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ANNUAL REPORT

PROGRAM ACTIVITIES

NATIONAL INSTITUTES OF HEALTH

1958

CLINICAL CENTER

NATIONAL INSTITUTE OF DENTAL RESEARCH

DIVISION OF B10LOG1CS STANDARDS

DIVISION OF GENERAL MEDICAL SCIENCES

DIVISION OF RESEARCH GRANTS

DIVISION OF RESEARCH SERVICES

NATIONAL INSTITUTES OF HEALTH
PUBLIC HEALTH SERVICE
U. S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE

tfeC"

ANNUAL REPORT

PROGRAM ACTIVITIES

Oi. NATIONAL INSTITUTES OF HEALTH

1958

CLINICAL CENTER

NATIONAL INSTITUTE OF DENTAL RESEARCH

DIVISION OF BIOLOGICS STANDARDS

DIVISION OF GENERAL MEDICAL SCIENCES

DIVISION OF RESEARCH GRANTS

DIVISION OF RESEARCH SERVICES

NATIONAL INSTITUTES OF HEALTH

PUBLIC HEALTH SERVICE

U. S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE

PUBLI "VICE KHATK

SUMMARY ANNUAL REPORT OF PROORAM ACTIVITIES
CLINICAL CENTER

OFFICE OF THE DIRECTOR .c':

Seri

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 0

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

Offic

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<

led
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)

1951

1955

1956

1957

1958

1,1*52

1,889

2,1*09

2,891*

3,300

lili

6i*,i*i5

96,1*05

109,71*3

120,131

11*0,151

176

261*

300

329

381*

1*0

1*8

1*1

hit

9,371

ll*,606

20,Uii6

21,1*87

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;.

Anat-

Diagnoetlc Radiol

Internal Medicine

Peychia
Ophthalmology

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 medic.il 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

volunteers

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
ne: 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 Uy 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 10g000 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 . .

1953

195:.

195<
1956 . .

;oo

00
6,100

^00

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

Publications

! 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"
"Cryptococcosis"

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

- Vol, W, N0o 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
Physician

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

Admissic

Inpatient lotters , . . . 3,108

Outpatient comspondonce pre: . 1,7^3

Physician referrals

Acknowledgement letters ••••••••••

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

Congressional contacts

Telephone calls ....... » 102

Letters answered . lhO

Cc.
PUBLIC HE ICE

SUMMARY ANNUAL REPORT OF PROGRAM ACTIVITIES

CLINICAL

ADMINI STRATI VE BR,"

PATIENT ACTIV ?ION

The Pctient
eetabli. 7. Dur olendar year it

continued to cixry out ita primary

nal
net

pat:

2. To provide a patient library service.

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

charged eea of t

Inst

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

Recreation

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 page 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,
NCI, NIAMD, and NINDB.

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 additional
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
PUBLIC HEALTH SERVICE NATIONAL INSTITUTES OF HEALTH

SUMMARY ANNUAL REPORT OF PROGRAM ACTIVITIES
CLINICAL CENTER
OFFICE OF THE DIRECTOR

CHAPLAINCY DEPARTMENT CC-OIX

Serial No.

PROTESTANT CHAPLAIN

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
staff.

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

1,296

Counseling

Rounds

l,02h

Criticalo

819

Baptisms

Funerals

Relatives

906

Weddings

. Pre-Oper.

521

Worship

Chapel Services

Attendance

3,982

Average

Holy Ccnsiunion

Chapel

60U

Bedside

198

Patient Referrals

Doctors

Nurses

771*

Social Workers

168

Clergy

Family

Patient

Others

Staff Relation*

Counseling

Conferences

1,072

Staff Mtgs.

Lectures, etc,

■ 19

Comrninity

Addresses

Meetings

Services

Professional

Conference a

Administration

Correspondence 752

Planning

737 brs.

Conferences

vith Chaplains 6l

Study & Research 739 hrs,

CATHOLIC UE/i?y:Ili

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.

SUMMARY OF SERVICES FROVIDEP BY THE CATHOLIC CBAPLAIN

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.

JEWISH CHAPLAIN

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.

SUMMARY OF SERVICE

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

PUHLIC HEALTH SERVICE, NATIONAL INSTITUTES OF HEALTH

SUMHART ANNUAL REPORT OF PROGRAM ACTIVITIES
CLINICAL CENTER
ADMINISTRATIVE BRANCH

EMPLOYEE HEALTH SERVICE BRANCH QO-12

Serial Number

Major Goals

The Employee Health Service acts ao the focal point for all natter*
relating to the health of employees 0 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 exandnationa0 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 0 We hop© to increase our efforts along
these lines during the next year, realizing that 60 to 80$ of ineffective
performance in industry i3 due to emotional problems* We can think of no
area that needs our concerted efforts more than this one0

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 pastB we stand ready to assist any NIH scientist in the
proper utilization of the employee population In suitable research
projects o

Major Problems

—*■! inr i ■mil mwjm 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 2S633

Immunizations ooo«oo««oo o 8,23u 8j,760 3^009 28lil3

Pre~empo Physical Scamlnationa 000 1*828 lp637 1&915 18351

Laboratory Examinations 0 „ 0 . « » 12,868* 6,528 5,9l<5 6CU75

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

* Revised system of accounting which now includes all procedureso

Serxal Mo0 CC~12

lo Clinical Canter

20 Employee Health Service

3o Bldg0 ID, Rmo B2«A06

PBS « HIE*

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

Objectives*

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 Baployedt

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

Patient Material*

The relatively stablee significantly large enough0 cooperative NIH
otrolcyee population was considered the ideal resource for con-
ducting this clinical research,. The vaccine was offered on a

-2-

Patlent Material: (Continued)

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

Major Findings t

■ ^wmmiim Kill

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 NIHt

Perhaps the greatest significance of this study is the denonstration
of the clinical research potential of occupational Kedicine0 Proper
utilization of the vast pool of clinical roaearch material found in
the NIH employee population is a major goal of the Employee Health
Service©

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 »

TABULATION OF REACTIONS TO INFLUENZA VIRUS VACCINE

PRELIMINART. ANALYSIS

% of individuals

reporting reactions «

1st. Inoculation

2nd Inoculation

Reaction*

1957-58

1*58-59

1558=59

Any Systendo Reatioa

29o2

20o6

12d

Fever, Unspecified

3oU

lo2

Fever* under 100° F

12o0

0o3 5°3

0008 203

Fever, orer 100° *"

1«6

loO

China

12 oh

6,2

3o6

Fatigue

21o2

15°9

8o9

Nausea

7o0

5a6

2o8

Any Local Reaction

l*2o2

aic6

27o5

Redness

23ol

26 o3

17 oO

Pain

25*5

22 o$

Hio5

Swelling

15o6

17ol

llo9

Other Reaction

13*1*

9o5

5»6

Ho Reaction

10.5

Ii0o5

51ol

Individuals Reporting Lost Tine

86 (l*oW)
158 (lo8 days,
mean)

28 (2o0Jt)

Total Days of Lost Time

Reported

Sh (106 days,
mean)

Number Vaccinated

2021

2675

2003

Number Reporting

H589 (SW)

2088 (78%)

1381 (68%)

Calendar Yea
PUBLIC HEALTH SERVICE ItATIONAL INSTITUTES OF HEALTH

SUMMARY ANNUAL REPORT OF PROGRAM ACTIVITIES
CLINICAL CENTER

CC-21

SOCIAL SERVICE DEPARTMENT Serial No.

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 program 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

- 1, 2 (sad. 3)

ie need sff

will rrucii needs would te ■' -related

\ced on
- House), tended that

:ho provide aome service to patier.'
the supervision and di
re uaiforaity of grade* quality of
speosibD

Participation. Again this year more

clir -ilising social data supplied "by mease

Ox the StStff Of

1 Health

in rosea; . Seurology, Cane.

and and :'/-'■ i sense are having increasing i

to supply 3ata frost their casewoxfc purposes.

nations, such as, the Heart Control Program,
voluntary health laiaats of other

hospitals have consul staff members about : larch plans

^ggestions, salnly about research on the
social ii ■:-, disern:

rtacsxt led a seminar oa "Social

auored by the Americsl Hospital
rtion of the Stoticaal Assoclr

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it the I&iversity of Cosiiecticirt. 2he

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rial baa "been geared particularly to
one to ub froa a variety of oattings.

indigent pet.i sities

iteaily Eeribera t£lo could cot pay for board
al phases of

his calendar year were $6,990*^3 and
deposits, larg taation and Welfare / m, Inc.,

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PUBLIC HEALTH SLRVICE NATIONAL INSTITUTES CF HEALTH

ANNUAL REPORT
CLINICAL CENTER

ANESTHESIOLOGY DEPARTMENT CC~3li

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 N3DR0

Administration of Anesthesia. 1958

Sponsoring

Institute

Procedures

Percent

NHI

1*72

ItO

NCI

261

22o2

NINDB

217

18 o5

NIDR

136

11 06

NIAMD

1*3

3o7

NIAID

3.7

NIMH

0o3

1\16

= 160o0g

Anesthesiology Department

Iln INHALATION THERAPY SECTION ACTIVITIES o During the year
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 Drf. J- W0 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, Fe 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„ G0 R, Christenson, directly participated
in a project sponsored by NIDR, namelys 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 drugp
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,
respectively.-

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.

