, SRI, DRS, BIH 

Han Tears (calendar year 1958): Patient Visits: 113 




1/5 Patients : 31 



Project Description: 

To devise an accurate "yardstick" or method of evaluating 
the result of periodontal treatment. 

Patient Material : 

During 1958, 31 adult patients were studied for malocclusion 
and temporomandibular joint syndrome. Criteria for inclusion 
in study were that they have twenty-eight teeth in occlusion, 
some degree of demonstrable malocclusion, and some evidence 
of periodontal disease. 

Methods Employed : 

1. The existing occlusion in patients under study is recorded 
on an anatomical articulator and all pretreatsnent records 
(i.e- mandibular movements, photographs, roentgenography, 
etc.) recorded for later reference. After functional 
analysis, the occlusion is corrected by selective grinding 
and the clinical course of the disease charted. 


part A (cc 

2, During the past two years it has been desaonstr: 

that the information obtained from laminagraphs and 
cephalocetric x-rays is of little value in this study. 
Since the degree of mucosal inflammation offers a 
good index of inflammatory periodontal disease, we 
believe that th3 problem could perhaps be evaluated 
by color ime trie methods. However, it was found t 
comparison of tissue color to a standard blank (e,g ., 
paint industry aheet guide) was inadequate. The use 
of standard eclor transparencies was also found to 
be inadequate due to inconsistencies of the emuls: 
In order to eliminate the variable of human 
judgment, work was begun <in conjunction with the 
photographic department) on a clinical camera . oe- 
signed to accurately measure changes in gingi 
color and contour. The camera, which is currently 

.g modified, holds much promise for successful 
application in che early future. 

3 An additional method for evaluating clinical progress 
' related to balancing of occlusion is to record the 
sounds of occluding teeth picked up from the mastoid 
area, forehead, and bridge of the nose sy a micro- 
phone ar *d on tape. Several variations of 
microphc been tested and at presen t t be 
techniqv :g standradized to record only th_ 
sounds essential to this study. 

Major Findings: 

The camera developed in this study is semi-automatic, 70- 
Tfte camera oevc * . . f<i Cer wheel containing 

millimeter, and equipped with a t-^f* ^ k 

.n-aT-o rhat ranse in censity ij.oa uga- -** ". 
seven reo filters cnat raujje *^ j«,j«,-.-- 4m.»u eiaht 

uh»n fltarted the camera automatically indexes itse~ eigne 
When atarted, the c ^filtered photograph of 

times: tbe ^"^^^he^emaining seven e.x **« 

tn e e filter whtel. Zach ex^auTe is lade through a different 
filter Siintaneously^ough a separate optical system, 
nicer. ai «" n ~ 1n it>A acoss the upper margin of -=acti 

a "gray scale" is P Linted "*®!!^ sca ie" provides insurance 
of the seven aegatives. Tne gray scale ^ . 

r^ative through ~'^^^*^ of the 

densitometer (by comparing the two e-p*-* 

" 3 • »eiiisjL Boa 

Part A (continued) 

"gray scale" versus the photograph) can give the 
negative's original reading. The purpose of the filter 
wheel is to find the color density of the tissue being 
photographed. When the filter density matches the color 
density of the tissue, the tissue "blends" with the filter 
color and cannot be separately distinguished on the 
negative. By means of the densitometer, it is believed 
possible to accurately assign a color index of the original 
tissue and of the tissue's subsequent changes. The c&aera 
may also have application in eye diseases, dermatology, and 

Significanc e to Dental Research ; 

1 It is generally accepted that malocclusions and mal- 
functions of the teeth are Important local causative 
factors in periodontal disease. It is usually noted, also, 
that in periodontal disease, the occlusal pattern is un- 
balanced and not in harmony with the temporomandibular ar- 
ticulation. TV. 8 affects not only the tissues adjacent to 
the teeth but also the facial muscles, ligaments, and othes 
allied structures. Thus, improved methods of examination 
and recording of occlusal disturbances should contribute 
to our better understanding of periodontal disease and 
treatment procedures. 

2. The camera developed in this study may have important 

application tc many fields in addition to dentistry, i,e^ 
dermatology, pathology and ophthalmology. 

Proposed Course of Project : 

It is hoped that tools being developed will aid in the 
measurement of the changes that occur in the teeth and 
surrounding structures (accurate findings in this study 
will aid in the evaluation of balanced occlusions in perio- 
dontal disease). We will then be able to more accurately 
determine the exact influence of occlusal balance obtained 
by selective grinding in the prevention and treatment of 
periodontal disease. 

Part B included: Ho 

Serial Ne NIDR-- 

Clinical Investig?. 
3o Bethesda 9 Maryl&rsd 
Individual Project Report 
Calendar Yea? 1958 

Part A 

Project Title: Twelve-Mcnth Changes in Claas II 

with and without Occipital Headgear Thejrapy 

Principal Investigator: Dr D D Ro Poulton 

Other Investigators: None 

Cooperating Units: Philadelphia Center for Research in Child 
Growth, Wo Mo Krogoan, Director 

Man Years Ualendar year 1958 - from January to August) 
Total: 1 1/2 
Professional: 3/k 
Other: 3/* 

Project Description: 

Objectives : 

To determine changes in fasial growth and tooth position 
occurring during orthodontic therapy 


Application of intend. ttant force from an occipital headeap 
to an appliance en the maxillary archo Evaluation ©f shanges 
and comparisons with untreated cases will be made via 
roentgenographie eephalometries,, facial phctogmpha and 
articulated models of the dentition,. 

Patient Material : 

Thirty-two patients ages 9 to 11 are under headoap therapy o 

Major Findin gs: 

lo Molar relationships were corrected and inoisor positions 
improved in nearly alii the treated sasess, largely by a 
substantial distal mevement ©f the upper dentlti©n Moat 
cases showed a marked distal position of the maxillary 
and unerupted tooth budSo 

Serial K© Nn>R~6ji 

Part A (contii' 

2© Comparison with the control group revealed some 
inhibition of forward maxillary growth and a slightly 
greater increase in lower face height The dentition 
in the control group moved forward more than the upper 

3o Regular daily wearing of the headgear 9 and the amount 
of mandibular growth , were shown to be the most important 
factors in obtaining a prompt treatment resp©nse 

ifo Analysis of the mechanics of the headgear force used 
here suggested vhat changing the attachment and direction 
of pu'.l in relation to the maxillary roots might improve 
the a<ticn of the appliance,, 

5o From comparisons with tooth movement reported in other 
studies 9 the occipital headgear appears to be the most 
effective appliance for moving upper teeth distally 

Significance to Dental Research : 

A study of response differences should produce knowledge 
that may enable the orthodontist to utilise the available 
applirances with more efficiency and to the greater benefit 
of the child with malocclusion,, A detailed knowledge of 
the type of changes possible with a given appliance will 
allow a more fruitful direotion of efforto 

Proposed Course of Project t 

Final reports were in progress at the time of Dr P©alt«n s s 
resignation from the service, September,, 1958« when he left 
to take a position in California,, Dr Poulton has planned 
to follow up the study for the second year during two 
return visits to NIH when all patients will be re-examined 
by himo 

Part B included: Yes 


Individual Project Report 
Calendar Year 1958 

Part B t Honcrs B Awards, and Publications 

Publications other than abstracts frcm this project: 

Foultcn g Do Ro» and Pruianaky, S Report of a case with 
supernumerary teeth and treatment involving extraction of 
three upper iniisora Jo of Dentistry for Children,, 
212-2J4 ?rd Quarter 9 1958, 

Pcu He Twelve-m nth changes in class II male><s©lu=> 

hsadgear therapyo This 
paper has been passed by the NIDR Editorial Board and 
has been submitted for publication 

Honors and Awards Relating to this Project: 

/sis of NIH Prograa Activities 
January-December 1958 

Division of Biologies Standards 

The Division of Biologies Standards is responsible for adminis- 
tering the provisions of the Public Health Service Act aa they pertain 
to the control of biological products and for the development of 
Regulations within the provisions of this Act. The legal description 
of the products is complex, but in effect it includes vaccines, serums, 
toxins, antitoxins, and related products, including human blood and its 
derivatives, which are offered for sale, barter and exchange in inter- 
state cornnerce or for export. 

Development of realistic standards for these products and the 
exercise of proper control over their safety, purity, and potency can 
be effected only if backed by an active research program of sufficient 
flexibility to provide information as it is needed for the formulation 
of such standard/ , One of the main objectives of the Division has been 
the development of a research program based on the premise that the 
control of biological products can only be successful when supported by 
such a program of research. 

The functions of the Division arc. implemented through the 
activities of its four operating laboratories; the Laboratory of Viral 
Products, the Laboratory of Blood and Blood Products, the Laboratory of 
Bacterial Products, and the Laboratory cf Control Activities. Detailed 
descriptions of the various projects being conducted by these laboratories 
and by the Office cf the Director are set forth in the following 41 
project reports. 

It has been considered desirable this year to break these projects 
down into greater detail than has been cone on previous occasions in 
order to give a better idea of the scope of the programs being followed. 

The substances of these individual reports may be summarized 
as follows: 

Laboratory of Viral Products ; 

The main preoccupation of this laboratory has continued to be 
poliomyelitis vaccine. The cumber of lets of vaccine submitted for 
test purposes and release as demonstrated by a number of additional 

factors, have entered the field to keep the more strictly routine 
aspects of the testing of polio vaccine a continuing activity of the 
laboratory. These have been tbe a n nounced entry of additional manufacturers 
into the field and ':he work entailed in the licensing c pie antigens 
containing polio vaccine. Two manufacturers; namely, Parke, Davis and 
Merck Sharp & Dohme, were on the point of being licensed for 3uch 
products at the years' end. 

During the year there has been an encouraging increase in the 
potency level of poliomyelitis vaccine. This has been sustained for 
most of the period in the case of Types II and III and there is an 
indication that Type I is also on the increase. Some studies carried 
out by members of the staff of the laboratory on the antibody rises 
produced by the injection of large amounts of poliomyelitis vaccine 
suggest thatvten vaccines of high potency become available, it will 
be possible to achieve good antibody levels even after a single dose. 

During the latter part of 1958 the DBS has turned its attention 
to the problems concerned with live polio vaccine and this will be an 
increasing activity during 1959. An advisory comnittee on this subject 
was appointed by the Surgeon General during the year and hold one meeting 

In anticipation of the development of practical immunizing agents 
for measles, the Laboratory of Viral Products has expanded its activities 
in the field of measles virus during the year and at the close of the 
year was in process of mounting one additional project (not listed) 
on the study of the development of antibodies in tv^nap subjects to 
measles. Dr. Rivadeneira, a Visiting Scientist, is engaged in this 

During the year additional studies have been conducted with 
the chick test for polio vaccine potency and with the antibody neutrali- 
zation test developed by Dr. Melnick, with a view to introducing these 
into manufacturing and testing practices. 

During the course of the year it has been possible to enter 
into collaborative studies with members of the staff of Walter Reed 
Army Institute for Research in the evaluation of the potency of dried 
smallpox vaccine. This work is still in process but the results ob- 
tained thus far seem to indicate that it will be possible to achieve 
a correlation between the testing methods which have been proposed; 
namely inoculation of chick embryos, tissue culture titrations, and 
the routine rabbit skin scarification methods. Following the outbreak 
of smallpox in Bast Pakistan, it was possible to obtain some of the 
dried vaccine used at that time and study these for potency on a 
comparative basis. 

- 3 - 

A matter of continuing interest which has been widely 
confirmed in other laboratories is the work reported by Dr. Eddy 
of the laboratory and Dr. Stewart of the National Cancer Institute 
on the polyoma virus. 

Laboratory of Blood and Blood Products : 

Control Program. The goals of the control program of the 
Laboratory of Blood and Blood Products are to raise the standards 
of achievement of approximately 400 operating units, each of which 
is concerned with the safety of blood and blood products. The 
rapid expansion of the number of producing laboratories, a rapidly 
developing awareness of the need for improvements in techniques 
and standards, and increasing variety of new products has resulted 
in a situation demanding technical and administrative coordination 
among blood banks . The divergent interests of the operating organi- 
zations have caused a pressing need for the NIH to assume leadership 
in this area. 

The goals of the program have not changed, but there is an 
increasing number of requests to broaden the program to include 
technician training programs, reference preparations, and increased 
Inspections and licensing. 

This program has defined problems of local organizations 
and provided a pattern for local groups desiring to Improve blood 
bank procedures in their areas. Rapid progress is being made in the 
development of technician evaluation and training programs and 
inspection programs that operate voluntarily generally on a state-wide 
basis. The staff of this Laboratory assists with the planning and 
operation of such local programs by consultation and lectures on 
request within the limitations of available staff. 

A major problem continues to be a lack of qualified staff 
but this is being met by internal training programs and closer 
coordination of the staff concerned with our control activities. 

During the last three months of 1958 a team of four professional 
workers have been relieved of administrative and control functions to 
develop a more active program of fundamental and applied research. This 
provides for the first time for more rapid application of research to 
the practical problems of control and production, but has not been 
operating long enough to permit an evaluation of the potential of the 
group or to develop a well -coordinated research program. 

Research Activities. The laboratory research program carried 
on by the Laboratory of Blood and Blood Products has as its goal 

- 4 - 

the investigation of properties of blood products to develop more 
effective control procedures applicable to the various blood products. 
The research in this field is being started. 

The investigation of the immunological and physico-chemical 
properties of blood proteins, including coagulation systems, is now 
possible by the addition of professional staff having interest and 
training qualifying them for such research. This new program of 
laboratory research is starting in an orderly way and should provide 
the research information necessary to support our control functions. 
The first phase of this new blood program provided the physic -"Chemical 
data needed to compare new blood products proposed to the National 
Research Council for consideration as shock units to be stockpiled 
for defense purposes. This particular project also served as an 
initial project for the research group which now seems ready to 
undertake more fundamental studies. 

This activity is too new to have developed program problems 
at this time other than to make the perennial space problem more 
acute. The advantage of additional professional resources has more 
than offset the disadvantage of crowding. 

Laboratory of Bacterial Products : 

The Laboratory of Bacterial Products is in the process of 
developing a research program bearing upon the continued or increased 
safety, purity and potency of bacteriological biologic products and 
on the development of methods for the control of new products. Products 
listed for early consideration are BCG vaccine, staphylococcal products, 
diphtheria and tetanus toxoids, typhoid vaccine, and allergens. 
Inability to obtain qualified staff members and shortage of space 
have delayed initiation of the total program. The main portion of 
the study on the components of diphtheria toxin was terminated 
due to retirement of the principal investigator. Studies relating 
to the etiology of cancer, to pertussis products, and to the selection 
of a standard and a better assay procedure for the Schick test toxin 
(diphtheria) were continued. A study of cholera vaccine was recently 
initiated. Coordination of the program with international standardization 
developments is carried out insofar as practical. 

In the study of the components of diphtheria toxin an extra 
component was demonstrated in preparations of gravis strains by means 
of electrophoresis and ultracentrifugation. It was composed of large 
size molecules; it induced more marked edematous reactions in guinea 
pigs than other fractions and was lacking in similar preparations 
of "PW8" strains. It is suggestive that this component may be 
related to the marked edematous lesions characteristic of gravis 
infections in man. 

- 5 - 

In further studies on fibroma -myxoma virus trans format ion 
it was found thac the transforming agent was destroyed by desoxyribonuclease 
and by photodynamic action in the presence of toluidine blue. An attempt 
is being made to study the developmental cycles of fibroma and myxoma 
viruses in tissue culture by means of electron microscopy. 

Studies relating to the control of pertussis vaccine and anti- 
pertussis serum have been continued. More than 100 lots were tested 
for both potency and freedom-from- toxicity. It has been determined 
that the time interval used between vaccination and challenge in the 
potency testing of aluminum adjuvant -containing vaccines, which had 
been questioned, was satisfactory; the same results were obtained with 
intervals of 14, 21 or 28 days. Collaboration in the assay of pertussis 
vaccines used in British field trials has provided information that 
gives assurance that the United States potency requirement is adequate. 
An antipertussis serum which will be designated as the United States 
Standard has been dried and assayed for potency. 

A method has been developed for the preparation of formalinised 
red blood cells which retain their original adsorptive properties. 
The cells are stable and can be maintained for long periods at 4°C or 
when freeze-dricl. They appear to be satisfactory for use in all 
types of hemagglutination teats and therefore may prove to be very 
useful in inmuno os,ical studies. 

In the continued study of the effect of combinations of 
antigens and of aluminum adjuvants on the Immunological response to 
the individual component antigen, a theoretical model of the effect 
of the interaction between antigens and between antigens and non-ant igenic 
substances in multiple antigen preparations on enhancing cr inhibiting 
the immunological response to an individual antigen has been developed. 
The new method for the preparation of formalinized red cells and 
modifications in the Ouchterlony and Oudin gel diffusion technics 
are contributing to the evaluation of the interacting factors. 

A project has been initiated to develop a quantitative 
protection test for the evaluation of the potency of cholera vaccine. 
It will be carried out in reference to and coordinated with the 
proposed World Health Organization international cooperative efforts 
towards the establishment of a standard of potency related to human 

L forms of bacteria have been isolated from 12 tissue 
culture lines. Neither L form3 nor PPLO (pleuropneumonia -like 
organisms) have been isolated from either primary monkey and rabbit 

kidney cells, tissue culture media, or medium supplements. Bacteria 
which developed jrrcm isolated L forms have been reverted to the L 
form by growing in the presence of tissue culture media containing 
penicillin and streptomycin. The conversion of bacterial contaminants 
in the presence of antibiotics to L forms which are not readily 
detected by ordinary sterility test procedures, present a real problem 
in the handling of tissue culture lines. 

Laboratory of Control Activities ! 

Tha activities of this laboratory are bast summarised in the 
attached project report No. 2 of tha Office of the Chief, Laboratory 
of Control Activities. It should be noted that during the first 
11 months of 1953, 3,183 lots of various products have been submitted 
for release by licensed manufacturers. Of these, 3,136 have been 
released by tha UH and 19 have been withdrawn by manufacturers 
and 28 rejected by the HI. 



hi' 1 " 

FiiS -NIB 

Individual Project Report 

Calendar Year 1958 

Serial No. DBS 

1. Office of the Director 

2 . Bethesda 

In the Division of Biologies Standards, program direction and 
administration is cantered in the Office of the Director, with 
centralisation of administrative procedures relating to personnel, 
travel, supply and accounting. Major objectives are determined and 
program responsibilities are assigned to the operating units within 
the Division. For each major area, staffing patterns are developed 
and are continually reevaluated in relation to program changes. 
The required financial support is determined for each area in rela- 
tion to the Division's total budget. A major activity of the Office 
of the Director is the maintenance of the proper balance between the 
control activities and the research programs, the latter being 
supported to the degree that such investigations are required in 
order to meet the control responsibilities of the Division. 

The Director working with members of his immediate staff, inclu- 
ding a legal ••sisL*iQl, conduct* a continuing review of existing 
regulations for the control of biological products, making revisions 
as necessary and al&o develops additional regulations as required 
for new products. 

Close and continuous liaison IS maintained with technical 
representatives of industry. Frequent meetings are held between 
members of the Division's professional and administrative staff 
and groups of manufacturers who are concerned with common prob- 
lems. In addition, 153 to 200 conferences are held each year 
with technical leoreeentatives of manufacturers vhc desire to 
discuss production and testing programs peculiar to their own 

The production of poliomyelitis vaccine continues to be 
closely monitored through the operation of the Technical Aid Pro- 
gram. A liaison scientist, under the direct supervision of the 
Director, is assigned to each of the establishments manufacturing 
this product. These individuals routinely visit the industrial 
plants and maintain close liaison with the technical and research 
staffs of the producers. 

The Office of the Director services the Technical Committee 
on Poliomyelitis Vaccine, the Public Health Service Committee on 
Live Attenuated Poliomyelitis Vaccine, as well as numerous Ad Hoc 
Committees which are appointed from time to time to consider 
matters relating to the control of biological products. 

The Director, or his designated representative, attends 
meetings of committee* and advisory groups of the World Health 
Organization,, as veil as those of the National Research Council, 
Armed Forces Epidemiological Board, etc. During 1958, Division 
scientists have taken an active part in the World Health Organi- 
sation's program for the development of international uniformity 
of biological products. 

Within the Office of the Director, the Information Office 
continues to be concerned with, (1) communication through such 
media as press, radio, television, publications and exhibits of 
information to the public on the Division's control and research 
activities, and (2) internal communication of such information 
through reports to the Director, M2E S and to the Surgeon general 
as well as to the Department and to the Congress. 

During 1953 „ considerable time was devoted to matters eoe- 
cemed with constructing, equipping, and financing the new DBS 


Control Activities 


Serial No. DBS - 2 thru 5 

Ettl—f d Obligation* for P.Y. 1959 

Total: 311,320 

Direct: 254,320 

Reimbursements: 57,000 

Serial No. DBS 

1. Control Activities 

2. Office of the Chief 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Office of Chief, Laboratory of Control Activities 

Principal Investigator: Dr. rf„ G„ Workman 

Other Investigators: None 

Cooperating Units: Three 

Man Years: Patient Days: None 

Total: 3 
Professional: 1 
Other: 2 

Project Description: 

Under the Director, Division of Biologies Standards, this 
project is responsible for the area of activity dealing directly 
with licensed establishments In relation to the licensing and 
control of biologic products. This activity is supported by 
sections on reference standards, on control tests, and on pyro~ 

The activities include: 

(a) Determination of eligibility of establishments and of 
individual products for license. This determination is 
made on the basis of a review of the integrity of manage 
ment, the physical facilities for manufacture and testing 
of products , the scientific and professional qualifica- 
tions of personnels and the evidence of continued safety. 
purity, and potency of licensed products and of any 
products for which an application for license is being 

(b) Supervision of annual and special Inspections of licensed manu- 
facturers and of establishments for which an application for 
license has been made,, 

(c) Release of individual lots of biologic products for distribu- 
tion on the basis of review of manufacturer's protocols, manu- 
facturer's and NIH tests „ and on other data relating to the 
quality of the individual lot of the product. 

(d) The establishment and distribution of physical biological 

(e) Review of requirements and Regulations now in effect for such 
constructive, revision as is needed and development of require- 
ments and Regulations required for new products, 

1 Maintenance of close working relations between this Laboratory 
and the other Laboratories of the Division in order to insure 
continuous knowledge of all activities and data needed for 
licensing of establishments and of products and of the testing 
and release of individual lots oi products. 

Cg> Serving on various committees, such as the Council on Drugs 
of the American Medical Association., the General Committee 
of Revision of the United. States Pharmacopoeia , and collabo- 
rating with other official or non-official agencies operating 
in fields of interest similar to or identical with those of 
this Laboratory. 

(h) Cooperation with the Department of National Defense and other 
laboratories not licensed for interchange of information relat- 
ing to the control of biologic products. 

Significance to D ivision of Biologies Standards: This project is 
directly related to the legal and administrative functions of the Division. 

Proposed Course of Projects It is proposed to continue the control 
testing activities at approximately the present level. It is desirable to 
improve the quality of testing wherever this can be accomplished, and to 
develop improved methods in order that the control activity may become more 
effective without necessarily increasing the volume of work being done. 
This Laboratory remains inadequately staffed and it is proposed to fill key 
positions, particularly the Chief of the Reference Standards Section, and 
of the Pyrogens Testing Section, as soon as they can be filled. 

Part B included Yes / 7 So /X~7 


Individual Project Report 
Calendar Year 1958 

Serial No, DBS_;_3_ 

Part A. 

Control Activities 
Control Tests 

Project Title: 

Control Testing and Investigations Related 
to Control of Biological Products. 

Principal Investigator: Mr. Robert W. Kolb 

Other Investigators: 

Cooperating Units: 

Mr. Otis L. Green 

Dr. Michael E. Barile (Laboratory of 

Bacterial Products) 
Dr. Ernest C Cutchins (Laboratory 

of Viral Products) 

Lt. Col. Abram S. Benenson, MC S USA 

Director, Division of Immunology 
Walter Reed Army Institute of Research 
Walter Reed Medical Center 

Man Years 
Total: 8.25 
Other: 6.25 


Patient Days: None 

Project Description: 

The general purpose of this project is to assure com- 
pliance of licensed biological products with standards estab- 
lished in accordance with the biologies control provisions of 
the Public Health Service Act. Recommendations for release of 
individual lots of biological products are made on the basis 
of evaluation of manufacturer's and NIH tests. The evaluation 
includes control testing for sterility and safety of all 
licensed biological products and control testing for potency 
and purity of antitoxins; therapeutic immune serums, blood 
derivatives, bacterial vaccines, toxins and toxoids, viral and 
rickettsial vaccines, diagnostic substances for dermal tests, 

antivenins and venoms. Such control also involves (a) evalua- 
tion of manufacturer's protocols of tests on lots of biological 
products for compliance with minimum standards (b) preparation 
of official releases and (c) maintenance of a permanent record 
and filing system for releases, protocols of manufacturer's 
tests, and samples of all biological products submitted to the 

In addition to the above* this project involves: (a) 
research designed to improve existing in vivo and in vitr o 
methods of tasting and to develop new methods for which there 
is a need; (b) investigation in cooperation with intra -agency 
laboratories, commercial establishments, domestic and foreign 
governmental agencies In which studies and assays are carried 
out on numerous biological and pharmaceutical preparations, and 
(c) training activities designed to instruct and orient new 
government personnel and visitors from both domestic and foreign 
health laboratories in biologies control testing procedures as 
carried out at the National Institutes of Health,, 

Major Fi ndings and Accomplishments,, 

(a) Biological products produced by laboratories 
holding U, So Government license, both here 
and abroad have been tested for conformance 

to National Institutes of Health requirements, 
A total of 4,624 control tests were conducted 
within this laboratory to insure sterility, 
safety, potency and purity of products submitted 
for release. In addition, 1,444 other control 
tests were carried out on biological products 
received as inspection samples from licensed 
establishments during the year, 

A permanent record and filing system for releases, 
protocols of manufacturer's tests and samples of 
all biological products submitted to the Division 
of Biologies Standards was maintained. During the 
first 11 months of 1958, 3 S 183 lots of biological 
products have been submitted for release by 
licensed manufacturers „ Of these, 3,136 lots were 
released and 28 lots were rejected by NIH and 19 
lots were withdrawn from consideration for release 
by the manufacturers „ 

(b) Participated in a special Influenza Virus Vaccine 
Control Program organized this year for releasing 
sufficient vaccine for Armed Forces needs. Engaged 

in cooperative testing to insure the sterility 
and safety of the vaccines released and partici- 
pated in the correlation and evaluation of the 
manufacturer's and NIH test data for determining 
acceptance of the vaccines submitted. 

(c) Cooperative service testing was rendered to 
several intra -agency laboratories and to a number 
of domestic and foreign governmental agencies, in 
which a total of 896 control tests were carried 
out on numerous biological and pharmaceutical 
preparations. Official reports of results were 
made to the laboratories or agencies concerned. 

(d) Seven complaints from the Armed Forces and other 
users of biological producta were investigated; 
reports and recommendations for action were made 
to the Chief, Laboratory of Control Activities „ 

(e) Numerous new human biological preparations were 
submitted for review and consideration of U. So 
Government license during the current year, Care- 
ful screening and testing of these preparations 
served as a basis for their licensure or rejection. 
Complete reports and recommendations were made 
following review of 11 establishment license 
applications and 30 product license applications, 

(f ) Assisted in the control inspection program by 
conducting 7 annual, 8 prelicensing, and 2 special 
trouble spot establishment inspections during the 
year . 

(g) The cooperative study in conjunction with the 
Laboratory of Bacterial Products on the establish- 
ment and preparation of an NIH Standard Diphtheria 
Schick Toxin was continued during the year. 
Experiments were completed with two mature 
diphtheria toxins comparing various laboratory 
potency test procedures to determine the toxicity, 
avidity and stability of the toxins. The results 
of the experiments are being evaluated and 
statistically analyzed for selection of the best 
toxin as the reference material . It is antici- 
pated that the project will be completed within 
the next year. Further investigations are to 

- 4 - 

(1) Studies on the effectiveness of freeze 
drying and stability of the material in 
the dried state. 

(2) Comparison of the dried reference to the 
International Standard for Schick Toxin 
and with Commercial Schick Toxins. In 
these tests significant variations will 
be determined and final report will be 
made a 

(h) A collaborative inter-agency study with Walter 
Reed Army Institute of Research involving both 
laboratory and clinical experimental trials to 
determine the stability and effectiveness of 
dried smallpox vaccine was initiated. Since 
there is only limited information concerning 
this type of vaccine and the military is 
intensely interested in obtaining such a vaccine 
with a more extended dating period than current 
market materials, it was felt that such a joint 
project would be of mutual value. This laboratory 
will participate in conjunction with Dr. Cutchins 
of the Laboratory of Viral Products in performing 
rabbit scarification potency tests and tissue 
culture titrations (plaque and tube) on samples 
of vaccine stored under varying temperature 
conditions over a 92 week period. It is too 
premature to evaluate the significance of results 
obtained to date. It is anticipated, however, 
that these studies will (1) provide sufficient 
and interesting basic information to extend the 
dating period for use of this type of vaccine, 
(2) furnish a correlation between in vivo and 
in vitro test effectiveness as well as data 
concerning comparative response in human and 
animal testing, and (3) serve as a basis for 
modifications of existing DBS requirements for 
the product. 

(i) Completed determinations of diphtheria and 

tetanus antibody levels on 21 samples of patients 
serums in connection with a study being carried 
out by Dr. V. K. Volk under the sponsorship of 
the American Public Health Association Subcommittee 
on Multiple Toxoids „ A report of the results was 
made . 

- 5 - 

(j) The limited study initiated last year to 

evaluate the stability of Rabies Vaccine was 
continued during the year. Both commercial 
and NIH Reference Rabies Vaccine preparations, 
following storage at various temperatures for 
varying intervals of time, were compared for 
antigenicity in accordance with NIH requirements. 
Results of experiments to date have demonstrated 
that dried rabies vaccine of duck embryo origin 
is relatively stabile for at least 6 months 
foil cuing storage at 37°C whereas dried rabies 
vaccine of rabbit brain origin shows instability 
after one month storage at the. same temperature. 
No significant differences or loss of stability 
was noted for either type vaccine when stored at 
5°C during this period. Final experiments in 
this study are scheduled for completion within 
the next year. 

(k) A potency test procedure has been proposed for 
the assay of Paratyphoid A and Paratyphoid B 
antigens in Typhoid and Paratyphoid Vaccine. 
A study to evaluate this method was initiated 
during the year in collaboration with several 
commercial biological laboratories. This in- 
vestigation seemed appropriate inasmuch as there 
is no official test method for the potency assay 
of these antigens, consequently extended dating 
cannot be given Typhoid and Paratyphoid Vaccine 
in the same manner as other biological products, 

Preliminary tests were carried out in order to 
establish the mouse virulence of the proposed 
strains of Salmonella paratyphi and Salmonella 
schottmuellerl cultures. Test results indicated 
that the proposed strains were satisfactory for 
use as challenge cultures in a mouse-protective 
potency assay of Typhoid and Paratyphoid Vaccine. 
Mouse LD50 values for these strains were established, 

Information gathered to date is too incomplete 
for conclusions to be drawn as to the suitability 
of proposed potency teat method, however, further 
experiments will be carried out in an effort to 
establish a sui table method. 

(1) Participated in numerous committee actions within 
the Division involving reviews, modifications, 
and revisions of existing regulations and require- 
ments for biological products. Also, assisted 

in the review of monographs pertaining to 
biological products for publication in the 
sixteenth edition of the U, S. Pharmacopeia. 

(m) Thirteen visitors representing eleven foreign 
countries were given orientation and training 
in control testing activities during 1958. The 
length of the training periods ranged from one 
day to two weeks. In addition, participated 
in a lecture series involved in a personnel 
training course given within the Laboratory of 
Blood and Blood Products for blood bank inspectors, 

(n) A medical student f assigned to this section for 
a 3 month tour of duty, completed a review of the 
current medical literature cm the problem of 
staphylococcus infections „ Further „ he engaged 
in a limited project to evaluate control methods 
used in the assay of staphylococcus toxoids and 
antitoxins. It is believed that contributions 
made will be helpful In providing basic back" 
ground information required for continuation in 
the development of adequate test procedures for 
the control of staphylococcus biological prepara- 
j|» tionsu 


& (o) Proposed course - To continue objectives and 

methods described above, and to further Increase 

effectiveness and research activities with 
acquisition of authorized additioas to the 

J* professional staff, 

f Part S included Yes / 7 Wo /X~7 

Serial No, DBS.i&_ 

1« Control Activities 
2 „ Pyrogens 
3 „ Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A . / 

Project Title: Section on Pyrogens 

Principal Investigator: Thomas F„ Probey (% year) 

Harry W. Burruss (Jj year) 

Cooperating Units: None 

Man Years: Patient Days; None 

Total: 2% 
Professional: % 
Other: 2 

Project Description: 

The major responsibility of this section is the tasting 
of certain licensed biological products to determine their 
safety, purity, and stability as defined in the control 
regulations of the Public Health Service Act, 

The safety of some licensed biological products, 
especially the blood products, of human and animal origin, is 
determined by the pyrogen test; purity is determined in part 
by total solids content; stability of dried products is meas- 
ured by moisture determinations. Principal products for test- 
ing include antitoxins, vaccines, blood albumin, globulin, 
fibrinogen, plasma and serums. 

All pyrogen test results reported in the manufacturers' 
protocols to the Laboratory of Control Activities were eval- 
uated by the principal investigator . On the basis of this 
evaluation and additional NIH tests on samples of the product 
submitted, recommendations for release or rejection were made 
Records of all tests were maintained and officially reported, 

Control work in this section involves research studies 
designed to adapt the above mentioned tests to new products, 
to study the factors influencing the test, and the adapt- 
ability of these tests as means of evaluating the safety of 
experimental pharmaceuticals, including radioactive Isotopes, 
under development by the NTH Pharmacy Department for clinical 
study in the N1H Clinical Center. Blood products under study 
in the NIK Blood Bank are also included. 

Coded records continue to be maintained on certain lots 
of poliomyelitis vaccine. Samples from this supply were de- 
livered periodically throughout the year to the Laboratory of 
Viral Products for testing. 

It is also the responsibility of this project to instruct 
visiting scientists from domestic and foreign health laboratories 
and from the licensed biological establishments in the techniques 
used in performing the work of this section. 

Samples of each lot of Normal Human Serum Albumin, released 
by the Laboratory of Control Activities, were distributed to 
various hospitals for clinical testing. Reports received on 
these lots of albumin confirm the reliability of the pyrogen 
test as a means of determining the purity and safety of the 

One member of this group serves as a consultant on problems 
relative to the control of yellow fever vaccine. Individual 
conferences were held with Dr. Pitfcman prior to her attending 
(as an observer) a meeting of the Expert Committee on fellow 
Fever of the World Health Organization. Other discussions were 
held with Br* Bottle and Dr. van loosler, of the Laboratory of 
Viral Products 8 on working out a suitable monkey safety test, 
to be adopted by the World Health Organization, for yellow 
fever vaccine secondary seed virus. 

Part B included Yes/"""T 

Serial Ho v DBS-.& 

L Control Activities 
2.. Reference Standards 
3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A , 

Project Title: Section on Reference Standards 

Principal Investigator: R. P. Miller 
Other Investigators: Elmer C. Russell 
Cooperating Units: None 

Patient Days: None 

Man Tears: 
Total: 4 
Professional: 1 
Other: 3 

Project Description: 

Objectives : Since biological products cannot be standardized s 
with few exceptions, by chemical or physical means, recourse mu3t 
be had to testing their effects in relation to physical reference 
preparations. It Is the function of this project to provide these 
standard preparations for the use of manufacturers and others 
engaged in standardisation.. In this way uniformity of potency ia 
established throughout the country. It is largely by the use of 
these standards that those portions of the biologies law relating 
to potency can be enforced, A great deal of painstaking and 
responsible work is necessary to Insure that standards issued are 
satisfactory as official standards for this country and bear a 
known relation to International Standards. 

Methods Employed : Old standards must be replaced as they becom. 
exhausted Standards are required for new products and for certain 
of the older ones for which it has not been possible to prepare 
standards heretofore. This is dependent in increasing knowledge in 
a number of fields, such as immunology, biology, biochemistry, and 
medicine Important features of this work are the improvement of 
methods of storing and preserving standards, such as by freeziag 
and drying so that their potency may be maintained, and to correla 
those standards where possible with those established in other 
countries or with International Standards. 

• 2 

I- Proposed Course of Project: It i« proposed that the project be 
thoroughly reevaluated from the point of view (1) replacing some 
standards Hearing depletion such ae toxins, vaccines, and serums: 
(2) reinvestigating certain Standards, a few of which are badly 
needed but not in existence. 

II -Proposed Project: Working with Dr. Bottle in the study of the allergic 
enccphalitic properties of rabies vaccine. (This includes measuring the 
potency in mice and the paralysis in guinea pigs.) 

Ill -Project in Progress: 

1. Preparing;, testing and distributing Standards and Reference 
preparations for use of manufacturers and others engaged in standardisa- 

2o The control testing of yellow fever vaccine; this vaccine is 
tested for potency,, safety and sterility. (Mice and guinea pigs are used 
for this testing.) 168 lots of yellow fever vaccine have been tested so 
far this year. 

3. Shipping vaccines and viruses in the frozen state. (This includes 
packing in dry and wet ice.) 

4 a Preparation of rabies reference vaccine for World Health Organise 

Standards and Reference Preparations: A Culture collection of organ* 
isms used in the production and testing of biologic products .is also main- 
tained* Official standards and reference preparations now maintained are 
as follows: 

Adenovirus Virus Type 3 

Adenovirus Polyvalent Vaccine 

Anti-A Blood Grouping Serum 

Anti«B Blood Grouping Serum 

AntioRh Typing Serum<>A-&«£h 

Antimeningococcus Serum 

Botulinus (A) Antitoxin 

Botulinus <B) Antitoxin 

Botulinus (C) Antitoxin 

Botulinus (A) Toxin 

Botulinus (B) Toxin 

Cholera Opacity Reference 

Coomb's Anti-Human Serum 

Diphtheria Antitoxin 

Diphtheria Antitoxin Flocculating Ses*sn 

Diphtheria Toxoid 

Dysentery Antitoxin 


H. Influenzae Antiserum 

Histolytica Antitoxin 

Hi&tolyticus Toxin 



Influenza Virus Vaccine for CCA Titration 

Influenza Virus Vaccine for Potency 

Ludox Nephelometry Standard 

Mumps Reference Serum 

Nitrogen Standard (Protein) 

Oedema t lens Antitoxin 

Oedematiens Toxin 

Reference Old Tuberculin 

Perfringens Antitoxin 

Perfringena Toxin 

Pertussis Opacity 

Pertussis Vaccine 

Plague Agglutinating Serum 

Plague Vaccine 

Poliomyelitis Control Serum 

Poliomyelitis Immune Globulin 

Poliomyelitis Vaccine 

Poliomyelitis Virus 

Purified Protein Derivative of Tuberculin 

Rabies Challenge Virus 

Rabies Production Virus 

Rabiea Vaccine 

Rabies Serum 

Scarlet Pever Streptococcus Antitoxin 

Scarlet Pever Streptococcus Toxin 

Smallpox Vaccine 

Sordelli Toxin 

Staphylococcus Antitoxin 

Staphylococcus Toxin 

Staphylococcus Antitoxin 

Staphylococcus Toxin 

Tetanus Antitoxin 

Tetanus Antitoxin Flocculating Serum 

Tetanus Toxin 

Tetanus Toxoid (Alum Precipitated) 

Tetanus Toxoid (Fluid) 


Typhoid Vaccine 

Typhoid Turbidity Standard 

Typhus Control Serum 

Vibrion Septique Antitoxin 

Vibrion Septique Toxin 

This Section, from January 1, 1958 up to December 6, 1958, has 
distributed to commercial houses, Health Departments, Universities and 
foreign countcies the following Standards and Reference Preparations: 

Antitoxins 290 vials Toxins 90 vials 

Cultures 325 vials Vaccines 1566 vials 

Serums 1265 vials Viruses 435 

Drying: This section during the year January 1, 1958 up to 
December 6, 1958, has dried the following: 

Cultures ----- — — — — - — 18X6 vials 

Serums — - — — — — ■ 73 vials 

Vaccines --.— ..^...1X82 vials 

Viruses ................ —1000 vials 

Part B included Yes /"*'" / Ho /X / 


Bacterial Products 


Sarlal No. DBS - 6 thru 16 

Batlmatad Obligations for F.Y. 1959 

Total: 166,420 

Dlract: 136,120 

Reimbursements: 30,300 

Serial No. DBS- 6 

1. Bacterial Product.: 

2. Bacterial Toxins 

3. Be the a da 

Individual Project Report 
Calendar Year 1958 

Part A . 

Project Title: I. Components of Diphtheria Toxin. 

Principal Investigator: Dr. Sara E. Branham (Matthews) 

Other Investigators: Dr. C. W. Hiatt 
Mr. Donald Riggs 

Cooperating Units: Biochemistry and Biophysics Unit, Research 
Section, LVP 

Man Years; 

Total; 11/12 
Professional: 7/12 
Other: 1/3 

Project Description: 

Objectives ; The objective was to determine if there 
were any differences between the components of the toxins 
produced by the gravis type of C. diphtheriae which induces 
an especially severe type of disease and those of the "PW8" 
toxin used exclusively in the preparation of diphtheria 
biologic products. 

Methods Employed : The methods were described in earlier 
annual reports. They include (1) electrophoretic separations 
of the components by paper strip, paper curtain and Tiseliuo 
methods, (2) immuno-electrophoresis in agar and (3) ultra- 
centrifugation separation of the components which were examined 
for toxicity, antigenicity and electrophoretic mobility. 

Major Findings : All strains of C. diphtheriae studied 
induced typical symptoms and pathological lesions in animals. 
The gravis strains, however, caused more marked reactions; for 
example, there occurred in guinea pigs a great deal more edema 
and free fluid and more frequent local hemorrhages and paralysis. 
In preparations of the gravis strains an extra component was 
demonstrated by means of electrophoresis and ultracentxifugation. 
(Hiatt) . This component found only in the bottom of the 

centrifugated material was composed of heavy large size 
molecules. It Induced definitely more marked edematous 
skin reactions in guinea pigs than other fractions and 
was lacking in similar preparations of PW3 strains. 
Although it is suggestive that this component may be 
related to the marked edematous lesions characteristic 
of gravis infections in man (sometimes referred to as 
bull's neck), it has not been proven. 

Significance to the Institute : If it is proven that 
the edematous component is related to the peculiar pathology 
of gravis infection and that it i» found to be antigenic, 
than it may be possible to develop an effective prophylactic 
agent . 

Proposed Course of Project : Work by the principal 
investigator has been terminated due to retirement. Results 
are ready for publication. 

Part B included Yes /X~7 No / 7 

Serial No. BBS 

Individual Project Report 
Calender Year 1958 

Part B : Honors, Awards, and Publications. 
Publications other than abstracts from this project: 

(Reported in 1957 as "In press.") 

Branham, S. E. Reference Strains for the Serologic Groups 
of Meningococcus. International Bulletin of Bacteriological 
Nomenclature and Taxonomy, 8: 1-15, 1958. 

Serial Mo. D3S-7 

1. Bacterial Products 

2. Haemophilus Studies 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A . 

Project Title: II. Study of the Protein Composition of 
Diphtheria Antitoxin Produced in the 
Guinea Pig. 