VI- PUBLICATIONS AND PRESENTATIONS,

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

Calendar
PUBLIC HEALTH SERVICE NATIONAL INSTITUTES OF HEALTH

SUMMARY ANNUAL REPORT OF PROGRAM ACTIVITIES
CLINICAL CENTER

CC-35

DIAGNOSTIC X-RAY DEPARTMENT Serial No.

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

Av>p:

j re obta

- 3 -

Major Problems Encountered:

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

.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

Honors , Avarda , and Publications :

The Dc; 2 received an award for her id

ty locking device used In cor.j
angle

vKis fonr
7 Radiology and h

"ccioty. He t ^ on the

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

rs published by
ditional

' on.

•adlological subjects van by st

Serial No. CC-35-1

1. Diagnostic X-ray Oept,

2. Clinical Center

PHS-NIH

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'

Branch)

-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_ :

ns.

-6

PUBLIC HEALTH SERVICE NATIONAL INSTITUTES OF HEALTH

SUMMARY ANNUAL REPORT OF PROGRAM ACTIVITIES
CLINICAL CENTER

DENTAL DEPARTMENT CC-36

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
C.C.

2409

). 956
Bxamina'

1230

Visits

6710

Treatments

8167

1957

Admissions to
C-C.

3339

Examinations

1782

Visits

9491

Treatments

10, £78

1958

Admissions to
C.C.

3292

ninations

1937

Visits

9157

Treatments

11.756

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.

CLINICAL PATLOLO:

Serial No.

Mission

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.

Pereonaal

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*

Operations

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

Manual

Atttoastie

C::Ic"iG3

$ 0.33

$ 0.06

Protein

0.33

0.13

Glucose

Co 32

0.11

Blood Brea Bitrogea

0.3S

0.11

Red Blood Count

2.22

0.19

vatic vs.

Manual Method

Manual

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.

Records

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 eni.no 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.

Awards

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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Calendar Year 1958

PUBLIC HEALTH SERVICE NATIONAL INSTITUTES OF HEALTH

SUMMARY ANNUAL REPORT OF PROGRAM ACTIVITIES
CLINICnL CENTER

REHABILITATION DEPARTMENT CC-33

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
response.

fo Self-care evaluation (Activities of Daily

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

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

aQ Superficial - hot packs (Hydrocollator)9

infrared, paraffin bath, whirlpool.

b. Deep - short wave diathermy, microwave 8
ultra-sound.

Co General body heat - Hubbard tank. Moistaire

cabinet o
3o Therapeutic exercises

a. General exercise ~ passive, active assistive9

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

techniques, mass movement patterns and

proprioceptive facilitation techniques o

c. Ambulation training, including elevation9
with and without parallel bars8 braces 8
crutches, canes 0

d. Posture exercises

-2~
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 activities0

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
instruction,,
4o Miscellaneous

a. General and local application of ultra-violet
lighto

bo Contrast baths.

c, Scientific massage*

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

f. Application of splints and casts to maintain

joint in good anatomical and functional

position,
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, splints8

prostheses.

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 0

-3-

Rehabilitation Department

5o Substituting constructive interests for the

insecurity ana anxiety which may develop during
the research study0

6n 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*uaT,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

studieso
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 vicinity0

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 has9 as mi?;ht be
anticipated „ slowed down. An additional mat table has been
purchased for the Physical Therapy area8 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 frame9 enabling patient to be turned from prone
to supine position and vice versae

-4-

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 0
'.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 areap
viz08 Arts and Crafts, has been returned to the
Rehabilitation Department 0 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 Chief8 Occupational Therapy
Service,, with assistance in the conduct of the program
from the Gray Ladies 0 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

-5-

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,
1959.

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 patients0
It is3 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#0
Dr. Tillye Cornman was at one time the Acting Chief
of the Rehabilitation Department and more recently a
clinical associate in the NINDBC 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 etcQ

-6-

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
services,,

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 area0
It will still be useful, with some simple screening,
as a locker room before 8:30 A.M. and after 5:00 P'.M0
A small expenditure of money can thereby provide
additional and essential treatment and office spaceQ

During calendar year 1957, Miss Helen Applebaum9
staff occupational therapists attended Miss Margaret Roodvs
course on "Neurophysiology in the Treatment of Neuro-
muscular Dysfunction"0 This year Mr. Otho Bell9 staff
physical therapist, took the same course a They are now
using thse new technics with what we consider to be
favorable results 0 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 Servicec

Calendar Year 195©

PUBLIC HEALTH SERVICE HA21CKAL IHSTTTUTES OF HEALTH

SUMMARY AHTOAL REPORT OP PROGRAM ACIIVITIBS
CLIHICAL CENTER

CC-^2

PHARMACY nSP/JRO£SOT

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
disturbing.

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 prescriptions from the
Pharmacy, vith only "Instructions for use" and prescription
numbers.

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.

CENTRAL STERILE SUPPLY SERVICE

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

PHARMACEUTICAL nsVELOBffiOT SERVICE

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 cooperation, 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 cooperate 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
lent]

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 mlcrocry 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 cooperating hospitals in Cancer Research. Products ere
prepared, bottled and packaged according to a randomisation schedule end
shipped to cooperating hospitals. The cooperating 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 preparations 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 Clinical 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 procurement 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. Iapropox 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
iaprovenents, which we feel are necessary.

- 5 -

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PUBLICATIONS, SPEECHES GIVEN, GR SIGNIFICAlflf ACCOMPLISHMENTS
BY PHARMACY DEPARTMENT PERSONNEL INCLUDE

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

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■•■ '

LJMff * ft

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

-9-

:T,: ANNUAL REPORT ( .-IIVITIES

CENTER

MEDICAL RECORD DEPARTMENT CC-1>6

JI'IVES:

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

:iinical
;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.

PSOGBESS MADE;

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
reports.

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
reports.

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.

1R PROBLEMS:

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
machine)-

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
Minor

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

PROPOSED ACTIVITIES DURING TEE K3XT CALENDAR YEAR

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
Registry.

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

l?i3_

222

.bients

patient

First Registration
Readmission

Ikjk
1826

33CO

1181
1713

289^

patient (First Registration)

1605

1588

falk-In

106

Emergency

REGISTERED PATIENTS

U613

5. Employee Registration

1727

1682

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

1951*

1955

1956

1957

lif52

1889

2I409

2894

1326

1875

2369

2865

6M15

96,1*05

109,7^3

120,131

53,188

90,625

96,879

116,931

89,278

134,066

158,301

182,679

176

300

329

7£

1^7

19^

2^5

139

I87

233

2?5".

3300

3275

11+0,151

142,394

188,31*0

384

278

269

: Department Fage 8
harged Inpatients by Institute and Georgraphic Origin, Calendar Year 1958
TKI NIAID NIAMD HHI NIRDB NIMH N333R Total. Patlentc

110

128

1*20

Central

**5

152

l Central

1*0

tic

65U

3>+5

206

610

387

105

2350

Central

130

1 Central

".torieo

1. S.

eots

893

1*09

321

888

556

160

3275

fliington, D.C. 146

215

rginia 171

La are included in reported figures under South. Atlantic Region

126

139

113

602

123

172

13^

810

163

11*

53*v

>rd Deparv

Page 9

^patient visits, Calendar Year 1958

.1953

l/oH

1955

1956

1957

1958

Janu2ry - Mexch

16W

2879

14-969

l»83fc

6282

•• June

2156

333^

5213

6195

6nk

•- September

262 "

27^2

U029

5184

5356

5921

jer - Decembe

IO85

2825

ktfk

5080

5052

616U

Total

13^7

9371

H»606

20Wt-6

211-87

21J-51*!

Calendar Year 1958
PUBLIC HEALTH SERVICE NATIONAL INSTITUTES OF HEALTH

SUMMARY ANNUAL REPORT OF PROGRAM ACTIVITIES
CLINICAL CENTER

NURSING DEPARTMENT CC-51

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. Development and Trends

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

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. Progress 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. Major 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
services.

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
problem.

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

PUBLIC HEALTH SERVICE NATIONAL INSTITUTES OF HEALTH

3MMARY ANNUAL REPORT OF PROGRAM ACTIVITIES
FINICAL CENTER

rrRITION DEPARTMENT CC-6l

Serial No.

I CONTRIBUTIONS TO THE CLINICAL RESEARCH PROGRAM

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
intakes.

(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
coagulation.

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
studies.

(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
loss.

.'- /

- ■- 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 many patients simultane-
cisly.

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
reitors.

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.

E. MAJOR PROBLEMS ENCOUNTERED

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
srgery.

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» we 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.

:vJurition 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

1951

1958

J auary
Druary
rch

Aril

Jne

Jly

^gust

Eptember

C tober

Ivember

Icember

-rerage 115-4

Reflects inventory adjustment.