Principal Investigator: Dr. Sara E Branham (Matthews) 

Other Investigators: Dr. C. W. Hiatt 
Mr. Donald Riggs 

Cooperating Units: Laboratory Unit No. 4 
Research Section. LVP 

Man Years : 

Included with Project Title I. 

Project Description: 

Objectives : To determine if diphtheria antitoxin 
produced in the guinea pig with the aid of Freund's adjuvant 
causes flocculation of diphtheria toxin as does antitoxin pro- 
duced in the horse. 

Methods Employed : The equivalence sone of flocculation 
will be determined by nitrogen analysis of the specific 
precipitates . 

Major Findings : To be determined. 

Significance to Institute ; Information obtained in this 
study is relevant to the general topic of antibody production 
in immunized animals. 

Proposed Course of the Project ; To carry out the study 
as proposed and if results are significant they will be published. 

Part B included Yes / 7 No /X~7 

Serial No. DBS- 9 

1. Bacterial Product* 

2. Cancer Products 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A . 

Project Title: Fibroma-rayxoma Virus Transformation. 

Principal Investigator: Dr. Lawrence Kilham 

Other Investigators: Dr. Joseph Shack (NCI) 

Dr. William G. Banfield (NCI) 
Dr. Edwin Lerner (NIAMD) 
Dr. C. W. Hiatt 

Cooperating Units: National Cancer Institute 

Man Years: 

Total : 1 

Professional; 1/2 
Other: 1/2 

Project Description: 

Objective : To discover the mechanism by which one 
virus can be transformed into another. 

Methods Employed : Myxoma virus, "killed" by use of 
heat and chemicals is employed as the transforming agent for 
live Fibroma Virus in tissue culture. Urea is used as a 
deproteinizing agent to remove the outer coat of the Trans- 
forming agent and to reveal the inner core of desoxyribonucleic 
acid (DMA). 

Mai or Findings : 1. Virus transformation can be produced 
in nearly 1007. of experiments using heat-killed, Urea-treated 
Transforming agent (TAM) . 2. Urea-treated TAM is destroyed 
specifically by desoxyribonuclease (DN-ase) . It is not affected 
by ribonuclease (RN-ase) . 3. Ether and detergents have also 
been used in preparing an active transforming agent. These 
types of TAM, however, are not destroyed by DN-ase. 4. TAM ia 
readily destroyed by photodynamic action in the presence of 
toluidine blue. 5. The transforming agent attaches to and/or 
enters tissue culture cells by itself without the presence of 
live fibroma virus. 

Significance to the institute ; Since nucleic acids 
are the basis of all living things, anything which can be 
learned about their nature and mode of action should be con- 
sidered as fundamental research which might, eventually, 
lead to a batter control and understanding of microorganisms. 

Proposed Course of Project ; 1 . Study the nature of 
the Urea-treated TAM in regard to size, behavior on high-speed 
centrifugation, and chemical properties. 2. Follow Myxoma 
Virus through the various stages of its treatment by physical 
and chemical agents--by use of electron microscopy. Present 
plan is to prepare purified virus by means of Fluorocarbon. 
3. In an effort to widen the biological base of this 
research, attempts are being made to transform vaccinia by 
means of Fowl Pox virus. The hope is to develop genetic 
markers and to produce hereditary changes among various mem- 
bars of the pox group of viruses. 

Part B included Yes /X~7 No /~~7 

Serial Ho. DBS 

Individual Project Report 
Calendar Year 1958 

Part B : Honors, Awards, and Publications 

Publications other than abstracts from this project: 

1. Kilham, Lawrence , Fibroma-Myxome Virus Transformations 
in Different Types of Tissue Culture. J. Nat. Cancer 
Institute, 20: 729, 1958. 

2. Kilham, Lawrence, Lerner, E. ; Hiatt, C. and Shack, J. 
Properties of Myxoma Virus Transforming Agent. 

Proc. Soc. Exp. Biol, and Med., 98; 689-692, 1958 

Serial Ho. DBS- 9 

1. Bacterial Products 

2. Cancer Products 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A. 

Project Title: Virological Study of Rat Tumors. 

Principal Investigator: Dr. Lawrence Kilham 

Other Investigators: Dr. C. W. Hiatt 

Dr. L. J. Olivier (NIAID) 

Cooperating Units: NIAID - Laboratory Tropical Medicine 

Man Years: Patient Days? None 

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

Project Description 

Objectives and Methods : The initial objective was to 
recover a virus from rat tumors employing the tissue culture 
methods used by Stewart and Eddy in the recovery of Polyoma 
virus from mouse tumors. 

Major Findings : An apparently new agent , termed Rat 
Virus (RV) has been recovered in Rat Embryo tissue cultures. 
It is recognisable by its cytopathogenic effect and its 
ability to hemagglutinate. In regard to a number of properties 
such as resistance to ether, effect of RDE, and type of CPE 
induced, it bears some resemblance to the S.E. Polyoma Virus, 
although there is no cross-neutralization. Circulating anti- 
bodies to RV are widespread among wild and laboratory rats. 
The virus has been isolated 3 times from 2 different sources, 
one being Fisher rats with Sarcomas induced by Cysticercus 
fasciolanis and the other an Osborne -Mendel rat bearing a 
transplantable leukemia. A paper on this virus is being 
submitted for publication. 

Significance of Program : Increased experience in field 
of viruses in relation to cancer. 


Proposed Course of Project ; 1 . Inoculation of new 
born rats of various inbred strains with RV to determine 
if virus does induce tumors or other pathogenic effect. 

2. Continued efforts to isolate virus from rat embryos due 
to suggestive evidence that virus is vertically transmitted. 

3. Serologic survey of Germ Free rats by Hemagglutination 
Inhibition and by T. C. neutralisation tests. 4. Electron 
microscopy — taking advantage that Rat Virus attaches to 
guinea pig Erythrocytes but does not elute from them. 

Part B included Yes / 7 Ho jjTJ 

Serial No. DBS- 10 

1 . Bacterial ?rodu< 

2. Haemophilus Studies 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Pertussis Vaccine. 

Principal Investigator: Dr. Margaret Pittman 

Other Investigators: None 

Cooperating Units : None 

Man Years: Patient Days: None 

Total : 1.6 
Professional: 0.2 
Other: 1.4 

Project Description: 

Objectives : The objectives are the attainment of 
optimal properties for pertussis vaccine processed by licensed 
manufacturers . 

Methods Employed : 1. The potency and toxicity of the 
pertussis component of all lots of products containing this 
antigen are kept under constant surveillance by reviewing 
protocols and license applications submitted by manufacturers, 
by conferences with representatives from manufacturer 8 s by 
testing the majority of the lots submitted (about 200 per year) 
for potency and freedom- from- toxicity, and the maintenance of 
running records of tests for each manufacturer's products to 
observe any trend away from consistency. 

2 . Determination of the optimum time between vaccination 
and challenge in the potency test of mineral adjuvant containing 
vaccine 8 containing various concentrations of adjuvants (alum- 
aluminum hydroxide, aluminum phosphate) and varying in potency 
are being tested to determine the protective response at 14, 21 
and 28 days after vaccination. 

3. Study of the cause of the change in the density of 
the Opacity Standard for Pertussis Vaccine. 

4. International contacts are maintained (a) by 
cooperating in the potency testing of vaccines used in field 
trials in England, (b) by cooperating in tbe evaluation of 
national and manufacturer's references , (c) by frequent 
visitors from foreign control laboratories and by partici- 
pating with WHO in the development of international recom- 
mended requirements. 

5. Determination of the effect of phemerol as a 
preservative on the toxicity and potency of pertussis vaccine 
to be used in combination with poliomyelitis virus vaccine. 

Major Findings ; 2. No significant differences have 
been found to date in the i&xmme response of mice challenged 
14, 21 or 28 days after vaccination. 

4. From two reports of the field trials in England 
(one report now in press), it has been demonstrated that pro- 
tective activity of pertussis vaccine against heme exposure 
to whooping cough is directly related to mouse protective 
activity of the vaccine. In the first report vaccines ranging 
in potency from 8 to 26 units (DBS values) afforded protection 
from 71 to 95% of children exposed in the home; in the second 
report the relative figures were 10 to 28 units (DBS values) 
versus 52 to 92% protection. The latter results are based 
upon fewer numbers of exposures. 

In a recent comparison of the potency of the International 
Standard for Pertussis with that of the U. S. Standard, the 
same value was obtained as was obtained three years ago in an 
international cooperative study. This substantiates the practi- 
cability of the assay for the potency of pertussis vaccine and 
its reproducibility. 

5. From limited experimentation it appears that vaccines 
preserved with phemerol are no more toxic than those preserved 
with merthiolate. 

(The above numbered paragraphs under Major Findings 
correspond to similarly numbered paragraphs under 
Methods Employed.) 

Significa nce, to... the. Institute ; The significance of the 
findings to DBS are that they provide information in relation 
to the regulations and improvement of pertussis vaccine. 

2. The interval of 14 to 17 days between vaccination and 

challenge, as now required in the potency test of pertussis 
vaccine, is a satisfactory time for those products containing an 

aluminum compound and needs not Co be altered. It has been 
claimed by a British scientist that the optimum interval is 
28 days. 

4. International contacts provide a broad base of 
information for use in improving pertussis vaccine. From the 
results of the British field trials, it can be deducted that 
the potency requirement of 12 units per total human immunizing 
dose of pertussis vaccine is adequate to afford protection to 
approximately 85% of children from home exposure; and that the 
lower limit of 8.0 unit permitted is too low. 

5. Insofar as toxicity is concerned, phemerol seems 
to be as satisfactory as merthiolate as a preservative for 
pertussis vaccine. The latter is detrimental to the potency 
of poliomyelitis vaccine, hence cannot be used in combinations 
with poliomyelitis vaccine. 

Proposed Course of Project : To continue along the 
same lines until a particular phase is completed, then to 
start new ones as indicated. The application of the results 
to fundamental observations on the immunological and physio- 
logical responses induced by Haemophilus pertussis , in vaccine 
or in infection, is kept under consideration. 

Part B included Yea / 7 No /5~7 

Serial No. DBS-- II 

1 . Bacterial Products 

2. Haemophilus Studies 

3. Betfoesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Antipertussis Serum Standardisation. 

Principal Investigator: Dr. Margaret Pittman 

Other Investigators: None 

Cooperating Unit : None 

Nan Years: 

Total; 0.7 
Professional; 0.1 
Other : 0.6 

Project Description: 

Objective a : The objectives are to establish a standard 
of potency for antipertussis serum. 

Methods Employed : 1. Development of a mouse passive 
protective test. 2. Preparation of a U. S. Standard for 
antipertussis serum and the assignment of a potency value. 

Major Findings : A mouse passive protective test in which 
reproducible results can be obtained within reasonable limits 
has been developed over a period of several years. 

A large lot of concentrated antipertussis serum (rabbit) 
has been desiccated. 

Significance to the Institute : Will enable DBS to 
establish potency regulations for antipertussis serum. 

Proposed Course of Project : The method of the protection 
test and the results of evaluation of lots of serum collected 
over a period of several years will be submitted to manufacturers 
of the product to obtain opinions and cooperation in the estab- 
lishment of a Standard of Potency for this product. 

Part B included Yes / 7 No /X_/ 

Part A, 

Serial Ho. DBS -12 

1. Bacterial Products 

2. Haemophilus Studies 

3. Bethesda 


Individual Project Report 

Calender Year 1958 

Project Title : Study of Methods of Preparation of 
Formal inized Red Blood Cells. 

Principal Investigator : Dr. Louis Csizmaa 

Other Investigator : None 

Cooperating Units; None 

Man Years: Patient Days: None 

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

Project Description: 

Objectives : The objective of this project was to 
develop a method of preparation of stable formalinized red 
blood cells that would be as sensitive in the hemagglutination 
test as fresh red blood cells. 

Methods Employed : 1. Red blood cells were treated 
with formalin under conditions that practically eliminated cell 
collision (dilution, agitation and gradual exposure to formalin) 
and that induced slight swelling of the cells. 

2. Improvement of a micromethod for hemagglutination. 

3. Preparation of diphtheria toxoid and tetanus toxoid 
sensitized red blood cells. 

Major Findings : 1. Stable formalinized red Mood cells 
vere prepared from Mammalian, reptile and avian bloods The 
cells showed no crenation, were evenly dispersed and retained 
the adsorptive properties' of fresh red cells. They were suitable 
for use in all types of hemagglutination tests i^.ng Sa Ik s 
pattern technic, and were stable after long periods au A C. or 

2. Sharper end-points for hemagglutination were 
obtained by the improved micromethod. 

3. Adsorption of purified antigens onto the formalinized 
cells after tannin treatment resulted in stable reagents. 

Significance to the Institute : With a simplification 
of the test and the use of stable antigen sensitized red cells, 
it may be possible to apply the use of hemagglutination tests 
as an aid in evaluating the potency of a number of biologic 

Proposed Course of the Project ; Elaboration of other 
adsorbed antigens and further studies of the mechanisms 
involved . 

Part 5J included Yes / 7 Mo jTJ 

Serial No. DBS- 

1. Bacterial Products 

2. Haemophilus Studies 

3. Betheeda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title : Effect of Combining Antigens and of 

Aluminum Adjuvants on the Antigenicity 
of the Individual Antigens. 

Principal Investigator : Dr. Louis Csismas 

Other Investigators: None 

Cooperating Unit i None 

Man Years: Patient Days: None 

Total : 1 

Professional: 1/2 
Other: 1/2 

Project Description: 

Objectives: There is a trend towards the incorporation 
of multiple antigens in one product. The objective of this 
project is to determine the influence of the presence of 
other antigens and also aluminum compounds upon the Immuno- 
logical response of the host to an individual component antigen^ 
also to try to elucidate the mechanism of action of "modifiers. 

Methods Employed : 1. Development of a theoretical 
model of protection against infection and of the effect of 
"modifiers" (inhibitory or enhancing) on immunological response. 

2 Hemagglutination titrations of mouse antisera with 
the use of diphtheria and tetanus toxoid-sensitized formalinized 
red blood cells. 

3 Simplification and improvement of the Ouchterlony 
and Oudin gel diffusion technics and the development of a nomo- 
graphical method of determining the concentration and diffusion 
coefficient of antigens with the use of two tubes. 

4. Simplification of the Spearman-K&rber method for the 

evaluation of D50 and generalization of itB use to include 
more than one type of response in evaluation. 

5. Preparation of aluminum hydroxide and aluminum 
phosphate gels. 

6. Determination of the effect of different strains 
of mice on the evaluation of the potency of diphtheria and 
tetanus toxoids and pertussis vaccine. 

Major Findings ; 1. Results of hemagglutination 
teste of sera from animals vaccinated with diphtheria and 
tetanus toxoids ran parallel to those obtained from animal 
protection tests. 

2. Pertussis vaccine was found to be a more effec- 
tive adjuvant than aluminum compounds. 

3. The effect of adjuvants differed when tested in 
different strains of mice. 

4. The mineral adjuvant effect in mice was greater 
if the vaccine were prepared in a concentrated form and then 
diluted before use, than if it were prepared in the final 
form for use. 

Significance to the Program of the Institute : The 
methods developed may have application in the evaluation of 
the potency of a number of biologic products. The influence 
of "modifiers" on potency evaluation is of both general and 
theoretical importance in the control of multiple antigen 
preparations . 

Proposed Course of Project; To complete the studies 
on the comparison of the effect of aluminum hydroxide, 
aluminum phosphate and alum and on the influence of combining 
antigens on immunological response and to prepare the perti- 
nent information for publication. Also, to attempt to 
determine whether the "modifying" effect is on the agent or 
on the host by using different combinations in vitro and 
in vivo . 

Part B included Yes / 7 No /X~7 

Serial No. DBS - 14» 

1 . Bacterial Products 

2. Haemophilus Studies 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title : Re-evaluation of Requirements for Cholera 

Principal Investigator ; Dr. John C Peeley 

Other Investigator : None 

Cooperating Units : None 

Man Years: Patient Days; None 

Total: 0.41 
Professional: 0.33 
Other: 0.08 

Project Description: 

Objective : The objective of this project is to 
attempt to define and/or establish conditions for the per- 
formance of a reliable quantitative potency test for cholera 
vaccine. Insofar aB possible, this study will be carried 
out in reference to and coordinated with the proposed World 
Health Organization international cooperative efforts toward 
establishment of a standard of potency related to human 

Methods Employed : Work on this project has just begun. 
It is anticipated that a thorough study of the variables 
involved in virulence of cholera vibrios for laboratory 
animals will be carried out with the aim of establishing 
and/or defining conditions for a suitable experimental test 
system for quantitative in vivo potency evaluation of cholera 
vaccine Current plans call for the eventual establishment 
of a reference vaccine to be tested in parallel with vaccines 
under test in order that results can be comparatively expressed 
in terms of the reference vaccine. 

Major Findings : There are no major findings at this 
time since this project has just been initiated. 

Significance to the Institute ; While Asiatic cholera 
is not at present considered to be a major public health 
problem in this country, increasing numbers of people are 
traveling into areas of the world where this disease is still 
endemic. Protection of the traveler to these areas is a 
desirable health measure. Cholera vaccine is a matter of 
concern to the Armed Forces for similar reasons. Even today, 
epidemics of cholera still occur in certain underdeveloped 
areas of the globe and vaccination is an important measure 
in their prevention and control . A potent cholera vaccine 
must be available to meet these needs. Since cholera vaccine, 
like other biologies, is subject to variation in potency, 
adequate control over this property is essential . 

The potency test for cholera vaccine that is currently 
employed in this country is essentially a qualitative procedure. 
Although it is capable of detecting vaccines of very low potency 
it lacks desirable quantitative features which should allow 
ready comparison between vaccines and reference preparations. 
It is anticipated that a quantitative test based on a reference 
vaccine and similar in design to tests currently employed with 
typhoid and pertussis vaccines would add continuity to potency 
control of cholera vaccine. 

At present, there is no general agreement concerning 
the type of potency test that should be employed with this 
product. Partly as a result of this, there is no adequate 
information dealing with the relevancy of potency tests in 
animals to the central problem of vaccine potency in man. 
Ultimately, it is hoped that information gained from potency 
tests can be correlated with the protective activity of 
cholera vaccines against human cholera. The present project, 
if successful, may contribute toward establishment of a 
standard of potency that will aid in the assurance that 
persons receiving cholera vaccine are being immunized with 
a product of suitable potency. 

Proposed Course of Project ; To follow through as 
indicated above . 

Part B included Yes / 7 Ho jTJ 

Serial No. DBS -IS - 

1. Bacterial Products 

2. Haemophilus Studies 

3 . Bethesda 

Part A. 


Individual Project Report 

Calendar Year 1958 

Project Title : The Schick Test Toxin (Diphtheria) 

Principal Investigator : Dr. Michael F. Barile 

Other Investigators : Mr. Donald B. Riggs 

Mr. Robert W. Kolb (LCA) 

Cooperating Units : Section on Control Tests, LCA 

Man Years: Patient Days: None 

Total: 1.3 
Professional 0.6 
Other : 0.7 

Project Description: 

Objectives : 1. To select a diphtheria toxin for the 
U. S. Standard for the Schick test toxin. 2. Re-evaluate 
the requirements of the Schick test toxin with reference to 
the selection of a more accurate potency assay. 

Methods Employed : Two commercial preparations of 
diphtheria toxin, previously selected from other lots, were 
examined by methods given below. 

Toxin potency was determined by means of both skin 
reactions and lethal action. Guinea pigs and rabbits were 
used for each test. 

Combining power (toxin-antitoxin binding avidity) of each 
toxin was determined in vitro , as Lf, and in vivo , as Lr 
and L+, at various levels (0.1, 0.01, and 0.001 units) of 

The velocity of neutralization (speed of toxin-antitoxin 
reaction) was determined by means of skin testa performed at 
various time intervals after combination of toxin and antitoxin, 
The intervals were less than 30 seconds, 30, 60, and 100 
minute 8 and 2 A hours. 

Several agents (Evans blue and trypan blue) were 
given intravenously to determine whether dye deposition 
at the skin reaction site provides a acre accurate method 
for delineating the reaction than erythema uhich is used 
in measuring the Minimum Reactive Dose (H.R.D.). In an 
attempt to find the most reproducible end-point for toxin 
activity, the "MRD" erythematous reading was compared with 
edema, necrosis and total score (summation of erythema, 
edema and necrosis) readings. 

Major Findings; toe toxin was 2.67 times more potent 
then the other, but the avidity of each was approximately 
the same. Comparative values for the toxins on potency, 
combining power and velocity of reaction showed & constant 
relationship and were independent of the animal (guinea pig 
or rabbit) or test system vLDa or M.R.D.) used. Either 
animal or test system may be used effectively if the test 
is standardised. 

The sensitivity of the evaluation of the MRD was not 
improved by using edema, necrosis and/or total score meas- 
urements. Dye localization also did not improve evaluation. 

Significance to the Institute : The establishment of 
a Standard Schick test toxin (diphtheria) and more precise 
method of assay will provide means for increasing the 
quality of the Schick test toxin. 

Proposed Course of Project ; Both toxins which appear 
to be suitable for a Standerd will be compared with the World 
Health Organization Standard Schick toxin (diptheria) and 
possibly with the standards from other countries. 

The toxin selected for the Standard will be preserved 
in the freeze-dried state. More than one menstrum may be 

Part B included Yes 7 No /X~7 

Serial Ho. DBS°1A 

1 . Bacterial Products 

2. Eaeoophilus Sfcu: 

3. Betheada 

Pare A 


Individual Project Report 

Calendar Year 1958 

Project T'.tle : L Forms of Bacteria from Tissue Culture. 

Principal Investigator : Dr. Michael P. Barile 

Other Investigators : Mr. Donald B. Riggs 

Mr. George D. Gardner (LVP) 

Cooperating Units : Tissue Culture Unit, LVP 

Man Tears: Patient Days: None 

Total: 0.2 
Professional: 0.1 
Other: 0.1 

Project Description: 

Objectives : To develop methods of purifying the LVP 
tissue culture lines contaminated with pleuropneumonia- like 
organisms (PPLO) and/or L forms of bacteria. 

Methods Employed : Appropriate media and environment 
for the cultivation of PPLO and L forms of bacteria was used. 

The population of L forms was determined by dilution 
and plate methods. 

Major Findings: L forms of bacteria were cultured from 
11 of 14 tissue culture lines. 

Repeated examinations for PPLO and/or L forms were 
negative from 1) primary monkey and rabbit kidney cells 
2) tissue culture media. Parker media No. 199 and 3) txssue 
cultural supplements (calf sera, ascetic fluids, etc.). 

The growth curves of the L form and tha L form infected 
viable tissue culture cell paralleled each other The popula- 
tion of the organism remained approximately 100-fold gre£ 
than the cell. The L form maintained its population as lc: 
the cells remained viable. 

The number of L forrae recovered from infected 
tissue cells was increased at least 100-fold by sonic 
disruption of cells. The organism was more resistant 
than the tissue cell to the action of sonic vibrations. 

Several bacteria which were developed from L forms 
isolated from the Lac cell line were induced to revert to 
L forms when exposed in vitro to tissue culture media 
containing penicillin and streptomycin. It is possible 
that L forms were induced in tissue culture lines by the 
action of antibiotics on bacteria. 

Significance to the Institute ; This study has 
demonstrated the presence of L form contaminants in tissue 
culture lines. Since the results indicate that the L forms 
were induced from bacterial contaminant® by the presence 
of antibiotics, it appears that reliance cannot be placed 
on antibiotics to completely control bacterial contamina- 
tion in tissue culture lines. 

Proposed Course of Project; To determine whether 
antibiotics can induce bacteria to produce L forms in 
tissue cultures. 

To determine the effect of chemical and physical 
factors, including antibiotics, on clearing tissue lines 
of L forms. 

To determine the effect of L form infection on the 
infectivity of certain viruses in tissue culture lines. 

Part B included Yes Hj No jjTj 


Viral Products 


Serial No. DBS - 17 thru 37 

Eatlaated Obligation for P.Y. 1959 

Total: 965,050 

Direct: 787,950 

RalaburaeoMnts: 177,100 

Serial Mo. DBS - 17 

1 . Laboratory of Viral Produe-; 

2 . Tissue Culture Unit 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: The Tissue Culture Preparation and Tissue Culture 
Testing of Killed Virus Vaccines. 

Principal Investigator: Dr. 6. A. Hottle 

Other Investigators: Dr. Paul Gerber 

Mr. George Gardner 

Cooperating Units: The monkey safety testing unit provides 
monkey kidneys . 

Man Years: 
Total: 11.0 
Professional: 2.0 
Other: 9.0 

Project Description: 

Objective : To prepare primary and continuous cell cultures for 
research and test purposes, and to test vaccine for safety by tissue 
culture methods. 

Methods : The methods are those outlined in the Regulations and 
tissue culture methods reported in scientific literature. 

Plod logs : (a) During the year 53 samples of poliomyelitis vaccine 
and 10 samples of adenovirus vaccine were tested for safety in tissue 


(b) From January 1 to Hovember I, 1958, the following tissue 
cultures were prepared: 

(1) Primary cell preparations - see attached. 

(2) Stable cell culture preparations - see attached. 
Part B included Yes / 7 So /X~7 

January 1, 1958 - December 1, 1958 

Primary Cell Preparations: 


Monkey Kidney 
Rabbit Kidney 
Dog Kidney 

. : akey 







2.jpz . 8 os. 

Total Vol . of packs c 

Gel la Prepared 

5,229 cc. 
981 cc. 
11 .5 cc. 
9.75 cc. 










for Othe 




.5 cc. 

January 1, 1958 - December 1, 1958 

•Stable Cell Culture Preparations: 





Monkey heart 






Drew RK 




Parker RK 




Hunan Aanlon 


Monkey Muscle 



Hep II 




Human Heart 

2 oz. 

8 oz. 
















32 oz, 








* Human breast carcinosis from Dr. Baron. 

Serial No. DBS-lg 

1. Laboratory of Viral Product! 

2. Section on Testing 

3. Bethesda 


Individual Project Report 

Calendar Year 1956 

Part A. 

Project Title: Experimental Studies with Viruses and Cell Culture. 

Principal Investigator: Dr. Samuel Baron 

Other Investigators: Dr. Eugene V. Barnett 
Mr. Richard J. Low 
Mrs. Alice M. Gochenour 
Mrs. Beatrice L. Burch 
Dr. C. V. Biatt 
Dr. Alan S. Rabson (HCI) 

Cooperating Units: National Cancer Institute 

Man Years: 

Total: 3 1/4 
Professional : 3/4 
Other: 2 1/2 

Sub-project A: Immunisation with Increased Amounts of 
Poliomyelitis Vaccine. 

Objectives : To determine a method of immunization of man with 
poliomyelitis vaccine which would result in an early antibody 
responsedwhen used during epidemics of poliomyelitis. 

Methods Employed : Immunisation of non- immune persons with 
multiple inoculation or a single large inoculation of poliomye- 
litis vaccine. 

Patient Material : Volunteers from the Employee Health Service 
at KIH (Dr. John M. Lynch) and children at the Children's Center 
of the Government of the District of Columbia at Laurel, Md. 
(Dr. Wilfred R. Ehrmantraut) . 

Major Pindingo : An early antibody response occurred in She 
groups which received a single intramuscular inoculation of 10 ml . 
of poliomyelitis vaccine ia comparison to the groups which 
received multiple doses of 1 ml. or a single dose of 1 ml. Wo 
reactions occurred. 

Significance to the Division : Development of a method for 
poliomyelitis immunisation under epidemic conditions which 
promises to be much more protective. Provides data for 
establishment of potency requirements for future poliomyelitis 

Proposed Course : A quantitative dose response curve in man 
will be dona with Or. Eugene V. Barnett for purposes of com- 
parison with animal potency tests. 

Sub-project B: Comparative Study of Ultraviolet Light 
Inactivation of Animal Viruses. 

Objectives ; Determination of the ultraviolet light iaactivation 
properties of a number of animal viruses for characterisation 
and classification. 

Bfethods Employed : Aqueous suspensions of various viruses are 
exposed to ultraviolet radiation. Samples are taken at appro- 
priate intervals and assayed for iaf activity. Plot3 are then 
made for each virus relating decline of infectivity with time 
of irradiation. Intensity of irradiation is measured with an 
Archer radiation seter and also by inactivation of a standard 
virus preparation (poliovirus Type III, Saukefct strain). 

Major Findings ; Iaactivation rate and slope are correlated with 
relationship of viruses as determined by other properties. 
Xaacti.vation rate is not proportionate to virus sisa as is true 

of bacteriophage. 

Sig nifica nce to the Division: Basic information on the physical 
property of ultraviolet light icactivaticn of viruses. Poten- 
tial application for identification &ad classification of viruses, 

Proposed Course : For the present it seems necessary to eospare 
the iaactivation rates of more viruses to determine whether the 
correlations are true. 

Sub-project C; Assay of Antibody to Poliovirus by Inhibition 
of Plaque Siase. 

Objectives : Development of a new and easier technique for the 
assay of poliovirus antibody by the use of a saefchod which does 
not require serial dilution of serum. 

Methods Employed : Monolayers of monkey kidney cell culture 
are infected with small amounts of poliovirus and then over- 
layed with agar containing i rename serum. Diameters of sesu 
ting plaques are measured in comparison to serum- free control 
plaques. Results are analyzed for correlation with known 
serum antibody titer, reproducibility and variation among 
serum from different species. 

Major Findings : Plaque size inhibition is wall correlated 
with polio7irus antibody titers as measured by accepted testa. 
Reproducibility appears somewhat better than the metabolic 
inhibition test. Antiserum from man, chicken, and monkey are 
indistinguishable by the plaque size inhibition test. In 
tests of over 300 sera, methods have been developed for rapid 
large scale application. 

Significance to the Division ; Potential application in the 
serology unit for ootency tests of poliomyelitis vaccine in 
animals and man. 

Proposed Course of Project : Application to chick potency test 
for quantitation of antibody response which may increase accu- 
racy and reproducibility. Write for publication. 

Sub-project D; The Use of Skim Milk in Maintenance Media 
for Viral Studies in Cell Culture. 

Objective a : To develop a cell culture maintenance medium which 
approximates the theoretically ideal medium sore closely than 
media now in use . 

Methods Employed : Observation of various cell cultures for 
viability and suitability for virus studies while exposed to 
experimental media. Modified neutralization tests for detection 
of inhibition of virus effects by media components. 

Major Findings : Studies completed last year demonstrated 
suitability of skim milk in maintenance medium in relation 
satisfactory cell culture maintenance and freedom from inhi 
to most viruses tested. The inhibitor sfcudy has been extended 
and has shown the medium to lock inhibitors to a wide range of 
viruses . Studies of the use of skim milk in plaque medium have 
indicated its value in virus plaque assay in both primary and 
continuous cell cultures. The plaque mediuia has been applied 
for the determination of the poliovirus sensitivity of various 
cell cultures (see project number 1) and for plaque asc- 
measles virus. 

- 4 - 

Significance to the Division ; Development of a new medium 

for virus assay which may be applied for vaccine safety testing. 

Plaque assay of heretofore unplaquable viruses. 

Proposed Course : Study of plaque assay of measles virus, 
adenovirus and influenza viruses with skim lailk overlay medium. 

Sub-project S: A Semiautomatic Machine for Serial 
Dilution of Serum. 

Objectives ; Development and evaluation of a device which will 
simplify dilution of sera and which may be applied to the large 
scale work done by the DBS serology laboratory. 

Methods Employed ; Twelve Svedmyr hand diluting pipettes have 
been assembled in parallel on a newly designed hand-CEanked 
machine. Twelve serial serum dilutions were done simultaneously 
by the use of the machine and compared to accepted methods for 
dilution. Similar dye-dilution and hemagglutination dilution 
comparisons were made, 

Major Findings ; The accuracy and reproducibility of antibody, 
dye and hemagglutination titers obtained with the machine seem 
at least as good as those obtained by hand pipetting. Appli- 
cation of the device for routine use in the DBS serology 
laboratory has resulted in marked simplification and time saving. 

Significan ce to the Division; A development which allows 
greater productivity by the serology unit. 

Proposed Course : To complete ccaaparisoa of hemagglutination titers 

and then to write the results of the study for publication. 

B included Yes /x"7 Ho /~T 

Serial No. DBS 


Individual Project Report 
Calendar Year 1958 

Part B : Honors, Avarda, and Publications 

Publication other than abstracts froa this project: 

Baron, 8. and Lov. R. J. Hew Maintenance Medium for Cell 
Culture. Science, 128:89-90. 1958. 

Rabsou, A. S., Legallaio, P. T. and Baron, S. Adaptation 
to Serum- free Medium by a Phogacytic Cell Strain Derived 
from a Murine Lymphoma. Nature, 181:1343, 1958. 

Serial No. _DBS-l9 

1 . Laboratory of Viral Prodi 

2. Section on Testing 

3 . Betheada 


Individual Project Report 

Calendar Tear 1958 

Part A. 

Project Title: Functions Associated with the Poliomyelitis Vaccine 
Safety Testing. 

Principal Investigator: Dr. Samel Baron 

Other Investigators: Dr. Gerald L. Van Hoosier 
Dr. Ruth Kirschsteln 
Dr. George A. Hottle 
Mrs. Alice M. Gochenour 
Mr. Richard J. Lov 

Cooperating Units: None 

Man Tears: 

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

Project Description: 

Sub-project A: Concentration of Poliomyelitis Vaccine and the 
Sensitivity of the Monkey Safety Test. 

Objective s : To Increase the sensitivity of the monkey safety test 
by increasing the effective volume tested in monkeys. 

Methods Employed : Experimentally infected poliomyelitis vaccines 
are concentrated 100-fold by the ultracentrifuge-gelatin method 
developed in this laboratory. The concentrate is inoculated iv 
spinal ly in ons group of monkeys and unconcentrated material is 
inoculated into an equivalent group of monkeys. Comparison of 
paralytic rates, pathological findings and virus Isolation is made 
between the two groups. 

Ms J or Findings : Monkeys inoculated with concentrated experimental 
vaccines were infected with poliovirus more frequently than those 
monkeys Inoculated with equivalent but unconcentrated material. The 

increased infectivity was correlated with the degree of concentra- 
tion. This monkey safety test using concentrated vaccine was 
3oore sensitive or at least equally sensitive to poliovirus in 
experimental vaccines than was the monkey kidney cell culture 
test when equal volumes of vaccine were used. 

Significance to the Division s The use of concentrated vaccine 
in the monkey safety test appears to be the most sensitive tech- 
nique developed for the detection of residual live virus in 
poliomyelitis vaccine. 

Proposed Course ; To use the test on a more routine basis for 
currently produced vaccine ia order to gain more experience. 

Sub-project B; Determination of Suitability of Vervet Itonkeys for 

Poliomyelitis Vaccine Safety Testing and Viral Studies. 

Objectives ; To determine whether vervet monkeys and vervet cell 
cultures are as sensitive as rhesus monkeys to poliovirus in 
experimental residually infected vaccines. 

Methods Employed; Comparative monkey and cell culture poliovirus 
infectivity titrations in vervet and rhesus monkeys and in 
monkey kidney cell cultures. 

Major Findings; Vervet monkeys and rhesus monkeys are equally 
sensitive to untreated or formal in- treated poliovirus. Vervet 
kidney cell culture is twice as sensitive to untreated poliovirus 
as is rhesus kidney cell culture. There is no indication that 
vervet culture is more sensitive to formal in- treated poliovirus. 
Kidney cell cultures are more sensitive than monkeys to untreated 
poliovirus^, but less sensitive or equally sensitive to fom&lin- 
treated virus. 

Signi ficance to the Division ; Development of alternate sethods 
of vaccine safety testing in the event that rhesus monkeys become 
unavailable. Search for mora sensitive safety test ssathods. 
Evaluation of cell culture aad monkey safety tests. 

Proposed Course; The project is completed. 

Sub-project Cs Sensitivity of Various Cell Cultures to Poliovirus. 

Objectives; Comparison of poliovirus sensitivity of various call 
cultures by roller tube and plaque infect ivlty titrations with 
rhesus asonkey kidney cell cultures. 

Determination of poliovirus sensitivity of rhesus monkey 
kidney cell cultures before and after use in the poliomyelitis 
vacc'.ne safety test. 

Methods Employed : Comparative roller tube and plaque titrations 
of polioviru8 in appropriate cell cultures. 

Major Findings : Many cell cultures derived from human and 
monkey tissue arc as sensitive as rhesus kidney cell cultures 
to poliovirus. Two cell cultures (H. Ep-2 and the DBS strain 
of HaLa) are 2-fold more sensitive to untreated poliovirus. 
Routine determination of the poliovirus sensitivity of cell 
cultures used for the safety test have demonstrated consistent 
high sensitivity. 

Significance : Development of alternate or improved methods for 
the cell culture safety teat. Control and evaluation of the DBS 
safety test. 

Proposed Course : Use of the newly discovered highly sensitive 
cell cultures to detect poliovirus in experimental residually 
infected vaecinen in comparison with rhesus kidney cell cultures. 
Continued control, of sensitivity of cell cultures used for the 
vaccine safety test. 

Part B included Yes / 7 Mo jTl 

Serial Ko. DBS- 2Q 

1. Laboratory of Viral Product 

2. Laboratory Bait Ho. 
3« Bethesda 


Individual Project Report 

Calendar Year 1953 

Part A. 

Project Title: Measurements of poliovirus antibodies by the 
metabolic inhibition test. 

Principal Investigator: Dr. Bugene V. Harnett 

Other Investigators: Mrs. Beatrice L. Burch 

Cooperating Units: None 

Man Years: 

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

Project Description: 

A continuing project which includes screening monkey sera 
of nonkeys used in the monkey potency teat, titrating post- 
vaccination immune monkey sera, screening chicken potency sera, 
guinea pig potency sera, and human sera collected in field studies. 

During 1958 human sera froo the Communicable Disease Center and 
the World Health Organisation have been titrated for poliovirus 

Vaccines of foreign and domestic manufacture have bean 
tested for potency in chickens, monkeys, and guinea pigs and 
the serologic response measured in this laboratory. 

Part B included Yes /~ Mo jTJ 

Serial Ho. DBS-£i 

1. Laboratory of Viral Produci 

2. Animal Testing Section 

3. Bethesda 


Individual Project Report 

Calendar Tear 1958 

Part A. 

Project Title: Monkey Safety Teat for Poliomyelitis Vaccine and 
Adenovirus Vaccine and Potency of Poliomyelitis 
Vaccine in Monkeys and Chicks. 

Principal Investigator: Dr. Gerald L. Van Hoosier, Jr. 

Other Investigators: Dr. Ruth Kirschsteln 
Dr. Eugene Bernett 
Dr. Paul Gerber 

Cooperating Units: None 

Man Years: 
Total: 11.3 
Professional: 0.3 
Other: 11.0 

Project Description: 

Monkey Safety Test for Poliomyelitis Vaccine and Adenovirus Va ccine 

Objectives : To test poliomyelitis and adenovirus vaccines for 
virus which will Infect monkeys and to test the potency of poliomye- 
litis vaccine. 

Methods Employed : Outlined in the PBS Regulations for the 
above vaccines. 

Findings : 1. Safety Tests: During 1958, 77 poliomyelitis 
vaccines and 5 adenovirus vaccines were tested for safety in monkeys. 
Among 1,560 monkeys, examined by Dr. Kirschsteln during this period, 
lesions of some kind were found in the CHS of only 10 monkeys. In 
3 of the animals,, the pathologic lesions were suggestive of e specific 
etiologlc agent and each of these was confirmed by virus isolation. 

- 2 - 

2. Potency testa: During the period January 1 to December 1, 
1958: 12 Potency tests in monkeys (*) 
129 Potency tests in chicks (2) 

(1) Bach test consisted of 12 monkeys. 

(2) Each test consisted of 45 chicks. 

The chick test is under consideration for replacement of the 
potency test in monkeys. 

Significance to the Institut es Integral part of the DBS program. 

Proposed Course : To cosstisiue in 1959 on about the saase scale. 

Part B included Yes /sT So / / 

Serial Ho. DBS 


Individual Project Report 

Calendar Year 1958 

Part 8 ; Honors, Awards, and Publications 

Honors and Awards relating to this project: 

In January 1958, 13 members of the monkey safety testing group 
received incentive awards for work done on poliomyelitis vaccine 
safety testing in 1956. 

Serial Ho. DBS -2 2 

1. Laboratory of Viral Products 

2. Animal Testing Section 

3 . Bethesda 


Individual Project Report 

Calendar Tear 1958 

Part A. 

Project Title: A Comparison of the Sensitivity of Rhesus and 
Verve t Monkeys to Untreated and Formaldehyde 
Treated Poliovirus. 

Principal Investigator: Dr. G. L. Van Hoosier, Jr. 

Other Investigators: Dr. Sanuel Baron 

Dr. Ruth Kirschstein 
Mr. 6. Rusten 

Cooperating Units: None 

Man Tears: 
Total: 0.3 
Professional: 0.1 
Other: 0.2 

Project Description: 

Objectives : To determine if Verve t monkeys are satisfactory 
test animals for the monkey safety testing of poliomyelitis vaccine, 

Methodo Employed : Routine. 

Major Findings : Vervet monkeys may be used instead of Rhesus 
for the safety testing of poliomyelitis vaccine. 

Significance to the Division : Vervet monkeys may be used for 
tests if Rhesus or Cynomologus are unavailable. 

Proposed Course: Project completed; report in progress. 

Part B included Tes / 7 No /x7 

Serial Ho. PBS-g3 

1 . laboratory of Viral Pr< 

2. Animal Xestiag Section 

3. Bethasda 


Individual Project Report 

Calendar Tear 1958 

Part A. 

Project Title: Studies on the Monkey Beuroviruleace of Attenuated 

Principal Investigator: Dr. Gerald L. Van Hoosier, Jr. 

Other Investigators: Dr. Joseph L. Kelnick 
Dr. C. P. Li 
Dr. Ruth Xirschstein 

Cooperating Units: None 

Man Tears: 
Total: 0.4 
Professional: 0.1 
Other: 0.3 

Project Description: 

Objective : To determine the neurovirulence of attenuated 
strains of poliovirus used experimentally for human immunisation. 

Methods : Intracerebral and intraspinal inoculation of monkeys . 

KM or Findings : All of the attenuated polioviruseo thus far 
examined are capable of producing signs of poliovirus infection af 
intraspinal Inoculation. All strains produce CMS lesions visible on 
histological examination when injected infcraspinally. Pew, if any 
signs of poliovirus infection are observed after inoculation by other 
routes although occasionally histopathological lesions of polionya- 
litis are found. 

Significance: This project at present provides s. basic oriea- 
tation for further studies should a license application be submitted 
for the use of attenuated poliovirus vaccines. It is anticipated 
during the next year, activities vlll be considerably expended. 

Proposed Course: A report of findings has been submitted; in 
as part of Dr. Melnick'u publication. Further studies are planned m 
soon as space permits . 

Part B included Yes / 7 Mo /X_/ 

Serial No. DBS -2$ 

1. Laboratory of Viral ?v< 

2. Animal Testing Section 

3 . Bethesda 


Individual Project Report 

Calendar Tear 1958 

Part A. 

Project Title: Studies on B-virua. 

Principal Investigator: Dr. G. L. Van Hoosier, Jr. 

Other Investigators: Dr. Ruth Kirschstein 
Dr. C. P. Li 

Cooperating Units: None 

Man Tears: 

Total: 0.2 
Professional: C.l 
Other : 0.1 

Project Description: 

Objectives : (1) To study the nature of B-virua infections in 
monkeys aa encountered in the routine testing of poliomyelitis vaccine 

(2) To study the characteristics of B-virua in tissue culture. 

(3) To determine if persons working with monkeys develop neu- 
tralizing antibody to B-virua and/or Herpes simplex. 