Clinical Center

Clinical Cente

Price Index

Ration Cost

Price Index
118.2$

Ration Cost

112. 8#

$1.42

$1.46

113-6

1.38

118.7

1.47

113.2

1.38

120.8

1.50

113.8

1.U6

121.6

1-57

Ilk. 6

1.50

121.6

1.58

116.2

1.62

121.6

*l.6o

117. h

1-51

121.7

1.63

117-9

l.*8

120.7

1.51

117.0

1-5U

120.3

1-57

n6.k

1.1+2

1.61

116.0

1.U2

116.1

1-52

1.47

(120.6)

(1-5^)

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.

CI. CHANGES AND IMPROVEMENTS MADE

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
staffs.

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
areas.

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

I. FUTURE OBJECTIVES

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 lSOOn3, 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-
scopy

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 con4,.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,
1958.

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
NIAMD and NCI)

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. BrecEer

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

Index

SUWjrtAK.

,rt oi; AraxKisiaariC/ri

KIDR No.

Project Title

Investigator

S. J. Kreshover

Prenatal Influences on Fetal
Development.

LABORATORY OF HISTOLOGY AK9 PATHOLOGY
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

Tissues.

8 P. H. Keyes Experimental Animal Carles.

PHS-N1B

Individual Project Reports

Calendar Year 1958

Index

LABORATORY OF MICROBIOLOGY

1UM SS» Investigator

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

10 S. E. Mergenhagen

11 H. W. ocherp &

S. E. Mergenhagen

W. Scherp

.i.

Howe)

Howell, Jr.

V. Jordan &

J. Fltzgera:

Shiota

R. Onata

6. Haopp

A. Kevin &

Go Hastpp

Project Title

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

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
Bacteria.

Studies on the Microorganisms As-
sociated with Periodontal Diseases,

Investigations In the Field of the
Spirochetes.

Studies of Interactions Between Oral
Microorganisms.

7IS-NIH

Individual Project Reports

Calendar Year 1958

Index

LABORATORY OF MICROBIOLOGY (Continued)
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.

LABORATORY OP BIOCHEMISTRY

24
25

Investigator
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
Disease.

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.

PHS-NIH

Individual Project Reports

Calendar Year 1958

Index

LABORATORY OF BIOCHEMISTRY (Continued)

NIDR No.
29

Investigator
R. H. Larson

Proiect Title

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

Zipkin

S. Levis

Blumenthal

34
35

R. C. Likins

S. A. Narrod

Biochemistry & Physiology of the
Salivary Secretions.

Non-dental Physiological Effects of
Fluoride.

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
Calcification.

Enzymological Studies in Relation to
Oral Tissue & Periodontal Disease.

EPIDEMIOLOGY AMD BIOMETRY BRANCH

NIDR No.
36

37
38

Investigator
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-
ship.

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

PHS-WIH

Individual Project Reports

Calendar Year 1958

Index

EPIDEMIOLOGY AND BIOMETRY BRANCH (Continued)

nidr No.

Investigator

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
Disease.

Relation Between Dental Caries and
Periodontal Disease.

NIDR No,
43-C

44-C

45-C
46-C
47-C
48-C

49 -C
50-C
51-C

CLINICAL INVESTIGATIONS BRANCH
Investigator

C. J, Witkop

C, J, Witkop

C. J. Witlcop

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.

PHS-NIH

Individual Project Reports

Calendar Year 1958

Index

CLINICAL INVESTIGATIONS
NIDk No. Investigator
A. D. Merrit-:

52-C

53-C

56-C
57-C

58 -C

65-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
Children.

General Anesthesia (Dental) Baseline
Data.

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

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
Occlusion.

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

Sumcf.ry - Research

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

Total

Direct

Reimbursements

$1/414,000

1,700

+32,300

NAT; _ .RESEARCH

SUMMARY STATEMENT

January-December 1958

aural Activities

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 (

Litative
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-
accrr^

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
nvesti-
£ regular fell jere na.de 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
liable.

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-
n.al factors in oral dices:
case oriented studies in the biophysical and t
i.cal 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; r.to-

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
radiocalclum.

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(PGc
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:

Hunan 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

I
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

jsly
been an found ir %noth'r inbred group

he succes
demo for the prediction and detection of

ills.

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

tion of , closely associated

tosoual

! 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 ■ironic Stomatitis. Recurrent aphthous stomatitis
painful disease of the oral mucous neubrar.es, 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 Facial Grew-h end Development. 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.

Dental 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

RATIONAL INSTITUTE OP DENTAL RESEARCH

R2K)RT ON ADMINISTRATION

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 aia.il 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 progr.no 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.

1-26-59

HATIOHAL IMSTITUTE OF DEMTAL RESEARCH

Individual Project Reports
Calendar Year 1958

OFFICE OP ASSOCIATE DIRECTOR:

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

Ottiem •£ A— odf r.i?Gc(;ar

Estimated Obllftat.'onc for F.Y. 1959

Total

Direct

Reimbursements

Project Kuabers;
KIDR-1

$63,800
$43,200
$20,600

Serial So. HIDR-1

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

PHS-NIH

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
malformations.

Methods 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
trimester.

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
occurred.

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

HATIOHAL INSTITUTE OF DEKTAL RESEARCH

Individual Project Reports
Calendar Year 1958

HISTOLOGY & PATHOLOGY;

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 &

Estimated Obligations for

Total

Direct

Reimbursements

$23!
$15<

Project Numbers:

»ir:-

HIDR-5

MDR-6

:< - 7

HIDE -8

[JCDR-2
lo Histology & Pathology
2c Bethesda, Kd„

PHS-
IndiviJual 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:

Objectives

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

b0 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
axes.

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 Research;

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
bOMt

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

Icctron

jponcnt of Dental Enamel.

Pre / (accepted

chlo projGc; :

Serial No„ NIDR-3

1. Histology & Pathology

2, Bechesda, Md.

PHS-HIH

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
copy.

Principal Investigator: Drc 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. Tomes1 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 Nos 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 0 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

PHS-HIH

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.

PHS-NTH

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

bona*

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
acid.

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-
trix.

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»
genesis.

Part B. included: Yes

- 4 - Serial No, HIPR-4

ras-iiB

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:
Hone

Serial No. KjgR°5

1. Histology & Pathc?

2. Bethesda, Hdc

PBS-NIH

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 Findings:

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

PHS-NIH

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:
None

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

PHS-NIH

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
Other:

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 Employed:

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
tissues.

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 No0 N1DR-6

PHS-NIH

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:
None

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

PHS-NIH

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,e0 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 0 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 No0 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 0 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
fibers*

Part Bo included; Yes

Serial No. NIDR-7

PHS-NIH

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:
None

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

PHS-NIH
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
process.

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-
culty,.

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
factors.

Significance to Dental Research;

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

PHS-NIH

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.
Res.)

Honors and Awards relating to this project:
None

KATIONAL INSTITUTE OF DENIAL RESEARCH

Individual Project Reports
Calendar Year 1958

MICROBIOLOGY:

<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. Scherp 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
Leptotrlchla.

15.

H. V. Jordan and R. J. Fitzgerald: Inhibitors and Dental
Carles.

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. Nevin 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

Total

Direct

Rain bur 5 assents

Project Numbers
SIDE -9
HIDR-10
NIDR-11
HIDR 11
NIDR-13
HIDR 14
HIDP
HID?
HIDR- 17
'•--18
NIDR-19
HIDR -20
NIDR-21
HIDR -22
NIDR 23

$292,200
$198,500
$ 93,700

Serial No, NIDR-9
1. Microbiology
3, Bethesda, Md

PHS-N1H
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
Dame

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
disease.

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

PHS-NIH
Individual Project Report
Calendar Year 1958

Part B°. Honors, Awards, and Publications

Publications otber than abstracts from this project:

None

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.

PHS-NIH

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.

PHS-NIH
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
mlcrobiota.

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 peats! 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.

PHS-NIH
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
cells.

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"
physiology.

Proposed 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

3J Betheade, Md,.

PHS-NIH
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:

10 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.

PHS-NIH
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
publication.

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

PBS-NIH
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:
None

Serial no. NIDR=15

lo Microbiology
3 Bethesda, Mdc

PHS-NIH
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

PHS-HIH
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-

None,,

Serial No. NIDR°16

lo Microbiology
3. Bethesda, Mdo

PHS-NIH
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

PHS-MIH
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'IM- »•• *■ P» Vitro Chemical .nd
Bacterial Changs. i„ ^liva. j. D.nt7 rST~37: 780. 1958.

Honor* and Awrdi relating to this project:

None

Serial no. NIDR-17

1. Microbiology
3. Bethesda, Md.

PHS-NIH
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

PHS-NIH
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:

None

Serial No. NIDR-18
lo Microbiology
3. Bethesde, Md,

PHS-NIH
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
sources.

- 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

PHS-NIH
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

nana

Serial No, N1DR-19

1 „ Microbiology
3. Bethesda, Md

PHS-NIH
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
route.

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^'foi0!!^1/6",10"8 ""P10*1** hematoxylin and eosin
stains for studying the progress of lesions.