Major Findings : (1) The intraspinal inoculation of poliomys> 
vaccine into monkeys occasionally activates a B-virus infection. 

(2) B-virua forms intramuscular inclusions inmmonkey kidney 
tissue culture as do several simian viruses. 

(3) Some individuals working with monkeys do have neutralising 
antibodies to B-virus with titers as high as 1:16. Human gamma 
globulin also neutralizes the virus. These antibodies are probably 
the result of a common antigen between Herpes simplex and B~virus . 

Significance : Helps to understand the nature and characters 
of an agent occasionally infecting humans. 

- 2 - 

Propo sed Course ; Report for publication in progress. 
Discontinued because of lack of proper safety facilities, 
technical difficulties with the neutralisation test for 
Herpes simplex. 

Part B included Yes / 7 Mo jjTJ 

Serial Mo. DBS •£$ 

1. Laboratory of Viral Products 

2. Animal Testing Section 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A, 

Project Title: Studies on B-virus. 

Principal Investigator: Dr. G. L. Van Koosier, Jr. 

Other Investigators: Dr. Ruth Kirschstein 
Dr. C. P. Li 

Cooperating Units: Hone 

Man Tears: 
Total: 0.2 
Professional: 0.1 
Other : 0.1 

Project Description: 

Objectives : (1) To study the nature of B-virus Infections 
in monkeys as encountered in the routine testing of poliomyelitis 
vaccine. (2) To study the characteristics of 3-virus inttissue 
culture. (3) To detercine if persons working with monkeys 
develop neutralizing antibody to B-virus and/or Herpes staples. 

Ma jor Pindings : (1) The intraspinal inoculation of poliomye- 
litis vaccine into monkeys occasionally activates a B-virus 
infection. (2) B-virus fores intramuscular inclusions in M.K.T.C. 
as do several simian viruses. (3) Some individuals working with 
monkeys do have neutralizing antibodies to B-virus with titers 
as high as 1:16. Buaan gamma globulin also neutralises the virus. 
These antibodies are probably the result of a cooson antigen 
between Herpes simples end 3-virus. 

Significance to the Division : Helps to understand the nature 
and characteristics of as agent occasionally infecting humans. 

Serial Bo. DBS -2 6 

1. Laboratory of Viral Prodi 

2. Laboratory Unit 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: The Pathogenesis of Viral Diseases. 

Principal Investigator: Dr. Ruth L. Kirschstein 

Other Investigators: Dr. George Hottle 

Dr. Gerald Van Hoosier 

Dr. Samuel Baron 

Dr. Lawrence Kilham (LBP) 

Dr. Alan Rabson (NCI) 

Dr. Bernice Eddy 

Dr. Robert Huebnar (MIAID) 

Dr. Wallace Rove (NIAID) 

Dr. Isidore Brodsky (NIAID) 

Dr. Hilton Levy (NIAID) 

Cooperating Units; Laboratory of Bacterial Products (DBS) 
Pathologic Anatomy Branch (NCI) 
Laboratory of Infectious Diseases (NIAID) 

Man Years : 

Total . 1 
Professional; 1 
Other : None 

Project Description: 

This activity represents the studies in the field of 
pathology carried out in relation to the various phases of the 
work of the Division where this is needed both in control and 
research activities. The various sub-projects with descriptions 

1. (With Drs. Hottle, Van Hoosier and Baron) The pathol- 
ogy of the central nervous system of monkeys in the safety test 
for poliomyelitis vaccine. 

As outlined in the Current Regulations for the monkey safety test 
for poliomyelitis vaccine, histologic sections of lumbar and 
cervical spinal cords of monkeys inoculated with vaccine are 

- 2 - 

examined for the presence of lesions. One thousand, five hundred 
and sixty monkeys were examined in 1958. Lesions were 
encountered on tan occasions . On three occasions the pathologic 
lesions were suggestive of a specific eticlogic agent and these 
were confirmed by virus isolation. It is expected that this 
endeavor will continue. 

1(a). The comparative pathology of intraspinally induced 
poliomyelitis in Rhesus and South African monkeys. It has been 
found that these animals are equally susceptible to She disease. 
This project is completed. 

1(b). The pathology of "B-virus" in monkeys as seen in 
the safety teat. The lesions differ from the naturally occuring 
disease as described by Keeble et al . and seem worthy of report . 

1(c). The comparative pathology of partially inactivated 
strains^ attenuated strains asjd fully virulent strains of polio- 
myelitis in monkeys. The value of the concentration technique to 
uncover trace amounts of virus has been shown. A method for 
standardisation of degree of attenuation of the viruses is being 
developed. It is expected that this endeavor will continue and 
increase in importance as the possible use of "Live Virus Polio- 
vaccine" is stressed. 

1(d). The pathology of the central nervous system of 
monkeys Inoculated intraspinally with ECHO 9 virus and adenoviruses 
3 P 4, and 7. These agents are apparently quite innocuous and seem 
to give limited, if any, lesions. It is not expected that this 
study will continue. 

2. (With Drs. ?an Hoosier and Baron) Induction of polio- 
myelitis in parakeets. Attempts to induce poliomyelitis in 
parakeets have been made in order to follow up a report of the 
isolation of the virus from a parakeet which was the pet of a 

child who died of bulbar poliomyelitis (Lancet P £;512 [Har. 8, 1938]) 
Inoculation of 12 parakeets intramuscularly, intraspinally, and 
intraperitoneal ly with high titers of all three types of polio- 
viruses has failed to cause any clinical or histologic evidence 
of poliomyelitis. Thus, it would seem that parakeets are probably 
resistant to infection with this virus. This will be so reported. 

3. (With Drs. Rabson and Kilhaxa) The pathology of the 
lesions induced in suckling squirrels and rabbits with the fibroma 
viruses. The viruses cause widespread disease and death in the 
suckling animals. The squirrel fibroma virus causes lesions in 
the lungs of suckling squirrels which resemble pulmonary adenoma- 
tosis is seen in man and other anitaals. This is the first time 

- 3 - 

that the lesion of pulmonary adenomatosis has been conclusively 
shown to be caused by a virus . 

4. (With Dr. Van Hoosier) The pathology of the lesions 
of the newly isolated "monkey tumor viru3." This agent has been 
isolated in Nigeria (Nature, July 19, 1958) and was sent to us 
by Dr. C. H. Andrewes, National Institute for Medical Research, 
England. The agent causes subsutaneous tumors, the cells of 
which contain large viral intracytoplasmic inclusion bodies. In 
light of the results indicated in No. 3, attempts to study the 
agent in suckling monkeys will be made as these animals become 

5. (With Dr. Eddy) Lesions in suckling rabbits induced 
by the polyoma virus . Histologically, they consist of inter- 
digitating bundles of proliferating fibroblasts resembling 
similar lesions in man known as "Fibromatoses." This report 
has been submitted for publication. 

6. (With Drs. Huebner, Rowe, Brodsky and Levy) The 
pathology of a mouse "leukemia-like" lesion induced by a^viral 
agent The filterable agent isolated from mouse Ehrlich's 
ascites cell tumor has now been serially passaged through several 
hundred passages. The pathologic picture is, perhaps, more 
clearly defined. In its earliest stages, the lesion is confined 
to the spleen, but later spreads to involve the liver, bone marrow 
and occasionally other organs. The process, in its late stages, 
appears histologically indistinguishable from a malignant process 
of hematopoietic tissue. The process does not seem to vary in the 
suckling animal. Transplantability of the "tumor" has thus far 
not been accomplished. Splenectomy allows the animals to survive 
longer, but does not vary the ultimate course of the disease. 
X-irradiation and chlorambucil can halt the process temporarily, 
but again, have no effect on the ultimate course of the disease. 

Continued efforts to transplant the tumor will be made. 

Part B included Yes /X_/ No / — / 

Serial No. DBS 


Individual Project Report 

Calendar Year 1958 

Part B ; Honors , Awards, and Publications 

Publication other than abstracts from this project; 

Kirschstein, R. L., Rabson, A. S., and Kilham, L.s Pulmnnary 
Lesions Produced, by Fibroma Viruses in Squirrels and Rabbits. 

Cancer Research. (In press) 

Serial No. DBS -27 
PHS-NXH L Laboratory of 

Individual Project Report Viral Products 
Calendar Year 1958 2. Laboratory Unit 

No, 4 
3. Bethesda 

Part A. 

Project Title: Studies of the Inactivation of Viruses, 

Principal Investigator: C W. Hiatt, Ph. D. 

Other Investigator: Jerome J. Helprin, Ph. D„ 

Cooperating Units: None. 

Man Years: Patient Days; None 

Total: 1.50 
Professional; loOO 
Other- 0-50 

Project Description: 

Oblectives . 

1„ To gain information about the fundamental nature of 
the various biological properties of viruses through the study 
of the mechanisms of inactivation of these properties by chemical 
and physical agents. 

2. To develop an understanding of processes of inactivation 
which are applicable to the preparation of viral vaccines. 

Methods Employed. 

Various animal and bacterial viruses are cultivated in cell 
cultures and the infective fluids from these cultures are subjected 
to treatment with graded doses of the inactivating agent, Samples 
taken at the different dose levels are assayed for infectivzty„ and 
the kinetic relationship between dose and survival ratio is estab- 
lished. From the characteristics of the survival curve, Inferences 
are drawn about the properties of the virus and the mechanisms of 
the inactivating process. 

- 2 - 
Ma for Findi ngs* 

1 . Re lative , susceptibil i ty of animal ^ viruse s. to. in- 
activation by photodynamic action . 

When tissue-culture fluids infected with various viruses 
are Irradiated with Intense polychromatic visible light after 
addition of trace amounts of a thiazine dye (e.g., toluldlne 
blue) there is a photosensitized oxidative reaction which des- 
tsoys infectivity. The species of viruses tested have been ob- 
served to vary greatly la their susceptibility to Isaac tivafc ion 
by this process „ Members of the enteric group (poliomyelitis, 
Coxsackie A9, ECHO 1) are very resistant » with half -Uvea under 
standard conditions of 20 to 40 minutes. BCBO 10 is distinguish- 
able by a shorter half -life of about 10 minutes . Vaccinia 
virus, on the other fcand s is highly susceptible to this treat- 
ment;, with a half-life of only 6 seconds. The several adeno- 
viruses tested are also rapidly inactivated, with halJi-lives 
of only a few seconds „ Information obtained in this investi- 
gation may prove useful in the classification of viruses and is 
also of interest in connection with the preparation of viral 
vaccines o 

2. The mechanism of >ii i^ctiva i t i< ion i of, coliphagesjpy 
ph otodynamlc action ., 

The photodynamlc action of toluldlne blue on T2 and T3 coll- 
■phages reveals that these two phage types are strikingly different 
in their uptake of the dye. T3 phage absorbs the dye very rapidly, 
and consequently the rate of ia&ctlvation is independent of the 
time of contact with dye before irradiation. The uptake of dye by 
T2 phage 9 in contrast , Is very slow. Its rate of photodynamlc in 
activation,, therefore is greatly Influenced by time of contact 
with the dye. The temperature dependence of the rate of dye uptake 
is pronounced (Q10b3»8) and confirms the existence of a permselective 
membrane around this phage. 

3 . Petenaiaatloa .of iii «cgion spectra, for the iaaetivation 
of r v irus es ^ by ni the , phofcpdyaa m ic i ac tion of various dye s , 

Equipment has been constructed for irradiation of stationary 
films of liquid with monochromatic light of precisely «tef;®rg»inad 
wavelength and intensity* This equipment, when perfected in ©per» 
ation 8 will be used to determine reaction rates and quantum yields 
for the iaaetivation of T3 coliphage with various dyes and at 

- 3 - 

cloaely spaced wavelength intervals between 2900 and 7500 A. 
By study of the action spectra thus obtained it may be possible 
to specify the state of excitation of the dye molecule in the photo- 
dynamic reaction 

4o Method for the microbiological assay of free form- 
aldehyde in biological products ,, 

A detailed method has been developed for the assay of micro- 
bicidal formaldehyde at various stages in the inactivation of viral 
vaccines with formalin. The method is based upon a series of dose* 
response curves for the inactivation of T2 coliphage with* 
hyde at 37 °C. The formaldehyde free to attack the phage can be es- 
timated with an uncertainty of less than _ 5% when the formaldehyde 
concentration is between 10 and 100 micrograms per ml. Present 
chemical methods for distinguishing between free and bound formal- 
dehyde suffer the disadvantage that the formaldehyde which is "free" 
for a chemical reaction may not be available for microbicidal action 
The bacteriophage method ab orjgine escapes this disadvantage, hut 
the data collected thus far are not sufficient to establish whether 
or not it will be useful analytical tool* 

5. The effect of transverse mixing upon the rate of 
jnactivatlon of microorganisms in continuously flowing thin films of 
liquid irradiated with ultraviolet light * 

A theoretical study of the distribution of light intensity in 
thin films of liquid in centrifugal-film irradiators was completed, 
and the influence of layer thickness and fluid absorbance was des- 
cribed for the two limiting cases: a) turbulent flow, with nearly 
perfect mixing, and b) laminar flow with no mixing. This study,, whic'r 
will be terminated upon preparation of a report for publication, will 
assist in defining the range of application of continouous-flow irra- 
diation equipment in sterilising biologicalSo 

6. Compilation of data concerning the kinetics of viral 
inactivation . 

Data from the literature pertaining to the kinetics of in- 
activation of any virus by heat, radiant energy, chemical agents., 
or any other method which is amenable to quantitation are being 
collected as part of a continuing effort to maintain a broad 
familiarity with the general subject. These data are being 
studied and compared to isolate patterns of response which may 
serve to define an integrating principle „ A review article on 
this subject is being prepared for publication. 

The continued Investigation of the mode of action of 
virus laactivants is expected to contribute to the fundamental 
basis for the control of viral vaccines. 

Proposed Course T o f Proje ct. 

The lines of investigation described under "Major Findings" 
will be continued and extended with the exception of item 5« which 
will be terminated upon completion of the published report and 
item 4, which will be brought to a close as soon as the necessary 
data have been collected,, 

(Part B included) Yes/TJ Ho / 7 

PBS-NIH Serial Ho. DBS- 

Individual Project Report 
Calendar Year 1958 

Part B: Honors, Awards B and Publications 
Publications other than abstracts from this project! 

Helprin, J. J., and Biatt, C. W., Photosenaitization 
of T2 coliphage with toluidine blue., J. Bacterid, <in publication. 

Honors and Awards relating to this project: None, 

Serial Ho. DBS- ZB 
FHS-NIH l. Laboratory of 

Individual Project Report Viral Products 
Calendar Year 1958 2. Laboratory Unit 

No. 4 
3. Bethesda 

Part A . 

Project Title: Physical and Chemical Characterization of Viruses 

Principal Investigator: C. W. Hiatt, Ph. D. 

Other Investigators: None. 

Cooperating Units: None. 

Man Years: Patient Days: None. 

Total: 0o83 
Professional: 0,33 
Other: 0.33 

Project Description: 

Objective . To obtain information about the physical dimensions 
and chemical composition of animal viruses and to correlate this 
information with biological properties. 

Methods Employed . Standard methods of physical and chemical 
analysis, including ultracentrifugation and electron microscopy, 
are supplemented by specialized techniques for concentrating and 
purifying viruses. 

Major Findings . A technique has been developed for rapid elec- 
tron microscopic visualization of hemagglutinating viruses. The 
virus is adsorbed on erythrocytes as in the customary hemagglutin- 
ation procedure, but with very low concentrations of erythrocytes 
to reduce the probability of agglutination. The erythrocytes are 
then simultaneously lysed and fixed by treatment with increasing 
concentrations of formaldehyde. The fixed stroma, with virus par- 
ticles still adherent to their surfaces, are then dried, shadowed 
with chromium, and observed in the electron microscope. This tech- 
nique, which is presently being applied to the study of several new™ 
ly isolated viral agents, differs from other methods employing stroma 
in that it does not require preparation of the stroma prior to ad- 
sorption of the virus. 

Significance to the Program of DBS . The development of methods 
for the rapid characterization of newly identified viruses will 
assist in the testing of viral vaccines for safety and purity. 

Proposed Course of the Project. This project will be contii 
and expanded in scope with emphasis placed upon rapid methods of 
characterization which may be applied to a wide variety of viruses. 

(Part B not included.) 

Serial No. mS-2.9 

1 . Laboratory of Viral Prodtsc 

2 . Laboratory Unit 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A 

Project Title: Respiratory Diseases 
Principal Investigator: Dr. Bernice E. Eddy 
Other Investigators: None 
Cooperating Units: None 

Man Years: 

Total : 8 
Professional: 1 
Other : 7 

The Unit is concerned with four primary projects and one 
minor project. These are: Influenza viruses, adenoviruses, 
vaccines for multiple respiratory agents, tumor viruses and 
pneumococci . 

Methods Employed : The methods employed for these projects 
consist of animal inoculation, propagation of viruses in animals, 
embryonated eggs and tissue culture made from various tissues from 
different animals, propagation of bacteria in culture media, 
centrifugation, differential filtration and serological methods. 

I. Project Title: Influenza Viruses 

Man Years: 

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

Objectives: ' The objectives of the project are to control 
and improve influenza virus vaccines and to improve methods for 
evaluating the safety, purity and potency of the product. 

The disease influenza, was a problem in the U. S. during 
the first part of the year and interest in ionization with 
influenza virus vaccines has continued. From January 1 to 
December 7 1958, 90 lots of influenza virus vaccine, some made 

up of as much aa 650 liters, were considered for release as 
f ol lows i 

Lots submitted in Lots submitted Lota released 
Va c cine 8 latter part of 1957 in 1958 in 1953 


(Asian strain) 1 5 6 

Polyvalent military 

(six strains) 18 17 

Polyvalent civilian 

(four strains) 21 45 55 

Total 22 68 78 

Three lots of vaccine for civilian use were withdrawn by the 
manufacturer, one lot of vaccine for military use failed and 
disposition of it has not been made, and eight lots are under test. 

A license was issued to one :-:ew manufacturer for the pro- 
duction of influenza virus vaccine. Strains of virus were supplied 
to this manufacturer and numerous consultations on problems 
connected with requirements and testing were held. 

Another manufacturer submitted two lots of diagnostic 
influenza antigens along with an application for a license. 

Major work on the project s A. Potency tests on influenza 
virus vaccines. Tests for viral content fry the chicken cell 
agglutination (CCA) method were made on six lots of vaccine sub- 
mitted for release and antigenic potency tests in mice were per- 
formed on 78 lots of vaccine submitted for release and on one lot 
of vaccine from Germany. 

8. Control of diagnostic influenza virus antigens. Ho 
licensed diagnostic antigens were submitted for release and of two 
lots submitted with an application for license which were tested, 
one lot was satisfactory and the other was not. 

C. Chicken cell agglutination (CCA) test for virus content. 
The CCA test for virus content of laonovalent influenza virus 
vaccine varies under different conditions and much tiras was spent 
in determining the cause of the variations and in working with the 
manufacturers in an effort to standardise the test. Laboratories 

- 3 - 

in other countries also had r.ifficulty with the test and CCA 
determinations were made on seven lots of influenza virus 
vaccine for Dr. Isaacs in England. Comparative tests of the 
CCA method with a simpler, less time consuming hemagglutination 
test are underway and 91 lots of vaccine were tested by both 
methods . 

D. Reference standards for influenza virus vaccines. 
Three new reference influenza virus vaccines were obtained, tested 
and distributed to the manufacturers. Two were adopted as official 
reference standards during 1958. 

(a) Vaccine, Lot 811, was adopted as a reference for the 
mouse antigenic tests for civilian formula influenza virus 
vaccine containing the PR-8, PR- 301, Asian and Great Lakes 

(b) Vaccine, Lot XI, was officially made the reference for 
the mouse antigenic tests for military formula influenza 
virus vaccine, containing the Swine, PR-8, PR-301, Asian, 
Lee, and Great Lakes strains. 

(c) Vaccine, Lot CCA6, was tested for CCA value under dif- 
ferent conditions for suitability as a reference preparation 
for the CCA test. Samples were also sent to the different 
manufacturing laboratories for comparative testing. When 
the results of all tests are available, a CCA value will be 
assigned and the vaccine will be used to replace the present 
reference preparation, Lot CCA5. 

Along the same line, a large pool of Jap 305 allantoic 
fluid was prepared in an attempt to make a stable dried 
reference standard for the CCA test. The material will be 
dya'. /;-.ed, dried, and extensive tests will be carrted out 
to determine the viral content of the preparation. 

(d) Studies of the antigenicity of current influenza virus 
strains. This is necessary to make certain that the strains 
cursently causing disease are similar to strains in the 
vaccines. One strain* supplied by the Armed Forces Comnissios 
on Influenza, is being studied for possible substitution for 
the PR 301 strain component of the military and civilian 

* Ann Arbor strain, AA/1/57, type B. 

- '■:■ 

The general aim of the project is to fulfill the obliga- 
tion of controlling and improving influenza virus vaccine. The 
oroposed course of the project will follow along the same course 
as in 1958 with core emphasis on studies of new strains of influ- 
enza viruses. 

XI. Project Titles Adenoviruses 

Man Years; 
Total: 1 2/3 

Professional : 1/6 
Other; 11/2 

Objectives ; The objectives of the project are to control 
and improve adenovirus vaccine and to improve methods for evalu- 
ating the safety, purity and potency of the product. 

Adenovirus vaccines have been of more interest to the 
military than to civilians, since new troops are particularly 
susceptible to adenovirus infections. During 1958, three lots 
were submitted for release by a licensed manufacturer and six 
lots were submitted by a manufacturer requesting a license for 
the product. 

Major w ork on the project; (a) Potency of adenovirus 
vaccines for types 3, 4, and 7. Different raafchods for determining 
the potency of adenoviruses vaccines were tried out and one method 
involving the immunization of guinea pigs and the use of their 
sera in a serum-virus neutralisation test in known numbers of HeLa 
cells was used to test all adenovirus vaccines that have been sub- 
mitted. Efforts are being continued to simplify and improve the 

(b) Studies of the susceptibility of tissue culture cells 
for the adenoviruses and inhibitors for the viruses in normal sera. 
These studies are of importance in safety tests of the adenovirus 


(c) Reference vaccine. A lot of vaccine used in a field 
study has been used as a reference vaccine for all potency tests. 
Plans are underway to obtain new monovalent lots of vaccine to test 
for suitability for pooling and use as a reference preparation. 

The general aim of the project is to fulfill the obligation 
of controlling and improving adenovirus vaccine. The proposed 
course of the project will fellow along the same course as in 1958. 

HI. Project Title: Vaccines for multiple respiratory agents. 

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

Objectives : This project i3 for the purpose of anticipating 
the need for the control of multiple antigen vaccines. (One manu- 
facturer is currently working on a vaccine that contains eleven 
different viruses.) 

Major vork on the project : Pools of adenoviruses of types 
other than 3, 4, and 7, HA and CA viruses are being prepared and 
tested for virus content. 

The production of humoral antibodies in animals and rapid 
testa for antibodies for each of the antigens is a projected 

It is important that information and tests, if possible, 
are in readiness before requests for the license of a new product 
are received. 

IV Project Title: Tumor Viruses. (This work has been done in 

conjunction with Dr. Sarah 
Stewart, MCI) 

Man Years: 

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

Objectives: The objectives of the project are to isolate 
a virus or viruses from human neoplasms and to find a way to prevent 
neoplasia in man. (The work was started in an effort to gain infor- 
mation on the etiology of lung cancer.) 

Ma lor F indings : Work has centered on a tumor virus 
(SE polyoma) isolated from three sources of strain AKR mice. The 
virus has induced over 20 histologically different neoplasia in 
mice; sarcomas and angiomatous lesions in hamsters and rats; and 
benign nodules in rabbits. The virus has been characterized by 
filtration, production of cytopathogenicity and plaques in tissue 
cultures, hemagglutination, drying, storage at different tempera- 
tures, antigenicity, etc. See part B. An attempt xs being made 
to use the SE polyoma virus as a model for the recovery of a 
virus or viruses from human neoplasms. 

- 6 - 

ung cancer or any cancer is an infectious disease, 
there are prospects that a vaccine might be effective in pre- 
venting it. The propagation of the etiological agent, is the 
first requisite for the preparation of such a vaccine. 

VI. Project Titles Pneumococci 

Man Years: Insignificant 

Objectives: The objective of the project is to control 
pneumococcus typing sera. This is a minor project; only four 
lots of pneumococcus typing sera were released in 1953 . 

Major Findings : The nomenclature of the Danish and 
American types were correlated and a paper jointly with 
Dr. ?. lauffmann and Dr. E. Lund is in preparation. 

Part B included Yes /TJ Ho / 7 

Serial No. BBS 


Individual Project Report 

Calendar Year 1958 

Part B: Honors, Awards, and Publications 

Publications otber than abstracts from this project: 

1. Eddy, B. E., Stewart, S. E., Young, R. and Mider, G. B. 

Neoplasms in hamsters induced by mouse tumor agents passed 
in tissue culture. J. Nat. Cancer Inst., 20:747-760, 1958. 

2 Eddy, B. E., Stewart, S. E. and Berkeley, W. Cytopathogenicity 
in tissue cultures by a tumor virus from mice. Proc . Soc . Exp. 
Biol, and Med., 98:848-851, 1958. 

3. Eddy, B. E., Rowe, W. P., Hartley, J. W., Stewart, S. E. and 
Huebner, R. J . Hemagglutination with the SE polyoma virus. 
Virology, 6 290-291, 1958. 

4. Eddy, B. E., Stewart, S. E. and Grubbs, G. E. Influence of 
tissue culture passage, storage, temperature and drying on the 
SE polyoma virus. Proc. Soc. Exp. Biol, and Med., 99;289-293„ 

5. Eddy, B. E., Stewart, S. E. Stanton, M. F. and Marcotte, J. 
The induction of tumors in rats by the SE polyoma virus mouse 
embryo tissue culture preparations. J. Nat. Cancer Inst., 
(In press) 

6. Eddy, B.E. and Stewart, S. E. Physical properties, hemagglu- 
tinating and cytopathogenic effects of the SE polyoma virus. 
Canadian Cancer Research Proc. 3. (In press) 

7. Stewart, S. E., Eddy, B. E. and Borgese R. Neoplasms in mice 
inoculated with a tumor agent carried in tissue culture. 

J. Nat. Cancer Inst., 20:1223-1243, 1958. 

8. Stewart, S. E., Eddy, B. E. and Stanton, M. F. Induction of 
neoplasms in mice by a tumor agent carried in tissue culture. 
Canadian Cancer Research Proc. 3. (In press) 

9. Stewart, S. E. and Eddy, B. E. Properties of a tumor inducing 
virus recovered from mouse neoplasm. Proceedings of Prospectives 
in Virology. Ruthers Univ. Press, New Brunswick, N. Y. 

(In press) 

10. Stewart, S. E., Eddy, 3. E. and Stanton, M. F. Neoplasms 
in certain mammals induced by a tumor agent, the SE polyoma 
virus, carried in tissue cultures. Proc. 7th International 
Cancer Congress (London) . (In press) 

11. Stewart, S. E., Eddy, B. E. and Irwin, M. L. Tumor induction 
by the SE polyoma virus and the inhibition of tumors by 
specific neutralising antibodies. APHA. (In press) 

12. Stewart, S. E. and Eddy, B. E. A review of the biological 
properties of the SE polyoma virus. Leukemia Symposium of 
Hematology Congress (Rome) . (In press) 

Honors and Awards Relating to this Project; 

1 . President of Washington Branch of Society of American 
Bacteriologists . 

2. Vice-President and Board of Managers of Washington Academy 
of Science. 

Serial Ko. DBS-3Q 

1. Laboratory of Viral Produc 

2. Research Section 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Tissue culture assay of smallpox vaccines. 
Principal Investigator: Dr. Ernest Cutchins 
Other Investigators: None 

Cooperating Units: Amy Institute of Research, 

Walter Reed Army Medical Center 

Man Years: 
Total: 3/4 
Professional: 1/4 
Other: 1/2 

Project Description. 

A Previous work has examined the susceptibility of 
vaccinia virus and smallpox vaccines of several tissue culture 
^sterna with the possibility in view that a tissue culture assay 
«i«ht replace the standard rabbit skin scarification test of 
smlllpox^ine. Cultures of primary rabb " «£jj "'ET22 
a compromise in susceptibility and general availability, have been 

^beccTlva: a 1 Tarn lesTf^smatlpox vaccine are comparatively 
assayed ^rabbit kidney tissue culture and by the standard 

of vaccine have been invited to participate in this program. 

B In collaboration with the Army Institute of Research, 
the stability of a dried smallpox vaccine is being studied 

• 2 - 

method (Section on Control Tests) and by egg infectivity and 
pock counts, by rabbit kidney tissue culture roller tube an.d 
plaque counts and human inoculation (Army Institute of Research) 

This project will provide information needed in the 
evaluation of tissue culture assay of smallpox vaccine and the 
setting of standards for dried smallpox vaccine. 

Part B included Yes jTJ Mo l~~l 

Serial No. DBS 


Individual Project Report 

Calendar Year 1958 

Part B : Honors, Award, and Publications 

Publications other than abstracts from this projects 

Cutchins, Ernest and Warren, Joel, Comparative susceptibility 
of cell cultures to vaccinia virus: Application to the 
standardization of smallpox vaccine. Proc. Soc. Exp. Biol, and 
Med., 97:456-462, 1958. 

Serial No. DBS -31 

1. Laboratory of Viral Pro 

2 . Research Section 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A . 

Project Title: Humoral antibody response to smallpox vaccination. 
Principal Investigator: Dr. Ernest Cutchins 
Other Investigators: Rone 
Cooperating Units: Hone 

Man Years: 

1. Total: 3/4 

Professional : 1/4 
Other: 1/2 

Project Description: 

A Conventional techniques utilizing animals, eggc, or 
roller tubes of tissue cultures to measure neutralizing antibody 
have not proved satisfactory for the demonstration of vaccinia 
neutralizing antibody following smallpox vaccination. A neutrali- 
zation test has been developed in which the antibody content of a 
serum is measured by the ability to reduce by 507o the mimber of 
tissue culture plaque forming units of a vaccinia virus suspension 
following incubation with the serum. In children, primary vacci- 
nation has resulted in vaccinia neutralizing antibody titers ol 
1-128-1-512. Among adults, geometric mean titers ranged from j.:Ji 
for those with 10 years or more since vaccination to 1:123 tor those 
who had been re-vaccinated 1 month to 1 year prior to testing. 
Since the above data represent a relatively small number of samples, 
additional sera from persons of varying vaccination histories will 
be tested. 

Objectives of the project are to study antibody response 
at various intervals after re -vaccination and to evaluate the use 
of antibody response to vaccination as a measure of vaccine potency. 

Part B included Yes / / Mo /X_/ 

Serial No. DBS-3Z 

1. Laboratory of Viral Products 

2. Research Section 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Preparation of Experimental Measles Vaccine 

and Collection of Measles Reference Antiserum. 

Principal Investigator: Dr. Ernest Cutchins 

Other Investigators: None 

Cooperating Units: None 

Man Years : 
Total: 7/8 
Professional : 3/8 
Other: 1/2 

Project Description: 

A. Measles virus is being adapted to chick embryo tissue 
culture. A number of procedures aimed towards producing a pre- 
paration having a high virus titer are being tested. Results 
thus far are inconvlusive. Virus adapted to chick embryo tissue 
culture will be used to prepare reference virus, experimental 
killed vaccine, lyophilized and other preparations. 

B. To obtain pools of standardized antiserum for use in 
the Division's research program on measles virus and vaccine, 
human sera are being screened for the presence of measles 
neutralizing antibody. Pour of forty sera tested thus far were 
positive for antibody in a roller tube neutralization test. 
Additional sera will be tested and positive sera will be further 
standardized by plaque neutralization and complement -fixation 

Part B included Yes / 7 No /x*7 

Serial Wo. DBS -33 

1. Laboratory of Viral Products 

2. Eesearch Section 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Antigenic content of poliomyelitis vaccine as 

measured by combination with neutralising antibody. 

Principal Investigator: Dr. Ernest Cutchins 

Other Investigators: Continuation of 

work of Drs. Melniek and Benyeah. 
Cooperating Units: Rone 

Man Years: 
Total: 2 1/8 
Professional: 3/8 
Other: 1 1/2 

Project Description: 

In the continuing effort to accurately measure the potency of 
poliomyelitis vaccine, an in vitro test has been devised. During a 
primary incubation, antigen in the vaccine is allotted to coabine 
with antibody in a standard antiserum. Live virus is then added 
to the mixture . In a second Incubation antibody not already com- 
bined with vaccine antigen is free to neutralize the live virus. 
Aliquots of the mixtures are plated on monkey kidney tissue culture. 
Prom the number of plaque -forming units present is the test prep- 
aration in comparison with those in the control, a ratio of the 
potency of the test vaccine to a control vaccine is obtained. The 
potency of 76 coenarcial and experimental vaccines has been obtained 
by this method. To follow stability, aliquots of a number of vac- 
cines stored under various conditions are being tested at six month 
intervals. The testing of current vaccines will continue. 

Should the test prove a satisfactory indicator of vaccine 
immunizing capacity, it can be used to supplement present polio- 
myelitis vaccine potency tests. 

Part B included Yes / 7 Bo /F7 

Serial No. PBS-,,? 1 *- 

1 . Laboratory of Viral Pi 

2. Laboratory Unit Ko. 3 

3. Bethesda 


Individual Project Report. 

Calendar Year 1958 

Part A. 

Project Title: I. Studies on variation in virulence of poliovirus. 
11. Studies of the virulence characteristics of 

attenuated poliovirus strains. 
III. Studies on mouse sarcoma 180 and human leukemia. 
IV. Studies of proteolytic enzymes produced by 

monkey kidney tissue culture. 
V. Sparing effect of abalone on mice infected x*ith 
VI. Susceptibility of host cells toward Simian virui 

Principal Investigator: Dr. C. P. Li 

Other Investigators: Dr. Robert Kramer 

Dr. C. W. Hiatt 

Dr. R. L. Kirschstein 

Dr. Benjamin Prescott (NIAID) 

Cooperating Units: Bacterial Biochemistry (KIA1D) 

Man Years: 
Total : 6.5 
Professional' 1.5 
Other: 5.0 

Project Description: 

I. Studies on variation in virulence of poliovirus. 

O bjective and Methods : These have been described in the 
annual report of 1957. The objective now is to see whether such 
studies would throw new light on the problem of chemoprophylaxis 
or chemotherapy. 

Work Accomplished : The third paper of this series of 
studies entitled. "Variation in virulence of poliovirus . Ill . On 
Type III by plaque method" was published. (See Part B) It was 
also oresented at the Federation Maeting in Philadelphia on 
Aoril"l5 1958 and an abstract was published in Fed. Proc 1958, 
Vol 17 Ho 1, Part I, 522(2041). The 4th paper entitled, 

"Studies on variation in virulence of poliovirus. IV. The effect 
of 5-bromouracil and stable cell lines" has been prepared and 
approved by the DBS Editorial Committee for publication. This 
paper vas also presented at the International Congress for Micro- 
biology in Stockholm, Sweden on August 7 , 1958 and an abstract 
was published in the Abstracts of the Confess, P273, 15N. 

Current Work : It was found that 5-fluorouracil inhibited 
directly Type III poliovirus both in tissue culture and in mice. 
The degree of inhibition depending on the concentration of the 
5-fluorouracil. Further study on the mechanism of the inhibition 
is in progress, hoping to throw some new light on the problem of 
chemotherapy of experimental poliomyelitis. 

II. Studies of the virulence characteristics of attenuated 
poliovirus strains. 

This is a side line of project I. It was found that 
5-fluorouracil inhibited the attenuated Type I virus (LS-C) much 
more markedly than it did for the virulent Type I virus (Mahoney) . 
The possibility of using the sensitivity to<4?ard inhibition of 
cytopathogenicity by 5-fluorouracil as a marker to differentiate 
virulent and avirulent strains of poliovirus is being studied. 

III. Studies on mouse sarcoma 180 and human leukemia. 

Objective s 1. To devise a simple and quantitative method 

for screening anticancer compounds using sarcoma 180 as a test tool , 

2. To screen a limited number of plant and animal tissue 
polysaccharides or extracts for their anticancer effect. (In 
cooperation with Dr. Prescott) 

3. To see whether sarcoma 130 can be induced by a filtrate 
of the tumor tissue or cells. 

4. To see whether a variant of sarcoma, cells and/or a 
human leukemia cells can be developed, 

Current Work ; Intramuscularly transplanted sarcoma 180 from 
a DBA mouse was passed serially by intracerebral route (IC) in 
newborn randomly bred Swiss mice. Passages were made with 10% 
suspension of minced mouse brain in Eagle's medium with 2% calf 
serum. When the sarcoma cell in the suspension was stained with 
crystal violet, they could be easily differentiated from the brain 
cells and 5 therefore ^, could be counted in the counting chamber. 
To date the sarcoma has been through a total of 30 IC passages. 

- 3 - 

It required about 400 sarcoma cells to give a 100% "take" in 
newborn mice by the IC route. When the brain suspension was 
inoculated subcutaneously into 3-4 week-old mice, it required 
about 400.000 sarcoma cells to give a 100% "take." The sarcoma 
was also adapted to hamsters by the IC route and it is now on 
the 8th serial passage. A strain of TC culture sarcoma cells 
also produced sarcoma in adult mice by subcutaneous injection 
but the strain seemed to be much less neurotropic as revealed 
by IC inoculation into suckling mice. The technique of sub- 
cutaneous injection into adult mice with a suspension containing 
a standard number of sarcoma cells is suggested for screening 
anticancer con-pounds. 

Among a limited number of substances tested for anti- 
cancer effect, 5-fluorouracil was found fairly effective against 
sarcoma 180 . 

Filtration experiments with sarcoma 180 are being conducted. 

IV. Studies of proteolytic enzymes produced by monkey kidney 
tissue culture. 

This work, done by Dr. R. Kramer, was presented at the 
April 1958 Federation Meeting in Philadelphia and en abstract 
appeared in Fed. Proc . , 17(1), Part 1, P521 (2035), 1958. Thin 
project was ended when Dr. Kramer left us and he is preparing a 
manuscript for publication. 

V. Sparing affect of abalone on mice infected with poliovirus. 

The details of this project w re reported in the 1957 
annual report. It is now ended and a manuscript is in preparation. 

VI. Susceptibility of host cells toward Simian virus. 

This project being carried out by Dr. Kramer io now suspends* 
Part B included Yes jTJ Ho /~7 

Serial No. DBS 


Individual Project Report 

Galendar Year 1958 

Part B ; Honors, Awards, and Publications 

Publications other than abstracts from this projects 

1. Li, C. P. and Jahnes, W. G. Variation in Virulence of 
Poliovirus. III. On Type III by Plaque Method. Proc. Soc. 
Exp. Biol, and Med., 98s330, 1958. 

2. An Outbreak of Influenza- like Disease in the Chinese Army 
Medical College in 1941. Am. J. Pub. Health, 48s760, 1958. 

Serial Mo. DBS -3S 

1 . Laboratory of Viral Products 

2. Laboratory Unit No, 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Laboratory Cultivation of Homologous Serum 
Hepatitis Virus. 

Principal Investigetor : Dr. Joseph P. O'Malley 

Other Investigators: None 

Cooperating Units: Hone 

Man Years : 
Total: 0.95 
Professional: 0.75 
Other: 0.20 

Project Description: 

Ptagnents of human liver obtained at laparotomy or by 
needle biopsy are grafted onto the chorioallantoic membrane of 
the developing chick embryo. These grafts are later exposed to 
human plasma known to produce serum hepatitis in human volun- 
teers, in an attempt to produce a cytopathogenic effect in the 
graft tissue. 

Trypsinized tissues from the developing chick embryo and 
various mammals are being screened for cytopathogenic effect 
following inoculation with known icterogenic human plasma. 

The laboratory cultivation of serum hepatitis virus remains 
one of the major unsolved problems of basic and medical virology. 
Its solution would permit the reestablishment of safe human plasma 
transfusions and a more widespread use of blood and blood products. 

Part B included Yes / 7 No /XT 

Serial No. DBS -36 

1. Laboratory of Viral Product 

2. Laboratory Unit No. 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Standardization of Measles and Mumps 
Neutralizing Antibody in Human Gamma 

Principal Investigator: Dr. Joseph P. O'Malley 

Other Investigators: None 

Cooperating Units: None 

Man Years: 

Total: 0.35 
Professional: 0.25 
Other: 0.10 

Project Description: 

Gamma globulin samples representing fractions from 
plasmas pooled in different years from various parts of the 
United States are being screened to determine their measles 
and mumps neutralizing antibody levels. The viruses are tested 
in tissue cultures only. This is the initial step toward the 
establishment of a national standard antibody pool for measles 
and mumps viruses. 

Part B included Yes / 7 No /TJ 

Serial No. Di 

1. Laboratory of Viral Produs 

2. Research Section 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title; Study of Measles Virus and the Serologic Response 
of Experimental Anitaalg to the Virus Grown in 
Tissue Culture. 

Principal Investigator: Dr. Miklos N. Dreguss 

Other Investigators: None 

Cooperating Units: None 

Man Years : 

Total: 1 year, 6 months 
Professional : 1 year 
Other: 6 months 

Project Description: 

Objective : The objective of this research project is to 
atudy measles virus in tissue culture and to develop serologic 
tests for measuring in vitro the antigenicity of the virus,, and 
the titer of the corresponding antibody. 

Course of Investigation : Pirat, measles virus strains were 
studied in tissue culture by using various cell types for propaga- 
tion. Then, the problem of immunologic response has been approach; 
by an investigation of antigenicity of the tissue culture grown 
virus both in experimental animcls and in vitro . 

Methods Used : The tissue culture method of Enders and 
Peebles has been utilized, with some modifications, for propa- 
gating the virus in a variety of primary and continuous -line 
tissue culture cells. Artificial immune sera were produced with 
such virus in experimental animals applying various routes of 
immunization. The correlation between antigen and antibody 
evaluated by serologic methods among which complement -fixation 
was used extensively as best fitted for in vitro studies. 

Findings r. In the preceding year, studies have been 
completed vith a number of primary and continuous cell lines 
suitable for growing measles virus in tissue culture, and 
immune sera were produced by injection of live virus into 
several species of animals. A method of reproducible complement- 
fixation test was worked out, and preliminary studies were con- 
ducted of other serologic methods . 

This year, the experimental work included the following? 
(1) Extending the number of cell types for propagating the virus 
in tissue culture; (2) Study of the immune sera produced in 
experisiental animals by the use of diversified antigens in 
CF-test; (3) Preliminary attempts in immunisation of guinea pigs 
and rabbits with measles virus adapted to grow in chick embryo 
tissue culture. 

(1) Beside the five cell types of human and simian origin 
as used last year, attempts were made to grow the virus in pri- 
mary chick embryo tissue culture, in ferret kidney, guinea pig 
kidney, and dog kidney cells. HeLa cells and monkey heart cells 
were regularly used for continuous subculture of the "Edmonston" 
strain of measles. A high titer tissue culture strain of this 
virus, grown in monkey heart cells, was utilized to initiate a 
chick embryo tissue culture series. In serial passage, the viru- 
lence (titer) of the virus in the latter type of cells seemed to 
decrease rather than increase as the appearance of suggestive 
cytopathic effect, described by Enders and co-workers, is concerned. 