L.™!™^^1"108 °£ •nlMl tl68ue8 to demonstrate and
determine the location of ,he spirochete, in tissue sections,

Major Findings:

It was found that pure cultures of the small oral trenonemeB
fn .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 ht0 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 toe4odr.8::18nL>have been recovered *&&&*£ i.: s

w?^ tyl 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^ni86r*ted ^T1^" ln Sundance in lesion.
d?7i f ^ hOUr" t0 4 d*ys of du«tion. The organisms

tat ««Lr:; T iD :hC aubeP^h*^«l connective tfssue
.««< ! COn!ln«d to the inoculation sites. Hematoxylin and
Jnf L^rl°ed 8eCti°n8 exhibited vnrlou. gradations^ .cute
«.!Jr^ yure8P0n" includi*S abscess formation. Preliminary
studies are being conducted in rabbits employing various »rm2

ijfh ;he°ILnd * *«—«**»» diphth^oiS iS co^ination
with the small oral treponeme. Large spreading lesions are

or'th^se •!*£" °ir1^ 8tUdlC8 - "»- -"ions
r cnxs pb**e of tQe problem have not been completed.

Significant to Dental Research :

le.i^i8 i8 5he/ir8t P°»itive evidence that reproducible
.Piroc8Jan beA1lni£i*ted bv 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
organisms-

2. To investigate the effect of spreading factors, such as
hyaluronide.se 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 fhi effects of the spirochetes on the various fixed
tissue elements of the rabbit sk;

6 To Investigate oral organisms that may act as aymfolo1
organisms in the production of experimental lesions in rabbits

Part 8 included Yes

- 4 Serial No„ NIDR-19

PHS-NIH
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:
None

Serial No. MIPR-20
1. Microbiology
3. Bethee<3a, Hd,

PHS-NIH
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 spirillac
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!

Serial

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 vincentii 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««s0 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 spirochetes4 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 Ireponemataceae has been
completely satisfied by cocarbosylase, coenzyme A3 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, Nr.DR-21
1. Microbiology

3. Bevhesca, Md»

PKS-NIH
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, dfc, 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
resistant.

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
chemicals.

- 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

PHS-NIH
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;
Mone

Serial no. NIDR-22
lo Microbiology
3. Bethesda, Md,

PHS-NIH
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
procedures.

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

PHS-NIH
Individual Project Report
Calendar Year 1958

Pert B; Honors, Awards, and Publications

Publications other than abatracta from this project:

Rogosa, M,, Fitzgerald, R. j0 , 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.

PHS-NIH
individual Project Report
Calendar year 1958

Part A

Project Title; The Rat as An Experimental Animal for Research
on periodontal DiseaseQ

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
flow.

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 no0 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
diet.

2. oral examination of live rata., 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
undertaken.

Part B included No

NATIONAL DESTITUTE OF DEHTAL RESEARCH

Individual Project Reports
Calendar Tear 1958

BIOCHEMISTRY:

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-
tions.

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-
cation.

35. S. A. Narrod: Enzysao logical Studies in Relation to Oral Tissue
and Periodontal Disease.

005 '16 $
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1, Biochemist
3. Bethesda.

MIH
^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 ) :
2%

•on:

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.

dinfis:

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
techniques.

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

PHS-NIH
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.

PHS-NIH
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 8rouP 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^^rr0'610^' "^ (dermU>» tendon» b^s,
thf« ?<« <*•»"»>• It may play a role in the function of

in! r%5 T8 a?d lQ P«»»»lo«ic«l ".tes which involve them
studL^f T<* Carl68 ^ Perio^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

cor8i?*J,fltUKle8- SPecifi""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

PHS-N1H

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„

FHS-NIH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: Nutrition and Dental CarleB with Smphasis on

Protein and Cereal Foods. Lysine, and Phosphate
Supplements.

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

Part 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.

Significance 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.

Proposed Course of Project ;

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
pl.ir.ning stage.

Part B included Yes

- 3 - Serial No. NIDR-26

PHS-NIH

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,

PHS-NIH
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
carboxypeptid7.se 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 Dental 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
enzyme.

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

PHS-NIH

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.

PHS-NIH

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 in 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
ttudies.

(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

PHS-NIH

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,

:
ntS.

: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 Research:

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.

jabpro ject B:

Objectives;

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 Research:

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
experiments.

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

PHS-NIH
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

iochemlstry
3. Bethesda, Md.

FHS-NIH

Individual Project Report

Calendar Year 1958

Part A.

Project Title: Biochemistry and Physiology of the Salivary
Secretions.

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.

PHS-NIH

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:

Objectlvej:

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.

Significance 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

PHS-NIH

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,
1958.

Serial No. NIDR-3?.
1. Biochemistry
3. Bethesda, Md„

PHS-NIH

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:
Objectives:

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.

Significance to Dental 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
conditions.

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.

PHS-NIH

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):
Total:

Professional
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-
selves.

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 hc.ve evidence that rat submaxillary

glands have little or no galactokinase activity, but do
ihow glucokiiase and ATPaee.

Significance to Dantal 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

PHS-NIH
Individual Project Report
Calendar Year 1958

Serial No. NIDR-34
7 , Biochemistry
3. Bethesda, Hd.

Part A

Project Title: Biochemical and Biophysical Studies of
Calcification.

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
strontium.

Methods Employed:

(a) Synthetic hydroxyapatites labelled with Ca*5 and Sr89
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 in 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 Sr89 in the formation of hydroxyapat ite from a solution
of labelled elements. The ratio of Ce^.'Sr89 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
1957).

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 tn 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

PHS-NIH

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.

PRS-HIH

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. &
Metabolism)

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.

Major Findings:

(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 histelytlcum
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 re.uct.cn.

Signiflcnr.ee 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
plcnned-

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.

NATIONAL INSTITUTE OP DENTAL RESEARCH

Individual Project Reports
Calendar Year 1958

EPIDEMIOLOGY & BIOMETRY BRAKCH:

36. A. L. Russell: Epidemiology of Oral Diseases Among Alaskan
Natives.

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
Disease.

atology & B

Estimated Obli^atlopa for FY. 1959

Direct

R e iabur s eaen t s

$130,000
$ SI

$ Ul

ct Numbers:

HIDR-36

KIOR-37

HIDR-38

MIDR-39

KIvR-40

HIOR-41

RX»R-£2

»1 No. S3
>gy & Bis

PHS-BXH
Individual Project R^
Calendar Year 1958

Proje gy of Oral Di3ea: es Among Alaskan

/es.

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:

PtOj

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 Enployed:

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
people.

Part B included: No

Serial Ho. HIIB^S?
1. Epidemiology & Biosietry
3. Be :hesda, Md.

PHS-HIH \
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 ,^ators: 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
Professional:
OCher:

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
caries.

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 Iteployed ;

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
supplies.

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, 60 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.

Significance 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
en.^i

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.

PHS-MH

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
•gca.

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.

Siail.tr data were collected on 577 male persons 11 - 17

s of age In this country for comparison with the India
idy

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';
areas.

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.

PHS-N2H

Indivi.d1 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:

Objectives:

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
children.

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.

Significance 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
severity.

Proposed Course 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.

JHS-NIE

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:

Objectives:

::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 Employed :

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.

Significance to Dental Research:

Studies among the Ethiopian natives allow an investigation
of oral diseases in groups showing varying degrees of
priniiiiveness.

;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
..ni India.

Part B included: Bo

al No, HIDR-41
1« Epidemiology & Biometry
3. Betheada, Md.

PHS-NIH

Individual Project Report
Calendar Year 1958

Part 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

Professional:

ot;. l i

Project ton:
:

To compile a descriptive epidemiology of the group of
ise entities commonly called "pyorrhea."

Methods Employed:

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, WIDE

- 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.

Significance to Dental Research:

After dental carles, the periodontal diseases are the
:nd most iaportant cause of tooth loss, and affect

7 0 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 Fro \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-
tinued.

Part B included: Ho

1. Epidemiology & Biometry
3o Be this da, Hd<

7HS-IXH

Individual Project Report

Calendar Tear 1953

Part A.

Project Title: Relation Between Dental Cables and Periodontal
Disease

Prinelpii Investigator: Mr. C. L. White

Other Investigators: Dr. A. L. Russell

Cooperating Units: 3ooe

Man Tears (calendar year 1958)
Tot.

Professional: 73

Other: 2 3

Project Description:

Objectives:

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 readings :

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.

Significance 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 -
Fropcised Course of Project:

ontinue and expand the outlined study.

Part B included: Ho

ITUTB OF DEWTAL RESEARCH

Individual -.eporta
Calendar Year L958

CLINICAL INVESTIGATIONS:

43-C. C. Jt Wltkop: 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 Steph"'

virooment in Relation to Dental t,
Dli • -

k. M. Stephan: Interrela'. Between

in the Etiology of Denta;
Rj:

'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
■.see-

56-Co I, I. Ship; Stress in the Dental Pati-

V, S Baer; Esterase Activity Associated with th?
--posits on Teeth.