(2) Extensive studies were conducted by complement -fixation 
test on inaaine sera produced in guinea pigs by a variety of methods. 
These findings, not suited for a brief review, will be published 

(3) A small number of guinea pigs and a few rabbits have 
been imcamized with measles virus grown in chick embryo tissue 
culture (early passage material) for cross-testing in CF-test the 
antibodies thus produced with virus as an antigen grown in cells 
of mammalian origin. So far, soma encouraging but also some 
conflicting results were obtained in serologic tests, to be 
pursued further. 

Significance of the Project in Relation to DBS Program s 
The experimental results as briefly outlined may serve as a basis 
for practical control methods the need of which may arise in 
connection with experimental^ or practical measles vaccines. 

Part B included Yes / 7 Mo /x"7 


Blood & Blood Products 


Serial No. DBS - 38 thru 41 

Estimated Obligations for P.Y. 1959 

Total: 497,900 

Direct: 406,500 

Reimbursements: 91,400 

Serial No. DBS-38 
1. Laboratory of Blood & Blc 

2„ Blood Bank 
3, Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Development and Application of Immunohematological 
Research Procedures 

Principal Investigator: Dr. Paul J. Schmidt 

Other Investigators: Miss Marietta Carr 

Mr. Webster C. Leyshon 
Mrs. Eleanor Morrison 
Mr. Joel Solomon and 
Dr. John T. Tripp 

Cooperating Units: NIDR, NINDB, N1AMD 

Man Years: 

Total: 2.4 
Professional: 1.0 
Other: 1.4 

Project Description: 

Objectives : The objectives of this study are to search 
for and stockpile the rare agents needed for research in blood 
immunology and to develop better methods for their discovery 
and use. 

Methods Employed : 1. Intensive study of large groups 
of blood cell and serum samples from blood donor and recipient 
populations are screened for rare antigens and antibodies using 
research serums prepared in this laboratory. These materials, 
both cells and sera, are maintained here and made available 
to other workers for reference studies. 

2. Application and use of these rare reagents and 
equipment is performed in large-scale genotyping of inbred 
populations in cooperation with other Institutes . 

3. Since donor samples are often received for study 
from other investigators, special sampling collection 
equipment developed here is being tested under field conditions 
of storage to determine its long-term utility. 

4. Serial determinations of antibody response to 
natural stimulation are being measured in volunteers and also 
in patients in cooperation with NIAMD. 

Major Findings; 1„ During the year, extensive studies 
have been made with a rare Anti-Cellano Serum found by this 
laboratory, which has peculiar properties making it very useable 
under field conditions. This serum contains a potent saline 
agglutinin especially useful for genotyping without the 
assistance of mechanical equipment „ Several rare blood factors 
in the Rh system are being investigated which are helping to 
chrow light on the intricacies of hereditary transmission and 
the serological complexity of the red cell. These and other 
materials have performed very well when tested after frozen 
storage and have been exchanged with other workers „- 

2„ The blood sample pilot tube developed in this 
laboratory has been evaluated in routine use and has proved 
extremely satisfactory in preserving clotted blood for 
laboratory tests for a period of at least 28 days, 

3„ Almost 2,000 complete genotypes for blood factors 
have been done this year on specia].Xy~selected populations 
in cooperation with HIDE, BINDS, and HXAMD. Each of these 
populations is under intensive genetic investigation and the 
overall attempt is to determine correlations between the 
transmission of the observed heritable characteristics. In 
addition, these studies have resulted in the discovery of 
families possessing extremely rare blood factors which have 
then supplied the blood cells needed in other studies. 

4, The antibody response project Is just beginning., 

Significance to t h e Institute: Any effective control 
is based upon adequate knowledge of production methods. This 
laboratory must therefore be aware of sources „ processing 
problems and utilization under field conditions of biological 
reagents which are presently fare, but which will undoubtedly 
be in routine use in a few years. 

Proposed Course ii of i Pro jject: 1. Attempts to streamline 
genotyping procedures without compromising accuracy will be 
continued in an effort to make routine the application of such 
procedures to genetics, blood transfusion, tissue transplantation 
and biologies control. In addition, the materials used are 
being made available to qualified research workers who need 
reference materials* This ultimately will provide reference 
standard preparations for control test methods. 

2. The intricate testing presently done in National 
Institutes of Health laboratories on samples received from field 
teams will be done in parallel by the field teams in an attempt 
to develop diagnostic biologies which would be useable in places 
entirely devoid of utilities and transportation. Such places 
still exist in inaccessible areas of the world and could also 
result from disaster on emergency situations, Results from 
these tests would permit more thorough studies of population 
genetics and blood transfusion problems by smaller laboratories 
throughout this country. 

3. Information gathered from the pilot tube study will be 
made available to manufacturers of biologies by publication,, Paralle 
studies by other workers on the viability of stored red cells, will 
eventually permit extending the routine safe dating period for Whole 
Blood (Human) from 21 to at least 28 days. 

4 . Any evaluation of the clinical importance in transfusion 
of the newly discovered blood factors depends on adequate in vivo 
measurements. A basis for long-term studies is being established 
by measuring the correlation between isoagglutinin titers and 
antibody-forming potential in both normal subjects and patients 
with various diseases. 

Part B included Yes / X / No / / 

Serial Eo„ ESS 

Individual Project Eeport 
Calendar Year 1958 

Part B; Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Schmidt, P. J., McGianiss, M. H„» Leysfeon, W„ C. and 
Kevy, S. V.: An Aati»k (Anti-Cellano) Serum with 
the Properties of a Complete Saline Agglutinin. 
Vox Sang. 3; December 1958. 

Serial No. DBS -39 

1 . Laboratory of Blood 


2. Blood Bank 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title: Stability and Potency of Formed Blood 

Principal Investigator: Dr. Paul J. Schmidt 

Other Investigators: Dr. -John S. Fordtran 
Dr. Shervin V. Revy 
Mrs. Eleanor G. Morrison 
Mr. Joel Solomon 
Dr. John T. Tripp 

Cooperating Units: National Cancer Institute 

Man Years: 

Total: 1.9 
Professional: 0.6 
Other. 1.3 

Project Description: 

Objectives : To delineate the conditions under which 
the formed blood elements can be stored and retain their potency 
for use in effective transfusion and to establish criteria 
for proper testing and handling of these materials to insure 
safe clinical transfusion. 

Methods Employed : 1. The antigenicity of the blood 
factor known as the Rh Q variant (D u ) is being evaluated by 
determining both its response to in vitro testing, and its 
effect in clinical transfusion. 

2. The addition of various blood preservatives is 
being evaluated for their effect on the physical properties 
of red cells from both normal donors and sensitized patients. 

3. A consistent search Is being made for rare human 
red cells which can be stockpiled in frozen storage for 
transfusion purposes, and for better Methods of nsaking them 
available for shipment to the point of need. 

4. The side reactions of patients to the transfusion 
of foraed blood elements are being critically evaluated using 
both clinical reports and tests for leuko-agglutinins and 

1 euko- pr ec i p i t ins . 

5. In cooperation with SCI, various blood fractions 
having hemostatic activity are supplied for £he treatment of 
bleeding in patients with acute leukemia. 

Major Fin dings 1. Evidence which is still prelimi- 
nary indicates that it may be easier to detect Sh variant 
(D u ) blood cells than previously expected, and that the trans- 
fusion of this material may have less clinical significance 
than anticipated. 

2. The sub-aero method of storage can be applied to 
sensitised red cells from patients with hemolytic disorders 

and is a practical and effective method to obtain comparative 
data on the course of their disease. The preservative 
properties of sntihistatninic drugs have been reinvestigated 
and found to have their effect directly on red cells without 
any effect on plasma proteins . 

3. A bank of rare human red cells for transfusion 
purposes has been established and plans for its operation were 
discussed at a national masting. This material is stored at 
-45 °C. and probably will have a five-year dating period. A 
patient at a neighboring hospital has been successfully treated 
with stockpiled frozen cells from this bank when this material 
was the only known source of needed negative blood. 

4. A study of over 10 ? 000 transfusions given at the 
Clinical Center has been made, and side effects from these 
transfusions have been categorized according to etiology. 
I>euko~ agglutinins have been confirmed to bs a rare cause of 
transfusion reactions. 

5. It was specifically determined and reported that in 
double blind experiments, fresh whole blood less than four 
hours from the donor had a favorable effect on thrombopenic 
bleeding episodes but this did not affect patient survival. 

Significance to the Institute ; The extremely 
perishable formed blood elements, red cells, white cells, 
and platelets, which come under the control of this 
Division can only be evaluated by active experience in 
determining the criteria for their stability and potency. 
A program of research in this field enables those involved 
in control to be kept aware of latest methods for product 

Proposed Course of Project ; 1. Many additional 
studies of the laboratory reactions and the transfusion 
effects of Rh Q variant (D u ) red cells must be made to 
establish the clinical significance of this factor and the 
need for the present practices and control procedures which 
are based on theoretical considerations. 

2. The stability of in vitro antigen-antibody bonding 
under conditions of sub-zero storage will be investigated to 
determine applications in control procedures. 

3. Continuing efforts will be made to expand the 
scope and size of the stockpile of frozen blood cells from 
rare donors for transfusion and to make this material 
readily available. 

4. Continuing efforts will be made to reduce the 
incidence and severity of side effects to transfusion by 
acquiring better knowledge of the causes of post-transfusion 
reactions . 

5. Attempts will be made to isolate the factor in 
fresh whole blood responsible for its efficacy in hemostasls 
and to obtain it in a more concentrated and stable form. 

Part B included Yes /x7 No l~l 

Serial No, 

Individual Project Report 
Calendar Year 1958 

P art B : Honors, Awards , and Publications 

Publications other than abstracts from this projects 

Zubrod, G., Levy, A., Williams, 6., Thomas, L„, Lynch, J., 
Schmidtf P„ } and McCulbugh, H.: Hepatitis Among Hospitalized 
Patients „ 
Hospitals., 31: 50-56, November 1957 

Eevy, S, V., and Morrison, E. 6.; Stability After Frozen 
Storage of an in vivo Antigen-Antibody Bonding on Red Blood Cells, 
Am. J„ Clin, 
Path, 30; December 1958 

Clemens., T., Jr», Breslow 8 A., Ebaugh, F. 6. 9 Jr„, Schmidt s 
P.J,, and Holly, P : Red Blood Cell Survival and Immunologic 
Studies in a Patient with Acquired Hemolytic Anemia Exhibiting 
an Autoimmune Antibody of Anti-e Specificity,. Proceed* 6th 
Congo Inter. Soc. Hemotology, Grime & Stratton, 1958,, pp. 874-875 

Freireich, E„ J., Schmidt, P. J a , Schneiderman, M.A, S and 
Freij, Emil, HI? A comparative Study of the Effect of Fresh and 
Preserved Whole Blood Transfusion on Bleeding in Patients with 
Acute Leukemia. 
Hew England J, Med.„ ia press 

Serial do. DBS-4-Q 
1. Laboratory of Blood & Blood 

2„ Blood & Blood Derivatives 
3 . Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A 

Project Title: Stability and Potency of Blood Fractions 

Principal Investigator: Dr. R. T, Suchinsky 

Other Investigators: Dr. David L. Aronson 
Miss Ann L. Dayton 
Mr. Paul 0. Fehnel 
Dr. John S. Finlayson 
Dr. Sandor S. Shapiro 
Dr. John T. Tripp 

Cooperating Units: Hone 

Han Years: 

Total: 2.0 
Professional: 1.0 
Other: 1.0 

Project Description: 

Objectives: To delineate the changes occurring in 

Z methanisn/of action, and methods of assay of blood 
coagulation components and related systems. 

^hods^£l£ved: 1. S-r^^J/S^l^ir 

— uS;^^^ 

changes in the proteins. 

2, Parallel samples of liquid and dried Normal Serum 
Albumin (Human) are stor-afd at 32°C, 5°C, and ambient room 
i:emperature» At regular intervals the materials are tested 
for stability by viscosity determinations, ultraeentrifugation, 
al.ectrophoresls, and uephelometry. 

3. The effects of bovine fibrinolysio. on fibrinogen 
are being investigated through the use of routine clotting 
methods such as one- and two-stage prothrombin time, 
thrombin time, and recalcification time. The possibility 
of this being applied as a fibrinolysin assay is also being 

!l£lS£,J!iS^ilSS. : *• ^*e investigation of the heat 
treatment: of plasma indicates, in general, that the most 
resistant plasma fractions are those which contain high 
£85V) albumin concentrations. The results of. these 
studies have been made available to the manufacturers who have 
released Che information to the National Research Council,. 
The information Is being used in the consideration of the 
type of material to be recommended for storage In the defense 

2. Viscosity determinations on albumin stored for 
four years Indicate that there may be a loss of stability 
at high temperatures of storage and that It may be necessary 

to maintain stockpiled material at controlled low temperatures 
or in other forms than the liquid state. 

SigBificance. to. the .Institute; All blood and blood 

products controlled by this Division are required to be 
stable and potent. The methods for evaluation and assay of 
these characteristics la X_tro. are being developed in order 
to p3.ace our control functions on a foundation of knowledge 

gained through research. 

Proposed, Course 0f_.Pg9J.gctj 1. Various fractions 
of plasma proteins will be isolated B and the effects on 
them of a variety of physical and chemical treatments 

2, The study of shelf Ufa of albumin will be 
continued in order to establish a definitive period of 
potency and the suitability of reworking outdated material. 

3. Determination of suitable assay conditions for 
fibrinolysin will be investigated and an attempt made to 
determine its mechanism of action. 

Part B included Yea/ ,7 $a tZIjkZJ 

Serial No. DBS-Vl 

1 . Laboratory of Blood & Blo< 


2. Office of the Chi«f 

3. Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 

Project Title Development and Application of Control 
Procedures for Blood and Blood Products 

Principal Investigator: Dr. J. T. Tripp 

Other Investigators. Miss Marietta Carr 
Miss Ann L. Dayton 
Mis b Emmy L. Denny 
Dr. Richard Suchinsky 
Mr. Joel M. Solomon 

Cooperating Units: None 

Man Years . 

Total: 1.9 
Professional- 0.7 
Other: 1.2 

Project Description 

Objectives : To develop improved control procedures and 
standards by evaluating the present control procedures and 
manufacturing methods as these affect the safety, purity and 
potency of biologic products. 

Methods Employed : 1. Samples of blood derived biologic 
products are sent as unknowns to manufacturers for comparative 
testing and evaluation by new and old methods or reagents. This 
type of project is conducted on a voluntary basis with the 
licensed blood banks. 

2. Study of bacterial contamination of blood and blood 
products is being approached in two ways; first, by following 
the rate of growth of recognized common contaminants of whole 
blood, and secondly c by a fundamental study of the mode of 
action of bacteriological filter pads and blood filters. The 

methods include a technique of preparing thin mounted 
sections of filter pads for microscopic examination to 
study trapped particles and bacteria. 

3. Shipping methods for labile blood preparations 
are investigated by simulated and actual shipments with 
various types of containers. 

4. The possibility of developing improved methods 
of identifying allergenic extracts is being studied by 
attempting production of specific anti-serums in rabbits. 
This is of secondary importance and is the first step in 
the investigation of the possible role of allergy as a 
cause of post-transfusion reactions. 

5. The methods applicable to control testing of 
blood diagnostic reagent serums and blood group specific 
substances have been evaluated and found to urgently need 
new test methods that lack the subjectivity characteristic 
of the present methods. Quantitation of blood group anti- 
bodies aad their corresponding antigens have been facilitated 
by the acquisition of special photomicrographic equipment 
designed to provide a graphic,, non-subjective, and permanent 
record of each experiment. By it® use, subjectivity has 
been nearly eliminated. 

The employment of the spectrophotometer with red 
blood cells sensitized with antibody followed by dye 
treatment has also been used to estimate reaction rates 
and equilibria. Ag&inj, the use of a s&echanical recording 
device has increased sensitivity while decreasing errors 
previously dependent upon judgment alone. 

Major Findings; 1 . The results reveal the needs for 

additional or revised standards and procedures to meet the 
following problems of blood banks i 

Blood label e, labeling procedures, and blood 

bank laboratory record forms. 

Methods of testing Anti-Human Serum for the 
Coombs test and other blood bank reagents to secure 
more reliable results in the hands of blood bank tech- 
nicians. (Part of the problem may require expanded 
programs of technician training as a direct approach 
to the problem of safety of blood for transfusion.) 

2. Both aspects of this study of contamination in 
blood products are in progress, but there have been no 
major findings to date. 

3. The shipping container developed by this 
laboratory under research contracts has been calibrated 
for the amount of refrigerant necessary in relation to 

the ambient temperature, internal temperature requirements , 
and the time in transit. 

4. Animals are under immunization with -whole and 
purified allergens, but there are no results to date. 

5. Results to date indicate that the method of 
linking antigens to carrier red cells is more accurate 
than other methods of evaluating the potency of blood 
group substances. Preliminary results in the case of serums 
indicate that quantitative tests of potency with more 
emphasis on avidity may be applicable to the control testing 
of blood diagnostic reagents and gives promise of resolving 
some of the present problems. 

Significance to the Institute ; This project consis- 
ting of a variety of parts related to a single objective is 
yielding information on which to develop new and improved 
control procedures. 

Proposed Course of Project : It is proposed to continue 
these projects as rapidly as time permits with the exception 
of No. 3, which is completed and will be published. 

Part B included Yes / 7 No /X~7 

Annual Report of Activities 

Division of General Medical Sciences 

Calendar Year 1958 


The Division of General Medical Sciences was established 
by Secretary Marlon Folsom on July 16, 195S. The purpose of the 
Division is to provide a new organizational framework for the 
conduct and continued development of basic non-categorical re- 
search, and research training and fellowship programs encompass- 
ing general medicine, public health, aging, and the clinical and 
pre-cllnlcal sciences. 

The Division is made up of the Center for Aging Research, 
transferred from the National Heart Institute, and the General 
Research Grants Branch and the Research Training Branch, both 
transferred from the Division of Research Grants. 

The Division was first assembled when 24 persons moved from 
Buildings T-6, 3 and the Colemont Building to Building 16 in Sep- 
tember 1953. By the end of the year there were 45 persons on duty. 

The Division is headed by G. Halsey Hunt, M.D., formerly 
Director of the Center for Aging Research. Frederick L. Stone, Ph.D., 
is assistant division chief and Chief of the Research Training Branch. 
The Research Grants Branch is under Richard R. Willey, Ph.D. The 
position of Director of the Center for Ayin^ Research has remained 
unfilled since Dr. Hunt assumed his new post. 

The growth of the programs managed by DGMS is indicated in 
part by increases in appropriations from approximately $5,000,000 
in 1956 to $24,637,000 in fiscal year 1959, with expenditures car- _ 
ried out or planned in FY 1959 as follows: 

Research Projects $15, 337 , 000* 

Research Fellowships 3,260,000 

Research Training 6.040,000 

Total $24,637,000 

♦Does not Include $1,284,000 in unprogramned funds, which, as of 
January 1959, had not been released by the Bureau of the Budget. 

- 1 - 

Pollowlng la a brief discusaion of the DGMS branches and a 
review of major program developments during calendar year 1953„ 


Projects supported by the General Research Grants Branch 
fall principally Into three categories: fundamental medical and 
biological research; problems of public health, including environ- 
mental health, medical care and nursing; and aging. The grants in 
these areas seek to support nev developmental and exploratory proj- 
ects in areas not covered by the Institutes' specific disease- 
oriented Investigations. 

During calendar year 1953 a total of 1,087 general research 
applications were reviewed, of which 792 were approved in the 
amount of $10,653,9 33, or approximately two-thirds of the funds 
allocated DGMS for Its research program during fiscal year 1959. 

Fundamental bio -medical research projects included work in 
biochemistry, pharmacology, metabolism, biophysics, physiology, 
cell biology, yenetlcs, embryology, endocrinology, pathology, ana- 
tomy, and hematology. In the clinical sciences there were grants 
in anesthesiology, pediatrics, surgery, orthopedics, obstetrics 
and gynecology, and dermatology. 

In addition, the General Research Grants Branch provides 
coordination and support for research projects of interest to more 
than one categorical program. 

Examples of work in DGMS -supported basic research projects 
during 1953 include: 

Dr. Sidney Pox, of Florida State University, is 
exploring the steps involved In the transition of amino 
acids to proteins and nucleic acids. The studies are 
providing clues as to the origin, structure and biosyn- 
thesis of the protein and nucleic acid moleculeSo By 
simulating the temperatures believed to exist at the 
dawn of lire, the investigators have been able to pro- 
duce a variety of amino acids — which are the building 
blocks of protein -- from a few selected biochemical 
•t«p'les. The results reveal a formation pattern that 
parallels the building-up processes of the living cell; 
and this suggests a possible pathway from non-living 
matter to the living cell. The tremendous biological 

2 - 

significance of the work lies in this hypothesis: 
If spontaneous generation of protein can be 
launched and provided with a mechanism of repe- 
tition, future developments of the resulting 
living matter may be subject to changes brought 
about by environmental factors. 

In another project, DGMS -grantee Dr. Robert 
Briggs, of the University of Indiana, has developed 
a technique for transplanting the living nuclei of 
one cell to another cell. For many years, scientists 
have been Intrigued by the fact that the nucleus of 
the individual Living cell dominates and apparently 
controls the life processes and chemical activities 
of the entire cell, Including the surrounding cyto- 
plasm. The new technique of transplanting nuclei 
will permit scientists to learn more about the 
interreacting roles of nucleus and cytoplasm -- 
and thus more about one of the most basic processes 
of life Itself. 

The work of the General Research Grants Branch in public 
and environmental health grows out of the Increasing number of 
problems involving the conditions and circumstances of America's 
social and industrial existence. The goals Include learning to 
control the biological, chemical, physical and radiological en- 
vironment in order to prevent or correct unheal thful conditions. 
In a related area, DGMS is supporting efforts which seek to deal 
effectively with the physical and psychological factors of indus- 
trial and occupational health, and to prevent accidental injury. 

During the past year, DGMS -supported projects in these areas 
included studies in: air pollution, water pollution and sewage 
disposal, food technology and contamination, occupational medicine, 
radiation, toxicology, accident prevention, medical administration 
services and facilities, nursing, rehabilitation, and epidemiology. 

An example Is the development, by Dr. James 
Pappenhagen of Kenyon College, of a new method for 
the quantitative measurement of nitrate ions in 
river water. This represents a major step in the 
search for improved criteria to assess more pre- 
cisely the extent of water pollution. 

Another example is the research on air con- 
taminants in the Los Angeles area by Drs. Paul Hotln 

- 3 - 

and Marilyn Thomas of the University of Southern 
California. These investigators have shown that 
siiiOg, produced synthetically, impairs the repro- 
ductive ability of mice and causes a high mortality 
rate among young mice. The possibility that air con- 
taminants may have an adverse biological effect on 
human beings at the low concentrations, which reg- 
ularly occur in urban areas, has been a matter of 
serious public health concern. 

Additional projects are supporting research 
into the toxicity of certain insecticides, radiation 
effects, means for detecting bacteria in drinking 
water and hazards in the use of certain food preser- 

The Ceneral Research Grants Branch makes grants for a vari- 
ety of studies on the problems of aging. 

Examples during calendar year 1953 included 
research in the aging of si. in, Including changes 
in sweat glands and hair follicles; changes in the 
lungs resulting from age, and the effects of aging 
on rats. This latter study has revealed that there 
are adverse effects on the fertility of rats born 
to older mothers and possible effects also on the 
longevity of the offspring. 

(These were in addition to grants made during the year by 
the Institutes for projects of value to research on aging, and in 
addition to the grants and work involved in the interdisciplinary 
research projects at Duke University and at the Albert Einstein 
College of Medicine, which are discussed more fully in the section 
on the Center for Aging Research.) 


The Research Training Grants Branch, established several 
years ago to help meet the nation's critical needs for research- 
trained manpower, has responsibility for the research fellowship 
programs; research training in the basic medical and biological 
sciences, and basic science training in special areas related to 
research dermatology, research anesthesiology, research pediatrics, 
research surgery and research obstetrics. 

- 4 - 

The growing Importance of these programs is indicated by an 
increase in research training funds from $2,962,000 in fiscal year 
1953 to $6,040,000 in fiscal year 1959, and in research fellowship 
funds from $1,498,000 to $3,260,000. During the past year an in- 
tensive effort thus has been carried forth to extend the scope and 
value of the training programs to meet the enlarging demands for 

A total of 155 training grant applications with a value 
of $4,297,687 were approved during the 12 months ending December 
31, 1953, and applications for an additional 97 grants were dis- 
approved or deferred. Fellowship awards totaled 521, out of 737 
reviewed, at a cost of $1,964,603 during calendar year 1958. 

Epidemiology and biometry constituted the nucleus of the 
original research training program. During the past year, train- 
ing committees have been established or approved for establishment 
in biochemistry, genetics, pathology, pharmacology, physiology, 
anatomical sciences and experimental embryology and development. 

Epidemiology and Biometry 

During calendar year 1958 four applications were approved in 
epidemiology with a value of $12\,652. This brought the total 
number of programs approved to 14, at a total cost, as recommended 
by the National Advisory Health Council during the past two calendar 
years, of $678,834. 

As of December 31, 1958, 12 programs were providing training 
for 89 students in approved schools of public health and one school 
of dentistry, in the epidemiologic approach to problems such as 
heart disease, cancer, mental illness, accidents, disorders of re- 
production, dental problems, air pollution, metabolic disturbances, 
radiological hazards, aging and communicable disease processes. 

In biometry, seven applications were approved during calendar 
year 1953 at a cost of $151,864. This brought to 20 the number of 

- 5 - 

programs at a Council -approved level of $366,725 for the past two 
calendar years. Instruction was being provided for 194 trainees 
in six medical schools, three statistical laboratories in univer- 
sity graduate schools and eleven schools of public health. 


Following the establishment of the Biochemistry Training 
Committee in the summer of 1953, a total of 22 research training 
applications were acted on favorably by the end of the calendar 
year, at a cost of $783,623. Seventeen programs were being sup- 
ported at 15 universities throughout the nation. Four of these 
had been transferred to the Division of General Medical Sciences 
from the National Heart Institute and others from each of the other 

More than 100 graduate students were receiving basic bio- 
chemistry training in a number of specific disciplines, including 
enzymology, cellular biology, radioisotope studies, intermediary 
metabolism, physiological chemistry, vitaminology, microbial bio- 
chemistry, protein structure, physical biochemistry and many re- 
lated fields of biochemistry, where a major shortage of research- 
trained manpower exists. 

An additional benefit of the program is the strengthening of 
the recipient biochemistry departments in teaching staffs, facil- 
ities and equipment. These departments thus are able to provide 
more and better training for undergraduates as well as for the 


With the establishment of the Genetics Training Committee, 
which held its first meeting in October 1953, a broad program was 
begun in all aspects of training in medical genetics. Thirteen 
applications in the amount of $235,274, were approved before the 
end of the year, providing instruction for 27 predoctoral and 10 
postdoctoral trainees. 

The grants are supporting training programs in many aspects 
of the science of genetics, including medical and human genetics, 
cytogenetics, virus and bacterial genetics, biochemical genetics 
and genetic ultrastructure. 

- 6 - 


Twenty new training programs were approved In pathology 
during calendar year 1953. With the assistance of the newly 
formed Pathology Training Committee, the National Advisory Health 
Council approved 26 applications at a cost of $732,729, providing 
instruction for 93 trainees, 65 of whom were at various levels 
of postdoctoral training. A substantial portion of this money 
was used for full or partial Bupport of research-oriented faculty 
members in order that they might devote more time to research 

Due to the essential role of the pathologist in the diag- 
nosis of disease, pathology training programs in the past have 
tended to neglect the development of the experimental pathologist. 
To bring this field into proper focus as a keystone of research 
and training for research, modern techniques must be offered to 
talented young pathologists. 

Through DGMS support, research training now can be offered 
in almost every type of disease process with the most modern tech- 
niques of investigation, including electron microscopy, modern 
biochemical methods, histochemistry and tissue culture. 


The formation of a Pharmacology Training Committee in the sum- 
mer of 1958 permitted the launching of an active program in this 
field, where an acute shortage of well-trained scientific personnel 
exists. By December 31, 1958, a total of 26 applications had been 
approved in the amount of $674,717. Stipends were provided for 
93 predoctoral and postdoctoral trainees in all phases of pharma- 
cology research, Including toxicology, drug metabolism, experimental 
therapeutics, chemical pharmacolosy, antimetabolites, chemotherapy, 
analgesics, cellular pharmacology, neuropharmacology, human phar- 
macology and others. 

The grants have helped to strengthen pharmacology depart- 
ments by providing funds for additional staff, supplies, equipment 
and other items Important to well-balanced programs., 


Eight training grant applications in physiology were approved 
during the year following the establishment of the new Physiology 

- 7 

Training Committee . The grants are providing Instruction for 34 
predoctoral and eight postdoctoral trainees at a cost of $269,593. 
Several of the programs will make training available to students 
from a wide geographical area or enable them to use certain unique 
facilities. The programs will serve to increase the rate of pro- 
duction of physiologists, by relieving the need for many students 
to support themselves as part-time research or teaching assistants. 
The stipends will help reduce the graduate study time to three or 
four years. 

Anatomical Sciences 

The new Anatomical Sciences Training Committee was formed 
during the year, and by the end of 1953, 16 applications had been 
approved at a cost of $451,305. The funds will go to 11 schools 
of medicine and one research institute, providing a broad spectrum 
of training and support for 52 predoctoral and 19 postdoctoral 
trainees. The training areas Include neurophysiology, electron 
microscopy, histochemistry and experimental embryology. The pro- 
gram will help relieve the present acute shortage of anatomists 
by enabling qualified Individuals to become competent teachers and 
investigators in this area. 

An award also was approved by the National Advisory Health 
Council supporting the 1960 International Anatomical Congress In 
New York. The Conggess should have a stimulating effect on teach- 
ing and research In the biological and basic medical sciences. 
It will provide a unique opportunity for formal and Informal Inter- 
national communication of scientific information in these fields, 
and should serve to bring into sharp focus the fundamental need 
for, and Importance of, the preclinical sciences in clinical med- 
icine and medical research. 

Experimental Embryology 

Six applications in experimental embryology were approved 
during the year at a cost of $127,615. These were the initial 
steps In a program of rapidly increasing significance. Just be- 
fore the end of the calendar year the Director of the National 
Institutes of Health approved plans for the establishment of an 
Experimental Embryology and Development Training Committee to re- 
view training grant applications. The program will support 
research training for post-residents or other postdoctoral and 

- 3 - 

predoctoral trainees in electron microscopy, genetics, develop- 
mental physiology, neonatal pathology, fetal biochemistry and 
other related basic science fields. 

Training for Clinical Specialists 

During the past year, programs were begun to provide basic 
science training to clinical specialists (non-resident or post- 
resident) in several fields, including anesthesiology, dermatology, 
pediatrics and obstetrics. 

In research anesthesiology, four applications were approved 
at a cost of $59,646. 

Two applications for training in dermatology were approved 
at a cost of $39,536, providing support for one predoctoral and 
four postdoctoral grantees. 

In research pediatrics, one application was approved at a 
cost of $32,000. 

The programs will aid individuals with M.D. degrees who have 
been hampered in entering an academic career in medical research and 
teaching either because they need support or lack opportunity to 
prepare themselves in the required fundamental scientific disciplines. 

Additional Training Grants 

Training grant applications approved during the year for 
other disciplines included: biology, $154,578; microbiology, 
$187,254; nutrition, $30,000; radiological health, $16,740; multi- 
disciplinary projects, $66,332, and other programs, $121,820. 

Experimental Training Grants 

The Experimental Training Grants program, begun in calendar 
year 1956 at a level of $500,000 a year for five years, achieved 
full operation in 1958 when the last five of the 13 approved med- 
ical schools began their programs of providing special research 
training for selected students to stimulate and equip the trainees 
for careers in academic medicine. These five were the Schools of 
Medicine at Western Reserve University, the University of Minnesota, 

- 9 - 

the University of Texas, Tulane University and Yale University. 
The other eight, which had Initiated their programs prior to 
January 1, 1953, were New York University, the University of 
Pennsylvania, the University of Rochester, Emory University, 
Washington University, Johns Hopkins University, the University 
of Virginia and the University of Washington. 

Also during 1958, the first meeting of Program Directors 
from each of the 13 schools was held for an exchange of ideas and 
information about the different approaches to the project. The 
meeting was considered of such benefit that a second such confer- 
ence has been scheduled for March 30, 1959. 

Toward the end of the calendar year firm plans were formu- 
lated for program evaluations, which will begin in the Spring of 

At the end of calendar year 1953, at least 172 students were 
receiving stipends of between $500 and $3,200 each from the grant 
funds. The magnitude of the program is better indicated, however, 
by the fact that between one-fourth and two-thirds of the total 
enrollment at each school are participating in research training 
on a part-time basis, opportunities for which are provided for 
students In the first three years at each school. 

Ten schools have used grant funds to strengthen and enlarge 
their summer research programs, and five are providing the means 
for students to continue research during free time throughout the 
academic year. Six schools have strengthened their programs for 
students who wish to drop out of medical school for one full year 
for research training in a basic science department. Nine schools 
have provided the students greater opportunities to improve their 
foundations in mathematics, physics, biometry, chemistry and be- 
havioral sciences. Four schools have developed decelerated curricula 
giving students great amounts of free time for research and basic 
science training. 

Other benefits of the program Include the provision of 
funds for faculty support and special courses. 

Generally the grant funds have given the schools' research- 
training programs coordination, coherence and special direction 
which otherwise would not have been possible. 



The Research Fellowship program, functioning as a part of 
the Research Training Grants Branch, acted on 737 applications 
during calendar year 1953. Of the total, 521 were approved at a 
value of $1,964,603 and 216 were deferred or disapproved. Funds 
for the fellowship programs grew from $1,493,000 in fiscal year 
1953 to $3,260,000 in fiscal year 1959. 

Senior Research Fellowships 

Senior Research Fellowships, provided to strengthen scien- 
tific areas in preclinical departments of medical, dental and 
public health schools, were awarded to 59 scientists in 35 schools 
located in 22 states and the District of Columbia during calendar 
year 1953, at a cost of $703,954. These awards were in addition to 
65 renewals, totaling $771,581. The fiscal year 1959 appropriation 
for the program was $2,000,000. 

The fellowships provided assistance to preclinical science, 
in departments such as anatomy, behavlorial sciences, biochemistry, 
biophysics, epidemiology, genetics, microbiology, pathology, phar- 
macology, physiology, etc. 

The achievements of two grantees during the year help illus- 
trate the benefits of the program. One scientist, as a member of 
a research team, discovered two new hidden viruses responsible for 
respiratory illnesses — a finding which may lead to the develop- 
ment of a preventive vaccine. Another Fellow, engaged In cancer 
research, discovered that the establishment of epithelial-like cells 
in continuous culture from human tissues provides a more effective 
means of studying the metabolism of human cells in vitro than the 
previously used tissue explant method. Re confirmed that the estab- 
lished cells may be maintained in a state of active multiplication, 
the rate of which can be measured accurately. 

Regular Research Fellowships 

These awards are for the full-time research training of 
scientists at the predoctoral, postdoctoral and special levels for 
careers in the fundamental biological sciences. During calendar 
year 1953 a total of 50 Regular Fellowships were awarded at a cost 
of $208,723. These went to 36 researchers in 24 schools located in 
14 states and England and Denmark, and to 14 nurses engaged in re- 
search training at 10 schools in eight states. 

11 - 

Part-time Student Fellowships 

Awards under this program went to 324 students during cal- 
endar year 1953, at a cost of $209,952. The grantees are working 
on projects in 42 institutions, located in 26 states and the Dis- 
trict of Columbia. This program, by stimulating at an early stage 
the research interests of promising students, has been successful 
in promoting a continuous flow of highly motivated recruits into 
medical, public health and nursing research fields. It increases 
the number of full-time researchers in the related basic fields: 
and it creates an Increased number of physicians and others with 
research orientation who can better apply basic research findings 
at the clinical level. 

Fostsophomore Research Fellowships 

A total of 23 postsophomore fellowships were awarded during 
calendar year 1953, at a cost of $65,393. These went to students 
in 17 institutions located in 14 states and the District of Columbia. 
This program permits superior medical and dental students to obtain 
one to three years of research training prior to the completion of 
their professional degrees* In general, these fellowships are 
awarded to candidates at the natural break between their preclinical 
and clinical course work. 

Foreign Fellowships 

This program, to provide postdoctoral training for citizens 
of foreign nations as a part of the Postdoctoral Research Fellow- 
ship Program, was reinstated during calendar year 1953 after 
having been discontinued in 1952. In the 12 months ending December 
31, 1953, 17 awards had been made at a cost of $90,797. These funds, 
all from the fiscal year 1953 appropriations, were provided by the 
different Institutes of the National Institutes of Health. 

For fiscal year 1959, the Division of General Medical Sciences 
was given $345,000 for foreign fellowships from the General Research 
and Services appropriation. By the end of the calendar year, ap- 
proximately 3 applications had been received. These applications 
and the 17 awards represent nearly all the countries of Europe and 
Scandinavia, England and Australia. The program is being extended 
to include nine countries in Central and South America, in addi- 
tion to Ceylon, India, Iran, Japan, New Zealand, Pakistan, Thailand 
and the Philippine Islands. 

- 12 - 


Under the reorganization effected by the Secretary's memo- 
randum of July 16, 1953, the Center foi Aging Research was trans- 
ferred from the National Heart Institute, where it had first been 
established in 1956, to the Division of General Medical Sciences. 
Its authorities and responsibilities havs been set forth by the 
Public Health Service Manual of Organize :ion and function, as fol- 

Section 7-15: Provides that DGMS Is responsible for 
"... fostering National Institutes of Health re- 
search and training activities in the field of aging." 

The section provides that the Center for Aging Research, as 
a component of the Division, is given responsibility for 

"directing a program designed to stimulate biological, 
psychological and social sciences research in the 
field of aging; providing direct financial support 
of such research; fostering other Nil: research sup- 
port of activities in this field; making studies in 
this field to identify research areae needing greater 
support and trained research manpower requirements, 
and to promote development of appropriate trainl&j 
activities; sponsoring conferences, symposia, and 
seminars in, and coordinating information on, aging 
research and research training problem i; maintaining 
liaison among the PHS Intramural research activities 
in the field of aging." 

In addition, in a memorandum of December 11, 1953, from the 
Director, NIH, to the Chief, DGMS, it was stated: 

"The Division of General Medical Sciences is hence- 
forth to be considered the focal point for providing 
desired consultation in the development, and review of 
NIH research and training grant programs" in the field 
of aging. 

Multidisciplinary Programs 

A principal accomplishment during the past year was estab- 
lishment of a second large interdisciplinary research program in 
aging at the Albert Einstein College of Medicine, Yeshiva Univer- 
sity, New York, New York. The Center for Aging Research provided 

- 13 

staff assistance to Dr. Irving London, Principal Investigator, 
and his associates, during the development of the plans for this 
program. The $4O0,(V.'O award for the first year's operation was 
granted by the Surgeon General during formal ceremonies at Albert 
Einstein. The remaining four years* commitment will involve about 
$300,000 per year. Financial support for the program is derived 
from the National Heart Institute, the National Institute of 
Mental Health and the National Institute of Arthritis and Metabolic 
Diseases (H-3333, M-2562, A-2965). 

The Duke University School of Medicine, the first to re- 
ceive a large NIH grant for an interdisciplinary research program 
in aging, was visited during October 1953. At this time, a year 
after the project had been initiated, most of the preparatory 
staffing and programming tasks had been completed and 22 collabo- 
rative projects had been activated. An oral report on this visit 
was presented to the November meeting of the National Advisory 
Mental Health Council by Dr. Stanley Mohler, Medical Officer of 
the Center. A supplemental written report was given Council mem- 
bers, and, upon request to Dr. Kenneth Endicott's 20-School Survey 
Task Force to help evaluate the impact of research and training 
grants on medical school programs. 

Also during the year, staff assistance was provided the 
Medical Branch of the University of Texas, at Galveston, in develop- 
ing a proposal for a large interdisciplinary research program on 
aging. The application has been considered by a Special Multi- 
disciplinary Study Section and will receive Advisory Council review 
in March 1959. 

Other Grants 

During the year, the staff of the Center for Aging Research 
reviewed all of the research and training grant applications made 
to NIH during 1953. Those applications which related primarily or 
secondarily to aging were followed through Study Section and Ad- 
visory Council review and filed according to the actions taken. A 
summary of the number and dollar values of NIH research and train- 
ing grants in aging, which were present In the Center for Aging 
Research active files during the calendar year, and listed by In- 
stitute and Division, is presented as an appendix at the end of 
this report. 


A series of conferences on the similarities and differences 
between physiologic aging and radiologic life-shortening were held 

- 14 - 

during 195 3 under the auspices of the American Institute of Bio- 
logical Sciences. Support for these conferences was derived from 
the National Institutes of Health (H-3650) and the Atomic Energy 
Commission. The conferences were organized around six scientists 
selected from a roster of eminent researchers who had demonstrated 
an Interest both in the problems of aging and In radiation studies. 
These persons served as panel chairmen and each held one or more 
conferences with experts of their own choice. The individual 
conferences covered such topics as irradiation-induced changes at 
the cellular level, irradiation-induced genetic changes, irradiation* 
Induced organ changes, and irradiation-induced whole animal changes. 
The chairmen met In October 1953 and plan to publish a combined 
summary of the major results of their conferences as a supplement 
of Federation Proceedings, the publication of the Federation of 
American Societies for Experimental Biology, as soon as editing can 
be completed. 

Information Activities 

Work in providing professional and public information on 
aging during the year included the following: 

The Excerpta Medica Foundation was given a 
joint grant by HHI and NIMH for support of a per- 
iodical, called "Gerontology and Geriatrics," a 
monthly publication in English of worldwide abstracts 
in the field of gerontology. The first issue appeared 
in July 1953. 

The Center for Aging Research prepared its annual 
multilithed booklet: Activities of the National 
Institutes of Health In the Field of Gerontoloty , 
January 1958 . The one hundred page booklet covers ex- 
tramural research and training projects which were 
active as of January 31, 1953, plus Intramural re- 
search projects conducted during calendar year 1957. 

It became apparent that there was also a need for 
a narrative summary, which would provide a more detailed 
account, Institute by Institute, of the research and 
training programs in aging being conducted with NIH grant 
support. Such a descriptive summary was prepared from 
reprints and progress reports which were provided by 
the grants branches of the various Institutes. In each 
case where progress report information was utilized, 
permission of the investigator concerned was obtained. 

15 - 

A second booklet entitled "Aging, A Review of 
Research and Training Grants Supported by the National 
Institutes of Health" was compiled during the year and 
went to press in December 1953 „ 

An article prepared by Dr, G. Halsey Hunt, entitled 
"Implications of Aging as Predicted by Population Changes" 
appeared in "Geriatrics," Volume 14, Number 1, January 
1959, pp. 1-7. 

By request of the editors of the journal Geriatrics , 
Drs. Hunt, Robert Akers and Stanley Mohler jointly pre- 
pared an article entitled "The Research Grant Program of 
the National Institutes of Health," which has been accepted 
for publication early in 1959. 

Or. Hunt published a paper entitled "The Importance 
of the Medical Record in the Treatment of Prolonged Illness" 
in the Journal of the American Association of Medical 
Record Librarians , Volume 29, No. 4, August 1953, pp. 133-134, 
The paper was based upon a talk presented in March 1953 at 
the Institute for Medical Record Personnel in Chronic 
Disease Facilities, Chicago, Illinois. 

A three-panel exhibit was prepared at the request of 
the Gerontological Society, and displayed during its Novem- 
ber meeting in Philadelphia. The exhibit depicts the 
activities of the Center for Aging Research and describes 
how the Center relates to other components of the Public 
Health Service which have specific responsibilities in 
the field of aging. Also, the exhibit contains a flow 
chart which demonstrates the research grant mechanism, 
and the role played by the Center for Aging Research in 
the overall scheme. 

16 - 

The following tables show the number and annual value of re- 
search and training grants in aging that were active on January 31, 
1959 and January 31, 1953. The fiscal year columns indicate the 
appropriation from which funds were derived to support these proj- 
ects. These tables essentially reflect calendar year activity. 
The date of January 31 was chosen in order to give full effect to 
the actions of the November meetings of the National Advisory 





















Primarily Related to Aging 

PY 1953 FY 1959 



$ 18,192 



$ 550,423 










30, 360 



































Secondarily Related to Aging 

FY 1958 PY 1959 




$ 32,000 

$ 268,910 

$ 300,910 


























- 17 - 


Primarily Related to Aging 
Institute Number FY 1955 FY 1957 FY 1953 





















$ 12,253 


















$463,534 $1,779,151 $2,266,371 

Secondarily Related to Aging 
Institute Number FY 1955 FY 1957 FY 1958 























$ 204,213 


























- 18 - 

Annual Report of Activities 

Division of Research Grants 

Calendar Year 1958 

Summary Statement of Extramural Program 

Of the total appropriation of $294 million afforded to the 
[ational Institutes of Health for Fiscal Year 1958, almost 70 percent-- 
201 million--wa8 allocated to the extramural program to support 
■esearch and research training in the nation's universities, hospitals, 
ind medical schools. Exclusive of these funds, an additional $30 
lillion supported a program to construct and equip health research 
'acilities . 

At the beginning of Calendar Year 1958, the Division of 
:esearch Grants was the administrative center for the entire program 
rith the additional responsibility of conducting a noncategorical 
esearch and training program in the basic medical and biological 

Administrative necessity dictated a reorganization in order 
o develop properly the noncategorical research and training program 
rhile simultaneously fulfilling the expanding needs for central DRG 
ervices in all the extramural areas. This resulted in the establish- 
lent of the Division of General Medical Sciences, and the transfer 
f programming responsibilities of noncategorical research and 
raining to the new Division. DRG retained management of the 
tealth research facilities construction program, and administrative 
esponsibility--including technical review of all grant applications, 
iscal control, and business services--for the entire extramural 
if fort. 

The reorganization enabled DRG to increase the number of 
tudy sections (to 31) and to make use of additional a_d hoc committees 
n order to sustain the high quality of review of applications, 
"urther, DRG expanded the review of applications on a disciplinary 
lasis, established a central program for providing a continuing 
itatistical analysis and evaluation of the progress and direction 
if each aspect of the extramural program, strengthened liaison with 
;rantee institutions, and accelerated the handling of applications. 

Accomplishments in the extramural effort are recorded in the 
iupport provided by Fiscal Year 1958 funds: 7,028 research grants 
imounting to $99,480,968 were awarded to individuals in 699 institu- 
:ions located in 48 States, the District of Columbia, two territories, 
md 28 foreign countries. A total of 2,329 research fellowships 
imounting to $6,430,551 were awarded to individuals in 211 institutions 
.ocated in 42 States, the District of Columbia, one territory, and 
.1 foreign countries . 

In the health research facilities construction program, DRG 
twarded, on a matching fund basis, 177 grants totaling $30,200,095 
:o help build or expand research facilities at 134 institutions. 

- 2 - 

Congressional interest in this program is reflected by the fact that 
in August, 1958, the Congress enacted Public Law 85-777 which 
extended the program to 1961. 

The individual administrative programs within the various 
DRG components are briefly described in the following report. 

Research Fellowships Review Branch 

With the reorganization of the Division of Research Grants, 
the Research Fellowships Section became the Research Fellowships 
Review Branch. As currently constituted, the Branch has responsibilit 
for review of predoctoral, postdoctoral and special research 
fellowship applications, and for administering on behalf of the 
various Institutes and Divisions the many details regarding their 
individual Fellows. 

Review Techniques 

Along with the reorganization of the Branch, the procedures 
for fellowship application review were changed. The Central 
Qualifications Board was divided into seven disciplinary panels: 
Anatomy and Physiology, Behavioral Science, Biochemistry and 
Nutrition, Clinical Research, General and Physical Biology, 
Microbiology, and Pharmacology and Endocrinology. These panels 
meet monthly. Panel members have been selected from Institute 
intramural personnel, with an additional member chosen from 
executive secretaries of the study sections in the Research Grants 
Review Branch. 

After review by a disciplinary panel, an application is referre 
to the appropriate Institute or Division for review by its Specialty 
Fellowship Boards This dual review is now comparable in many aspects 
to procedures used in considering research grant proposals „ 

New Stipends and Allowances 

During this year both the National Institutes of Health and 
the National Science Foundation explored needs for higher stipends 
and allowances. This exploration on the part of both agencies 
extended over several months. As a result of these surveys, 
increased stipends and allowances went into effect January 1, 1959. 

Statistical Analysis and Evaluation 
of the Extramural Program 

During 1958, the Division of Research Grants established 
for the first time a statistical research program devoted to the 
quantitative analysis of the NIH extramural program and its impact 
on nation-wide research and training activities in the medical 
and biological sciences. 

Designed to be flexible, responsive, and extensive in 
scope, the program will furnish a wealth of new data to help 
guide medical research planning on the local, national, and inter- 
national levels. 

Centered in the Statistics and Analysis Branch, DRG, the 
program strengthens the Division's capacity for providing to NIH 
administrators, other Government agencies, and the Congress 
necessary information on the status, progress, and direction of 
every phase of the NIH extramural effort. Serving as an 
"intelligence center," the Branch provides DRG the framework 
for conducting continuing statistical analyses of both intra- 
and extra-NIH data in order to determine changing patterns, trends, 
needs, accomplishments, and dynamics of extramural research and 
training programs throughout the nation. From this data, the 
Division will evaluate and report on the status of grant- 
supported programs in relation to NIH plans and objectives, and 
on the impact of grant programs upon substantive research and 
training problems in this country—including the impact upon 
the research and training community and its resources. 

The Division of Research Grants will also be responsible for 
determining (and for taking corrective measures in) areas that are 
weak in data concerning national resources for medical research 
under the NIH program. 

Under the aegis of a newly-appointed chief of Statistics 
and Analysis Branch, DRG plans more extensive utilization of the 
facilities of the Bio-Sciences Information Exchange which serves 
as a clearing-house for information concerning all research grants 
made by seven Federal agencies and 90-odd private foundations „ 
The Exchange provides quantitative analyses of the NIH extramural 
program, and special reports of research by field, specialty of 
the investigator, geographical distribution, etc. 

In prospect, the expanded and reinforced statistical 
analysis and evaluation program will, figuratively speaking, 
position DRG as the check point "on the pressure and the pulse" 
of the nation-wide research and research training program 
administered at NIH. 

Internal Operations Branch 

The Internal Operations Branch was established in July 
of 1958, A variety of central service and housekeeping functions 
were placed in this branch under the direction of the Administrative 
Officer of the Division. These services include: 

Office of Branch Chief 

Budgeting, personnel planning, space, etc. 

Purchase and supply; inventory controls 

Property accountability 

Coordination of all Division administrative activities 

Miscellaneous Services Section 
Personnel recruiting 
Time and leave 
Mail and files 
Travel clerks 
Typing pool 

Grants Finances Section 

Processing payment of all NIH research grants 

Duplicating Section 

NIH duplicating plant 

Effective March 1, 1958, research and training grants of 
$5,000 or more were paid in two installments at about six-month 
intervals. Prior to that date, 90 percent of all grants were 
paid in full at the time of their activation. The revised procedure 
for paying grants i9 expected to provide substantial savings to 
the government in interest. 

Recruitment of new personnel for the Division was somewhat 
slower than planned: full-time filled positions increased from 
216 on January 1, 1958, to 239 on December 31, 1958. Many of the 
remaining 51 vacancies will not be filled until additional office 
space becomes available. 

The Duplicating Section processed 23,992 requisitions 
involving 152,437 offset masters and stencils, producing 19,498,370 
impressions. In comparison, total impressions during 1957 were 
15,925,214, and 11,680,182 in 1956. A plan, developed in Calendar 
Year 1958, is pending clearance of the Congressional Joint Committee 
on Printing. It proposes to establish duplicating facilities in 
the new Robin Building in Silver Spring in order to service the 
reproduction requests at that location when it opens for NIH 
use in the spring. This will permit the present DRG plant to 
continue meeting NIH-proper duplicating demands in 1959 without 
additional equipment and related space. 

During the last six months of calendar year 1958, the break- 
down of duplicating requests between extramural and other-than- 
extramural NIH activities were as follows: 

5 - 

No. of 


No. of 


No. of 


Average Copy 
per Plate 









Other than 

















Plans were developed, and necessary purchase orders were 
initiated, to convert the DRG Mail and File Unit into an open-shelf 
filing system in preference to four and five-drawer files. The 
open-shelf system requires less floor space, and will speed up 
filing activities. 

Health Research Facilities Branch 

The Division continued in its responsibility for the admini- 
stration and implementation of the health research facilities 
construction program. Members of the Health Research Facilities 
Branch reviewed all applications for construction and equipment for 
such facilities, and--in conjunction with Council members-- 
conducted all necessary project site visits. 


The enactment, on August 27, 1958, of Public Law 85-777 
had both positive and negative aspects: it extended the program 
until June 30, 1962, authorizing $90 million in expenditures 
during the additional three years; however, as a simple extension 
of the original act, no provision was made for constructing 
urgently needed facilities for training researchers in the health 
related fields 

Previously, and again in the 1958 Annual Report, members 
of the Council recommended to the Congress enactment of legislation 
to authorize a program for construction of training facilities. 
The Council further recommended that the training facilities and 
research facilities programs be administered as one, with review 
and advice by the Council. 

The Program 

The program authorizes expenditures not to exceed $30 million 
yearly. During 1958, the Health Research Facilities Branch received 
203 completed applications, 29 supplemental requests, and 84 
notices of intention to file. Grants were approved for 134 
institutions in 39 States, the District of Columbia, and Hawaii, 
representing expenditure of the $30 million appropriation for 
the year . 

- 6 

At year's end, however, 136 completed applications, and 
seven supplemental requests were still on hand for processing and 
submission to the Health Research Facilities Advisory Council. 
These represented an estimated expenditure of $35,237,886. 

Council Activities 

During the year, the Health Research Facilities staff and 
Council members conducted more than 200 project site visits. 

The Council held two instead of three meetings during 
the year, each lasting three days. The May meeting was 
postponed to late September in order that project site 
visits could be made to all institutions that had submitted 
applications prior to the deadline date of June 30. 

Research Grants Review Branch 

At the end of Calendar Year 1958, the Research Grants 
Review Branch consisted of 31 study sections. 

An important administrative development within the Branch 
was the refinement of the procedure for reviewing the increasing 
number of applications requesting large sums of money to support 
long-term, multidisciplinary research projects. In order to 
accelerate the activation of research in problem areas such as 
antibiotic-resistant staphylococcal infections, viral etiology 
of tumors, etc., the Branch initiated a series of interdisciplinary 
discussions through ad hoc inter-study section advisory panels. 

The success of these conferences is reflected in (a) the 
many research approaches currently utilized s and (b) the increasing 
number of applications for the conduct of research along the 
various avenues proposed by the inter-study section conferences. 

Significant activities of the study sections during the 
year follow. 

Bacteriology & Mycology Study Section 

As a result of a division of the Microbiology Study Section, 
the Bacteriology and Mycology Study Section was established and 
held its first meeting on October 7-8, 1958. The separation 
(which led to the simultaneous establishment of the Virology and 
Rickettsiology Study Section) was made principally because the 
increasing work-load jeopardized the high quality of scientific 
review. Its timeliness was attested by the fact that 67 research 
grant applications were reviewed at the first (October 7-8) 
meeting of the new study section, and 71 research grant appli- 
cations and two applications for research training grants were 
appraised at the second meeting held in January 1959. 

- 7 

In recognition of a general need for augmented research 
efforts in the face of an emerging problem with apparent national 
significance, an Inter-Study Section Conference on Staphylococcal 
Infections, sponsored by the Division of Research Grants, was 
held at Bethesda, Maryland on August 28-29. 1958. At this 
conference, 24 representatives of 13 study sections discussed and 
evaluated one another's views on staphylococcal research needs „ 
General and specific recommendations for projects on basic 
and applied research were formulated and communicated to the 
Institutes. At the time of this conference, three multidisciplinary 
research grant applications dealing with staphylococcus problems 
were reviewed by a Special Study Section (Staph) composed of 
individuals selected from the group of conferees „ This panel is 
potentially available for service on an ad hoc basis as the need 
may arise for review of multidisciplinary projects. 

Following this conference, the Bacteriology and Mycology 
Study Section became the focal point for review of research 
grant applications concerned with varied aspects of staphylococcus 
problems, and received for review the majority of applications in 
this area. At its first (October 1958) meeting, the study section 
reviewed 16 applications relating entirely or partly to 
staphylococcus infections and/or disease; of these, 13 were 
recommended for approval. 

Radiation Study Section 

The study section continued its consideration of the 
possibility of cooperative studies in radiotherapy. The formal 
report of the conference held at Highland Park, Illinois on May 
10-12, 1957, titled "Research in Radiology" was released as 
National Academy of Science-National Research Council Publication 
No 571. 

An informal meeting was held in Bethesda on May 17-18, 
1958 at which approximately 40 outstanding U„S. radiotherapists 
were in attendance. It was again agreed that studies of this 
nature are highly desirable, not only because of the direct need 
for statistically valid data on the effectiveness of radiotherapy, 
but also because of the potential beneficial impact on the practice 
and teaching of this speciality. It was agreed that immediate 
attention should be paid to the possibility of cooperative studies 
in carcinoma of the lung and of the cervix, and that studies on 
disease in other sites would be held in abeyance. Coordinating 
committees were established to proceed with the organization of 
cooperative research in these two areas. Additional meetings of 
the two latter committees were planned in order to complete detailed 
action on protocols, and to make the necessary administrative 
arrangements for initiation of the studies, 

The study section also sponsored an August 1958 conference 
in Burlington, Vermont, on "Tissue Oxygen Tension and Radiotherapy." 
Recent work indicates that radiosensitivity of both normal and 
tumor tissue is markedly dependent upon the tissue oxygen tension,, 
The purpose of this conference was to explore the basic information 
available at that time, determine areas needing further study, and 
evaluate the significance of these findings for clinical radio- 
therapy „ A major portion of the basic studies and clinical work 
to that date had been conducted in the United Kingdom; therefore, 
representatives from the major active groups in that country 
were present at the conference. It was agreed that the informa- 
tion concerning the inter-relationship between tissue oxygen 
tension and radiosensitivity has potential significance in clinical 
therapy, but that much additional research on the basic level and 
in animals is needed before techniques are developed for routine 
use in humans. 

The Executive Secretary continued his active participation 
in extramural activities on radiation protection, particularly with 
reference to activities of the Executive Committee and the Sub- 
committee on Wide-spread Radioactive Contamination of the National 
Committee on Radiation Protection and Measurements, as well as the 
Committee on Units, Standards and Protection of the American 
College of Radiology. In addition, he continued to serve as a 
voting member of the U.S. Atomic Energy Commission Radiological 
Physics Fellowship Board. 

Virology and Rickettsiology Study Section 

The ever-increasing workload of applications in the Micro- 
biology Study Section led to a June 1958 reorganization which 
resulted in the establishment of the Virology and Rickettsiology, 
and the Bacteriology and Mycology Study Sections, with some appli- 
cations going to the Genetics and the Biochemistry Study Sections. 

During the January and May meetings of the Microbiology 
Study Section, training grant applications were reviewed for the 
(then) Training Grants Branch of the Division of Research Grants. 
With the establishment of the Division of General Medical Sciences, 
this review became a function of the new Division. 

On August 28-29, a full-scale inter-study section meeting 
was held to explore the broad field of staphylococcal infections 
in hospitals, and to make concrete recommendations to the 
National Advisory Council on Allergy and Infectious Diseases. 
Members of 13 study sections and several ad hoc consultants each 
gave their opinions as to where knowledge was lacking and what 
could be done about it. The method, was very successful and it 
is hoped that this type of meeting will result in a stimulation 
of applications in the field of staphylococcus infections. 

- 9 

As a result of a Congressional recommendation in the last 
appropriation bill for cancer research, the National Cancer 
Institute asked the Virology and Rickettsiology Study Section to 
provide advice to the Institute and its Council for program use 
in the tumor-virus field. Since the previous round-table 
discussion session on staphylococcus infections was so successful, 
the method was again employed in an inter-study section meeting 
on tumor-viruses held on September 16, 1958. This meeting brought 
together scientists from all the fields related to viruses and 
malignancies. Plans were made for a long-range program which 
will be of invaluable use to the National Advisory Cancer Council 
in programming future activities 

Although no formal meeting was held by the special Inter- 
Study Section Committee on Influenza Research, each member was 
asked to discuss the problem with his study section at its fall 
meeting, and to report back on items of interest and recommendations 
for research that may be forthcoming. A preliminary report was 
submitted to the National Advisory Council on Allergy and 
Infectious Diseases and the National Advisory Health Council at 
their November meetings. It is anticipated that an augmented 
report will be made to the Councils at a later date following 
discussions to be held in the spring of 1959. 

Cell Biology Study Section 

At the June 1958 meetings, in which 185 applications were 
presented for review, it became obvious that the work load of the 
Morphology and Genetics Study Section had become excessive. As 
a result, the Section was split into a Genetics Study Section, and 
a Cell Biology Study Section. 

The Cell Biology Study Section continued its interest in 
promoting additional research effort on the study of the cell. 
This includes various investigations of the cell: its parts and 
their significance; its differentiation; functional activities; 
its products and their effects on the producing cell and its 
neighbors; the organization of cells into tissue; and the near 
and remote influences on proliferation. In the main, only rudimentary 
information is available on these problems. 

In recognition of the experimental nature of this program, 
other study sections and the Councils were asked to comment on a 
plan designed to contribute to the security and research effective- 
ness of talented cell biologists, and to provide for the inspiration 
and training of more investigators in this specialty field. 
Elements of the program included: (a) long-range support of highly 
qualified investigators in amounts to meet all their needs and 
to be spent within broad limits at their own discretion, and (b) 
establishment of a committee advisory to the study section, and 
exclusively concerned with the Cell Biology Program. The second 
phase of the proposal—the inspiration and training of more students 


of cell biology--was envisaged as primarily dependent upon the 
establishment of the above recommendations — especially that 
concerning funding. 

The phase was approved by the Cell Biology Study Section 
at the September 27, 1958 meeting, and the first draft of the 
proposal submitted to an ad hoc group for consideration. 

Cardiovascular Study Section 

The Cardiovascular Study Section reviewed 245 applications 
during Calendar Year 1958. 

The section considered the possibility of applying electronic 
computer techniques to physiologic problems „ Dr. Otto Schmitt, 
a section member and chairman of the Joint Executive Committee on 
Medicine and Biology, was supported through the study section 
(with Heart Institute funds) in conducting a Conference on 
Electrical Techniques in Medicine and Biology,, The conference was 
expected to result in a series of applications submitted by 
collaborators representing the medical and electronic fields. 
The study section strongly favors encouraging the welding of these 
disciplines . 

Genetics Study Section 

During Calendar Year 1958, the Genetics Study Section was 
established in order to handle the increased number of research 
applications in this field, and to give appropriate recognition to 
this important area of research. 

The first meeting of the study section was held in September, 
at which time 31 applications were reviewed and study section members 
were oriented to the NIH programs and apprised of the various 
Institutes" program interests in genetics. 

The workload for the second meeting (scheduled for January 
1959) doubled, with a 200 percent increase in new applications 

A major concern of the study section is that of strengthening 
research and training in medical and human genetics. It is anticipat 
that specific plans for study section activities toward this end 
will be crystallized at an early date. 

Physiology Study Section 

Although most of the year was spent in the review and 
evaluation of research grant applications, a considerable amount 
of time at the regular meetings was spent discussing other problems 
in physiology and the biological sciences. As a result of these 
discussions, the Physiology Study Section presented to the Councils 
at their November 1958 meeting a resolution recommending support 


for longer than five years, and a resolution recommending the 
creation of NIH-supported tenure positions at academic institutions. 
In addition, the study section worked on a proposal to recommend 
the establishment of special merit training grants in physiology 
in a few of the most noteworthy physiology departments. 

During Calendar Year 1958, the Physiology Study Section 
held no conferences or symposia. 

Tropical Medicine and Parasitology Study Section 

During 1958, there were approximately 500 active grants in 
the Tropical Medicine and Parasitology Study Section. Within the 
same period, 176 applications were reviewed for scientific merit; 
of these, 12 7 were recommended for approval. Twelve applications 
of the 16 received from 10 foreign countries and territories 
were recommended for approval . 

The influence of the Public Health Service in: (a) stimulating 
basic and applied research in state health departments and among 
young investigators in small colleges, and (b) in expanding the 
scope of research in academic and research institutions, was 
evidenced in the types of applications received. Examples of 
projects of a comprehensive nature include that of the Bishop 
Museum of Honolulu (being sponsored by PHS, NSF, ONR, ana UNESCO) 
for the compilation of a catalog of the insects of the Pacific 
area. In addition to the scientific information that will be made 
available to specialists throughout the world, this project is 
basic to the control of vector-borne diseases in the Pacific. 
These data are vital to military forces. 

With support from PHS, the Gorgas Memorial Laboratory, 
Panama, is undertaking two projects of major importance. The 
first deals with the transmission of leishmaniasis. During the 
initial stages of the first project, a potential reservoir host 
for a strain of Leishmania which is infective for human beings 
was found.. Techniques were later perfected for rearing infected 
sandflies by feeding them cultures of Leishmania . The larval 
habitats of the sandflies were found for the first time in Central 
America . 

The other Gorgas project deals with the ecology of sylvan 
yellow fever. It is an investigation of the factors which normally 
prevent — and occasionally permit — the passage of the virus to the 
areas west and north of the Panama Canal. The data obtained from 
this study will have direct application to problems of yellow fever 
now facing this hemisphere. 

The need for expansion of research in disciplines of tropical 
medicine other than parasitology was explored. As a guide for such 
expansion, an evaluation of all of the research reviewed by the 
TMP Study Section and sponsored by the PHS was initiated. Consulting 
specialists accepted the task of evaluating the projects in 


particular fields, e.g., schistosomiasis, bat rabies, trematodes , 
nematodes, arthropod vectors, malaria and Anopheles , and insecticide 
resistance. Completion of the evaluation of all projects reviewed 
by the TMP Study Section is anticipated during Fiscal Year 1959. 

Planning was initiated for a 1959 special conference on 
bat rabies to be sponsored by the TMP Study Section. The Virology 
and Rickettsiology Study Section is expected to participate. 

The study section' s meetings were held at NIH during 
January and October of 1958. The meeting in April of that year 
was held in San Francisco, California, following project site 
visits in California and adjacent states, A total of 36 project 
site visits were made during 1958 . 

Endocrinology Study Section 

Through the research grant mechanism, the Endocrinology 
Study Section began preparing radioactive steroids for free 
distribution throughout the world. This program was greatly 
enhanced by a gift of 460 millicuries of barium carbonate C 
through the efforts of Dr. Charles L. Dunham of the Atomic 
Energy Commission. The preparation of these steroid compounds 
neared completion during the year, with distribution scheduled 
to begin in early 1959. 

Aided by a National Heart Institute research grant, the 
Endocrinology Study Section held an International Work Conference 
on Hormones and Atherosclerosis at Brighton, Utah, on March 12-14, 
1958. The papers and ensuing discussions were sent to press and 
the volume was scheduled for distribution and sale in January, 1959. 

Through a research grant recommended by the study section, 
three-year support was given to a project that will prepare human 
urinary gonadotropins. The project was begun, and source material 
was obtained through the cooperation of the State of Kansas. It 
is anticipated that large amounts of standard material will be 
made available for distribution for research purposes, and for 
combining with materials from other countries for the purpose of 
establishing an international preparation. 

Another research grant recommended by the study section led 
to the establishment of an International Work Congress on the Assay 
and Chemistry of Pituitary and Urinary Gonadotropins. Papers 
resulting from this meeting, to be held in Gatlinburg, Tennessee 
on December 3-5, 1959, will be published in an independent volume. 

The study section continued to progress in preparing the 
several pituitary hormones. During 1958, a total of 143 requests 
for pituitary hormones and 60 requests for radioactive steroids were 
processed. The requests were received from biological laboratories 
throughout the world. 

13 - 

The Executive Secretary of the Endocrinology Study Section 
received an invitation to submit an original chapter titled, 
"Paradoxical Ovarian Secretions" for a book, "The Ovary." The 
volume is being edited by Sir Solly Zuckerman of Birmingham, 
England, and will be published by Academic Press in early 1959 , 

Hematology Study Section 

A subcommittee of the Hematology Study Section worked out 
detailed plans for a Conference of Fundamental Problems and 
Techniques for the Study of the Kinetics of Cellular Proliferation 
to be held at Salt Lake City, January 19-21, 1959, With support 
to be provided by the National Heart Institute, the subcommittee 
planned to publish the conference papers in a monograph. 

Among the topics to be discussed are DNA labeling techniques 
which are useful in assessing the span and fate of hemopoietic 
cells, regulation of leucocyte and platelet production, and the 
mechanisms that regulate the rate of red cell production. 

Human Embryology and Development Study Section 

The Human Embryology and Development Study Section sponsored 
joint ly--with the Association foi the Aid of Crippled Children, 
and the Collaborative Project on Cerebral Palsy and Other Neuro- 
logical and Sensory Disorders of the National Institute of 
Neurological Diseases and Blindness--a Conference on Epidemiological 
Aspects of Pregnancy Wastage. The conference was held March 
23-25, 1958, at Arden House, Harriman, New York, 

Because of the increasing number of applications presented 
on research problems dealing with the use of radioactive isotopes 
in pregnancy, the study section became increasingly more concerned 
with the radiation hazard involved. Plans for holding an inter- 
study section conference were placed under consideration, and 
appropriate researchers in the field concerned were invited to 
express their opinions regarding the desirability of holding such 
a conference. The information received will be given to the study 
section for consideration at the first meeting during 1959. 

Dental Study Section 

At the January 1958 meeting of the Dental Study Section, 
the panel conceived a symposium titled, "Clinical Estimation of 
Caries Incidence and Increments." The symposium was held in 
two annual sessions beginning in September 1958. The program dates 
were finalized at the Mav 1958 meeting of the study section. 

The first session was held at Ohio State University on 
September L. and 5, 1958 , The final session is to be held September 
21-24, 1959 at the same university. The symposium is composed of 
approximately 55 participants. At the termination of the study 


the findings will be published, probably in the form of an appendix 
to an established dental research journal. 

Applications for grants-in-aid of dental research continued 
to average approximately 65 each trimester. 

During the year, the study section formed several committees 
for the purpose of planning annual extracurricular activities. 
The first such activity will be an International Work Congress 
dealing with cleft palate and its associated phenomena. It 
will be held December 6-9, 1959, at the Mountain View Hotel, 
Gatlinburg, Tennessee. Each dental school was invited to send 
two observers for which partial subsidy will be available. 
Formal papers, together with the ensuing discussion, will be 
published in an independent volume. It is anticipated that five 
or six additional annual congresses will be held wherein all 
fields of dental research and education will be given minute 
consideration. The results of each congress will be published, 
each being a part of a general title concerning oral health and 

Sanitary Engineering & Occupational Health Study Section 

Increased interest in environmental health research was 
reflected by the fact that during 1958 a total of 152 research 
grants supported investigations in the four major areas of interest 
to this study section: air pollution, food technology, water supply 
and water pollution, and contamination by radiation. Activity in 
each area exceeding that for Calendar Year 1957. 

The major item of interest in the field of air pollution 
was the recommendation of the National Conference on Air Pollution 
(held during November 1958 in Washington, D. C.) that the Federal 
Air Pollution Control Act be extended beyond its planned expiration 
date of 1960. The recommendation reflected the growing national 
concern of industry, science, and the public on the problem of 
air pollution. During Fiscal Year 1958, 52 research grants were 
active in this field. 

In food technology, 13 research grant projects were active, 
as opposed to four such projects during 1956. Some of the neglected 
areas in which research was initiated during 1958 included the 
effects of treating foods by radiation, the growth of certain 
biological agents in foods, and studies of various phases of food 
production in order to minimize toxicological hazards from contami- 

Eighty-four water supply and water pollution research grants 
were made; 75 were continuation grants, and nine were for new 
studies . 


Three grant-supported projects in the area of environmental 
health contamination bv radiation dealt with radioactive fallout 
in the Pacific Northwest, radioactivity in marine organisms and the 
marine environment, and radioactive accumulation limits in edible 

General Medicine Study Section 

The General Medicine Study Section reviewed 228 applications 
requesting support of a broad spectrum of clinical studies in the 
fields of arthritis, connective tissue, diabetes, gastroenterology, 
cystic fibrosis, metabolism and dermatology. 

The section sponsored a January 6, 1958 Conference on New 
Frontiers in Ulcerative Colitis, held at the Woodner Hotel in 
Washington, D. C. Objectives of the conference were to: (a) 
re-evaluate the problem of ulcerative colitis in its fundamental 
aspects; (b) develop new ideas and research approaches to studies 
of this condition; (c) stimulate the interest of younger investi- 
gators, and encourage the investigators already engaged in 
research on this problem, and (d) to disseminate the conference 
proceedings through a publication for distribution to interested 
investigators and physicians. 

Pharmacology & Experimental Therapeutics Study Section 

The study section sponsored a symposium on catechol amines 
in order to integrate current information on the rapid advances in 
research on catechol amines, and to clarify seemingly conflicting 
research findings that tend to cloud the understanding and clinical 
significance of these amines. The symposium was held at NIH on 
October 16-18, 1958, and papers were presented by 14 U.S. and six 
foreign scientists. 

Among the findings presented was a new theory regarding the 
biochemical transmission of neural impulses. While the parasym- 
pathetic transmitter has been definitely identified as acetylcholine, 
the nature of the sympathetic transmission remains a major question. 
One researcher presented evidence that the substance released at 
sympathetic nerve endings is not epinephrine, but its precursor-- 
norephinephrine . 

Pathology Study Section 

The Pathology Study Section sponsored two conferences: 
(a) a workshop conference held June 16-20 at Western Reserve 
Medical School for the purpose of encouraging the use of experi- 
mental pathologic techniques in undergraduate teaching of 
pathology; and (b) an August 21-23 seminar at the University of 
Wisconsin for the purpose of attracting and stimulating pathology 
students and those in ancillary disciplines into experimental 


Jointly with the World Health Organization, the Pan American 
Sanitary Bureau, and the Food and Agriculture Organization of the 
U.S., the study section provided support for a June 1-5, 1958, 
International Conference on Human Nutritional Diseases at Princeton, 
New Jersey. The symposium was planned by a subcommittee of the 
study section. 

Further, the section participated in a working Conference 
on The Responsibility of the Chairman of the Department of Pathology 
for Training, Research, and Hospital Practice in the Field of 
Nuclear Energy. The conference was held February 6 and 7, 1958 
at Brookhaven National Laboratory. 

Behavioral Sciences Study Section 

The Behavioral Sciences Study Section met in four series 
of meetings, one of which was a September 1958 meeting held 
jointly with the Mental Health Study Section. 

Proposals considered at the various meetings and endorsed 
by the study section for submission to the Council included 
extension of the Russian translation activities to include total 
translation of the "Pavlov Journal of Higher Nervous Activity," 
and an expansion of coverage of relevant Russian literature in 
the Psychological Abstracts. Further, the section endorsed the 
support of a Conference on Objective Tests of Personality tenta- 
tively scheduled for May 1959. 

Some other areas for discussion at the meetings included: 
(a) the relevance of anthropological training to mental health 
research, and the insufficiency of ordinary fellowships and 
training funds for financing the necessary anthropological 
training, and (b) consideration of the Bayne-Jones report 
proposals for "program grants" and "base grants." 

Mental Health Study Section 

At its first meeting of Calendar Year 1958, the Mental 
Health Study Section expressed considerable interest in securing 
translations of Russian literature dealing with the behavioral 
sciences. Later, it formally endorsed a statement on "Need for, 
and Translation of, Russian Scientific Literature" prepared by 
the Behavioral Sciences Study Section with respect to: (a) trans- 
lation of the Pavlov Journal of Higher Nervous Activity, and (b) 
possible expansion of Psychological Abstracts to include more 
comprehensive coverage of Soviet literature. 

Biochemistry Study Section 

The rapidly increasing volume of research grant applications 
received in the Biochemistry Study Section reached the point 
where the quality of review of such application was jeopardized. 

17 - 

As a result, a new panel--Physiological Chemistry Study Section— 
was formed, composed in part of incumbent members of the original 
study section. Both panels were then brought up to operational 
strength through the appointment of new members, and were scheduled 
to meet separately during January 1959. 

Although in a major degree the two panels image one another 
in scientific qualifications and competence, the Biochemistry Study 
Section is responsible for evaluation of the projects oriented 
tot^rd organic chemistry, while the Physiological Chemistry panel 
is identified with the biochemistry proposals that embrace elements 
of clinical biochemistry. 

The Biochemistry Study Section continued its interest in 
the activities of the Committee on Biological Chemistry of the 
National Research Council. The committee's program includes 
developing criteria, specifying physical characteristics, and 
recommending methods of analysis for the guidance of laboratory 
users of commercially available biochemicals . 

During 1958, the study section was concerned with studying 
the many problems involved in the recruitment of graduate students. 
General discussions pointed to the need for development of stimuli — 
at pre-university levels — that would hopefully result in motivation 
of greater numbers of candidates for advanced degrees in biochemistry; 
and, better preparation in the physical sciences—at the undergraduate 
level— of those motivated to seek postgraduate training in anticipation 
of careers in research and teaching. 

The study section was fully aware of programs initiated and 
planned by the National Science Foundation, the American Institute 
of Biological Scientists, and other organizations to replenish and 
increase the scientific manpower pools for fundamental research in 
the future. It was, nevertheless, concerned with the specific 
measures that will assure appropriate training of future investigators 
in the life sciences, particularly in biochemistry. The study 
section believes that one area of opportunity, in particular, is 
being neglected by the programs extant for strengthening our future 
potential in this discipline. Namely, most of the departments of 
chemistry and biology in the smaller liberal arts colleges provide 
little or no teaching of the fundamentals of biochemistry, and do 
not bring to the undergraduate an understanding of biochemistry and 
related life science disciplines as areas of career research and 
intellectual opportunity. 

After discussion of all aspects of the problem, the study 
section directed that a subcommittee develop recommendations for 
constructive and workable measures designed to provide or enhance 
the motivation of undergraduates to seek and enter careers of 
research in the life sciences. The subcommittee will present its 
recommendations in the spring of 1959. 

D I V I S I N 



Serial No„ __PRS-1 


Summary of Program Activities 
Calendar Year 1958 

Established in 3.955, the Division of Research Services 
provides the scientific, technical, and engineering services re- 
quired to support and further the complex and expanding research 
programs of the National institutes of Health. 

The Division, which employs more than 8C0 persons with 
approximately 200 individual occupational skills, is comprised of 
six branches: Plant Engineering, Research Facilities Planning, 
Scientific Reports, Laboratory Aids, Sanitary Engineering, and 

DRS Programs — 1958 

Operated and maintained all buildings, utilities, and 
grounds within the NIH plant and installed and helped main- 
tain various types of equipment. 

Provided design, engineering, and construction services 
for the comprehensive NIH building and remodeling program < 

Coordinated the planning of all construction and remodeling 
work, and conducted engineering and architectural studies 
to assure the orderly development and growth of NIH 

Provided photographic, art, library, translating, writing, 
and editorial services to all of NIH. 

Translated and disseminated Russian material on medical, 
biological, and related sciences. 

Produced and supplied laboratory animals and media for use 
in research. 

DRS, Summary of program Activities--! 958, cont'd: 

Provided veterinary services, including diagnostic, surgi- 
cal, and x-ray, to NIH scientists. 

Designed and fabricated special scientific instruments, 
apparatus, and glassware that were unobtainable from 
commercial sources. 

Designed, maintained, and tested mechanical apparatus for 
research, clinical, and office needs. 

Designed, installed, maintained and tested electrical, 
water, and waste disposal systems throughout NIH. 

Supervised the completion of an NIH Waste Chemical 
Disposal Plant. 

Cleaned and repaired NIH laboratory glassware. 

Studied and improved all aspects of environmental health 
and sanitation at NIH, including food service facilities 
and air pollution. 

Expanded and improved the Gem-Free Animal Facility 
established at NIH in 1956, and instituted a project of 
developing transportable germ-free animal chambers for use 
in aboratories. 

Provided biometric consultative services on the design, 
execution, and analysis of experiments and studies con- 
ducted by the Institutes and DBS, and conducted independent 
research investigations within the Biometrics Branch. 

Operated an expanded central statistical data processing 

Programmed and processed administrative and research data 
on the new IBM Type 650 Electronic Computer and related 
data processing equipment. 

DRS, Summary of Program Activities— 1958, cont'd: 

Conducted and participated in review, refresher, and job- 
related courses designed for animal caretakers, laboratory 
technicians, and clerical personnel. 

Conducted independent Division studies—over and above 
those specifically requested— for the purpose of antici- 
pating research needs and improving the quality of the 
services provided by the Division. 

Accoirrp. .ishments 

In addition to discharging its routine program responsi- 
bilities, the Division worked toward improving and broadening 
services in specific major areas. Some of these include: 

Construction The Research Greenhouse, the 

Biologies Standards Building, and 
an extension to the Powerhouse were under construction during 1958, 
and extensive projects were in the planning stage. Significant 
progress was made in planning for the Dental, Office, and Surgical 
Facilities Buildings for which funds have been authorized. 

Improved Management In line with recommendations of 

Practices, Plant the Kearney Report of 1957, the 

Engineering Branch Plant Engineering Branch shops 

consolidation and transfer to the 
first floor of Building 13 was 60$? completed. Engineering design 
personnel from all sections were consolidated, with more effective 
utilization of engineering talent. Hew procedures adopted during 
the year include improved management of shop stores, with a reduc- 
tion in the number of items, the use of repeating requisition card 
forms, and the application of E.O.Q. 'Economic Order Quantity) 
system of replenishment of stock. 

Ivtester Utility Study The planning phase of a study 

designed to develop a master 
utility plan for the reservation has been completed and is ready 
for contract by Public Buildings Service, Completed plans will 
complement the master site plan developed in 1957 and vail provide 
essential data for the orderly planning of future construction. 

DBS, Summary of Program Activities— 1958, cont'd: 

Instrumentation Space New quarters have been provided 
and Program for the instrument Section, in- 

cluding adequate space in adjacent 
areas for the Engineering and Design and Fabrication Sections. 
Improvements in facilities, successful recruitment of young 
professional personnel, and the completion of significant reseai'ch 
instruments during the year have contributed to an improved 
climate for professional personnel. 

Animal Production Several nev/ species of animals 

were added during the year* There 
was an increase in animal production of 6.6% over 1957, most of 
which resulted from a reallocation of space, A IjS increase in the 
number of usable animals is attributable to the training of care- 
takers. An Assistant Chief of the Laboratory Aids Branch was 
recruited during the year, and his services are expected to 
strengthen the professional guidance of the Animal Production and 
Animal Pathology Sections. 

Staff. Office of Chief Additional personnel, including 

an Executive Officer and an 
Administrative Assistant, were added to the Office of the Chief 
in line with plans to strengthen the Division's budget and train- 
ing programs. 

Statistical Processing Requests for data processing, 

involving utilization of the 650 
Computer and other statistical processing equipment, developed 
rapidly during the year. Arrangements were made for the procure- 
ment of additional equipment and the accelerated employment and 
training of personnel. 

Staff. Research Facilities Improvements in staffing for this 
Planning Branch program through the recruitment of 

four engineers will permit more 
efficient management of the construction program. A system of 
assignments has been devised whereby an individual engineer is 
assigned to each building through its various stages of design 
and construction, thus providing continuity in communications with 
the Institute concerned and broad experience for young engineering 
personnel . 

Program and Budget A major step in long-range plan- 

Forecasts, ning was made possible through 

. Summary of Program Aetiv: 058, cont'd : 

participation in activities of th* Task Force on Intramural 
Research. The resulting guidelines will he of considerable value 
to professional and admir istrative personnel responsihle for 
future planning and development of programs. 

Training Training activities sponsored by 

the Branches and Sections were 
increased slightly and the Division continued to make use of 
specialized technical and professions; training courses sponsored 
hy colleges and universities. The Technical Writing Course for 
t'RS technolog'. °t:' ; and the course for animal caretakers were 
considered especially effective. 


Recruitment Adequate recognition in terms of 

professional opportunities, 
grades, and salaries continues to be the major and complex 
problem in fi3.1ing key DRS jobs. Intricately related to this 
problem is the DRS role in the research program. It should be 
recognized that in a central service activity scientists must 
have opportunities for development on a professionally acceptable 
plane. The DRS ability to provide services and materials of high 
quality and in ine with advanced research needs is dependent 
upon opportunities for continued development and upon professional 
recognition for its personnel. Progress is continuing in this 
important area. 

Training C os<? y related to recruitment 

and of equivalent value in the 
provision of efficient and high qua ; scientific services and 
materials is a well planned and continuing training program. 
Although instances of progress have been reported in this area, 
neither the efforts nor the results meet recognized needs. 
Additional leadership and impetus are being provided for this 
activity with plans for early emphasis upon job-related training 
and supervision. 

3udget Procedure The Division continues to be 

hampered in its efforts to respond 
to research needs due to the inflexibility of budgeting procedures, 
resulting in a lag between demands and services. Efforts to 
relate fees to services have further delayed the solution of this 
problem except when financing is through the Revolving Fund. 

Serial No. DRS-1.1 



Calendar Year 1958 

Budget Data ; 

Actual obligations for Calendar Year 1958: 
Total: $103,467 
Reimbursement: Office of the Chief, DRS--$103,467 

Serial No. DBS-1.2 



Ca .endar Year 1958 

Budget Data : 

Actual obligations for Calendar Year 1958: 
Total: $6,446,527 
Reimbursement: Total services, DRS — $6,446,467 

Pebruary 1959 Calendar Year 1958 


Summary of Branch Activities 1. DEIS- 2 

Serial Number 


Branca Chief 

Objectives and Programs 

The primary objectives of the Plant Engineering Branch are to 
provide three major central services in the oust effective and eco- 
nomical way. These major central services are: 

Operation and Maintenance , which consist of operating, main- 
taining, and repairing the physical mechanical, electrical, archi- 
tectural and landscaping facilities of the NIH; and maintaining and 
repairing laboratory and research apparatus. 

NIH facilities include 306 acres of land and 42 laboratory, 
administrative and service buildings, totaling over two and one-half 
million square feet of floor space. These facilities represent a 
capital investment of more than $100 million. 

Engineering Services , which consist of developing and prepar- 
ing plans and specifications for alterations, repairs, and exten- 
sions to the existing facilities, administering and inspecting con- 
struction work performed by private contractors, and providing con- 
sultant and technical engineering services for development of 
facilities and research apparatus. 

Construction Services , which consist of installing and fabri- 
cating facilities and equipment for research programs. In Shis 
capacity the Branch furnishes mechanical, electrical and architec- 
tural crafts services. 

Progress and Accomplishments 

In the interest of meeting the needs of NIH on an efficient 
and economical basis, major efforts were devoted to the following 
management improvements during the year: 

Physical rearrangement .and consolidation of PEB 
construction and maintenance shops. 