RATIONAL INSTITUTE OF DENTAL RESEARCH

Individual Project Reports
Calendar Year 1958

CLIWICAL INVESTIGATIONS (Continued) :

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
Prosthesis.

63-C. R3 S. Lloyd: 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

Cit:i..cal 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.
PHS-NIH

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
isolates.

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
States.

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<^ Research,

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-
t

To continue to ccumulat- Information on the hereditary
diseases present in these Isolate populations.

Part B ircluded: fes

- 6 - Serial Wo. NIDK-43 (C)

PHS-HIH

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
report)

Honors aud Awards relating to this project: Done

Serial Ho. Bg»-44r.,,CC)

1. Clinical Ifiiveatigatii

2. Human Genet ice

3. Bethesda, Maryland

FBS-HIB

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
dentin.

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
planned.

Part B Included: Yes

- 5 - Serial Ho. HIDR-44 (C)

EBS-MXB

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
PHS-HIH

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:

Objectives:

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
illness.

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
selection.

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
material.

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-
tained.

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 Research;

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
defects.

Proposed 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)

PHS-NIH

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

PRS-NIH

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; i0e„ 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
line.

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. «

ll!

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<

fhs-nth

Individual Project Report
Calendar Year 1958

£art_A.

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
Delaware.

Part B included: No

PHS-NIH

Individual Project Report

Calendar Tear 1958

Serial Hc0 KEDH-

lo Clij&cai^nvestigf

20 Human Genetics

3o Betheeda9 Maryland

Part A

Project Title: Sickle Cell Anemias and Other Inherited
Hematological Disorders

Principal Investigator: Dre D* L« Rucknagel

Other Investigators: Dr& C« Jo Witkop, Jre

Cooperating Units: Drub Jo EQ Seegmiller, Bo LaDu, Lc Laster9
NIAMD

Man Years (calendar year 1958): Patient Bays: 730
Total: 1 2/3
Professional: 2/3
Other: 1

Project Descriptions

Objectives:

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 dieease0

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 gene0
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 Noc

Part A (continued)

Relatives of propositi are studied hematologically using
standard hematologic t£chniques9 paper and starch block
electrophoresis and0 when indicated, oxygenation kino
studieBo

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 Drse 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 al-.io 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 oxygenatr.cn investigations have been perform* <
eighteen individuals with sickle .jell anemia0 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 time0
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 se9 and not with associated
systemic disease9 i0 e08 pulmonary insufficiency^
cardiac anomalies? etc0

The patient with pernicious anemia was found to excrete
large amounts of formimi.no glutamic acids 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, amethopterin0 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 anemia0

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 severity0 However 0
preliminary tr'.als on a small numbeT* of sickle cell
anemia patients have shown no significant results to
dateQ

Significance 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
mechanisms*

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
investigation.

2) Formyl metabolism in pernicious anemiae

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.
PHS-HIH

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.
Spuhler.

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
Tti_V0TV °*children "*><> ^ve moved fro* the two study
Ultl f 11£5f0rt 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 blring 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?tJ,f?d? Carfied 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<Jutry- ***** 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>&ya
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.

Proposed 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.

ras-m

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
Diseases.

Principal Investigator: Dr. R. M. Stephan

Other Investigators: None

Cooperating Units: Dental Clinic, Clinical Center, NIH

Years:

Total :

1 1/3

Professional:

1/3

Other:

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,
RHXI.

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 Dental 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 situ1' 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
cases.

Part B included: Ko

Serial No. NIDR-51 (C)
1. Clinical Investigations
3. Bethesda, Hd.
PHS-NIH
Individual Project Report
Calendar Year 1958

Part A„

Project Title: Interrelationships between Local and Systemic
Factors in the Etiology of Dental Diseases in
Rats.

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
substance.

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 of 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 No0 jSnibSLifil
I. Clinical Investigations
3. Betaesda, Maryland
PHS-HIH
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
disease.

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.

PHS-NIH

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

jit

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
categories.

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
palate.

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
nation.

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
speech.

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„
PBS-HXH
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
surgery,

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
barbiturates.

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)
activity.

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) .

,11*

- 3 - Serial Bo. Mg^JliJLSl

Part 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<>

flU

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 to 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
research.

•'i

- 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

ras-Bia

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:

Objectives :

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-
tions.

- 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-
raphy.

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
diseases.

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
symptoms.

4. Double-blind clinical trial of agents showing
promise for prophylactic administration to prevent re-
currences.

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
available.

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-
sults.

- 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

PHS-NIH

Individual Project Report

Calendar Year 1958

Serial No. NIDR-56 (C)

1. Clinical Investigations

3o Bethesda, Md.

Part A.

c
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
surgery.

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
visits.

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
cortisone.

Part B included No «J)

;;;>

Serial Ho. HIDR-57 (C)
1. Clinical Investigations
3. Bethesda, M,
PHS-HIH
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
activity.

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

prs-hih

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.
Sokoloff.

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}

Ptrt A (cor.fciausd)

(b) maxillary width (from first molar to first molar),

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/HS
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 of Project:

To continue the study of genetic influences on periodontal
disease.

Part B included: Yes

- 4

Serial Ko. HZBR-58 (C)

PES-HIH

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$ -

-•jrt

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
1:
1

Lou:

Objectives;

1. To det amine whetl.er apical nigr&ticn of periodontal
tissues occurs with age if annuals are kept under

conditions.

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
established.

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.

EBS-HH

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

PHS-N1H

Individual Project Report

Calendar Year 1958

Serial No0 NIDR~60 (C)

lo Clinical Investigations

3o Betheada, Maryland

Part A

Project Title: Histopathology of the Human Pulp

Principal Investigator: Dr0 He Rc Stanley, Jr0

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

2C To further evaluate the critioalness of the remaining
dentin thickness beneath cavity preparations in order to
compare pulp studies of other investigators0

3o To determine the mitotic frequency of odontoblasts and
all other less differentiated cells of the human pulp0

kD 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 pulp0

60 To determine what characteristics of pulp pathology might
be the result of pressure changes per se when frictional
heat is not presents

- 2 - Serial No0 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

8a 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 eosin0 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 predentin0 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 techniques9
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 predentin0 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,,

40 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«0) 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 turbines0 ""

Conventional speeds only utilize about 25 ec/ndno as '*"

compared to the turbines using close to 100 or more cc/mino ,„,

60 Apparently certa:in characteristics of the pulp are \Z\

not due to pathology but autolysis, as, for example, the '»

well-known characteristic of reticular atrophy0 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 examinatlon0

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 No0 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 studies0

20 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 studies0

3a Various investigators have suggested that dental caries
does not necessarily provoke severe alteration of the pulp0
Therefore, to leave behind some decay in the cavity prepare^
tion is preferred tc exposing the pulp0 A survey of
carious teeth is needed to determine the veracity of such
a suggest ion0

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.

20 We have accumulated over 500 human teeth, all of which
were prepared under local anesthesia0 All the maxillary
teeth were prepared under local infiltration and all the
mandibular teeth by block anesthesia0 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 Ko0 NHJR-60 (C)

PES.NIH
Individual Project Report
Calendar Tear 1958

Part Bs Honors, Awards, and Publications

Publications other than abstracts from this projects

Swerdlow, H,, and Stanley, Hs9 Jr0 Reaction of the human
dental pulp to cavity preparation, I, Effect of water
spray at 20,000 rpm, J,A0D0A, j>6: 317=329 Mar, 1958o

Stanley, H«, Jr0, and Swerdlow, Hc Aspiration of cells
into dentinal tubules, JQ Oral Surge, Oral Hed09 and
Oral Path,, 11:1007-1017 Sept, 1958.

Swerdlow, He . and Stanley, H0 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) 0 (In manuscript form for
the J0A0D0A0)

Honors and Awards Relating to this Project:
None

Serial Bo.

1. Clinical Gentei?

2. Dental Separtssat

3. Betheada, Md.

*as-KiB

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
prognosis.

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 r.p.su 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 Findings :

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

PHS-HIH

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-62mi {p
lo Clinical Center
2o Dental Depsrtasafc
3c Beehesda, m0
FHS°HIB
Individual Project Report
Calendar Tear 1958

Part A

Project Titles Cephaloaetric Evaluation in Full Denture Prosthesis

Principal Investigator : Dr0 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;

Objectives:

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
area.

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.

Significance 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»
PHS-HIH

Individual Project Report
Calendar Year 1958

Part A

Project Title: Maxillofacial Prosthetics

Principal Investigetor : Dr. R. S. Lloyd

Other Investigators: Ko.ie

Cooperating Units: Hone

Man Years (calendar year 1958): Patient Visits: 120

Total :

1/4

Professional:

1/4

Other

Hone

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-
structed.

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 So0 JEBLJ&JSQ,

1. Clinical Center

2. Dental Department

3. Bethesda, Md.

FHS-NIH

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

Total

Professional

Other

1/5 Patients : 31

1/5

None

Project Description:
Objectives:

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.

Eg.