DRS, Plant Engineering Branch, cont'd: 

Reorganization of PEB shops stores .management, 
physical consolidation and rearrangement of shops 
stores and establishment of a stock control system. 

Development of a central point of control for plan- 
ning, estimating and scheduling PEB construction, 
maintenance and related activities. 

REARRANGEMENT AND CON- A study of this program was initiated 
SOLIDAXION OP SHOPS in 1957 with the assistance of the 

consultant firm of A. 7. Kearney and 
Company. As a result of the study it vas decided to physically con- 
solidate the PEB construction and maintenance shops on the first 
floor of Building 13. These shops were originally located on four 
different floors of the building. 

Implementation of the shop consolidation plan began in 1953. 
The plan for consolidating PSB shops included moving the Instrument 
Shop and other activities of the Instrument Section, LAB, to the 
third floor. Building 13. Plant layout studies were completed for 
new arrangements of offices, machine tools and equipment for both 
the PEB shops and the LAB activities- Two PEB shops originally lo- 
cated on the third floor of Building 13 were temporarily relocated 
to the second floor, and the third floor area vas completely reno- 
vated for LAB activities. Concurrently, space originally occupied 
by the Plumbing Shop adjacent to PEB offices on the ground floor was 
renovated to provide additional PEB office space. The Plumbing Shop 
was temporarily relocated to space on the first floor- originally 
occupied by the Instrument Shop. 

Plans and specifications for the renovation to complete the 
consolidation of the PEB shops were completed during the year. The 
new arrangement is designed to increase the efficiency of PEB shop 
operations and permit increased service to the Institutes and 
Divisions of NIH. 

Completion of the consolidation of the shops is scheduled for 
August 1, 1959. 

REORGANIZATION 0? A study completed in 1957 determined 

PEB SHOP STORES that the Branch had in stock an average 

of 17.4 months supply of shops stores 
material, representing an investment of $439,800, and occupying 
16,300 square feet of space. It was estimated that inventories could 
be reduced to an average six months supply, representing a $242,000 
reduction in investment. Floor space could be reduced in proportion. 

DRS, Plane Engineering Branch, cont'd: 

In January 1958 (management of the PBB shop store* was con- 
solidated into a single organisation (stores were previously managed 
by the respective shops to which they were attached). In June 1958 
the A. T. Kearney Company was retained as consultant to assist in the 
development and installation of new stock control procedures. 

A more detailed analysis of stock material was made, which 
indicated that existing stock consisted of 12,590 separate items, 
having a total Inventory value of $435,550. New guide lines and 
control factors were developed to permit reduction in the number of 
items to 6,550, having an average Inventory of $116,600; a total in- 
ventory reduction of approximately $318,950, and a savings of approxi- 
mately $32,000 per year in carrying coats. (The above figures ex- 
clude construction materials financed through the NIH revolving fund). 
Considerable savings in labor are anticipated, but the exact re- 
quirements have not yet been determined. In addition, controls are 
built into the system which will greatly reduce if not eliminate 
outages of normal stock. 

By the end of 1958 approximately fifty percent of the new 
stores program was completed, with all major control factors deter- 
mined and procedural decisions reached. 

Portions of the program yet to be completed consist of the 
following: the bundling and bagging of stock in appropriate control 
quantities; the placement of stock in one storeroom; the preparation 
of formal procedural instructions; the acquisition of necessary 
office equipment, printed forms, and supplies; the determination of 
new personnel requirements and the completion of staffing. 

Present target date for completion of the transition to the 
new system is August 1, 1959. 

DEVELOPMENT OF A CBN- During the year the development of a 
TRAL CONTROL POINT central control point for planning, 

estimating, and scheduling the work 
of the Branch was started. The organizational function, called the 
Project Control Office, reports directly to the Branch Chief. 

All construction and alteration work (maintenance excluded) 
was processed through this office during the year. By planning, 
recording, estimating, and scheduling, a reduction was made in the 
average time between receipt and completion of projects. 

Also valuable data collected and experience acquired during 
the year have greatly assisted the Branch in reaching the conclu- 
sions that the following should be done as soon as possible; 


DBS, Plant Engineering Breach, cont'd: 

Maintenance. Bring control of maintenance work ae well as 
construe tion work under this office by developing standing job 
order* for routine maintenance work, and scheduling all other work. 

Determine areas and equipment to be inspected for preventive 
maintenance, the individuals who will make the inspections, and 
schedule the inspections from this office. 

Scheduling . Estimate man-hours required for each specific 
Job order in advance of scheduling. 

Develop e better method of obtaining man-hours expended on 
each specific Job order. 

Use the foregoing to improve work planning and scheduling. 

Engineering Design . The personnel and functions of the three 
units (Mechanical, Electrical, and Construction) located in the re- 
spective sections of the Branch and the Drafting Unit located in 
Project Control Office should be combined and made an integral or- 
ganisational section of the Branch to permit better control and co- 

The above methods and procedures will be developed as soon as 

OTHER ACCOMPLISHMBJRS Shop Work . The shops completed 6,177 

alteration and/or construction projects 
for the NIH Institutes and Divisions during the year. This work in- 
volved 144,800 man-hours and included materials costing over $201,500 
(cost of used materials is not included). 

The total number of specific maintenance projects completed is 
not available since this category of work has not been placed under 
work authorization control. Materials expended on maintenance work 
performed by the shops cost approximately $182,000. 

Engineering Design Projects . In addition to the engineering 
devoted to plans for work performed by the shops and plans for new 
building construction, the Branch designed end prepared plans and speci- 
fications for 62 alteration and/or construction projects during the 
year. The construction cost of these 62 projects is estimated at 

Construction Projects (Contract) . The Branch inspected and 
administered 72 NIH construction contracts during the year. These 
projects cost a total of $393,316. 

DRS, Plane Engineering Branch, cont'd: 

In addition the Branch assisted fcha Regional Office, Public 
Buildings Service, with the inspection and administration of 22 
repair and Improvement projects contracted for NIH. The cost of 
these 22 projects was $516,000. 

Training . The following training was provided for personnel 
of the Branch during the year: 

Air Conditioning at Carrier Corporation, Syracuse, New York, 
(6 weeks) * 2 mechanical engineers attended. 

Executive Development and Human Relations at Army Map 
Service Corps of Engineers (2 weeks) - 1 mechanical 
engineer attended. 

Air Conditioning Clinic at Trene Company, Washington, 0. C. 
(10 night sessions) ** 2 mechanical engineers attended. 

Technical Writing Course at NIH (IS night sessions) - 6 
supervisors attended. 

Bearing Maintenance at NIH by Central Armature, Inc. 
(8 hours of Instruction) - 130 operating engineers and 

Orientation of Sanitary Engineering Officers at Sanitary 
Engineering Center, Cincinnati, Ohio (1 week) - 3 sanitary 
engineers attended. 

Instrument end Pneumatic Control at Republic Flow Meter 
Company, Chicago, Illinois (2 weeks) - 2 operating engi- 
neers attended. 

Instruction on Analyzing Vibration at International 
Research & Development Corporation, Columbus, Ohio (3 day 
session) - 2 mechanical engineers attended. 

Supervisory Training in Personnel Management at NIH by 

PMB and PEB (12 two hour sessions) - 15 supervisors attended. 

In addition to the foregoing three separate schools were con- 
ducted for treining operating engineers. Glasses 2 hours in duration 
were held once weekly for the Laboratory & Miscellaneous Buildings 
Unit, the Heating Plant Unit, and the Clinical Center Unit of the 
Mechanical Engineering Section. The classes were taught by the re- 
spective unit supervisors with the occasional help of others. 

Personnel Recruitment . Recruitment of professional engineers 
during the year was difficult but reasonably successful. Six pro- 
fessional engineers GS-11 or above ware employed. 

Serial No, DRS-2.1 


Calendar Year 1958 

Budget Data : 

Actual obligations for calendar year 1958: 
Total: $3,532,962 

Reimbursement: $3,532,962 

February 1959 Calendar Year 1958 


Summary of Branch Activities l c DRS-3 

Serial Number 


Branch Branch Chief 


Branch objectives are to provide central services to the 
Institutes and Divisions in areas related to scientific reports and 
communication. These services are indicated by the Branch's organi- 
sational units, which are as follows: Library Section (including 
the Translating Unit and the Russian Scientific Translation Program), 
Medical Arts Section, Photography Section, Editorial Section, Pub- 
lications and Reports Section, and the Office of the Chief. 


Library Section 

Provides a prompt and comprehensive supply of pertinent infor- 
mation to NIH scientists and to investigators elsewhere. 

Assists the individual scientist in keeping currently informed 
not only on the antecedents of their experimental work but also in 
the rapidly accumulating knowledge in their own fields. 

Provides educational materials for formal and informal on- 
the-job training and education. 

Translating Unit 

Translates and loans translations from 19 languages into 
English as well as translations from the English language into French, 
Spanish, German, and Italian. 

Russian Scientific Translation Program 

Reports on current medical research in the USSR through ab- 
stracts and reviews, and in extenso translations of Russian journals 
and books. 

DRS, Scientific Reports Branch, cont'd: 

Provides background information on Russian medical research 
to assist American scientists in evaluating status and quality of 
Russian medical research. 

Medical Arts Section 

Provides a variety of central graphic services for the NIH 
scientific staff. 

Designs, constructs, schedules, and ships research exhibits. 

Prepares general, technical, and medical illustrations for use 
in lectures, scientific publications, and exhibits, and for adminis- 
trative, informational, and teaching purposes. 

Develops visual and functional tools, such as plastic and wax 
models, to assist the scientific staff at NIH. 

Photography Section 

Provides black-and-white and color photographs, photomicro- 
graphs, slides, transparencies, and motion pictures to illustrate, 
document, and validate research, both in the presentation of scien~ 
tific reports and in the provision of general information concerning 
NIH research accomplishments. 

Investigates potential photographic techniques that may con- 
tribute to the solution of special research problems. 

Designs special photographic equipment or adapts existing 
types to fit specific production or research problems. 

Editorial Section 

Offers broad editorial services and individual editorial anal- 
ysis (on request) to NIH scientists in preparing papers for presentation 
or publication, and to information officers and others associated with 
the preparation and use of NIH publications. 

Serves as a focal point for recording and approval (in the name 
of the Director, NIH) of all scientific papers, speeches, abstracts, 
press releases, announcements, and written radio and television mate- 
rials, exhibit and publication copy, and all other written informational 
materials directed to the professional and general public. 

Screens the scientific literature, retrieving NIH titles for 
the use of information personnel. 

Prepares the annual NIH bibliography. 

DRS, Scientific Reports Branch, cont'd? 

Publications and Reports Section 

Provides a variety of central information services for NIH S 
the Division of Research Services, and other NIH components.. 

Publishes the MH Record and the Calendar of Events . 

Gives prepublication assistance and training in writing and 
editing of scientific and technical reports. 

Prepares special articles, scripts, speeches, and press re- 

Through central photo, slide, and publication libraries, sup= 
plies published or illustrative materials for lectures, various 
professional publications, and for newspaper, magazine, television, 
and other popular media of communication. 

Performs binding, distribution, and vari typing services for 
the NIH staff. 

Progress and Accomplishments 

LIBR, RY StiCTIOI Three developments occurred during the year 

which combined to reformulate the basic 
service policies of the Library,. In January a memorandum from the 
HEW Director of Administration clarified the relation of the NIH 
Library to the Department Library and to PHS headquarters offices,, 
In Deoember a position paper was drafted to clarify the Library* s 
future relationship to the National Library of Medicine, and, through- 
out the ye^r, a seiies of decisions were ~iade clarifying the amount 
of service the Libr. ry can give to other than NIH employees without 
effecting the efficiency of service to NIH research,, 

As the result of a review of NIH collection responsibilities 
as they relate to the National Library of Medicine, the Department 
Library, and libraries of other federal agencies, a start was made 
on a large-scale withdrawal and transfer program., United States, 
state, and foreign health reports prior to 19kS were offered to the 
National Library of Medicine to complete its holdings; and state 
reports were returned to state governments.! Pamphlets and reprint 
files were weeded; 6,000 catalog cards representing public health 
reprints were weeded from the public catalog; and in connection with 
a daily review of Journals received, many hundreds of decisions were 
made to eliminate second copy files. 

Steady progress was made in revising the Library's list of 
acoepted subject headings in the public catalog,, 

DRS, Scientific Reports Branch, cont'd: 

Installation of a flexowriter has aided in catalog and repro- 
duction and procurement of journals. The Library now has flexowriter 
cards for 2,100 Journal titles currently received. 

Security measures established during the year included the 
installation of panic locks on doors, special drives for the return 
of overdue books, continuous scheduling at the circulation desk, 
restriction of student use of the Library, and the general tightening 
of property clearance procedures. Plans were drafted to augment these 
security measures in 1959. 

Circulation Services . The Library's Circulation Unit loaned 
66,009 pieces in 1950, an increase of 11. k percent over 1957. Inter- 
library loans were less (5,209 in 1958 and 5,81*8 in 1957), and the 
index of items borrowed from other libraries to total loans dropped 
from 9.8 in 1957 to 7.9 in 1958. This may be interpreted as demon- 
strating the improving effectiveness of the Library's selection policy 
and procedures.. 

Total shelving, which includes loans, items used in the Library, 
and new items added, showed a 30„6 percent increase over 1957° The 
internal use was 38° 8 percent greater than in 1957. The Circulation 
Unit set an all-time high in annual and monthly loans t 66,009 and 

Reference Services , Reflecting staff vacancies and increased 
time devoted to book selection and administrative problems, reference 
services showed a quantitative decline for 1958. The Reference Unit 
answered 1,7U2 requests for biographical data and 2,557 requests for 
bibliographic verification (1,882 and 2,725 in 1957) plus 2,65U re- 
quests for other information. The staff also produced 262 selected 
lists of references by scientific subject; conducted hi intensive 
literature searches on behalf of NIH scientists; and prepared J>2 bib- 

Comprehensive bibliographies reproduced and distributed related 
to, among other subjects, maternal disorders, psychological aspects of 
atomic bontoing, mathematical models for breathing, and medical research 
in Russia, A detailed subject index to PHS Intramural Research, 1957 
was also prepared by the Reference Unit, which during the year was 
reorganized to reflect a division of responsibilities by scientific 
subject area rather than form of literature. 

Processing ,, The Library added 32,780 serial issues and 3,069 
monographs, of which 2,088 monographs ware purchased, 257 came by gift, 
and 31 by exchangee In addition 693 volumes were bound in 1958, and 
31*3 new serial titles were establishedo 

Concurrent with these additions, the Library withdrew 5,21*7 
titles (29,1*07 individual pieces) from its collections (largely older 

DRS, Scientific Reports Branch, cont'd: 

titles no longer related to current 1,1 H research) and catalogued 
U,09U titles. A total of 29,130 cards were filed in the public cata- 
log, and 3U.897 were removed as part of the withdrawal program, making 
a net reduction of 5,767 cards in the public catalog. 

Translating Unit The Translating Unit prepared a total of 

7,937 typed translations from English into 
French, Spanish, German, and Italian, including papers and speeches 
for presentation by KIH scientists at various international congresses.. 
The Unit devoted more than 1? work-weeks to oral translations— a 
service that is expected to increase in 1959. 

Related assignments included interpreting for visitors who 
speak French and Japanese and assisting members of NIH's clinical 
staff in obtaining case histories from patients who do not speak 

The Unit prepared, distributed, and circulated a monthly list 
of translated articles available on loan to the NIH scientific staff. 
A total of 3>U02 translations whose availability was announced in this 
fashion were loaned in 1958, as compared with 1,635 in 1957- The list 
is sent regularly to Technical Translations , a semi-monthly publication 
issued by the Office of Technical Services, Department of Commerce „ 

Russian Scientific Liaison was maintained throughout the year 

Translation iT-ogram with the National Science Foundation, the 

Commerce Department's Office of Technical 
Services, and other agencies, and with individuals concerned with the 
conrmnication of Russian science to the American scientific community. 

Administration of grants activities under the program was re- 
assigned to the Division of Research Grants. 

Funds derived from the sale of agricultural surpluses and co- 
ordinated by the National Science Fo inaction were made available for 
scientific translation.. 

Eighty-seven issues (10,582 pages) of eight journals translated 
from Russian were published and distributed to 320 medical libraries 
and to 80 government installations, bringing the cumulated production 
of the Russian Scientific Translation Program to 13,600 pageso A ninth 
journal, the Pavlov Journal of Higher Nervous Activity, was added during 
the year; papers for two l9i>8 issues have been translated, and the first 
issues will be published early in 1959 » 

Abstrac ts of Soviet Medioine published U,786 abstracts in eight 
issues,""a 58 percent increase over "1957 . Arrangements for exchange 
with Joint Publications Research Service, which translates the Russian 
abstract journal Biologiya , will increase this total still further,. 

HIS, Scientific Reports Branch, cont'd: 

Ei,;ht hundred pag3s of translations -were reproduced under the 
title of The Central Nervous System and Human Behavior for distribution 
at a Macy Foundation symposium in February 1959 as well as to UOO 
libraries. Twenty-five individual scientific papers were selected by 
editors of American scientific journals for translation and republication. 

Publication of a monthly bulletin announcing new translations 
was approved, and a first issue went to press for distribution early 
in 1959- 

The 1958 revised edition (2,000 copies) of the Directory of 
Medical and Biological Institutes of the DSSR has aided materially 
in the planning or scientific missions and has achieved an international 
(including Russian) approval. Work is proceeding on further revision,, 

The Russian-English Medical Dictionary , compiled by the National 
Library of Medicine and published by the Academic Press, was distrib- 
uted to U00 medical libraries. Another National Library of Medicine 
publication, the Ouide to Russian Medical Literature , was distributed 
to 7,000 PHS grantees, and A List of Russian Review Papers in Biology 
and Medicine was distributed to American journal editors 

Projects scheduled for early completion include publication 
of Brucellosis in the USSR and of The Academy of Medical Sciences , 
USSR , five translations to be published by the Pergamon Institute, 
and five others ( Public Health in the USSR , Organization of Public 
Health , Problems of Etiology and rath'ogenesis of Tumors , Problems""of 
Infectious Pathology in Expe r iments on Monkeys , and Curricula of tfie 
Higher Medical SchoolsJ intended for program use. 

MEDICAL ARTS SECTION The Medical Arts Section acquired space in 

the sub -base rert of Building 1 for a new 
studio to construct and store exhibits. Nineteen obsolete exhibits 
were eliminated from the number currently available for NIH-PHS use. 
An inventory and scheduling system has been devised for determining 
the present and future availability of current exhibits, and a picture 
file of exhibits is maintained for prospective users. The file has 
color slides, black-and-white photographs, specifications, and his- 
tories of the use of each exhibit available. 

Twenty-seven new scientific exhibits illustrating progress in 
NIH research were designed and constructed (several on contract) during 
the year, and nine exhibits were redesigned to present current scien- 
tlfio findings. A portable exhibit unit, weighing 7 pounds and costing 
about $15, was designed especially for transporting by airplane , 

Plastics Research . New developments in plastic research con- 
ducted by the Section are yielding numerous benefits to NIH in the 

DRS, Sclent iflc Reports Branch, cont»d: 

form of plastics models, plastic heart valves., museum mounts 3 eirtbedded 
pathological specimens, and fabrication of new and economical items. 
Two hundred forty-tic such items were completed in 1958. 

The Plastics Unit has developed a plsstic filter holder and is 
now developing a plastic funnel liner. New types of plastic museum 
jars were exhibited at an international pathology and bacteriology 
conference. Plastic models of standard psychological testing materials 
were developed and turned over to a reliable manufacturer for com- 
mercial fabrication. 

Illustrations . The Section filled 216 requests for medical 
illustrations and* U16 other requests for general and technical 
illustrations, including graphics, cartoons, and designs for bro- 
chures. Among the major projects was a series of illustrations 
depicting hereditary characteristics in the iris of 25 NHNDB oph- 
thalmology patients, all members of the same family. The Section 
has illustrated surgical techniques to correct heart anomalies and 
to demonstrate mitral stenosis insufficiency and intraventricular 

Drafting . The Section aupolied 5,512 illustrations for charts, 
graphs, maps, varltyped tables, and chemical formulae plates. 

Training . A program to instruct requesting NUi scientists and 
others in drafting techniques, instruments, and materials was given 
a trial in the fall of 1953 and is expected to become a regular service 
in 1959, The program will be expanded to familiarize scientists with 
type-size reduction methods and with the kind of photographic quality 
required for publication, 

Moulage Models . Eleven moulage models were made from original 
casts. Two were for a continuing project of followup impressions of 
NlAMD patients, (The jatients return periodically, and each time a 
new moulage is made to record any physical changes that may have oc- 
curred. ) Three wax and two plaster models were prepared from an original 
cast of a normal human heart. One of these was used to create the large- 
scale hei'rt displayed by the American Heart Association at the Inter- 
national Congress of Cardiology (September 1958, Brussels), Many more 
moulage s will be made from the original cast, 

PHOTOGRAPHY SECTION Requests for photographic assistance to NIH 

medical, scientific, and information staffs 
continue to increase. In 1958, a total of 7,113 requests were filled 
in patient and clinical photography, motion pictures, photomicrography, 
and gross pathology. Fluorescent photographic techniques previously 
developed by the Section are now routinely requested. 

DRS, Scientific Reports Erancb, cont'd « 

Deadlines have been speeded up from an average seven days to 
four days for all routine photographic activities, Effective use of 
space, supply purchases, and equipment trade-ins has saved tine and 
money o 

Patient Photography , Seven hundred ninety=three requests were 
processed for patient and clinical photographs. Among the major pro- 
jects were the collaborative photographic recording of six cancer 
patients undergoing skin transplant, and the refinement of color, 
black-and-white, and infrared techniques devised for studying the 
effects of radiation burns. 

A long-term skin graft study begun by klAMD in 1956 required 
U2 appointments with one patient; 17 were in 1958, 

Another long-term study begun in 1958 involved continued 
photographio study of color changes in a patient suffering from 
hypersensitivity of subacute capillaries, 

A cooperative genetic study by HIKDB and I'HR required approxi- 
mately 200 shots of more than 80 people, including a field trip to 
demonstrate photograpnically the correlation between eye tumors and 
the maleruption of teeth, 

Photordcrography and Gross Pathology Eighty NIH scientists 
made use of the Section's photomicrographic services in 1958 j 571 
requests were received, and 12,301 photomicrographs were produced. 
The Section has assisted in all phases of NIH germfree studies, in- 
cluding autopsy records and photomicrography of sacrificed aniroalso 

Photographio techniques developed for NIAID and MAMD studies 
to show the extent of colloidal change within agar and starch gel 
plates have aided scientists engaged In virus research. 

At the request of and in cooperation with NINDB, the Photography 
Section has developed photographic techniques to provide serial patho- 
logical records of brain seoti^ns, including photographs of actual 
brains as they develop from fetal to adult si*e, A similar technique 
was used in NIMH research to photograph 100 serial sections of a 
porpoise head. All in all, 288 requests were processed for photographs 
of gross pathological conditions. 

Cinema tography Records , The Motion Picture Unit has documented 
progress of patient rehabilitation; portrayed heart surgery techniques; 
and cooperated in recording various experiments in KIH's closed tele- 
vision circuit and in obtaining images directly from the TV screen,, 

Total motion oicture footage in 1958 came to 3,950 feet in 
black-and-white film' and 13,950 feet in color. In addition 1,800 feet 
of sound recordings were made, and ?1,850 feet of film was edited and 
prepared for presentation, iiighty-three requests were made for this 

DRS, Scientific Reports Branch, cont'd* 

EDITORIAL SECTION During 1958 the Editorial Section, acting 

for the Director, NIH, cleared 1,935 scien- 
tific papers, abstracts, and speeches; 119 press releases; and 17 
exhibits. The Section alerted the Chief, Office of Research Information, 
to 92 papers, whose subject matter was of interest to the Office of 
the Director, NIH, and to the Surgeon General. 

Clearance policy on NIH information material intended for 
Voice of America Broadcast was established during the year. Besides 
NIH, approval must be obtained from PHS Chief of Information Services 
on this material. 

In November, the Section completed an interim report on the 
time lag between NIH clearance of scientific papers and publication 
in the journal of first choice by NIH researchers. The survey will 
continue in 1959 to include papers published in 1957 and 1958. The 
results of this study will be prepared for publication. 

The Section has prepared a first draft of a brochure on the 
Division of Research Services and another first draft of a commentary 
to accompany orientation lecture slides on the Division's activities. 

Transmittal of Informational Materials to NIH Printing Officer , 
with the cooperation of NIH administrative officers, steps were taken 
to reduce the Section's workload in preparing printing and duplicating 
requisitions for transmit -al to the NIH Printing Officer o During the 
year, copy was reviewed and printing requisitions were processed for 
111 pamphlets, brochures, health education leaflets, and other pub- 

Preventive Editing . The preventive editing program begun in 
1957 was continued for individual scientists requesting editorial 
analysis of their papers prior to publication. Preventive editing 
was applied to all press releases submitted for clearance and to 18 
scientific papers. 

Literature Reporting . The reporting service initiated by the 
Section in 19$7 is based on a screening of almost 200 scientific journals 
on arrival in the NIH Library, Daily reports were discontinued and in 
their place a twice-weekly distribution was made to Institute directors, 
research directors, laboratory chiefs, information officers, and others. 
During the year the Section issued 63 reports listing 1,063 titles of 
NIH scientific papers and 76 reports listing 2,U12 papers on NIH grant- 
supported projects. 

In 1958 the Section distributed 50 copies of Current Contents , 
(a periodical reoroducing the contents pages of nearly 250 primary 
science journals) to routing lists suggested by the NIH Scientific 
Directors. Late in 1958 the Scientific Reports Branch relinquished 

DRS, Scientific Reports Branch, cont'd: 

its role in introducing this periodical,, Subscription purchases are 
now made by the individual Institutes, but the Section lias arranged 
to combine all purchases on a master order and to distribute the in- 
coming issues to the subscribing Institute or Division.. 

NIB Bibliography , a total of 1,000 copies of the 1957 NIH 
annual bibliography ( The Published Scientific Papers of NIH. 1957 ) 
were published in September. In October an announcement describing 
the usefulness of the bibliography was prepared for Public Health 
Reports . "' 

The bibliography featured an author index for the first time. 
Several procedural changes have been initiated with the aim of ac« 
complishing earlier publication in 1959. 

Collected Reprints and Archives . The Section's files provide 
the periodic listing in the MH Record" of scientific papers received 
for clearance,. The Section maintains a central repository and index 
for all papers submit ed for clearance and has similar systems for 
all reprints it receives. All reprints (791) received for 1957 have 
been indexed, collated, and are ready for binding. Five hundred 
ninety -five reprints were indexed for 1958. 

Correspondence and Inquiries . The Section responded to 2,360 
pieces of correspondence and telephone calls from the general public, 
NIH staff, and other government officials. Inquiries covered the 
whole range of MH activities and required routing to infornation 
officers of every Institute, to departments and individuals in HEW 9 
and to other government agencies. 

PUBLICATIONS AND During 1958, the Publications and Reports 

REPORTS S&CTIO:. Section wrote a narrative report for the 

Office of Research Information detailing 
the Section's information program, and wrote statements on the 
Division of Research Services' long-range goals, several sections 
in the DHEW 1957 annual report, and the Division's 1957 and 1958 
Highlights of Research Progress ; assisted in preparing background 
statements for budget hearings; and participated in the final editing 
of DRS Branch reports for inclusion in the 1957 Analysis of Program 
Activities . 

The Section wrote and published a DRS flyer designed to interest 
electronics engineering graduates in NIH career opportunities. It 
revised the NIH section in The Public Health Service Today , a PHS 

The Section continued its work on updating Tfce National Insti - 
tutes of Health brochure «, Completion of this assignment was delayed 
by the NIH organisational changes that took place in the latter part 
of the year 

DRS, Scientific Reports Branch, cont'd! 

Final copy on a recruitment brochure for the NIH Board of 
Civil Service Examiners was prepared for 1959 publication. 

Periodicals ,, The Section has drcfted proposals for a revised 
format for letterpress printing of the NIH Record , at a slight increase 
in reproduction cost per issue. The changeover is planned for 1959 
pending approval by the PHS Chief of Information Services, The Section 
published 26 issues of the Record and issued the weekly Calendar of 
invent S o 

Special Writing Projects . The Section prepared a feature story 
on NIH for the Portland (Maine) Sunday -Telegram , a statement on the 
extent of research in several disease areas for the California State 
Legislature, a description of KIH for the World Book encyclopedia, 
information on NIH training opportunities for the annual education 
issue of the Journal of the American Medical Association , and a re- 
vision of the NlH section in the Pethesda -Chevy Chase Community Guide . 

Speeches were drafted for the Director, NIH, for his introduction 
of the speaker at the 7th annual NIH lecture and for his welcome of 
guests to the 8th annual instrumentation symposium and exhibit. The 
Section handled publicity and press relations for both events, and will 
continue this assignment in 1959<> 

Fifteen press releases were prepared and distributed, including 

two announcing the availability of the NIH translation of Russia's 

5-year olan in pharmacology and of the revised edition of the Directory 

of Medical and Biological Rese rch Inst i tutes of the USSR . 

Scripts were written for use in narrating a DRS slide presen- 
tation for the MH "compass" course and for demonstrating a new sound- 
slide projector. Script assistance was given in preparing a revision 
of the NIH information filn and of a fire prevention week demonstration. 
A script for a 20- minute sound-color film depicting DRS research services 
was begun. 

Central Audiovisual Services . Ihe Audiovisual Unit greatly 
increased its service to NIH staff members by expanding and improving 
its visual photofile. The Unit supplied 5$5 photographs for infor- 
mational use, 

A total of 662i slides were loaned to NIH staff members who 
lectured in all sections of the United States and in Europe and Asia. 

The Unit assisted in a number of NIH picture -taking projects 
for professional and general press use, including the taking of more 
than 300 pictures for U. S. News and fcorld Report . The Unit also 
assisted with the filming of NIH scenes for such nationwide TV projects 
as the CBS show "Conquest" televised November 16. 

DRS, Scientific Reports Branch, cont'd: 

Central Photocopy Preparation , Through its publication planning 
and varityping service, the Photocopy Preparation Unit produced a bro- 
chure for i:iAi1D, two editions of an NIMH newsletter, a booklet in French, 
the major portion of the Journal Current Research in Cancer Chemotherapy , 
a booklet on on insulin conference, three issues of Mental Health Ref ~ 
erence Guide , a booklet on r.edical and psychiatric social work, as well 
as numerous circulars, urograms, announcements, posters, graohs, and 
other items. 

Di stribution and Binding . The Section arranged for the binding 
of 279 volumes of various publications and distributed 13,685 NIH 
reports and publications during the year, including 1,535 copies of 
the 1957 Highlights of Research Progress , 6,632 copies of the Bayne- 
Jones report ( Advancement of ILdicaT Research and Education ), 238 copies 
of Data Relating to M H, and 1,355 copies of International''Medical 
Research , A flexible addressograph system to facilitate large-scale 
distribution of future oublications is being developed for use in 1959= 

Corres )ondence and Public Inquiries . During the year the Section 
wrote lC5 letters, answered 1,07? telephone inquiries, and distributed 
large quantities of printed materials in response to requests from public, 
legislative, and administrative sources, including frequent requests 
from scientists and organizations in foreign countries,, 

OFFICE OF THE CHIiSF Improvement in ths coordination of Medical 

Arts and Photographic sections has helped 
to solve problems arising from requests for services requiring the 
use Oi personnel in both sections. A marked reduction in the average 
of photographic items requested by individual requisitioners of charts 
and drafts, has been accomplished. During calendar year 1957 the 
average individual requests for chert and graph work required an 
average of 18 photographic items. This has now been reduced to an 
average of 10 prints and 2 slides for each chart request. This work- 
load reduction is reflected in the decline in total production by 
the Photographic Section. 

Library Plans for Security . The Scientific Reports Branch 
developed plans for security of the Library to protect its book 
collection and to improve services to NIH research investigators. 
Cooperation was obtained from the Plant Safety Branch. Provision 
of a book-charge clerk for night service, a monitor for the 3a te ■ 
evening hours to orotect the collection, and architectural changes 
to centralize traffic during the evening are the basic factors in 
the plan. Initial phases were acted on late in 1958 and other phases 
will be completed in 1959. 

Central Roster for Information Personnel . On behalf of the 
Chief, Office of Research Information and of the Information Officers 

DRS, Scientific Reports Branch, cont'dt 

of the Institutes and Divisions, the Office of toe Chief maintains 
(through interview) a central roster on applicants for future KIH 
positions. In 1958 more than 100 applicants for KIR" information 
positions were interviewed; seven were placed in MH information 
office a,, 

DRS Editorial Board , The Scientific Reports Branch administered 
the editorial review and clearance jrograms for the DRS Kditorial 
Boards Twenty -five scientific papers were reviewedo 

Promotions , nineteen Branch employees were promoted during 1958 
(20 inTOTC: 

Workload Report S o ."he Branch developed a uniform workload 
reporting system vfaich is ed by all sections with the excep- 

tion of the NIH Library, The Library's workload report provides the 
same basic data in more detail. 

Technical V.riting . The Scientific Reports Branch administered 
toe DRS technical writing CD-rse. 

Calenda r of Events .- The Branch accomplished a speedup in the 
delivery of the weekly Calender of Events by obtaining cooperation 
from its own .-Publications and He oris Section, Communications Section, 
DBO, and the Duplicating Section of DRG. The Calendar is now reaching 
the desks of MH personnel the week prior to the events scheduled.* 
Complaints have ceased. 

Coordination ., The Scientific Reports Branch developed a plan 
for coordinating requests for multiple services from more than one 
SRB Section at a time. This plan has enabled the Branch and its Sections 
to develop full understanding of the request and its ultimate usee The 
new procedure has Improved the quality of service to the requestor and 
has reduced the number of redo jobs. 

Overtime. The Eranch accomplished a reduction in the amount 
of overtime used in its Sections during 1958, from $7,317 to $5»282. 
The reduction of overtime reflects improvement in management and 
scheduling. The actual overtime used by sections is as follows: 

Overtime Comparison, Calendar 1957 and 1958 

Sections 1957 1958 


Medical Arts 



Publications and Reports 

Office of Chief 

Totals $7*317 $5,282 * 

DRS, Scientific Reports Branch, cont'd j 

« Does not include $1,500 overtime paid by Scientific Reports 
Branch to Library staff for indexing PHS intramural research 
projects reports as contained in the Analysis of Program 
Activities . Also does not include overtime for the Special 
Events Section, which was transferred on January 1, 1958, 
to the Clinical Center Information Office* 


LIBRARY SECTION Administration . Efforts to recruit a full- 

time assistant to the Librarian were initiated 
in the last quarter of 1958 and are continuing. 

Maintenance, The shelving load of 193,355 items is an increase 
of 30 6 percent in one year<> Manpower shortages a year ago resulted 
in shelving backlogs which summer employees and overtime helped to 
alleviate. An additional stack attendant is needed; although funds 
were made available in the latter part of 1958, recruitment for this 
position has been slow because of the shortage of good GS-1 and GS-2 

Binding. The backlog in binding remains at 12,000 volumes, 
but recruitment of a bindery assistant in 1959 and additional funds 
far binding will help reduce this backlog, A program planned to wipe 
out the bindir.g backlog within five years is in its first phase 
Binding increased from U9U to 693 volumes in calendar year 1958. 

Reference Service . Requests for bibliographies and for con= 
tinuing bibliographical service cannot all be met by the Library's 
Reference Unit. The addition in the last half of 1959 of a reference 
position to the Library staff (five are now assigned to the Reference 
Unit) and successful recruitment to fill an existing vacancy caused 
by transfer among the staff should result in some increase in service. 
Recruitment here is difficult because of a shortage of medical li- 

Reporting and Replacing of Lost Books, OverdueSf and Longer 
Service Hours . The Library has failed to develop a system that will 
improve the reporting and replacement of books found to be missing 
from the shelves. The absence of the books is a serious handicap 
to scientists and a fre4uent reason for complaints. This problem 
is related to the overall security problem of the Library and to 
a need for developing i '.proved procedures. 

Needed is the employment of a CB-3 library aide so that the 
Library staff can establish systematic recall of books on a weekly 
basis. At present, recall of books is infrequent. 

DR5, Scientific Reports Branchy cont'd: 

Implementation of plans developed during the latter part of 
1958 to improve library security and possibly reduce book losses 
binges on certain needed architectural changes and on the successful 
recruitment of a college student who could be employed part time to 
monitor the Library in the evening hours and on half days during we«k<= 

Library Extension Service , Needed is a clearly defined policy 
for NIH library extension service to NIH field units such as those 
established by NIMH at the Clinical Neuropharmacology Research Center 
in St. Elizabeths Hospital and at the Mental Health Study Center in 
Prince Ueorges County. 

Translating Unit Typing Backlog ., Increased use by the NIH 

siaff of the translation loans program has 
increased the Translating Unit's correspondence load to such extent 
that one clerk-typist, who formerly spent most of her time typing 
finished translations, is now doins this work. To keep production 
at maximum capacity, a ^S-3 clerk -typist is needed. 

Production Level . The translation production of the Unit is 
not up to its usual level because one translator was detailed to the 
Russian Scientific Translation Program. 

Russian Scientific Monograph Evaluation . A mechanism is needed 

Translation Program to take advantage ot informed scientific 

opinion in the selection of monographs for 


Other Languages . The scientific literature in certain areas 
of the world is growing. The translation of the literature of the 
Far East (China, India, Indonesia) and Poland is most pressing,, Some 
of tais will be done under Public Law USO funds, but evaluation is 
needed to dete.raine how far to extend this part of the scientific 
translation program. 

MEDICAL ARTS SECTION Long-Term Projects. The Medical Arts Section 

has experienced an increasing number of requests 
from NIH research staff which require the continuous service of personnel 
for long periods of time, ranging from two months to eight months per 
project o One such project, for example, is a laminography study that 
required at least $0 percent of one illustrator's time in 1958 in order 
to provide illustrations for a projected atlas. Another project in- 
volves cartographic drafting requiring minute topographic delineation 
and two to three months for one draftsman to complete. The Section, >s 
staff attempts in such cases to assist the scientific investigator 
but staff limitations and the pressure of requests for other services 
make it necessary at times to recommend contract services. This does 
not always neeTwith favor, thereby affecting the Section's good relations 
with those involved. 

DRS, Scientific Reports Branch, cont'd* 

PHOTOGRAPHY SECTION Space , The plan to provide space for part 

of the Photography Section's activities in 
the new CC Surgical Wing will relieve the Section of some long-standing 
problems. At the present time motion picture production suffers most 
from space limitations. 

S taffing and Automation . The three additional photographers 
requested for fiscal I960 would improve services in clinical and 
patient photography, color iiotography, and motion picture production. 
However, the need for more staff will be assessed carefully against 
a decline in requests for certain photographic services© Newly de- 
veloped office -copying equipment and other equipment for mass production 
of duplicates has reduced the Section's workload in routine photographic 
copying. The use of Polaroid equipment to produce simple slides for 
one-time use has also cut down the number of requests for slideso The 
Polaroid camera has met with some success in photomicrography, enabling 
the scientist to make quick observations without the necessity of using 
the Section's services. 

EDITORIAL SECTION Space . Additional space is needed to al- 
and levlate the crowded conditions in the 
PUBLICATIONS AND Building 8 quarters assigned to the Editorial 
REPORTS SECTION Section and the Publications and Reports 

Section. Administrative privacy is cur- 
tailed because both Section Chiefs are now sharing their offices with 
staff members* 

Form No. ORP-2 (Attachment 1) 

Serial No. DRS-3ol 

1. Scientific Reports Branch 

2. Medical Arts Section 
3« Bethesda 


Individual Project Report 

Calendar Year 1958 

Part A. 


Principal Investigator: Philip R. Joram 

Other Investigators: Helen G. Orem and George P. Marsden 

Cooperating Units: Moulage and Exhibit Units of the 

Medical Arts Section 

Man Years (calendar year 1958) Patient Days (calendar 1958) 
Total: 1.1 
Professional: 1.1 
Other: - 

Project Description: 

Objectives : Investigations are conducted in the 

industrial field of plastic materials 
and methods that can be used in the 

production of various scientific aids needed to solve medical 

research problems. 

Major Findings : The technical literature of the 

plastics industry is being reviewed 
and studied. A comprehensive file 
of materials and methods has been established. The project 
has produced the following developments: a specially designed 
and fabricated plastic filter holder for blood studies; psy- 
chological tasting materials for infants; a vacuum=forming 
technique used in the production of research exhibit acces- 
sories; a flexible molding technique for reproducing casts 
with severe undercuts; and an economical plastic museum con* 
tainer for gross specimens. 

Part B included Yes x No 

(Attachment 1) 

Serial No. DRS-3.1 

lo Scientific Reports Branch 


Individual Project Report 

Calendar Year 1958' 

Part B. : Honors, Awards, and Publications 


Joram, Philip R„; Grogan, Helen T., and Pruaansky, 
Samuel: Flexible Rubber Molds for Accurate Multiple 
Reproduction of Plaster Casts. Journal of Prosthetic 
Dentistry , Vol. 8, No. 1, (pp. 100-106), January 195B„ 

Joram, Philip R.: A Disposable Plastic Box for Paraffin 
Embedding. Stain Technology , Vol. 33, No. h 9 July 1958. 

Joram, Philip R. : Notes on the Meiller Antimony Trioxide 
Method for the Preservation of Gross Specimens in Color. 
American Journal of Clinical Pathology , Vol. 30, No. U, 
October 1950. 

Honors and Awards 

Philip R. Joram was elected to membership in the Society 
of Plastics Engineers in May 1958. 

Serial No„ DRS-3.2 

lo Scientific Reports Branch 

Calendar Year 1958 

Budget Data : 

Actual obligations for calendar year 1958: 
Total: 5661,927 

Reimbursement: $661,927 

Serial No., DRS-^ 

lo Scientific H ports Branch 
2o Russian Trar lation Program 

Calendar lear 1958 

Budget Data : 

Actual obligations for calendar year 1958: 
Total: 5328,555 

Reintoursement: $3 28,555 

January 1959 

Calendar Year 1958 


Summary of Branch Activities 


1. DRS - 4.1 

Serial Number 

3. Dr. Willard H. Eyes tone 
Branch Chief 


To provide scientists at NIH with various scientific services 
which are essential to individual research projects but which require 
central operation for effectiveness and economy. These include animal 
production, instrumentation, glassware preparation, media preparation, 
animal hospital services and comparative pathology. 


Programs of Laboratory Aids Branch are as follows: 

The production of high quality laboratory animals for research. 

The design and fabrication of unique scientific instruments 
not commercially available. 

The washing, sterilisation and processing of NIH laboratory 

The provision of types of culture media required by NIH 

The provision of surgical, radiologic, quarantine and holding 
facilities for larger animals, i.e., dogs, cats, monkeys, horses, 
sheep, goats, etc. 

The provision of comparative pathology services for the con- 
trol of diseases of laboratory animals. 

DRS, Laboratory Aids Branch, cont'd: 

Progress and Accomplishments 

ANIMAL PRODUCTION Reallocation of space in the animal 

colonies has allowed an increase of 
over 62,000 animals during 1958. 

Several exotic species have been added to the colony includ- 
ing the merione and the gerbillinae. Three mutant strains of hamsters 
have undergone differentiation tests to determine specific suscepti- 
bilities and are now useful research tools. The development of 
laboratory uses for new species has been a facet of animal production 
activities for many years. 