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<iCer 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 **«

tnee filter whtel. Zach ex^auTe is lade through a different
filter Siintaneously^ough a separate optical system,
nicer. ai«" n~1nit>A acoss the upper margin of -=acti

a "gray scale" is PLinted "*®!!^ scaie" 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
pathology.

Significance 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 Ne0 NIDR--

Clinical Investig?.
3o Bethesda9 Maryl&rsd
PHS.NIH
Individual Project Report
Calendar Yea? 1958

Part A

Project Title: Twelve-Mcnth Changes in Claas II

with and without Occipital Headgear Thejrapy

Principal Investigator: Dr0 DD 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 therapy0

Methods

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 Findings:

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©n0 Moat
cases showed a marked distal position of the maxillary
and unerupted tooth budSo

Serial K©0 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 height0 The dentition
in the control group moved forward more than the upper
face0

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©nse0

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
studies9 the occipital headgear appears to be the most
effective appliance for moving upper teeth distally0

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 Dr0 P©alt«nss
resignation from the service, September,, 1958« when he left
to take a position in California,, Dr0 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

phs

Individual Project Report
Calendar Year 1958

Part Bt HoncrsB Awards, and Publications

Publications other than abstracts frcm this project:

Foultcng Do Ro» and Pruianaky, S0 Report of a case with
supernumerary teeth and treatment involving extraction of
three upper iniisora0 Jo of Dentistry for Children,,
212-2J4 ?rd Quarter9 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:
None0

/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 announced 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
work.

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
protection.

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.

Iff*"
,L

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
organisation.

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, M2ES 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
building.

DIVISION OF BIOLOGICS STANDARDS

Control Activities

Laboratory

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

PHS-NIH

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~
gens*

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
made.

(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
standards.

(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 Division 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

2.
3,
PHS-NIH
Individual Project Report
Calendar Year 1958

Serial No, DBS_;_3_

Part A.

Control Activities
Control Tests
Bethesda

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, MCS USA

Director, Division of Immunology
Walter Reed Army Institute of Research
Walter Reed Medical Center

Man Years
Total: 8.25
Professional:
Other: 6.25

2.00

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
Division,

In addition to the above* this project involves: (a)
research designed to improve existing in vivo and in vitro
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 Findings 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, 3S183 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
include:

- 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

0 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

PHS-NXH

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
product*

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 Hov DBS-.&

L Control Activities
2.. Reference Standards
3. Bethesda

PHS-NIH

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 standardizeds
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-
tion,,

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 Organise0
tiotto

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-&«£h0

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

Fibrinogen

H. Influenzae Antiserum

Histolytica Antitoxin

Hi&tolyticus Toxin

Histoplasmin

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)

Thrombin

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 /

DIVISION OF BIOLOGICS STANDARDS

Bacterial Products

Laboratory

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

PHS-KIH
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

PHS-NIH
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

PHS-NIH
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

PHS-NIH
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

PHS-NIH
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

PHS-NIH
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

PHS-NIH

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.)

Significance, 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

PHS-NIH

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

PHS-NIH

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
freexe-drying.

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
products.

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

PHS-NIH

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

PHS-NIH

Individual Project Report

Calendar Year 1958

Part A.

Project Title: Re-evaluation of Requirements for Cholera
Vaccine.

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
protection.

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.

PHS-NIH

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
antitoxin.

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
investigated.

Part B included Yes 7 No /X~7

Serial Ho. DBS°1A

1 . Bacterial Products

2. Eaeoophilus Sfcu:

3. Betheada

Pare A

PHS-NIH

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

DIVISION OF BIOLOGICS STANDARDS

Viral Products

Laboratory

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

PHS-N1H

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
culture.

(

(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:

Bottlea

Monkey Kidney
Rabbit Kidney
Dog Kidney

. : akey
Rabbit
Dog

vs..*.

79,666

6,206

500

Processed
2,589
304

2.jpz. 8 os.
87,999
13,909
135

Total Vol . of packs c

Gel la Prepared

5,229 cc.
981 cc.
11 .5 cc.
9.75 cc.

OS.

4,410

Vol.

Cells

Prepared

for Othe

571

cc.

.5 cc.

January 1, 1958 - December 1, 1958

•Stable Cell Culture Preparations:

Bottles

Tubes

HeLa

18734

Monkey heart

4380

868

LAC

7766

Drew RK

1894

MK-Stable

6336

Parker RK

624

III

285

Hunan Aanlon

3329

Monkey Muscle

150

FRK

Hep II

6698

S-18*

Human Heart

2 oz.

8 oz.

8896

504

658

138

2314

150

1886

32 oz,
436
3

121

254

* Human breast carcinosis from Dr. Baron.

Serial No. DBS-lg

1. Laboratory of Viral Product!

2. Section on Testing

3. Bethesda

PHS-N1H

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
vaccines.

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.

Significance 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.

Significance 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

PHS-HIH

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

PHS-HIH

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.

Objectives : 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 Divisions 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.

Significance 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

PHS-KIE

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
antibodies.

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

PHS-N1H

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 Vaccine

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 Institutes 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

PHS-NIH

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

PHS-NIH

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

PHS-HIH

Individual Project Report

Calendar Tear 1958

Part A.

Project Title: Studies on the Monkey Beuroviruleace of Attenuated
Poliovlruses.

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

PHS-HIH

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 -

Proposed 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

PHS-MIH

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.

Major 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

PHS-NIH

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
follow:

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
3P 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
available.

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

PHS-NIH

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 Findings*

1 . Relative, susceptibility of animal^ viruses. 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 fcands 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 >iii^ctivaiti<ioni of, coliphagesjpy
photodynamlc 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 phage9 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 viruses^ byni the , phofcpdyaamic i action of various dyes ,

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»
ation8 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 formal.de*
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 Tof Project.

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

PHS-NIH

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
Vac cine 8 latter part of 1957 in 1958 in 1953

Monovalent

(Asian strain) 1 5 6

Polyvalent military

(six strains) 0 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 projects 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
strains.

(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
vaccines.

* 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 work 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
test.

(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

vaccines.

(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
study.

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 Findings : 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

PHS-KIH

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„
1958.

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

PHS-NIH

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

^on^^
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

PHS-NIH

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

PHS-NIH

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

PHS-NIH

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
teats.

Part B included Yes / 7 No /x*7

Serial Wo. DBS -33

1. Laboratory of Viral Products

2. Eesearch Section

3. Bethesda

PHS-NIH

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

PHS-NIH

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
poliovirus.
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.

Objective 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.

Objectives 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 and5 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

PKS-HIH

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

PHS-NIH

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

PHS-NIH

Individual Project Report

Calendar Year 1958

Part A.

Project Title: Standardization of Measles and Mumps
Neutralizing Antibody in Human Gamma
Globulin.

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

PHS-NIH

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
soon.

(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 Programs
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

DIVISION OF BIOLOGICS STANDARDS

Blood & Blood Products

Laboratory

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

Products
2„ Blood Bank
3, Bethesda

PHS-NIH

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 the 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 iiofi 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

Products

2. Blood Bank

3. Bethesda

PHS-NIH

Individual Project Report

Calendar Year 1958

Part A.

Project Title: Stability and Potency of Formed Blood
Elements.

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 RhQ variant (Du) 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 Findings 1. Evidence which is still prelimi-
nary indicates that it may be easier to detect Sh0 variant
(Du) 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 0 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
evaluation.

Proposed Course of Project; 1. Many additional
studies of the laboratory reactions and the transfusion
effects of RhQ variant (Du) 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,

PHS-NIH
Individual Project Report
Calendar Year 1958

Part 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», Breslow8 A., Ebaugh, F. 6.9 Jr„, Schmidts
P.J,, and Holly, P0: 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. Ja, 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

Products
2„ Blood & Blood Derivatives
3 . Bethesda

FHS-HIH

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
investigated.

!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
stockpile.

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 isolatedB and the effects on
them of a variety of physical and chemical treatments
evaluated,

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<

Products

2. Office of the Chi«f

3. Bethesda

PHS-NIH

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

SUMMARY STATEMENT

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„

GENERAL RESEARCH GRANTS BRANCH

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-
vatives.

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.)

RESEARCH TRAINING GRANTS BRANCH

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
researchers.

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.

Biochemistry

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
Institutes.

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
graduates.

Genetics

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 -

Pathology

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
training.

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.

Pharmacology

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.,

Physiology

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
1959.

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.

RESEARCH FELLOWSHIPS

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 -

CENTER FOR AGING RL SEARCH

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-
lows:

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.

Conferences

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
Councils.