Intensive retraining and refresher courses, combined with the 
development of an operations manual for caretakers, has reduced the 
number of unusable animals by 1 percent in the past year. (Since 1 per- 
cent of 1,000,000 animals is 10,000 animals, this is a very real benefit), 
Improved husbandry practices have also resulted in increased production 
and favorable comments from users. 

Operations during the last six months of 1958 were carried out 
under the revolving fund and the experience has not imposed serious 
handicaps on our operations. 

Animal Issues, as compared to 1957, are as follows: 

1957 1958 





164, 100 


Guinea pigs 






Hamsters and other 






Several areas have expressed an Interest in an inbred strain 
of rabbit. It is anticipated that the 10,000, 0GQ tij mouse produced 
since the inception of the colony will be born sometime during June 1959. 

Improved husbandry practices are intended to duplicate the 
results obtained in specific pathogen-free colony management in con- 
ventional animal quarters. Accelerated germfree activities in the 
Laboratory Aids Branch has bean tentatively scheduled for fiscal 1961 . 

DRS, Laboratory Aids Branch, cont'd: 

Development of standardized housing and caging equipment has 
permitted the salvage and reissue of approximately $45,000 worth of 
used caging equipment during the past year. 

INSTRUMENT SECTION Engineers and instrument makers at NIH 

found both their imaginations and their 
abilities stretched to the utmost by requests for instrumentation. 

"inly 9 of the 178 engineering jobs accomplished this year 
have been listed. Many of the remainder are, doubtless, of equal 
importance to the scientists for whom they were performed. Those 
listed, however, are typical of the requirements made upon the staff 
of the Instrument Section. 

1. An apparatus, developed for Dr. C. L. Li, NINDB, generates 
electrical pulse trains to stimulate cortical areas. The resultant 
cell discharges are displayed on an oscilloscope and both signals are 
automatically photographed. 

2. Developed for Dr. Glen Morrow, NFJ, was an apparatus which 
performs the functions of the heart and lungs of a patient undergoing 
cardiac surgery. This arrangement provides the surgeon with a dry 
operating field and as much time as he may require for the repair of 
damage or correction of malformations. Over 100 patients were operated 
on, using this apparatus, during 1958. Improvements and modifications 
continue, as experience dictates. 

3. An electrical system was devised frr Dr. G. Z. Williams, 
Clinical Pathology, whereby sequential functions could be televised 
and photographed automatically at precise time intervals. 

4. A long-term project requiring continuing developmental work 
is bcins performed for Dr. John Calhoun, NIMH, in support of hie studies 
in habits and functions of the rat. 

5. Considered impractical by several leading optical companies 

a specialized type of projector has been developed for Dr. John Van Buren, 
BINDB, by NIH engineering staff. The projector permits vibration- free, 
undistorted magnification and projection of tissue, from 5x through 90s, 
onto a 3-foot-square horizontal surface, and the sketching of details- 

6. Another unique device, on which outside contractors declined 
to bid, permits both macro and micro photographs to be taken with the 
same piece of equipment. This device, as well as others, will provide 

a wealth of data to optical firms as well as to Fred Meiller, SINDB, for 
whom it was developed. 

DRS, Laboratory Aids Branch, cont'd: 

7. Designed and constructed for Drs, Harry Eagle, NXAID, and 
Karl Piee, HIDR, was a continuous recording nuclear ion exchange column 
employing a unique, newly designad scintillation counter and new 
scintillation plastic tubing. The new apparatus can effectively monitor 
low radiation samples as well as very small volume flow. 

8. An automatic sampling and collecting apparatus for the 
determination of steroids in urine, designed for Drs. E. Heftmann and 

D. Johnson, NIAMD, is approximately SO percent completed and is expected 
to be in operation in 1959. 

9. Continuing tests of a rectilinear coordinate injection 
control for dual reagents, designad for use with an automatic sampling 
tnd collecting apparatus, have shown maximum variation from selected 
ratios of less than 1.5 percent, proving the soundness of principal in 
utilising this method of proportioning. The allowed variation from 
•elected ratios was 5 percent. 

10. A complex intercommunicating system, developed for 
Dr. Malt land Baldwin, NINDB, allows oral communication between all of 
the members of a surgical team even though they may be situated in 
Laboratories some distance from the actual surgical suite. The system 
Lb always under the control of the Chief Surgeon. 

In addition to these projects in direct support of research, 
•everal others have resulted in savings in time and materials „ 

1. Automation oK the machine which cuts and forms animal 
drinking tubes from stainless steel has reduced labor costs on this 
operation by 90 percent. 

2. Testing of new designs and materials has produced an all- 
nylon pipette carrier capable of handling 66 percent more pipettes per 
Hashing cycle with substantially decreased breakage. 

In support of Institute Research Programs, fabrication Unit 
:ompleted 4,061 projects with a value of $352,000. 

A more tightly knit organisation, permitted by combining 
Engineer ing and fabrication Units at a single physical location,, 
should provide more effective utilisation of personnel. 


PREPARATION SECTION The efficiency of pipette-washing machines 

was materially increased by adoption of a 
carrier constructed entirely of a synthetic material. Th® lighter 
weight reduces fatigue on the part of the worker and the resilience 
of the material reduces accidental breakage of pipettes during loading. 
The carrier itself is less susceptible to damage and will bold 66 per- 
cent more pipettes than the old style. 

- 5 - 

DBS, Laboratory Aids Branch, cont'd: 

Workload statistics for the year, as compared to 1957, are 
shovn be low: 

1957 1958 

Number of pieces processed 3,456,000 4,480,000 

Value of pieces processed $1,723,000 1,801,500 

Pieces repaired 43,000 34,000 

Savings effected by repairs $ 22,000 $ 16,000 

Caging equipment washed 422,000 415,000 

Replacement of inoperative automatic detergent controls will 
result in fever rejects, thus increasing operational efficiency. 

Renovations will allow safer and more satisfactory storage of 
dirty glassware. This is required because of present insufficient space 
for picking and sorting soiled glassware prior to washing. 

MEDIA SECTION Although the number of requisitions for 

bacteriologic and tissue culture media in- 
creased only 3 percent over 1957, the volume of media issued Increased 
20 percent. 

Because of their devotion to duty and their ability to cope 
with unusually heavy demands, the employees of the Section were the 
recipients of a departmental citation for superior service. 

Workload statistics Indicate the following: 

1957 1958 

Number of Requisitions 7,300 7,500 

Total volume Issued 32,000 liters 38,500 liters 

Installation of equipment to more efficiently perform certain 
filtration operations is planned. 

Consolidation of Glassware and Media Sections, planned for 
1958, will take place during 1959. 


SECTION The Animal Hospital became associated with 

the American Animal Hospital Association 
during the year, provided facilities for four courses in advanced 
clinical techniques in veterinary medicine and surgery, and was pre- 
sented with several research animals by conscientious persons and 

- 6 - 
DRS, Laboratory Aids Branch, cont'd; 

Workload for the year showed: 

Conditioning, and Quarantin ing 

Cats 2,200 

Dogs 2,900 

Primates 3,000 

Surgery and X-ray 

Operations performed 410 

Surgical assistance (hrs.) 1,300 
X-rays taken 1,100 

Long-term Holding 

Average number of animals held 301 

Although the incidence of tuberculosis among incoming monkeys 
increased 65 percent over 1957, deaths due to other causes were reduced 
by over 50 percent. Since the majority of deaths result from diseases 
other than tuberculosis, the number of usable monkeys was greatly 

Improvements in feeding and watering are under evaluation. 

Interesting observations were made on cattle afflicted with 
cancer of the eye. Reports of findings will be published in the coming 

Constant disease-control measures detected the presence of a 
bar tone 11a- like infection (feline infectious anemia) in the cat colony.. 
An a valuation of the current and potential danger of this disease to 
the colony is being made. 

In the fields of radiology and surgery, further studies are 
being made in contrast radiography and in the surgical lengthening of 
long bones which had been shortened by old, untreated fractures. Eypo- 
physcctomy and pineal body removal have been perfected to a high degree. 
Clinical and other significant data will be published in 1959. 

The large barn at the Rockvllle Farm is now occupied by KHfiS. 
All remaining facilities are being utilised to their full capacity. 

The Section will continue to investigate new sources of supply 
of research animals and methods designed to promote greater efficiency 
and economy of operations. 

- 7 - 
DRS, Laboratory Aida Branch, cont'd: 


SECTION Completing ita firat full year aa a central 

service operation, the Section found many 
opportunitiee to further man* a knowledge of animal diseases, thus 
indirectly contributing to reaearch at NIH. 

During 1958, the Section developed methods designed to control 
infectious catarrh, middle-ear infection, and endemic pneumonia in 
experimental rats, and to eliminate oxyuride from female mice and their 
offspring. The Section isolated viruses suspected of being causal 
agents of salivary gland disease of guinea pigs and infantile diarrhea 
of mice; and collaborated in studies in which evidence was found indi- 
cating that salivary gland disease and pregnancy toxemia in guinea pigs 
do not have a common etiologlc agent. 

Intensive studies were alao performed to try to determine the 
causes of death of monkeys from the time of trapping in India to the 
time of delivery to the research laboratory. 

In cooperation with the Animal Production Section a survey of 
the prevalence of bartonelloala in laboratory rats was undertaken. 

Intensive atudy will be devoted to identifying end developing 
methods of controlling the etiologlc agents of infantile diarrhea, 
pregnancy toxemia, salivary gland disease, mucoid enteritis, snuffles 
and other injurious diseases in laboratory animals. 

OFFICE OF THE CHIEF Continued increases in requirements for 

services by the Institutes have been 
reflected in the administrative workload of this office. In addition, 
the growth of the NIH has precipitated Increased demands for infor- 
mational reports from higher management echelons. 

Responsibility for the management of caging equipment for new 
production areas and genetics reaearch waa added to the office during 
the year in the Interest of improved financial management for those 

The position of Assistant Chief waa established during the 
year and filled by en experienced career veterinarian. This addition 
to the staff has added considerable strength to the professional and 
administrative capabilities of the Branch. 

- 8 - 
DBS, Laboratory Aids Branch, cont'd: 



SECTION The everpresent problem of insufficient 

space has prevented satisfactory supply 
of required numbers and strains of animals to investigators. Due 
to changing program requirements, this problem will continue to 
exist, to one degree or another. Lack of adequate facilities has 
prohibited the housing of amphibia, reptiles, and insects. 

INSTRUMENT SECTION Shortages of qualified persons in the fields 

of electronics and glassblowing have caused 
recruiting problems. A considerable amount of dissatisfaction has 
been expressed by employees vho receive appointments under the "TAPER" 
regulations. They are primarily concerned with the restriction of 
retirement and status privileges, so long as they continue under these 
regulations. Indefinite retention of these employees may eventually 
require that steps be taken to either establish registers of eligibles 
In shortage categories or waive restrictions regarding employee bene- 
fits to allow accumulation of retirement and reinstatement credits. 


PREPARATION SECTION A shortage of laboratory glassware presently 

exists. This shortage is primarily due to 
the hoarding of pipettes and other small items by individual labora- 
tories. If the laboratories restricted supplies to their daily needs 
and returned soiled glassware promptly to be processed, shortages in 
most items would be minimized. Several other factors have contributed 
to the shortage. In the past four years, glassware issues have 
increased from 1.1 million places to 4.5 million pieces. Funds for 
replacement, on the other hand have Increased only one-third (from 
$6,000 per month to $6,000 per month). In this same period, data 
show an average price increase of 16.25 percent in cost per item. 
Shortages caused by insufficient funds for replacement of broken or 
discarded glassware will assume significant proportions unless this 
facet of the problem receives attention. 


SECTION The facilities at the Roekville Farm are 

depreciating rapidly, under the original 
rental agreement, the owner agreed to provide certain services. 
For the past year, the owner has failed to provide these services. 
Since it is necessary to provide suitable habitation for the caretaker 
and his family, NIH has bsen obliged to furnish men and materials for 
emergency repairs. During 1958 these items were requested: 

1. Replacement of frozen water pipes. 

2. Replacement of leaking hot water heater. 

3. Repair of sagging and broken plaster in kitchen ceiling. 

- 9 - 
DBS, Laboratory Aids Branch cont'd: 

Th* soil at the farm has become parasitised because the limited 
space does uot provide for rotation of exercise areas. On the whole, 
this farm is rapidly becoming unsuitable for NIK needs. 

(Attachment 1) 

Serial Ho. 4.2 

1. Laboratory Aids Branch 

2. Comparative Pathology Section 

3. Betheada 

Individual Project Report 
Calendar Tear 1958 

Part A. 


Principal Investigator: Dr. Willard H. Eyestone 

Other Investigator: Dr. Roy P. Kinard 

Cooperating Units: Animal Hospital Section 

Man-years (Calendar Tear 1958): 
Total: 3 
Professional: 2 
Other: 1 

Project Description : 

Objectives : To be well informed, through experience and 

experiment, concerning the pathologic character - 
iotico of the diseases of these animals and to 
cooperate with other scientists of the NIH staff in the interpretation 
of related data. Data obtained from disease studies are of prime 
importance in planning disease control procedures used in the care of 
the animals in the csatral facilities of NIH prior to their use in 
laboratory experiments. 

Method Employed : Consists of a continuing survey of the gross 

and histopathologic characteristics of diseased 
tissues in animals. Animals for autopsy and 
tissues for examination are obtained from the NXH Animal Hospital, 
the National Zoological Park and from practicing veterinarians. 

Through contacts with other scientists of the institutes, 
special interest fields are selected and experiments are planned 
so that m*xf"q«p benefit may be obtained from the materials sub- 
mitted for pathological evaluation. 

Fart A. cont'd: 

In cooperation with the Animal Production Section, a survey 
of the prevalence of bartonellosis in laboratory rats was undertaken. 

Two visits to India were made by Dr. Kinard, for the National 
Advisory Committee on Rhesus Monkey Requirements, to try to determine 
the causes of death of monkeys during trapping and holding in India 
and transit to the U.S. Autopsies were made in flight and arrange- 
ments were mode with the University of Utrecht, the Netherlands, for 
autopsy of monkeys dying on KLM, Royal Dutch Airlines flights. 

A visit to Okatie Farm of the National Foundation was also 
made by Dr. Kinard to determine the cause of the high rate of 
tuberculosis among monkeys there. 

Major Findings : A utudy of the tissues obtained through autopsy 
of monkeys dead of natural causes or euthanasia 
from the Primate Unit of the Animal Hospital 
Section was maoe soroughout the year. Of 595 such monkeys received 
for autopsy, 523 were examined and 67 were discarded or were not 
examined completely because of advanced postmortem degenerative 
changes. Lasioas found indicate that the major cause of death was 
inflammation of the intestine (enteritis was seen in 37, cecocolitis 
in 134, and colitis in 192). The etiologic agents of this inflam- 
mation are not definitely known* but Shigella and Salmonella spp. 
have been Isolated from such cases. 

Several Intestinal parasites of monkeys, especially 
Strongyloldes and Esophagostomum , were identified but were not 
usually considered a cause of death. Other parasites identified 
include Castrodlscoldes (fluke of the large intestine of the 
rhesus monkey), P neumonyssus (lung mite of rhesus monkey), P ej&ciaua 
longicepa (the common monkey louse), Prosthenorchis (a round worm 
from the colon of the squirrel monkey) and Dipetaloaema (a filarial 
worm of the peritoneal and pleural cavities of squirrel monkeys). 

A total of 30 tuberculous monkeys were examined, 26 of which 
ware tuberculin reactors. Not all tuberculin reactors from the 
Animal Hospital were submitted for autopsy because of the danger 
of spreading infection upon incision of the lesions. Two cases of 
skin tuberculosis and one case of tuberculosis of the vertebral 
column were examined. 

Pneumonia, other than tuberculous, was seen in 87 monkeys, 
but was usually considered a complication following prolonged 
Intestinal inflammation. 

A. cont'd: 

Ho neoplastic lesions of monkeys ware observed. 

Summary of Results of Autopsies of Monkeys from Animal Hosoital 

Submitted for autopsy —.———. 595 

Examined 528 

Discarded due to degeneration — 67 
Major findings: 

Intestinal inflammations 

Bnteritis — 37 

Cecocolitis 134 

Colitis - 192 

Tuberculosis ————— 30 
Pneumonia — — 87 

In the bartonellosls survey , 77 rats of 13 strains were 
examined and no Bartonella were found except in a few rats 
purposely infected to check our technique. Twenty-one mice of 
four strains wera examined and no Bartonella wore found. 
Sperythroaoe ware demonstrated in two strains of mice. 

The visits to India yielded considerable first-hand infor- 
mation on the supply of rhesus monkeys from that country. A full 
report of the findings has been given to the committee. The 
committee members expressed the belief that the findings will b« 
of considerable value in improving the quality of imported donkeys* 

The direct causes of death in transit are the same as those 
mentioned above, i.e., intestinal inflammation and pneumonia. It 
is believed that Indirect or contributing causes of death are stress, 
exposure, altitude, etc. These contributing factors have not yet 
been thoroughly evaluated. 

The visit to Okatla Farm also yielded valuable information 
concerning the handling of imported rhesus monkeys. 

In addition to the autopsies mentioned above, others ware 
performed on animals of many species of sub-hustaa primates from 
the Rational Zoological Park and various laboratories at HIH ff and 
many diseases and lesions have been studied. The results of these 
autopsies have proved helpful to the scientists concerned. 

Fart B included Tea 

(Attachment 1) 

Serial Ho 4.2 

1 o Laboratofy~Aias Jsranch 

Individual Project Report 
Calendar Year 1958 

'art B; Honors, Awards, and Publications 

Publications other than abstracts from this project; 

Allen, Anton M., and Kinard, Roy F* Primary Cutaneous 
Inoculation Tuberculosis in the Macaca Hilatta Monkey c 
American Journal of Pathology; 34; 337-347; 1958„ 

Honor 8 and Awards relating to this project: 

(Attachment 1) 

Serial No. 4.3 ^ 

1. Laboratory Aids Branch 

2. Comparative Pathology Section 

3. Bethesda 

Individual Project Report 
Calendar Tear 1958 

Part A. 


Principal Investigator: Dr. Robert T. Habermann 

Other Investigator*: Dr. Carlton M. Herman 

Fletcher P. Williams, Jr. 

Cooperating Units: None 

Man-years (Calendar Tear 1958): 
Total: .25 
Professional: .125 
Other: .125 

Project Description : Fifteen raccoons and three red foxes were 

brought to the National Institutes of 
Health by Dr. C. M. Herman of the Maryland 
Cane and Inland Fish Commission. Six other raccoons ssere subsequently 
obtained from various areas in Maryland, Pennsylvania, and Kansas. 
All were ill and from areas where suspected rabies occurred. Micro- 
scopic and macroscopic examination of tissues, performed concurrently 
with the mouse rabies test, indicated that none ©are rabid. (Fourteen 
of the raccoons and two of the foxes wore positively diagnosed for 
distemper). Deaths of the seven other raccoons and the one fox were 
due to other diseases of a non-episootic nature. 

The Section cooperates actively with other agencies in the 
Interest of the Public Health of the Nation. 

Part B included Tea x Ho. 

(Attachment 1) 

Serial Bo. 4.3 

1. Laboratory Aids Branch 

Individual Project Report 
Calendar Year 1958 

»art B: Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Habermatm, Robert T. , Herman, Carlton Ho, and Williams, 
Fletcher P. , Jr. Distemper in Raccoons and Fox Suspected 
of Having Rabies. Journal of the American Veterinary 
Medical Association; 132, 1; 31-35; 1958. 

Honors and Awards relating to this project: 


(Attachment 1) 
Serial Bo. _4.4 __ 

1. Laboratory Mds Branch 

2. Comparative Pathology Section 

3. Bethesda 

Individual Project Report 
Calendar Tear 1958 

Part A . 


Principal Investigator: Dr. Leon Sokoloff, NIAHD 

Other Investigator: Dr. Robert T. Habermann 

Cooperating Units: NIAMD 

Man-years (Calendar Tear 1958): 
Total: 1.0 
Professional: 1.0 
Other : Hon© 

Project Description : Localised areas of spontaneous, apparently 

aseptic necrosis of bone have been observed 
in 31 of 670 mice during the second year of 

life. The etiology is obscure, but the lesions appear to be influenced 

by genetic factors and sex. Isolated instances occurred in rats sad 

a hamster. 

Because of the pathologic similarity to the several types of 
aseptic necrosis of bone that occur in man, these animals may provide 
a tool for future studies of these diseases. 

Part B included Tes x Z!e_ 

(Attachment 1) 

Serial Ben 4.4 

1. Laboratory Aids Branch 

Individual Project Report 
Calendar Tear 1958 

Part B: Honors, Awards, and Publications 

Publications other than abstracts from this project; 

Sokoloff , Leon, and Habermarm, Robert To Idiopathic 
Necrosis of Bone in Laboratory A.MoA. Archives of 
Pathology; 65; 323-330; 1958, 

Honors and Awards relating to this project: 


(Attachment 1) 

Serial No. 4^5 

1. Laboratory Aids Branch 
2„ Animal Production Section 
3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A . 


Principal Investigator: Samuel M. Polley 

Other Investigators: None 

Cooperating Units: None 

Man-years (Calendar Tear 1958): 
Total: .06 
Professional: .06 

Project Description : This project encompasses all aspects of 

research on the development and design 
of housing and caging equipment for 

laboratory animals. 

During the past year, new types of caging equipment were 
designed and tested at the request of, and in collaboration with 
scientists from various Institutes. In addition, some of the 
existing types of cages were redesigned to permit better utili- 
sation of the equipment and accessories. The new items and 
redesigned items are as follows: 

New Items 

Rat maternity cage feeding device 

Standard NIB magnetic cage latch 

Squirrel monkey cage 

Animal exhibit cages 

Walk floor and pan for small animal cage 

Magnesium shelf truck 

Magnesium platform truck 

Revisions of Cage Designs 

Mookay observation cage 

Plastic cages 

fiberglass cage 

Suspended cages 

Infant monkey cage 

General purpose cage - Mark V 

Guinea pig breeding cage 

Work on this project 1» continuing from year to year- Though 
there are standard items of caging equipment «t NXH, they are in 
constant need of revision, either for use in the Animal Production 
Section or In the various research areas. 

Salvage, revision, repair, and reissuance of caging equipment 
to laboratories. 

Part > Included Tes No_ 

Serial »o. J^ji _ 

1. Laboratory Aid© Branch 

2. Animal Production Section 

3. Betbesda 


Individual Project Report 

Calender Tear 1958 

Part A. 


Principal Investigator: Animal Production Section Staff 

Other Investigators: None 

Cooperating Unit*: Comparative Pathology Section 

Man-year* (Calendar Tear 1958): 
Total 0.32 

Professional: 0.17 
Other: 0.15 

Project Description : Husbandry practices and procedures in the 

care end maintenance of laboratory animals 
are always subject to changes end revisions.. 
Consequently, research studies on housing, feeding, cleaning, breedings 
sanitary procedures, etc., are an Integral part of any production 
colony activitlea. The present project is concerned with matters of 
this type. 

During the past year, typical subjects of study have been as 
follows: the use of antibiotics and other drugs in the control of 
disease, optimal population numbers in cages, use of various bedding 
materials, testing of guinea pig rations, testing of various dietary 
supplements, and handling procedures for mothers and litters, 
comparison of commercial animal caging, protective coating for 
permeable materials, evaluations of commercial mouse production 
methods, the uses of various types of insecticides, and production 
methods in large animal colonies. 

The results of these experiments, applicable in many cases 
to NIB alone, have not been published. Some produced negative 
results, while others provided sufficient information to warrant 
altering certain routine procedures. 

taxi. A. cont'd, 

Current studies include methods and procedures to be used 
in day-to-day activities, with a vieu tovard controlling sad 
limiting contacts between animals and probable contaminated 

The Section on Comparative Pathology is assisting in 
evaluating this activity. 

Part B Included Tes_ Mo 

SAttachmeat I) 
Serial No. 4,7 

1. Laboratory Aids Branch 

2. Animal Production Section 

3. Bethesda 


Individual Project Report 

Calendar Tear 1958 

Part A. 


Principal Investigator: Dr. George E, Jay, Jr. 

Other Investigators: None 

Cooperating Units: Hone 

Man-years (Calendar Year 1958): 
Total: 0,05 
Professional: .025 
Other: .025 

Project Description: This project is a continuation from 

calendar year 1957. 

Infantile dlarrhee In suckling mice is probably the most 
serious health problem confronting a mouse production program,, 
All production colonies ere subject to the disease, Random-bred 
mice and some Inbred strains show it only sporadically whereas 
other closely inbred strains shov it constantly. The etiology of 
the disease is unknown despite considerable past research. 

A genetic susceptibility and/or resistance has been suspected, 
since differences in incidence have been noted between strains and 
even between substrains. Because of these observations, it seems 
possible that heritable resistance to the disease could be promoted 
and fixed in a strain by genetic manipulation. The present project 
was undertaken to investigate this possibility in C3H mice. 

Animals for this project have been taken from the substrains 
of Cffl available in the Rill colonies. By appropriate breeding and 
testing methods, it is hoped that a C3H strain can be developed that 
will continue all the desirable characteristics. 

Part A. co 

The project so fat has shown promise,, but it will require 
store work to actually demonstrate substantial progress. If the 
program is successful, it will os&ri a very substantial saving in 
the cost of Cm mice, as well as a saving in tias and effort by 
the scientific staff. 

Some progress in the reduction of the incidence of this 
condition hao been noted However, additional studies concerning 
spontaneous tuaor incidence, and the hybrid animals' ability to 
grow hi Biocompatible tumors are indicated. 

Part B Included Tes Mo_ 

(Attachment 1) 

Serial Ko. 4.8 
1. Laboratory Aids Branch 
2o Animal Hospital Section 
3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A. 


Principal Investigator: William I Gay, D.V.M. 
Other Investigators: None 

Cooperating Units: Staff, Animal Surgery Unit, 
Animal Hospital Section 

Man-years (calendar year 1958): 
Total: 0.20 
Professional: 0.05 
Other: 0.15 

Project Description : 

Major Findings: The midshaft of several long 

bones of the dog has been 
successfully replaced with a 
prosthesis. Titanium devices used in previous experi- 
ments proved too v;eak for use in the femur., A stain- 
less steel device was developed that proved very 

The stainless steel device was used in dogs 
whose femurs had been shortened over one inch by 
previous injury. These bones were restored to their 
original length. 

- 2 - 
Pa rt A . (Cont'd) 

The use of an organic bone did not hasten new 
bone formation around the prosthetic device. Other 
materials will be tested for this purpose as they 
become available. 

Part B included Yes x No 

(Attachment 1) 
Serial So. 4.8 

1. Laboratory Aids Branch 

Individual Project keport 
Calendar Year 1958 

Part B: Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Gay, William I. A Method for Surgical Lengthening of the 
Femur of the Dog. Military Medicine; 123, 4j 1958. 

Honors and Awards relating to this project: 

(Attachment 1) 

Serial Ho„ 4,9 

1. Laboratory Aids Branch 

2. Animal Hospital Section 
3o Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A. 


Principal Investigator: William I. Gay, D.V.M. 

Other Investigators: Dr. James 0. Davis, HHI 

Cooperating Units: Staff, Animal Surgery Unit 

Man-years (calendar year 1953): 
Total: 0.20 
Professional: 0.05 
Other: 0.15 

Project Description : 

Major Findings : Puppies which were hypophy- 

sectomized for an earlier 
study (see 1957 project report) 
were found to have considerable variation in the growth 
potential of their tissues. When this was brought to 
the attention of Dr James 0. Davis, HHI, and Dr 
William Tullner, SCI, they requested that more animals 
be operated on for them to study* 

The animals were operated on and followed 
radiographically and clinically. They were then 
turned over to Dr. Davis to study their physiology 
and organ growth. 

Par;: 3 included Tea no 

(Attachment 1) 

Serial No. 4.10 

1. Laboratory Aids Branch 

2. Animal Hospital Section 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A . 


Principal Investigator: William I. Gay, D.V.M. 

Other Investigators: None 

Cooperating Units: Staff, Animal Surgery Unit 

Man-years (calendar year 1958): 
Total: 0.20 
Professional: 0.05 
Other: 0.15 

Project Description : 

Objectives : In this study contrast radiography 
techniques are evaluated as a way 
of demonstrating experimental sur~ 
gical results without sacrificing the experimental 


Course of Project : An exhibit of radiographs from 
this study was shown at the 
American Veterinary Medical 
Association annual meeting in Philadelphia this year. 

Attempts are being made to devise a safe and 
satisfactory technique to outline the pancreas and 
the ventricles of the brain of the dog. 

Part B .included Yes No x 

(Attachment 1) 

Serial Ko. 4oll 

lo Laboratory Aids Branch 

2. Animal Hospital Section 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A . 


Principal Investigator: Dr. James 0. Davis, HHI 

Other Investigators: William 1. Gay, D.V.M. 

Cooperating Unite: Staff, Animal Surgery Unit 

Man-years (calendar year 1958): 
Total: 0.20 
Professional: 0.05 
Other: 0.15 

Project Description : 

Mijor Findings : The pineal body was removed 

in its entirety by Dr. 
William I. Gay so it could 
be demonstrated histologically before valuable time 
was spent by Dr. James 0. Davis in extensive physio- 
logical studies. Formerly it was necessary to wait 
urjtil the completion of studies and demonstrate at 
autopsy that the pineal body had been completely 

Part B included 3tes Ho x 

(Attachment 1) 

Serial Bo. 4.12 

1. Laboratory Aids Branch 

2. Animal Hospital Section 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part Ao 


Principal Investigator: Mr. Roland Faulkner 

Other Investigators: None 

Cooperating Units: Dog and Cat Quarantine Unit 

Man-years (Calendar year 1958): 
Total: 0.20 
Professional: 0.05 
Other: 0.15 

Project Description : 

Course of Project : Feline Infectious Anemia 

has been found to occur 
frequently in our cat 
colony. A study has been undertaken to evaluate the 
danger of this infection to the colony. 

'art B included Yes No x 

(Attachment 1) 

Serial Bo. 4,13 

1. Laboratory Aids Branch 

2. Animal Production Section 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A, 


Principal Investigator: Samuel M. Poiley 

Other Investigators: Bone 

Cooperating Units: Staff, Comparative Pathology 

Man- years (calendar year 1958): 
Total: 0.55 
Professional: 0.30 
Other: 0.25 

Project Description : This project was begun 

during calendar year 1957, 
with a view toward creating 
a strain of rat resistant to chronic respiratory 
disease. The backcrosses of the hybrid animal to the 
NIH Black Rat have been completed, and the selected 
albino animal 3 are now being inbred. The nonresistant 
and partially resistant lines are being discarded. 

Offspring of the third filial generation have 
been submitted to several laboratories for trial, 
with favorable reactions from investigators. 

All retired breeders are autopsied. Subsequent 
reports have indicated no lung lesions, or lesions at 
a low level, 

Part A. (Cont'd) 

This colony is now receiving, and will continue 
to be medicated with a sulpha compound through the 
fifth filial generation. Subsequent generations will 
be untreated, and family lines that do not exhibit 
clinical symptoms, and are determined to be uninfected 
at autopsy, will serve as the nucleus for the new 

The objective is to develop a homozygous re- 
sistant animal o Genetic calculations indicate that 
50Z are homozygous and 50% are heterozygous resistant 
animals. Differentiation will be accomplished via 
bacteriological laboratory studies . 

Part B included Yes No x 

Serial No. 4 14 

1. Laboratory Aids Branch 

2. Comparative Pathology Section 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A. 


Principal Investigator: Dr. Robert T. Habernann 

Other Investigators: Fletcher P. Williams, Jr. 

Cooperating Units: None 

Man- years (calendar year 1958): 
Total : . 25 
Professional: .125 
Other: . 125 

Project D escription : Various chemical substances 

were mixed with ground animal 
feeds and drinking water to 

determine the efficacy of the chemicals in the control 

of internal parasites. 

Oxyurids were effectively removed from female 
mice and their litters by the administration of pipera- 
zine adipate to the pregnant mice and later to the 
weaned litters. 

Provides a means by which such animals can be 
made usable, thereby increasing the number of avail- 
able animals and lowering the unit cost of production. 

Part B included Yes x No 

(Attachment 1) 
Serial So. 4.14 

1. Laboratory Aids Branch 

Individual Project Report 
Calendar Year 1958 

Part B; Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Habermarm, Robert T. , and Williams, Fletcher P., Jr. 
The Identification and Control of Helminths in Laboratory 
Animals o Journal of the Rational Cancer Institute; 20, 5s 
979-1005; 1958. 

Honors and Awards relating to this project: 

(Attachment 1) 

Serial Ho. 4.15 

1. Laboratory Aids Branch 

2. Comparative Pathology Section 

3. Bethesda 

Individual Project Report 
Calendar Year 1958 

Part A o 

Principal Investigator: Dr. Robert T. Habermann 
Other Investigators: Pletcher P. Williams, Jr. 
Cooperating Units: Animal Production Section 

Man-years (calendar year 1958): 
Total : o 50 
Professional : . 25 
Other: .25 

Project Description : Various methods of sanitation 

and care of laboratory animals 
were employed to determine 

their efficacy for controlling diseases. 

Salmonella infection did not occur in our animal 
colonies where rigid standards of sanitation and care 
were observed. Experimentally induced infections in 
isolated animals did not respond to available thera- 
peutic measures. 

While other colonies have suffered deciminatlon 
due to salmonellosis, the National Institutes of Health, 
by maintaining its closed colonies and highly disci- 
plined husbandry practices, produces animals free of 
this disease, enabling investigators to distinguish 
between experimentally induced and naturally occurring 
diseases . 

Part B included Yes x No 

(Attachment 1) 

Serial Mo 4.15 

1. Laboratory Aids Branch 

Individual Project Report 
Calendar Year 1958 

Part B; Honors, Awards, and Publications 

Publications other than abstracts from this project: 

Habermann, Robert T. , and Williams, Fletcher P* , Jr„ 
Salmonellosis in Laboratory Animals . Journal of the 
Rational Cancer Institute; 20, 5; 933-945; 1958* 

Honors and Awards relating to this project: 

(Attachment 2) 

Serial Ho. 4.16 

1. Laboratory Aids Branch 

Calendar Year 1958 

Budget Data ; 

Actual obligations for Calendar Year 1958: 
Total: $1,032,587 

Reimbursement: $1,032,587 

(Attachment 2) 

Serial No. 4ol6 

1. Laboratory Aid* Branch 

2, Animal Production Section 

Calendar Year 1958 

Budget Data : 

Actual obligations for Calendar Year 1958: 
Total: $501,588 

Direct: $501,588 

Reimbursement : 

February 1959 Calendar Tear 


Suoeary of Branch activities 

Serial Buaber 

*• BKHETBICS 3. Harold F. Dora 

Branch Branch Chief 


The object Ives of the B lose tries Branch are: 

To provide expert bloo m tri e consultation concerning 
the deelgn e execution, and aaalyaia of experlxeate and studies. 

To plan and conduct research investigations , usually 
in collaboration with other scientists. 

To provide central statistical processing services 
by electrical accounting and etatistlcel awchiaes and by electronic 
data proceeding equlpaeat. 

The pragma of the Biosetrlcs Branch is to carry out these 
objectives. Branch ectivities continue fro* year to year and are 
not readily classifiable into progress with e definite beginning and 


Consultation The Branch engages in a great deal of 

consul tat ion with esperistsaters, 
statisticians, and administrators in the various Institutes and Divisions 
as wall as with persons free outside the Rational Institutes of Health. 
Bono of the sore iaporteat projects during 1958 were: 

DRS, Biometries Branch cont'd: 

Janet Pitsssator, CC: involving a 
study of the efficiency of selecting surgical instruaants by aurgle&l 
nurses depending on the kind and caouat of information the nurse Is 
given about the patient. Tale study was accepted In partial fulfilment 
of the requirement for a wester'* decree for Silas Fitswater by the 
Catholic Onlvaralty of America. 

Dr. Leon Sokoloff, HIAMD: analysing 
the consldarabla data of experiments studying various factors Involved 
In arthritis In animals. Sans of the factors are genetic, diet, ses B 
obesity, etc. 

Dr. Paul H. Beer, HIDE: continued 
studies of genetic and diet effects on periodontal disease. 

Dr. Anatole S. Dekabaa, NBSDB: 
studies Involving pregnancy wastage and abnormal neurological findings 
due to diabetes of Bothers end pregnancy complications . 

Dr. Bathan B. Eddy, HX&HD: continued 
consultation oe cooperative studies of analgesics in various hospitals. 

Dr. Kelsey C. Kline?, HXA2D: Dr. 
miner Is a member of Rocky Mountain Laboratory. A personal visit was 
made to the Laboratory to assist the staff in designing experiments. 
Other consultation has been by nail. 

Considerable tine is taken up In 
refereeing ataanscrlpts of BIB scientists, particularly frost 3L2JSD. 
The review of papers recoaasoded for revision or rejection involves such 
■ore work end tins than the review of acceptable stanuscripts due to the 
necessity of explaining the basis for the te cesmen dation. 

E£ab2«a«o£ kldits have been used to analyse 

Mafoe— tlcal Theory certain data involving a subjective 

scoring system. Sobs scientists at 
SIH have found this method useful and expect t© make more extensive use 
of it. However, there ere certain problems connected with this method, 
particularly those involving the use of the proper error variance. This 
problem is now being investigated. 

Jgyjugee Harold Dora gave e lecture oe population 

problems at the School of Hygiene and 
Public Health, Johns Hopkins university. 

Jerome Cornfield was an instructor at 
the special summer school for bismetrieiaas at the School of Public 
Health, university of Michigan from 16 June to 11 August. 

DOS, Biometries Branch, cont'd: 

Harold Bern presented a papas, 
"The Epidemiology of Primary Carcinoma of the Liver," at the Strasburger- 
Perella Conference, / 

Harold Dora presented papers oa tba 
mortality of smokers and nensmokers at tha Seventh international Cane*? 
CoagraaOe London, England and at tha annual meeting of tha American 
Statistical Association, Chicago, Illinois. 

visiting Scientists Professor Jazzy Heym&n, Director of 

tha Statistical Laboratory, University 
of California, opant six weeks with tha Biometrics Branch. In addition 
to giving a aarlao of three lectures, ha worked on mathematical models 
for the reletionshlp of ionising radiation and carcinogenesis. 

Dr. Anthony Palrbalrn of the London 
School of Hygiene and Tropical Medicine, cane to the Blone tries Branch 
during October 1958. Bis tints has been spent in familiarising himself 
with epidemiological studies of chronle respiratory and cardiovascular 
diseases with the general objective of preparing plans for a joint p rogram 
of research to be carried out by the Sat local Heart Institute and the 
London School of Hygiene and Tropical Medicine. 

Conferences Harold Dora participated In the 

following conferences: 

Symposium on the Geographical Pathology 
of Gastrointestinal Cancer, held tinder the auspices of the International 
Union Against Cancer in Copenhagen, Denmark, 14*17 July 1958. 

The Interrelations of Demographic and 
Economic Changs sponsored by the Ualvereitles~S8tieaal Bureau Ceamlttee 
for Economic Research at Princeton University, 5*7 December 1958. 

Participation In Activities Harold Dorn mas reelected General 
of Stations! and International Secretary of the International Union 
Professional Organisations Against Cancer. During 1958 the 

Union organised too international 
symposia and the Seventh International Cancer Congress. The more impor- 
tant activities of national professional organisations ware those of the 
Beseereb Advisory Council and Statistics Committee of the American Caneer 
Society, the Board of Directors and the Executive Committee of the Social 
Science Keseareh Council, and the Committee en Preventive Medicine and 
Social Science Eesearch of the Social Science Besesreh Council. 

D&3, Biometrics Branch, cont'd: 

Interagency ^— » fl»» a The Interagency Committee on Medical 

^Udlol Bocords Records of which Barold Dora is 

Chairmen, acted on severe! matters 
referred to it by tfaa Bureau of the Budget. It also prepared revision* 
of several of the nodical records used in Federal hospitals. 

Statistical Processing The IBM Type 650 electronic computer 

Activities and the Tape Sate Selector were turned 

over to the HIE during March. The 
training program for programmers and operators continued throughout the 
year. In addition to sending individual members of the Statistical 
Processing Section to courses organised by the International Business 
Machines Corporation, several special courses were given at the HXH to 
acquaint other personnel with the uses and operation of the computer. 

By the end of December two major 
administrative activities— accounts and payroll'— had been transferred frost 
BAH equipment to the computer. These, In combination with oue<»time 
competing and date processing jobs, and preventive maintenance, require 
about 115 hours per month or about 60 percent of regular one-shift time. 

During the last six months of the 
year a records system mas set «p to provide information concerning the 
number of hours of work done on each Job as a basis for a fee-for-service 
operation. This required some modification and expansion of the produc- 
tion records previously kept. 

Meier Problems 

The principal unsolved problem of the 
Branch is the lack of a satisfactory 
plan for financing tee Statistical Processing Section. The demands for 
service from the Institutes end Divisions have continued to increase at 
a foster rate than the expansion of processing facilities. This iastel- 
lotion of tho IBM Type 650 computer has not resulted in a decrease in 
the workload for BAM equipment. The number of new jobs received during 
the last six months of 1958 far exceeded those transferred from IAM 
equipment to the computer. 

It is estimated that by July 1959 
about 263 hours of computer time each month mill be required to meet the 
existing demand. This will require a two-shift operation. Approximately 
two-thirds of this workload will represent new jobs. At the same tine 
s corresponding increase is anticipated in jobs that can be dons most 
efficiently em SAM equipment as & result of program expansion in tho 
Division of ftesearch Grants, the Peychopharmaeology Service Center 
Institute of Hsurological Diseases and Blindness 9 and elsewhere. 

She increasing workload esssaot fca 
coapleted by working overt iae. Throughout the yaw the asefeer of hours 
of overtlse has avasvaged froa 20 to 23 perceae of segalar tisa.. I 
of this, a backlog of work has acetoaalated oo that seas jobs have 
delayed for several Booths, A decision either to expand she personnel 
and cculpnent of the Statistical Processing Sections in order to asset 
the existing and anticipated deaand or to curtail the deoaad so that it 
can be serviced by existing facilities, is overdoe. 

Pora Bo. 0A?°2 

Serial Ho. Da8-5,1 

lo Biese tries 

3. Bethesda 
Individual Project Report 
Calendar Tear 1958 

Fare A. 



Principal Inveatigator: Lucia Dunhaa, mi-, Hsxold W. Dora, 

Chief, BiOEStriss Branch; Harold L. 
Stewart, SCI 

Other Investigators: 

Cooperating Unite: Laboratory of Pathology, national 

Cancer Institute 

Man Tears (calender year 1958) : 
Total: 0.1 
Professional: 0.1 

Project Description: 

Objectives : (1) To deteraiae the incidence of 

cancer of the cervix and corpus in 
Jewish and non-Jewish woman; (2) To investigate factors that stay be 
associated with any observed differences in incidence. 

Method Begloyed : The study was carried on in Hew Tork 

City and Israel. In Hew York City 
newly diagnosed esses of cancer of the uterus were reported to the study 
headquarters. In addition, an independent seerch was aade in the reeerda 
of hospitals for all cases diagnosed during calendar year 1952. Trained 

Part B included Tea . So JL 

DBS, Bioasa tries Branch, cont'd: 

social workers ^terviewed the reported cancer cases and an equal at^fee? 
of control case* without cancer . The eas* procedure wad followed £a 
Israel except that. In addition, eases of cancer of the breast and ©vary 
ware reported and Interviewed . 