NIH RESEARCH AND TRAINING GRANTS IN AGING
ACTIVE AS OP JANUARY 31, 1959

Institute

Number

General*

NCI

NHI

NIAID

NIAMD

NIDR

NIMH

NINDB

TOTAL

Primarily Related to Aging

PY 1953 FY 1959

197

Total

$ 18,192

532,231

$ 550,423

120,021

424,407

544,428

227,338

,034,709

1,262,597

20,350

10,010

30, 360

37,148

453,958

491,106

38,735

60,774

99,559

340,344

514,716

355,060

109.706

190.741

300,447

$912,434

.221,546

$4,133,980

Institute

Number

General*

NCI

NHI

NIAID

NIAMD

NIDR

NIMH

NINDB

Secondarily Related to Aging

FY 1958 PY 1959

Total

TOTAL

207

$ 32,000

$ 268,910

$ 300,910

130,556

263,353

443,909

441,079

1,192,136

1,633,265

24,333

81,558

106,391

50,344

523,042

578,386

143,867

152,760

301,627

43,499

53,397

96,896

17.239

144,508

161.747

$938,417

$2,684,714

$3,623,131

*DGMS

- 17 -

NIH RESEARCH AND TRAINING GRANTS IN AGING
ACTIVE AS OF JANUARY 31, 195S

Primarily Related to Aging
Institute Number FY 1955 FY 1957 FY 1953

General*

NCI

NHI

NIAID

None

NIAMD

NIDR

NIMH

NINDB

TOTAL

131

$23,686

$23,636

$ 12,253

40,353

133,700

41,179

29,821

162,972

37.751

249,419

99,991

747,329

132,434
30,122

440,641
79,215

Total

261,677
140,344
836,029

173,613

59,943

627,299

116,966

$463,534 $1,779,151 $2,266,371

Secondarily Related to Aging
Institute Number FY 1955 FY 1957 FY 1958

General*

NCI

NHI

NIAID

NIAMD

NIDR

NIMH

NINDB

Total

TOTAL

143

$122,262

31,951

$ 204,213

90,207

141,437

231,644

349,597

703,632

1,053,279

2,501

60,574

63,075

J2.131

192,470

274,651

121,426

143,872

265,293

33,293

35,706

74,004

34,035

83,304

117,389

$840,557

$1,443,496

$2,284,053

*DRG

- 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
ciences

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
Req.

No. of
Plates

No. of
Impressions

Average Copy
per Plate

Extramural

10,884

68,100

7,785,254

114

Other than

904

5,833

2,188,760

375

Extramural

Totals

11,788

100

73,933

100

9,974,014

100

135

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.

Legislation

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 utilizeds 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 0

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
disease.

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-
nation.

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
shellfish.

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
pathology.

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 0 N

0 F

RESEARCH SERVICES

Serial No„ __PRS-1

DIVISION OF RESEARCH SERVICES

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
Biometrics.

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
facilities.

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
unit.

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.

Prohlerns

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 ;..it.y 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
1. OFFICE OF THE CHIEF,

DIVISION OF RESEARCH

SERVICES

PHS-NIH
Calendar Year 1958

Budget Data;

Actual obligations for Calendar Year 1958:
Total: $103,467
Direct:
Reimbursement: Office of the Chief, DRS--$103,467

Serial No. DBS-1.2
1. OFFICE OF THE CHIEF,

DIVISION OF RESEARCH

SERVICES

PHS-N1H
Ca .endar Year 1958

Budget Data:

Actual obligations for Calendar Year 1958:
Total: $6,446,527
Direct:
Reimbursement: Total services, DRS — $6,446,467

Pebruary 1959 Calendar Year 1958

PUBLIC HBALTH SERVICE - NATIONAL INSTITUTES OP HEALTH
DIVISION OF RESEARCH SERVICES

Summary of Branch Activities 1. DEIS- 2

Serial Number

PLANT ENGINEERING BRANCH 3. R. R. Holliday

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;

■0 JAJTT

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-
ordination.

The above methods and procedures will be developed as soon as
possible.

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
$531,350.

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
electricians.

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

1. PLANT ENGINEERING BRANCH

PHS-NIH
Calendar Year 1958

Budget Data:

Actual obligations for calendar year 1958:
Total: $3,532,962

Direct:
Reimbursement: $3,532,962

February 1959 Calendar Year 1958

PUBLIC HEALTH SERVICE - NATIONAL INSTITUTES OF HEALTH
DIVISION OF RESEARCH SERVICES

Summary of Branch Activities lc DRS-3

Serial Number

2o SCIENTIFIC REPORTS BRANCH 3. Taft S. Feiman

Branch Branch Chief

Objectives

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.

Programs

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 NIHS
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-
leases.

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
6,767.

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-
liographies.

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
English.

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 »

Abstracts 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 0

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 Experiments 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 mounts3 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
defect.

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 0 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
service.

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-
lications.

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 HEW9
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
publication.

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 year0

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 So

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 Institutes 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.

Distribution 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 MH, 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 So ."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.

Calendar 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

Library

Medical Arts

Photography

Editorial

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*

Problems

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
candidates.

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 phase0
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-
brarians.

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<=
ends.

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

translation.

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.

Staffing 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

PHS-NIH

Individual Project Report

Calendar Year 1958

Part A.

Project Title: PLASTICS RESEARCH AMD DEVELOPMENT

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

PHS-NIH

Individual Project Report

Calendar Year 1958'

Part B. : Honors, Awards, and Publications

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. h9 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

PHS-WIH
Calendar Year 1958

Budget Data:

Actual obligations for calendar year 1958:
Total: 5661,927

Direct:
Reimbursement: $661,927

Serial No., DRS-^

lo Scientific H ports Branch
2o Russian Trar lation Program

PHS-NIH
Calendar lear 1958

Budget Data:

Actual obligations for calendar year 1958:
Total: 5328,555

Direct:
Reintoursement: $3 28,555

January 1959

Calendar Year 1958

PUBLIC HEALTH SERVICE - NATIONAL INSTITUTES OF HEALTH
DIVISION OF RESEARCH SERVICES

Summary of Branch Activities

2. LABORATORY AIDS BRANCH
Branch

1. DRS - 4.1

Serial Number

3. Dr. Willard H. Eyes tone
Branch Chief

Objectives

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

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
glassware.

The provision of types of culture media required by NIH
scientists.

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

Mice

724,100

794,000

Rats

164, 100

156,000

Guinea pigs

32,700

27,200

Rabbits

10,500

10,800

Hamsters and other

17.600

23,700

animals

949,000

1,011,700

Several areas have expressed an Interest in an inbred strain
of rabbit. It is anticipated that the 10,000, 0GQtij 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.

LABORATORY GLASSKARE

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.

ANIMAL HOSPITAL

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
organisations.

- 6 -
DRS, Laboratory Aids Branch, cont'd;

Workload for the year showed:

Conditioning, and Quarantining

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
increased.

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
year.

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:

COMPARATIVE PATHOLOGY

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
activities.

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:

Problems

ANIMAL PRODUCTION

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.

LABORATORY GLASSWARE

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.

ANIMAL HOSPITAL

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

PHS-NIH
Individual Project Report
Calendar Tear 1958

Part A.

Project Title: A STUDY OF THE GROSS AND HISTOPATHOLOGIC
CHARACTERISTICS OP DISEASES IN LABORATORY
DOMESTIC AND CAGED WILD ANIMALS

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), Pneumonyssus (lung mite of rhesus monkey), Pej&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
Section:

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 HIHff 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 Ho0 4.2

1 o Laboratofy~Aias Jsranch

PHS-HIH
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:
Hone

(Attachment 1)

Serial No. 4.3 ^

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda
PHS-NXH

Individual Project Report
Calendar Tear 1958

Part A.

Project Title: DISTEMPER. IN RACCOONS AND FOX SCSPECT89 0?
HAVING RABIES

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

PHS-HXH
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:

None

(Attachment 1)
Serial Bo. _4.4 __

1. Laboratory Mds Branch

2. Comparative Pathology Section

3. Bethesda

PHS-B2H
Individual Project Report
Calendar Tear 1958

Part A.

Project Title: IDIOPATHIC HECROSIS OP BONE IH LABORATORY KSGB

Principal Investigator: Dr. Leon Sokoloff, NIAHD

Other Investigator: Dr. Robert T. Habermann

Cooperating Units: NIAMD

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

PHS-NIH
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 tti.ee. A.MoA. Archives of
Pathology; 65; 323-330; 1958,

Honors and Awards relating to this project:

Hone

(Attachment 1)

Serial No. 4^5

1. Laboratory Aids Branch
2„ Animal Production Section
3. Bethesda

FHS-NIH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: BOUSING AND CAGING RESEARCH AND DEVELOPMENT

Principal Investigator: Samuel M. Polley

Other Investigators: None

Cooperating Units: None

Man-years (Calendar Tear 1958):
Total: .06
Professional: .06
Other:

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 _

2. Animal Production Section

3. Betbesda

PUS -NIB

Individual Project Report

Calender Tear 1958

Part A.

Project Title: RESEARCH IN ANIHAL HUSBANDRY PRACTICES AND
PROCEDURES

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
substances.

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

PUS -NTH

Individual Project Report

Calendar Tear 1958

Part A.

Project Title: BREEDING POR IMPAHTILB DIARRHEA DISEASE
RESISTANCE IN MICE

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

PHS-HXH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: SURGICAL LENGTHENING OF LONG BONES
IN THE DOG

Principal Investigator: William I0 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
suitable.