Major Findings ; The Incidence rates for cancer of the 

cervix SBong Jewish feaalea are about 
one-fourth of those aneng non-Jewish fesalee In Sew York City. Ths 
Incidence rates are the saaa for Jewish fesales in Sew York Slty aad In 
Israel. In contrast, the Incidence of corpus cancer is essentially the 
saaa for Jewish and non-Jewish faoales. Within the Jewish population 
in Israel the Incidence of cancer of the breast, corpus, and ovary is 
higher for Aehkenaslc than for nen'4shkena>ic Jews, but no difference 
was observed for the incidence of cancer of the cervix. 

Course of Prelect : The field work has been csapleted end 

the analysis of the data is under **ay„ 
Preliminary results were presented before the Seventh International Cancer 
Congress, London, England, July 1958. A ponograph Is now in preparation* 

Pom No. ORP-2 CAttaefawant 1) 

Serial Bo. DBS-5.2 

lo Bio—tries' Branch 


3. Bethesda 


Individual Project Sapor t 

Calender Year 1958 

Part A. 

Project Title: CAHCK& Off THE OTBS0S 33J BB6S0 PKHALBS 

Principal Investigator: Lucia Dunham, MCI; Harold ». Dorn„ 

Chief, Biometrics Branch; Harold L. 
Stewart, HCI 

Othar Inveatigators : 

Cooperating Unlta: laboratory of Pathology, Rational 

Cancer Institute 

Han Tears (calendar year 1953) : 
Total: 0.1 
Professional: 0.1 

Project Deacrlption: 

Objective* : (1) To deteradne the incidence of 

cancer of the cervix and corpus in 
negro fancies; (2) To investigate factors that oay be associated with 
the develop— nt of these fores of cancer. 

Methods S aployad: All aevly diagnosed eases in Hew 

York City and Washington, 9. C. are 
interviewed by trained social workers. 

Course of Pro lac t : The interviewing of patients has been 

eoapleted in Sew Tork City but is still 
continuing in Washington. The interviewing will continue until a suffix 
elect number of cases for analysis has been interviewed. 

Part B included 

;,m Ho. OftP-2 (Attaeteant 1} 

Serial Bo. DaS-S.3 
1. Masetr&ea 

3. Bethesda 
Individual Project Report 
Calendar Tear 1958 

art A. 


Principal Investigator: Harold P. Dorn, Chiefs Biometrics 


Other Investigators : R. P. Kaiser 

Cooperating Unit : Field Investigat'Ons and Demonstrations 

Branch, MCI 

Men Teara (calendar year 1958): 
Total: 11.2 
Professional: 0.2 
Other: 11.0 

Project Description: 

Ob jectives : To determine differences in mortality 

from specific causes of death among 
persons with varying habits with respect to the use of tobacco; to inves- 
tigate factors other than tobacco—for example, occupation and reeideace°- 
vhlch may also account for the observed differences in mortality. 

Methods employed : Information concerning residence, 

occupation, and the use of tobacco 
is being collected from about 290,000 persons tfeo held U.S. Ceversaent 
Life Insurance policies in December 1953. As each person included in 
the study terminates a policy, the Biometrics Branch is notified by the 
?eterans Administration. If the policy is terminated as a result of 
death, information is collected from physicians and hospitals concerning 
the causes of death and the methods of establishing these diagnoses. 

Part B included Tea 

DBS, Bioaetrice 3roach, eoat'd: 

Major ^ladings: An analysis of the aorfcelity 
..,__, . saokers and noneaekers freWjuiy 

beceaber 1956 was presented before the Seventh International Cancer 
Congreos, London,, Bnglaad, July 1958 end before the annual sssatias ©f 
the Aaericen Statistical Association, Chicago, 111., Seeesbar 195S. She 
death rate froa ail causes of persons «a© have used tobacco is 32 peseent 
greater than that for persons eho have never stacked, Peree&e 
regularly aaoked only cigarettes haw a death rate 58 percent 
than the rate for nonsaokers. The excess aortality for cigaretts 
is greater for heavy than for light saokers. The aortality rate * 
regular cigarette-only saokers is 10 tines that for nonsaokero. 

Coarse of Project: It is espected that aortality records 

will be collected for about one aor® 
year after which an analysis of the results for the period 1957-59 will 
be aade. Plans are now being aade for a pathological study of the 
cancer deaths. 

?IS -V&X 

Individual Project Report 

Calender Tear 1958 

(Attachment 1) 

Serial to.J&fe&A 
1. Biometrics Branch 

3o Bethasda 

Part B.: Honors, Awards, and Publications 


Beer, Paul I. and Llebermaa e Jacob B. s '^Observations on Seme genetic 
Characteristics of the Ferlodontluea in Throe Strains of Inbred Kiea 9 " 
Oral Suraery. Oral Medicine and Oral Pathology. Dee. 1958 . 

Basse, Byron J., Jr., Liebernon, Jacob B. and Lusted, Lee 3., "Kidney 
Site in Acute Leukemia," J. ef Roentgenology. Radi ation, Therapy, and 
nuclear Medicine , p. 611, Oct. 1958. 

Cramer, Billot M., "Creation of Pitch through Binaural Interaction," 
J. of the Acoustical Society of America. Vol. 30, Ho. 5, pp. 413-417 
May 1958. 

Born, Harold P., "Darwin Revisited," Busanlcs Q uarterly Vol. 5, Be. 3, 
Sept. 1958. 

Born, Harold P., "Cancer Mortality Trends in the United States of 
America," Cancer. Vol. 3, 1958. 

Geever, B. P., Leoae, B.C., Gelser, P.B., and Lieberman, J.3., 
Studies la Man After Prolonged Ingestion of Fluoride in Drinking Hater. 
I. Kecropsy Findings in e Community with e Hater Level of 2.5 ppa," 
JADA . Vol, 56, pp. 499-507. 1958. 

Geevar, B. P., Leone, B.C., Gelser, F.B., and Lieberaaa, J.B., "Pathologic 
Studies in Man After Prolonged Ingestion of Fluoride in Drinking Hater," 
Pub, Health Reports . Vol. 73, pp. 721-731, 1958. 

German, James L. with the statistical assistance of Liebesman, J.S., 
'•The Glucose Tolerance Test After Cortisone Administration in Obese end 
Bonobese Men," Diabetes . Vol. 7, pp. 26l«268, July 1958, 

DaS, Bioae trice Branch, cont'd: 

Bonore and Awards 

Harold F. Porn 

(1) Elected President of the Washington Statistical Society 

(2) Reelected Geuerel Secretary, International Union Against Cone** 

(3) Elected Trustee of the Population Reference Bureau 

(4) Elected naaher of Exacutlw Coceeittee, Board of Directors, 
Social Science Research Council 

Calendar Year 1958 

Budget Data ; 

Obligations for Calendar Year 1956; 
Total: $551,155 
Direct : 
Reiabursenent: $551,155 

Serial Bfo. BRS-5. 5 im , 
1. ioael ss B2 

February 1959 Calendar Year 1958 


Summary of Branch Activities 1 DRS-6 

Serial Humber 


Branch Branch Chief 


Objectives of the Sanitary Engineering Branch center on 
maintaining and raising the standards of quality of the HIH physi- 
cal environment o The Branch program is designed primarily to 
study and control those factors at the HIH which affect laboratory 
and patient care environmental health and sanitation Another 
principal objective is to improve the quality and acceptibility of 
various utilities and research services „ 

The mechanism for meeting these objectives is found in the 
Branch' 8 four Sections: Insect and Rodent Control, Environmental 
Health, Equipment Testing and Quality Control* and Germ-Free 
Services. Each Section is held responsible for a particular pro- 
gram area and each la staffed with trained professional and 
technical personnel 9 Their personnel complements provide a broad 
range of related disciplines within the Branch without duplicating 
functions between the Sections o 


Insect and Rodent Control Section 

Surveys and assesses extent of pest prevalence* including 
animal ectoparasites and escaped rodents at the NIHo 

Develops pest control and eradication procedures founded 
on sanitation practices* such ass improved methods of animal room 
housekeeping; proper food and wa«te handling and storage practices! 
and introduction of rodent-proof design and construction features » 

Conducts continuing preventive -type pest control program 
including residual insecticide* trapping* and baiting activities » 

Insect and Rodent Control Section (conto) 

Prepares educational materials and lectures to selected 
employee groups on insect and rodent control procedures ■> 

Environmental Health Section 

Routinely inspects and/or tests principal environmental 
areas of health and sanitation concern as waste handling and dis- 
posal procedures and equipment; operation of patient and employee 
food services, including snack bar concessional and water., infants" 
formula and patients' liquid diets o 

Conducts special environmental surveys of laboratory 
buildings and patient care areas, such as operating rooms; studies 
and recommends measures for controlling air pollution, including 
chemical fumes and odors, and other industrial hygiene hazards, 
Including noise s 

Arranges for outside assistance and consultation to further 
supplement its surveys, such as in the fields of acoustics and 
vibration control ■> 

Equipment Testing and Quality Control Section 

Studies and recommends measures for improving quality of 
various products of central services, including special utilities, 
and for improving the performance of selected equipment o 

Evaluates methods, materials and equipment for removal of 
soil from laboratory glassware, animal cages, floors, and other 
surfaces . 

Investigates and improves the quality of distilled, deionized, 
and other special process waters used in research* 

Tests new sterilization equipment and methods, and determines 
effective and safe criteria for their use* 

Studies corrosion and related problems, and tests materials 
and plant processes for effectiveness in reducing corrosion o 

Germ-Free Services Section 

Provides Institute investigators with engineering, bacterio= 
logical control and mechanical maintenance services for investigations 
using germ-free animals <. 

Plans germ-free facilities for the Institutes • 

Designs, develops and tests new germ-free apparatus, sterile 
procedures and auxiliary apparatus 

Investigates and develops processes and equipment for con° 
trolling and/or containing microbiological contamination o 

Progress and Accomplishments 

IBSECT AHD RCDEHT COHTRCL SECTION Progress in insect control 

during the pest year has 
been difficult to measure. Up until 1958, an unusually high degree 
of insect eradication at minimum expense was achieved using 
chlordane, a chlorinated hydrocarbon* as the insecticide of choice , 
With the natural processes of species selection, the developing 
resistance of roaches to chlordane has reversed this favorable 
situation during the latter half of I9580 

To meet this challenge, this past year the pest control crew 
has been augmented by two new employees to a total of five*, One 
of the more experienced employees was shifted to evening duty to 
inspect and treat selected areas and handle requests for assistance, 
such as from nursing units » 

In spite of the Section's improved surveillance activities, 
and an increased rate of application of substitute residual 
insecticides, there was an increase in the number of insect com- 
plaints to 196 in 1958 Q8 compared with 152 in 1957° This is 
viewed as primarily due to the less effective residual insecticides 
currently available, That the increase was not greater is viewed 
as an indication of the generally ^ though not uniformly, favorable 
level of sanitation at the HIH combined with the general effective = 
ness of the Section's preventive type of insect control programo 

Ho outstanding problems in rodent control were experienced 
this yearo 

EHVTRQHMS]fEAL hratth SECTION The Section's sanitary 

inspections of food service 
areas and concessions have indicated a generally acceptable but not 
a completely satisfactory level of sanitation-, Special studies and 
recommendations have been made for improvements to and, in some 
instances, extensive remodeling of inadequate facilities o Under 
this category the following inspections were made; 

Barber shop ^ 

Routine food service 21 
Followup food service 11 

The water supply was routinely and randomly sampled each week^ 
with 8 total of kk8 water samples being examined for presence of 
coliforms as well as total count . The coliform index indicated 
satisfactory water quality at all times = The total bacteria count 
was generally acceptable except for buildings located in one areao 
Chlorination of the water distribution system in this area was 
followed by a satisfactory reduction of total bacterial counts « 

A total of 226 samples from the Clinical Center formula and 
tube feeding preparation kitchen were examined for coliform count . 
Samples collected and e xamine d weekly showed that the ingredients 
and preparation techniques were satisfactory except during one 
short periodo The cause of the high coliform count was traced to 
one vendor's product . Changes in the source of supply corrected 
this condition. 

A lecture series for foodhandlers was prepared and presented 
by personnel of this Section in cooperation with the Clinical Center 
attrition Department. Twenty classes were held on sanitary food 
handling practices for the personnel of the Hutrition Departanent. 

Marked Improvements made in the handling of infectious 
wastes during the year have considerably reduced the health hazards 
previously associated with the disposal of these wastes at the main 
incinerator. The number of cans of unbagged or Improperly bagged 
infectious wastes received at the incinerator has been sharply re- 
duced through tracing them back to their sourceo 

A variety of requests were received for assistance with 
problems of acoustics, ventilation* ultraviolet radiation equipment 
selection and installation, and building and area sanitation. 
These requests resulted in a number of reports and recoomendations 
designed to eliminate or substantially reduce environmental health 
hazards. The Section is spending an increasing proportion of time 
on problems in this general area. 

An extensive report was prepared, based on bacteriological 
sampling of operating room air, concerning methods for Improving 
air quality in the proposed surgical facilities for the Clinical 
Center. Ventilation details resulting from these recommendations 
will be Incorporated in the new surgical facilities. 

EQUIPMENT TESTTHG AHD QUALITY The Section's full author- 
CCHTRQL SECnCB ized personnel strength of 

four positions for calendar 
1958 was achieved when a physical chemist and an assistant sanitary 
engineer were recruited in March and June, respectively, enabling 
the Section to perform urgently needed equipment testing and quality 
control functions. 

A routine conductivity testing service for centrally dis- 
tilled water was instituted for the Clinical Center and other HIH 
laboratory buildings. This check facilitates prompt detection of 
trouble in the distilled water systems and forms the basis for the 
Section's weekly distilled water quality reports now being circu- 
lated to the Institutes. More recently, a weekly performance check 
of each of the six stills In the Clinical Center distilled water 
system was established to minimize distilled water shortages in 
this building. 

Various equipment layouts and specifications covering special 
distilled and deionized water systems were prepared. One of these 
vas for an HBffl micro-analytical laboratory requiring a high quality 
water free from metallic and fluorescent contaminants <> Shis dis- 
tilled water system will utilize a rechargeable mixed bed water 
deionizer in combination with a special single distillation still 
to produce the required high purity water „ 

Detergents were tested for compliance with HIE specifications 
covering chemical, physical and performance requirements. Annual 
savings obtained through purchase of such detergents through continu- 
ing contracts based on competitive bidding are estimated to be over 

Hew machines for washing animal cages were installed in 
Buildings k and 6, in accordance with SKB recommendations o Shle 
represents partial accomplishment of the Section's long-range goal 
of replacing outdated, inefficient equipment and providing mechan- 
ized cage -cleaning facilities in each of the original BIB laboratory 

A laboratory glassware -cleaning machine, deionizer, heat 
exchanger, and storage tank for hot deionized water were installed 
in Building 5 to improve the DBS glassware cleaning facility <> 
This equipment will be moved to the new DBS Building when it is 
completed , 

A variety of steam pressure sterilizers, gas sterilization 
equipment, water stills and deionizers, machines for cleaning 
laboratory glassware and animal cages* devices for cleaning refuse 
containers, and related equipment were tested for performance » 

A functional layout of the central glassware cleaning 
facility for the new Dental Research Laboratory Building was pre- 
pared. Specifications for water stills, sterilizers, and machines 
for washing cages and glassware in the new DBS and Dental Research 
Laboratory Buildings were also furnished to the Research Facilities 
Planning Branch* 

At the request of the Plant Engineering Branch, a study was 
made of corrosion conditions in the incinerator ducts and flue gas 
scrubber systems in Building 11, As a necessary measure towards 
solving the problem, special procedures were developed to deter- 
mine the acidity of the flue gases prior to entering the scrubber 
and after leaving the scrubber. 

Hew animal caging equipment, machines for washing cages and 
glassware, water stills, sterilizers, detergents, and related 
equipment and materials were inspected for compliance with 

GERM-FREE SERVICES SECTICH Germ-free research was 

solidly established at the 
HIH during the past year. That this was possible vas in large 
measure due to a number of significant accomplishments by the Germ- 
Free Services Section, including the following; 

Three additional germ-free facilities were placed in opera- 
tion at HIH during 1958. In addition to the extensive germ-free 
ai-ea in Building 8, two single laboratory -size, germ-free research 
areas are now in use by HIDR and H3AMD. Facilities and services 
for these areas were designed by SEB. The central germ-free area 
in Building 10 also continues in operation. 

With the above added space, 20 germ-free chambers are now 
in operation . In 1957 only nine chambers could be operated. 

Several procedures were changed which were aimed at making 
routine chamber operations less susceptible to chance contamination. 
Before these changes were made at midyear the contamination rate 
was about one per month. During the last half of 1958, there have 
been only two contaminations reported even though more chambers 
are in use. 

During 1958, eight shipments of germ-free rats and mice 
were received by HIH from Lobund Institute for use by four 
Institutes and DRS . In addition, three germ-free breeder rats of 
the Lund strain were brought from Sweden by Dr. Bengt Gustafsson 
for HIDR and HIAMD. 

A field trial of the Trexler flexible -film isolator was 
successfully completed during 1958. An evaluation study contrib- 
uted materially to the Germ-Free Services Section's increased 
competency in the use of plastic germ-free chambers and chemical 
sterilization techniques. These techniques have made the intro- 
duction of a wide variety of heat sensitive materials into the 
germ -free environment both feasible and convenient, and they are 
now a routine part of HIH germ-free operations. 

A rigid plastic germ-free chamber, capable of being assembled, 
sterilized, and used in an individual laboratory, was developed at 
HIH during 1958c 

A Gustafsson -type germ-free chamber was obtained in I&rch 
1958 and Bince then has been in use by HIDR. Its operation is 
being observed so that evaluation of its suitability to HIH 
conditions can be made. 

A continuing study was started in Ifey 1958 to determine the 
optimum breeding conditions for germ-free rats. Germ-free rats 
are being maintained under controlled conditions of noise, tempera- 
ture, humidity, and length of day-night cycle. Food is sterilized 
at the lowest practicable temperature, and bedding is provided. 

A formal agreement was consuma-fced with the University of 
Notre Dame for supplying NIH with germ-free animals at the initial 
rate of about 500 per year, with gradually increasing quantities 
in ensuing years. 

OTHER ACCOMPLISHMENTS In cooperation with the 

Research Facilities 
Planning Branch, manuscript material was prepared for a chapter 
on "Hospital Laboratories" to be included in the second edition of 
the Notional Research Council's textbook on "Laboratory Design". 

Assistance, including various informational materials, were 
furnished for an independent review and analysis of the Branch 
program, its impact and relations with other NIH staff and operating 
functions o Branch review and action on various recommendations in 
the report were in progress at the close of the year. 

Major Problems 

ALL SECTIONS NIH Space Utilisation s 

Equipment crowding in lab- 
oratories, corridors and other work areas throughout the NIH is 
being noted increasingly. As this condition continues to develop, 
such health and sanitation-related activities as insect and rodent 
control and routine cleaning will steadily become less effective * 
More hazards and, thus, opportunities for accidents will also 
develop. How to reverse this trend is viewed as an immediate and 
urgent problem. 

Financing Environmental Improvements . On various occasions 
the Branch has been confronted with how to finance immediately-needed 
health and sanitation related projects it has proposed. The present 
budgeting procedures whereby these items must be deferred normally 
until late in the fiscal year is uncertain and unsatisfactory. 
Greater budget flexibility is needed, with recognition of the neces- 
sity to take action towards expenditures of this type as they oevelop. 

NIH Staff Relations . Parallel and common interests in health 
and safety matters frequently develop between such groups as the 
Employee Health Service Branch, the radiological and general safety 
programs of the Plant Safety Branch, and the Sanitary Engineering 
Branch. While a forum for some of these problems has been developed 
in the "Environmental Health Advisory Group", there is need for some 
clearcut administrative mechanism through which these interests may 
Jointly take concerted and more effective action. 

Personnel Recruitment and Office Space. , The several posi- 
tions for which three of the Sect ions are recruiting are in 
professional categories. These positions are, at best, difficult 
to fill and with our existing inadequate space in which to place 
these professional personnel, inducements for their recruitment 
are thus limited. Section Chiefs do not now have separate offices 
which are needed for effective administration o Further, it has 
heen our experience that if three professional personnel are placed 
in an average -size office saodule, there are numerous disruptions 
not conducive to high-quality performance . We presently have this 
crowding in one office and will increase this problem as further 
positions are filled. 

GBHM-FK5E SEK7ICES SECTION Animal Supply. As the 

Branch '8 previous reports 
indicate y the problem of germ-free animal supply has not been 
solved. Only 91 germ-free rats were received from Lobund this 
year; 35 were contaminated upon arrival or during transfer to 
experimental chambers. Some limited breeding was successfully 
done at HIH* and an agreement has been reached with Lobund for 
supplying germ-free rats to KHo However, it is believed that 
large scale breeding of germ-free animals at HIH would best fill 
the needs of the investigators. Some general plans have been 
made in this direction, but until this proposal is an actuality, 
germ-free animals will be in short supply and severely restrict 
research In this area. 

Serial Ho d 

lo Sanitary Engineering Branch 
2o Equipment Testing and 

Quality Control Section 
3» Bethesda lk s Maryland 

PH5 — NIH 
Individual Project Report 
Calendar Year 1958 

Part A< 


Principal Investigator: Dr. Lawrence Lo Reed, Bacteriologist, GS~11 

Other Investigators: Harry Stierli, Sr» San. Engineer 

Dr. Irwin Ho Billick, Sr» Asst. Scientist 
J. Paul Davies, Engineering Aid, GS-7 
Frank Morelli, Bacteriologist, GS-7 

Cooperating Units: None 

Man Years (Calendar Year 1958) J 

Total: 0o7 
Professional: 0»5 
Other: 0o2 

Project Description: (See attached material) 
Part B included: Yes No 

Individual Project Reports 

Objectives ; 

To study recently developed gas sterilization equipment and 
methods^ and to determine criteria for their safe and effective 
use in various applications o 

Methods Employed ; 

Gas sterilization equipment ranging in size from a one -liter 
capacity portable unit to a large walk-in type chamber of about 
1*25 cubic foot capacity are under study. A variety of materials 
and apparatus together with representative test organisms are ex- 
posed to a range of conditions to determine the effect of the gas 
on mlcroorgaD-eRS and exposed materials . A rapid and fairly 
accurate method of determining ethylene oxide concentration of 
small samples of gas withdrawn from several locations within the 
chamber was developed using gas chromatographic apparatus o 

Major Findings ; 

Evaluation of an automatic control type commercial gas 
sterilizer utilizing an ethylene oxide-Freon mixture has been 
completed. The ethylene oxide-Freon mixture was found to be a safe 
and satisfactory sterilizing agent „ A minimum exposure period of 
2 hours sterilized all surfaces easily accessible to the ethylene 
oxide gas in a concentration of approximately 1^000 milligrams per 
liter at chamber temperatures of 120 9 Fo to 130*F. and relative 
humidity between 30# and 50^. A 4-hour exposure period was also 
found to be satisfactory for the above conditions with an ethylene 
oxide concentration of approximately 500 milligrams per liter. 

Significance to the Program of the DBS ; 

Commercially available equipment for gas sterilization of 
heat sensitive materials has been evaluated. This study has 
indicated that with certain precautions this type of equipment can 
be used safely and reliably in hospitals and laboratories to steri- 
lize the increasing number of heat labile materials and instruments 
being used. 

Proposed Course of Project; 

A portable one -liter capacity gas sterilizer using an 
ethylene oxide -Freon mixture is under study for kill of resistant 
microorganisms on small laboratory and clinical devices o 

The large walk-in steam pressure sterilizer* equipped for 
use with an ethylene oxide -carbon dioxide mixture s has not yet 
provided reliable bactericidal performance » Additional study and 
possible modification are required before routine satisfactory 
sterilization of contaminated oxygen tents and other large items 
can be achieved.. It is planned to conduct a series of tests with 
this large sterilizer checking microbicidal effectiveness s ethylene 
oxide concentration* temperature and humidity for various locations 
in the chamber » 

Serial No„ og ,,___ 

1 Sanitary Engineering Branch 
2<, Equipment Testing and 

Quality Control Section 
3» Bethesda 1^ dryland 

Individual Project Beport 
Calendar Year 1958 

Part A. 


Principal Investigator: Harry Stierli, Sr» San« Engineer 

Other Investigators: Dr. Irwin Ho Billick^ Sr» Asst. Scientist 
George Elmore s Assto Sane Engineer 
Harry Ettinger, Assto San Engineer 
Jo Paul Davies, Engineering Aid 5 G5=7 

Cooperating Units: Rone 

Man Years (Calendar Year 1958) 

Total: 0o5 
Profe88ional: OA 
Other: 0»1 

Project Description: (See attached material) 
Part B included: Yes No 

Objectives ; 

To continue investigation of distilled and deionized T-rater 
production, storage, and distribution problems with the aim of 
Improving the quality of centrally produced waters so that they 
can be used with a high degree of confidence In medical research » 

Methods Qnployed : 

A variety of distilled and deionized water systems in oper- 
ation at the HIH are under study. During Calendar Year 1958 studies 
were conducted toward development of rapid tests for detection of 
traces of copper in distilled water. 

Major Findings ; 

Central KIH distilled and deionized water systems were found 
to contain water of not less than 500,000 ohms specific resistance, 
except for occasional breakdowns in equipment . 

A rapid spot test, specific for copper, was developed for 
detection of traces of this contaminant in distilled water with a 
sensitivity in the order of 0.1 PPM. A more quantitative spectra- 
photometric analysis was also developed to determine amounts of 
copper in distilled water in the range of 0.01 to 0.1 PPM. 

Particulate matter obtained from the aluminum distribution 
piping of the Clinical Center distilled water system was found to 
be an insoluble aluminum hydroxide floe caused by air entering the 
system when the demand exceeded the supply. 

Significance to the Program of the DBS ; 

The Improvement of centrally distilled and deionized waters 
is essential for the support of the increasing number of investi- 
gat ions involving sensitive biochemical and biological systems. 
Routine conductivity checks serve as the first step in quality 
control of these special process waters. The rapid tests for 
detection of traces of copper will help to guide the DBS in re- 
tinning, or replacement, of components in the central distilled 
water systems. 

Proposed Courae of Project ; 

Sources and amounts of trace contaminants in water from 
various still and deionizer systems will continue to be studied* 
Determination of allowable amounts of these contaminants in such 
process waters will be made using known sensitive biological 
indicators such as tissue cultures and enzymes » Da ionizer ~isster 
still combinations, together with other special treatment^ will be 
further explored for use where there are unusually stringent 
requirements . 

Serial Koo 3 ^ 

Xo Sanitary Engineering Branch 
2 Equipment Testing and 

Quality Control Section 
3° Retbesda lh s Maryland 

Individual Project Report 
Calendar Year 1958 

Part A< 


Acting Principal Investigators Harry Stierli s Sr° San<> Engineer 

Cther Investigator: None 

Cooperating Units: Hone 

Man Years (Calendar Year 1958) : 



Project Descriptions (See attached material) 
Part B included: Yes Ho 

Objectives ; 

To investigate the nature and quantity of trace surface con~ 
taminants remaining on laboratory glassware cleaned with alkaline 
detergents and acids. To determine the criteria for "clean" 
laboratory glassware for various types of research work at the EXE. 
To study new methods of removal of soils and to improve existing 
procedures. To develop special and routine tests for control of 
quality of cleanliness of laboratory glassware surfaces. 

Significance to the Program of the DBS; 

Laboratory glassware is utilized by almost every type of 
research activity. Precise knowledge concerning the nature and 
quantity of trace surface contaminants and their effects on various 
types of research is required to reduce variables for analytical 
work and for study of sensitive biochemical systems. The study of 
new methods of soil removal and the development of precision methods 
for detection of trace amounts of interfering substances will im« 
prove the quality of cleanliness of laboratory glassware used in 

Proposed Course of Project : 

Analytical chemical procedures will be used to determine 
quantitatively the various trace contaminants found on laboratory 
glassware cleaned with alkaline detergents and/or acid followed by 
thorough rinsing. The effect of rinsing with ordinary water, 
softened water, deionizad water, and distilled water, used singly 
and in combinations, will be evaluated. Both hand and machine 
methods of cleaning will be investigated. Microchemical techniques 
and biochemical indicator systems will be developed to accurately 
determine presence of extremely small quantities of interfering 
substances on the surfaces of "clean" laboratory glassware. 

Serial Ho 

1« Sanitary Engineering Branch 
2c Equipment Seating and 

Quality Control Section 
3o Bethesda lk 9 Maryland 

Individual Project Report 
Calendar Year 1958 

Part A. 

Acting Principal Investigators Harry Stierli^, Sr<> San., Engineer 
Other Investigator: None 
Cooperating Units: None 

Man Years (Calendar Year 1958): 



Project Description: (See attached material) 
Port B included: Yes No 

Objectives : 

To determine the quality of steam required for various 
laboratory and hoapital uses. To test, evaluate and develop new 
methods of removal of particulate matter, oils, amines, and other 
eubstancea in steam supplies which interfere with certain labora- 
tory and hospital procedures . To investigate snd develop new 
boiler feed water treatment materials and methods for possible 
production of a cleaner, noncorrosive steam for general use at the 

Significance to the Program of DRS s 

Sterilization of laboratory glassware, culture media and 
supplies for tissue culture studies and other sensitive laboratory 
and clinical work required a "clean" steam for autoclaves . Parti- 
culate matter from pipe scale or corrosion, oil from lubricated 
pipe expansion joints and chemicals from boiler feed water treat- 
ment, including amines, each contribute to contamination of the 
steam supply. Development of new methods of removal of these 
contaminants and/or production and distribution of a better quality 
of steam are needed to eliminate interference with sterilization 
procedures . 

Proposed Course of Project : 

The contaminating substances in the HIH steam supply will 
be determined quantitatively using random samples from a represent- 
ative number of collection points in the steam supply system. 
Known concentrations of these contaminants on surfaces of labora- 
tory glassware and in media will be studied for effects on tissue 
culture work and other sensitive procedures used in medical research. 
New equipment for removal of particulate matter, for separation of 
entrained liquid and vapor phases, and for absorption or adsorption 
of amines in steam will be tested, evaluated and, if feasible, de- 
veloped for practical use. Boiler feed water treatment will be 
investigated, using new materials and/or methods, with emphasis 
on production of a noncorrosive steam which is fully acceptable 
for laboratory and hospital use. 

Serial No„ __ .5 

1. Sanitary Engineering Branch 
2 = Germ-Free Services Section 
3o Bethesda lk a Maryland 

Individual Project Report 
Calendar Year I958 

Part A< 


Principal Investigator: John L« S. Hickey 

Other Investigator; None 

Cooperating Units: NIDR, NIAMD, and NIAID are cooperating 
in a consulting capacity in field trial 
evaluation of equipment developedo They 
have no serial numbers for this. 

Man Years (Calendar Year 1958): 

Total: 0„3 
Professional: 0.2 
Other: 0»1 

Project Description: (See attached material) 
Part B included: Yes No 

Objectives : 

To design and develop a germ-free system capable of supplying 
germ-free animals in chambers which are conveniently transportable * 
easily operated,, economical and adaptable to the unique and varied 
research requirements of NIB* investigators within their own 
laboratories . 

Methods Employed ; 

(a) Operation and evaluation of existing types of equipment 
to determine their shortcomings * (b) consultation with NIH invests 
gators to determine their particular needs in equipment* and (c) 
design and fabrication or procurement of equipment* plus field 
evaluation under research conditions. 

Major Findings during 1958 ; 

Three germ-free chambers and two portable autoclaves built 
to NIH design were obtained and placed in use in May* September* 
and December, 1958 in investigators' laboratories. One* a portable 
stainless steel chamber, was constructed commercially. The others* 
made of rigid plastic and also portable* were assembled at NIH 
from parts purchased outside and made in the NIH Instrument Section. 
All the chambers are proving practical. It is expected that pro- 
longed use and observation will indicate the need for modifications^ 
which will be incorporated into later designs. 

Significance to the DRS Program ; 

Development of inexpensive* safe* and convenient equipment 
for use by the scientist is a requisite to expansion of research 
utilizing germ -free animals. Chambers of this type will eliminate 
the present requirement of a centralized experimental area by 
making available to the investigator germ-free chambers designed 
for use in a laboratory module. The decentralization and the 
adaptability of such chambers will allow more investigators in a 
greater variety of research areas to use the germ-free animal. 

Proposed Course of Project ; 

The prototypes of improved germ»free chambers which are now 
in use by investigators are being observed and their features 
critically analyzed. Modifications will be made to these chambers 
to suit investigators' needs* and standard specifications will be 
compiled when the design becomes final. 

Serial No* ,.6 

lo Sanitary Engineering Branch 
2 Germ-Free Services Section 
3<, Bethesda lk 9 Maryland 

Individual Project Report 
Calendar Year 1958 

Part A. 


Principal Investigator: John L» So Hickey 

Other Investigator: Dr. R. E. Horton 

Laboratory of Germ-Free Animal Research 

Cooperating Units: Dr. Ho L. Andrews a Radiation Branchy NCI 
Mr. E. G. McDaniel* LHE, NIAMD 

Man Years (Calendar Year 1958) : 

Total: 0.3 

Professional; 0o2 
Other: 0.1 

Project Description: (See attached material) 
Part B included: Yes No 


Objectives ; 

To develop a nutritionally well=defined diet which will 
produce a healthy germ-free guinea pig, with growth and other 
characteristics approaching those of the normal animal* 

Mat hods Employed ; 

Activitiea this year have been toward the study of irradi- 
ation aterilization of guinea pig diet, and included; (l) 
preparation of a complete practicable diet, (2) development of 
techniques for irradiation sterilization of diets, (3) development 
of control techniques to insure sterility of diets, and (h) testing 
of diets on germ-free animals both from a sterility and nutritional 

Major Findings in 1958 ; 

Techniques were developed for large-scale preparation, 
packaging, and irradiation sterilization of diets in convenient 
form, including fresh vegetables as well as dry diets. 

Proper irradiation doses were determined and control measures 
developed to insure uniform sterility of diets. 

late in the year, diets sterilized by irradiation were fed 
to kQ germ-free guinea pigs and rats and to control groups. No 
conclusive results have been obtained yet. 

Significance to DBS Program ; 

Development of a germ-free guinea pig diet which will produce 
guinea pigs as similar as possible to their conventional counter- 
parts will increase the probability that experimental data obtained 
with these pigs will be reliable and reproducible. Such a diet 
would make work with germ-free guinea pigs more practical to 
investigators and to any Central Service in DRS undertaking to 
provide germ-free guinea pigs to investigators. 

Proposed Course of Project ; 

The search for a suitable germ-free guinea pig diet will 
continue. Factors which will be investigated further are; 

1) alternate methods or procedures in diet sterilization, 

2) supplementing diet with essential vitamins, and (3) development 
of a synthetic diet. 

Serial No„ .7 

lo Sanitary Engineering Branch 
2o Environmental Health Section 
3o Bethesda lk s Iferyland 

PHS -«. NIH 
Individual Project Report 
Calendar Year 1958 

Part A 


Principal Investigator; Dr. Lloyd G„ Herman, Bacteriologist^, GS-11 

Other Investigators: Mr. John Lo S. Hickey, Sr» San<> Engineer 
Mr« Frank Morelli 8 Bacteriologist a GS-7 

Cooperating Units: None 

Man Years (Calendar Year 1958) i 

Total: 0.08 

Professional: o 04 
Other: OoOU 

Project Description: (See attached material) 
Part B included: Yes No 


Objectives ; 

To study the bacterial flora of the air in certain selected 
areas at the NIH, and to determine methods for reducing bacterial 
contamination of the air to the lowest possible levels . 

Methods Employed ; 

Since each area has specific functions, a variety of methods 
and air sampling equipment will be required to obtain the most 
informative data. For example, these will include; (l) a long- 
plate slit sampler, (2) a round-plate slit sampler, and (3) open 
plates of both types. 

Major Findings in 19 58; 

Significance to the DRS Program ; 

The results of this project should lead to the improvement 
of and optimum methods for maintaining the working and patient 
care environment o 

Proposed Course of Project ; 

A study of the airborne micrococcal population including 
factors in their production and dispersion will be carried out in 
typical patient care and research areas of the Clinical Center and 
other NIH buildings. Various types of air samplers designed for 
sampling over extended periods will be employed together with 
special selective media . 

The project will be limited initially to a period of 
preliminary screening to establish the range of bacterial popu- 
lation of the air in representative areas. 

Although not all micrococci present in the air are potential 
pathogens, one phase of the study will characterize the proportion 
of hemolytic coagulase positive, mannitol positive, egg yolk 
positive, antibiotic resistant and bacteriophage types present in 
the air samples. 

Representative areas to be studied include surgical areaB 9 
nursing units., animal rooms, and laboratories as may be necessary.. 
Air flov characteristics in the above areas will also be critically 
examined and adjusted to insure proper relationships of their air 
pressures with respect to areas external to them<> 

As the study develops, skin and nasal swabbing of pertinent 
staff members and patients may be found necessary. This phase 
will require the assistance of other departments or laboratories 
within the Clinical Center of the Institutes. 

A further evaluation of this project is anticipated as soon 
as the initial data outlined above can be obtained. 

Serial Bo„ «8 

lo Sanitary Engineering Branch 

Calendar Year 1958 

Budget Data ; 

Actual Oblications for Calendar Year 1958: $ 156,708 

Reimbursement: $ 156,708 


Summary of Branch Activities 1„ DRS-7 

Serial Number 


Branch Chief 


To plan and construct new research facilities as required by the new 
and expanding needs of NIH. 


To serve as the focal point at NIH for the assembly and correlation 
of all information required in programing and planning the construction of 
all new and additional research facilities that require the participation 
of the Public Buildings Service, General Services Administration and of all 
new and additional temporary construction of research facilities. 

To provide the major contact point for coordination between the 
Public Buildings Service or contract architect and the appropriate NIH 
division during the planning and construction of a research facility until 
it is occupied by NIH. 

Progress and Accomplishments 

The new Research Facilities Planning Branch was actively operating 
at the beginning of calendar 1958. During the year the activities of the 
Branch were divided into two sections, Construction and Design and Develop- 
ment, to improve operational standards. 

Staff Status - Considerable effort was devoted during the year to recruiting 
professional staff , The present staff consists of the Branch Chief, two 
Section Chiefs, five junior engineers, two secretaries , and an additional 
secretary who was employed during the year to provide stenographic services 
for the present professional staff. 

Building 8 - Germ free Area. The construction contract for converting the 
north half of Che top floor of Building 8 to a germfree animal facility 
was completedo Considerable difficulty was encountered with the construc- 
tion contractor, One major defect is to be corrected before the Public 
Buildings Service will close the contract. The NIAID section on Germfree 
Animal Studies moved into the new facility on May 21, 1958. 

Bull ding 10 - New Surgical Wing,. Completed working drawings for the 
Clinical Center's new surgical wing are scheduled for February 4, 1959. 
Because present funds are limited, deduct alternates have be.^i developed 
permitting the procurement of the basic facility within the available 
funds. The construction contract is scheduled for award on May 22 8 1959, 
and construction will be completed in December 1960. 

Building 11 - Extension, The final plans and specifications as completed 
in 1957 for the extension to Building 11 contained an alternate plan for 
an extension to house two compressors with current installation of one 
compressor. The contract award, with acceptance of the alternate plan, 
was made on May 14, 1958, to Grunley, Walsh and Blanche for $341,400. 
The estimated completion date is July 15, 1959 „ 

Building 28 - Wings . Wings C and D to Building 28 were made available on 
February 10, 1958, to the National Cancer Institute. Final acceptance by 
NIH of the entire project was made February 13, 1958. 

Building 29 - Biologies Standards Building. On release of funds by the 
Bureau of the Budget, the base contract for the new biologies standards 
building was advertised for bids on February 26, 1958. The contract was 
.iwarded to the Norair Engineering Corporation for $2,267,000. Construe- 
tion was begun May 26, 1958, and is progressing satisfactorily. The 
estimated completion date is October 6, 1959. Award of the laboratory 
equipment contract is scheduled for April 1959. The estimated occupancy 
date is January 1, 1960. 

Building 30 - Dental Research Laboratory . Final working drawings for the 
base contract for the new dental research laboratory were completed on 
October 22, 1958 Contract award is scheduled for February 24, 1959, and 
completion of the base contract for October 21, 1960. Plans for the lab- 
oratory equipment contract are being prepared. 

Building 31 - General Office Building . The scope of the construction 
project for the new general office building was materially enlarged in 
1958. The present design of the building will double the office space 
originally contemplated. The proposed facility will house 1900 persons. 
The size of the building was increased because of the rapid expansion 
of NIH extramural programs. 

The contract architect was directed to proceed with the development 
of the drawings and specifications for the enlarged project, which is now 
in the 75% working drawings stage. The contemplated schedule is to com= 
plete the drawings and specifications by March 30, 1959, and to award the 
contract on June 9, 1959. The estimated completion date of the contract 
is July 9, 1961 

Saint Elizabeths - NIMH Cooperative Research Project . On June 30, 1958, 
the National Institute of Mental Health occupied the renovated first and 
fifth floors of the William A. White Building and the new animal building 
at Saint Elizabeths Hospital. 

Waste Chemical Disposal Plant, Improvements to the Waste Chemical 

Disposal Plant were completed and the plant was put into operation by 
the Plant Safety Branch of the Division of Business Operations in August 
1958 . 

Cage Washing Facilities. Plans and specifications were completed in 1957 

for the construction of a cage washing building adjacent to Building 14, 
Wing F. This project was approved for construction when funds become 
available — at a later date. 

Greenhouse . Plans and specifications were prepared by a contract architect 

in the spring of 1958 for the construction of a greenhouse. Contract award 
was made to the James L. Partello Company on June 27, 1958, for $113,000 = 
Construction progress has been satisfactory. The estimated completion date 
is February 9, 1959. 

Land Utilization Study . During 1957 a group of consultants were retained 
to study NIH operations and to prepare a master site plan presenting the 
basic guidelines for orderly, future expansion of NIH, The study and the 
master site plan were completed on March 31, 1958. 

Topographic Survey „ In March 1958, the Research Facilities Planning Branch 
Initiated the planning for a photogrammetric survey of the NIH reservation. 
Specifications and drawings were prepared, and with the close guidance of 
the United States Geological Survey, the topographic survey was completed 
on August 23, 1958. 

Study on NIH Water and Sewerage Systems. A consultant was retained in 
1958 to study NIH water use practices and to evaluate the capacity of the 
sanitary sewerage system, An Interim report was received in June 1958. 
the final study is scheduled for distribution by February 28 8 1959c 


Personnel s The Research Facilities Planning Branch has encountered 
difficulty in recruiting experienced personnel though it has not had 
trouble in recruiting engineers without work experience. A senior 
engineer (6S-12) detailed to the Branch for the balance of fiscal 1959, 
is cov.' acting as Chief of the Construction Section* Another engineer 
(Sanitary Engineer, PHS Commissioned Corps) is serving as Chief of the 
Design and Development Section. A vacancy exists for a mechanical 
engineer (GS-12). 

Workload , Some inequalities in workload became apparent as the Branch 
was being organized. However, with the division of the Branch into two 
sections, assignments are more evenly distributed. 

Space Deficiencies . Office space for the Branch in Building 13 became 
inadequate as staff was increased. Present plans for relocation within 
the existing area will help alleviate space deficiencies . However „ 
conference space and storage space are not fully adequate. 

'eraonncl Training . TWo junior engineers attended a PHS orientation 
course at the Robert A. Taft Sanitary Engineering Center and the frane 
Jompaay's evening course in principles of air conditioning. A junior 
engineer attended the DRS technical writing course. It is hoped that 
mdget and workload in 1959 will permit more of the young engineers to 
it tend industry^ sponsored and university short courses. 


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