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 -
Part 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

PHS-HIH
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:
Hone

(Attachment 1)

Serial Ho„ 4,9

1. Laboratory Aids Branch

2. Animal Hospital Section
3o Bethesda

PHS-NIH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: COMPLETE SURGICAL HYPOPHYSECTOMjr
IN DOGS

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 Dr0 James 0. Davis, HHI, and Dr0
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

PHS-NIH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: CONTINUED STUDY IN CONTRAST RADIOGRAPHY

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

animals.

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

PHS-NIH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: COMPLETE REMOVAL OF THE PINEAL BODY
OF THE DOG

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
removed.

Part B included 3tes Ho x

(Attachment 1)

Serial Bo. 4.12

1. Laboratory Aids Branch

2. Animal Hospital Section

3. Bethesda

PHS-NIH
Individual Project Report
Calendar Year 1958

Part Ao

Project Title: SURVEY OP FELINE INFECTIOUS ANEMIA
IN THE NIH CAT COLONY

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

PHS-NTH
Individual Project Report
Calendar Year 1958

Part A,

Project Title: BREEDING POR RESPIRATORY DISEASE
RESISTANCE IS RATS

Principal Investigator: Samuel M. Poiley

Other Investigators: Bone

Cooperating Units: Staff, Comparative Pathology
Section

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
colony.,

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. 4014

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda

PHS-NIH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: IDENTIFICATION AND CONTROL OF HELMINTHS
IN LABORATORY ANIMALS

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 Description: 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

PHS-NIH
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:
Hone

(Attachment 1)

Serial Ho. 4.15

1. Laboratory Aids Branch

2. Comparative Pathology Section

3. Bethesda

PHS-NIH
Individual Project Report
Calendar Year 1958

Part Ao

Project Title: SALMONELLOSIS IN LABORATORY ANIMALS
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 Mo0 4.15

1. Laboratory Aids Branch

PHS-HIH
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:
Hone

(Attachment 2)

Serial Ho. 4.16

1. Laboratory Aids Branch

PHS-NIH
Calendar Year 1958

Budget Data;

Actual obligations for Calendar Year 1958:
Total: $1,032,587

Direct:
Reimbursement: $1,032,587

(Attachment 2)

Serial No. 4ol6

1. Laboratory Aid* Branch

2, Animal Production Section

PHS-NIH
Calendar Year 1958

Budget Data:

Actual obligations for Calendar Year 1958:
Total: $501,588

Direct: $501,588

Reimbursement :

February 1959 Calendar Tear

PUBLIC HEALTH SS&FICS - HaTSOKaL ZHSTZTDTSS 0? KBALSH
DX7I9Z0S GT BBSEaftCH 8BOTCES

Suoeary of Branch activities

Serial Buaber

*• BKHETBICS 3. Harold F. Dora

Branch Branch Chief

Qpftctfr

The object Ives of the B lose tries Branch are:

To provide expert bloom trie consultation concerning
the deelgne 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
terolnation.

Aeeeanlishaants

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 0 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, sesB
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 tecesmendation.

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 program
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 0
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
2.

3. Bethesda
PHSHHXH
Individual Project Report
Calendar Tear 1958

Fare A.

Project Title: CAHCE3. Of TOE DTESU7S 18 J3HISH AHD BGS-JSWISE

rmiuu

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
Other

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

PHS-HIH

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
Other:

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 Saployad: 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
2.

3. Bethesda
PHS-9IH
Individual Project Report
Calendar Tear 1958

art A.

Project Title: BX1ATWWSHIP OP THE OSS OP TOBACCO TO HEALTH

Principal Investigator: Harold P. Dorn, Chiefs Biometrics

Branch

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:

Objectives: 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
1.

3o Bethasda

Part B.: Honors, Awards, and Publications

Publications

Beer, Paul I. and Llebermaae Jacob B.s '^Observations on Seme genetic
Characteristics of the Ferlodontluea in Throe Strains of Inbred Kiea9"
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. Radiation, 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 0
May 1958.

Born, Harold P., "Darwin Revisited," Busanlcs Quarterly 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

JUS-HIE
Calendar Year 1958

Budget Data;

Obligations for Calendar Year 1956;
Total: $551,155
Direct :
Reiabursenent: $551,155

Serial Bfo. BRS-5. 5im ,
1. ioael ss B2

February 1959 Calendar Year 1958

PUBLIC HEALTH SERVICE - HATIOHAL 3BSTTTUTES OF HEALTH
DIVISION CF RESEARCH SERVICES

Summary of Branch Activities 10 DRS-6

Serial Humber

2o SANITARY EHGIHEERIHG BRAHCH

Branch Branch Chief

Objectives

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 0 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

Programs

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 0

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 examined 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
$6,000,

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
buildings.

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
specifications

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 Utilisations

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 Ho0 d

lo Sanitary Engineering Branch
2o Equipment Testing and

Quality Control Section
3» Bethesda lks Maryland

PH5 — NIH
Individual Project Report
Calendar Year 1958

Part A<

Project Title: EVALUATION QF GAS STERILIZATION EQUIPtEHT

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

Ac EVALUATION OP GAS STEBILIZATICH EQUIPMENT
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 1209Fo 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 ,,___

10 Sanitary Engineering Branch
2<, Equipment Testing and

Quality Control Section
3» Bethesda 1^ dryland

PBS ~ HIH
Individual Project Beport
Calendar Year 1958

Part A.

Project Title: STUDY CF DISTTT.TKD WATER AND DEICHIZED
HATER QUALITY

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 San0 Engineer
Jo Paul Davies, Engineering Aid5 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

B. STUDY CF DISTILLED WATER AHD DEICB1ZEU HATER QUALITY
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 03 ^

Xo Sanitary Engineering Branch
20 Equipment Testing and

Quality Control Section
3° Retbesda lhs Maryland

PHS — HIH
Individual Project Report
Calendar Year 1958

Part A<

Project Title: LABORATORY GLASSWARE SURFACE C0HTAM1HANTS
ABD THEIR REMOVAL

Acting Principal Investigators Harry Stierlis Sr° San<> Engineer

Cther Investigator: None

Cooperating Units: Hone

Man Years (Calendar Year 1958) :

Total 0

Professional: 0
Other: 0

Project Descriptions (See attached material)
Part B included: Yes Ho

Co IABGRATORV GLASSWARE SURFACE COmMmMS AHD THEIR REMOVAL
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
research.

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 lk9 Maryland

PHS — NIH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: HIGH QUALITY STEAM FOR LABORATORY AND HOSPITAL USIs
Acting Principal Investigators Harry Stierli^, Sr<> San., Engineer
Other Investigator: None
Cooperating Units: None

Man Years (Calendar Year 1958):

Total: 0

Professional: 0
Other: 0

Project Description: (See attached material)
Port B included: Yes No

D* HIGH QUALITY STEAM FOR LABORATORX AHD HOSPITAL USE
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
HIH.

Significance to the Program of DRSs

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 lka Maryland

PHS — NIH
Individual Project Report
Calendar Year I958

Part A<

Project Title: GERM-FREE SYSTEMS RESEARCH AHD DEVELCE>MENT

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

Eo GERM-FREE SYSTEM RESEARCH AND DEVELOPMENT
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
20 Germ-Free Services Section
3<, Bethesda lk9 Maryland

PHS — HXH
Individual Project Report
Calendar Year 1958

Part A.

Project Title: GERM-FREE GUIHEA PIG DIET DEVEL<S>MEBE?

Principal Investigator: John L» So Hickey

Other Investigator: Dr. R. E. Horton

Laboratory of Germ-Free Animal Research
NIAID

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

F. '. ?IG DIET DETEKPiEHT

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
standpoint.

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 lks Iferyland

PHS -«. NIH
Individual Project Report
Calendar Year 1958

Part A

Project Title: EVALUATION OF THE SIGNIFICANCE OF NUMBERS OF
AIRBORNE MICROCOCCI IN PATIENT CARE AND
RESEARCH AREAS OF THE HIH

Principal Investigator; Dr. Lloyd G„ Herman, Bacteriologist^, GS-11

Other Investigators: Mr. John Lo S. Hickey, Sr» San<> Engineer
Mr« Frank Morelli8 Bacteriologist a GS-7

Cooperating Units: None

Man Years (Calendar Year 1958) i

Total: 0.08

Professional: 0o04
Other: OoOU

Project Description: (See attached material)
Part B included: Yes No

Go EVAHIATION OF THE SIGNIFICANCE OF NUMBEES CP AIRBORNE MICROCOCCI
IN PATIENT CARE AND RESEARCH AREAS OF THE NIH

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 1958;
None.

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 areaB9
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

FES — BIB
Calendar Year 1958

Budget Data;

Actual Oblications for Calendar Year 1958: $ 156,708
Total:
Direct:

Reimbursement: $ 156,708

PUBLIC HEALTH SERVICE - NATIONAL INSTITUTES OF HEALTH
DIVISION OF RESEARCH SERVICES

Summary of Branch Activities 1„ DRS-7

Serial Number

2, RESEARCH FACILITIES PLANNING BRANCH 2. A. E. Williamson

Branch Chief

Objectives

To plan and construct new research facilities as required by the new
and expanding needs of NIH.

Programs

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 - Germfree 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

Problems

Personnels 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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