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3

A STUDY OF THE RESPONSE OF TWO
PATENTS WITH PARKINSONISM
TO 00518 01‘ ASCORBlC- ACID AND
THIAMIN!

Thus. {a r!» om o: M. s.
MICHIGAN STATE unmeasmr
Ruth Wafkins Do V00
1956

A STUDY OF THE RESPONSE OF TWO PATIENTS
WITH PARKINSONISM TO DOSES OF
ASCORBIC ACID AND THIAMINE

By

Ruth Watkins DeVoe

A THESIS

Submitted to the College of Home Economics of Michigan
State University of Agriculture and Applied Science
in partial fulfillment of the requirements
for the degree of

MASTER OF SCIENCE

Department of Foods and Nutrition
School of Home Economics

1956

ACKNOWLEDGEMENTS

The writer is appreciative of the help and coopera-
tion extended to her by Mrs. Beth Bates, R.N., Mr. Face,
Dr. Fink, Dr. Dunn, the staff of the Ingham County
Rehabilitation Center and of the cooperation of the
patients.

It is with a sense of real gratitude that acknowledge—
ment is made of the patient guidance and assistance given
by Dr. Dena C. Cederquist and Dr. Evelyn M. Jones, and of
the help received from Dr. Wilma D. Brewer and Dr. Margaret

A. Ohlson.

ABSTRACT

The nutritional status of two patients with
Parkinson's disease at the Ingham County Hospital was assessed.
Daily food records were obtained. Urinary excretions of
ascorbic acid and thiamine, and blood serum ascorbic acid
concentrations were determined by chemical analyses.

The three-month study was divided into four successive
periods during which the subjects were given daily oral
administrations of (a) a placebo, (b) 400 mg. of ascorbic
acid only, (c) 400 mg. of ascorbic acid and 3 mg. of thiamine,
(d) 3 mg. of thiamine only. Prior to period (a) the subjects
had received 200 mg. of ascorbic acid per day for one month
or longer.

Evaluation of the daily food records suggested that both
subjects consumed a low-iron diet. Their mean daily intakes
of iron were 31 to 35 percent below the 12 mg. per day
recommended by the National Research Council. The diets of
both subjects furnished adequate amounts of protein, calcium,
vitamin A, and riboflavin, but on the average supplied less
ascorbic acid and niacin than the amounts recommended by the
National Research Council. For one subject the mean daily
intake of thiamine was below 1 mg. except during the final
month of study. The caloric intake of one subject increased
at the rate of approximately one-hundred calories per month
throughout the study, while the other subject had a

relatively constant intake.

-2-

Blood serum ascorbic acid values were 2.10 and 1.86 mg.
per 100 ml. for the two subjects during period (a). An
increase in blood serum ascorbic acid occurred during the
periods when 400 mg. ascorbic acid were given daily.

Values were decreased to 0.83 and 0.85 mg. per 100 ml. six
weeks after termination of the ascorbic acid supplement.

The percentage responses to a test.dose of ascorbic acid
were 15.0 and 15.9. These values are below the range of
values observed for healthy women on self—selected diets.

The percentage responses to a test dose of thiamine
were 12.5 and 21.9, which is within the range of values
observed in healthy women.

There were no significant alterations in the subjects
well-being or physical strength, or psychological status
attributable to the therapy as assessed from the clinical
examinations of a physician, a physical therapist, and a
psychologist. .

The data on these two subjects do not permit the
establishment of definite conclusions, but they appear to
justify additional investigation of the ascorbic acid

nutrition of Parkinson's subjects.

A STUDY OF THE RESPONSE OF TWO PATIENTS WITH PARKIN-
SONISM TO DOSES OF ASCORBIC ACID AND THIAMINE

Thesis for the Degree of M.S. by
Ruth Watkins DeVoe

I.
II.

III.

V.
VI.
VII.

TABLE OF CONTENTS

INTRODUCTION . . . . . . . . . .
REVIEW OF LITERATURE . . . . . .

Parkinson's Disease . . . . .

Methods of Assessing Nutritional

Ascorbic Acid Analysis . . .
Thiamine Analysis . . . . . .
EXPERIMENTAL PROCEDURE . . . . .
Experimental Plan . . . . . .
Analytical Procedures . . . .
Estimation of Food Intake . .
Administration of Vitamins . .
RESULTS AND CONCLUSIONS . . . .

Evaluation of the Food Records

Serum Ascorbic Acid Concentrations .

Responses to Test Doses of Vitamins

Clinical Evaluations . . . . .
SUMMARY . . . . . . . . . . . .
LITERATURE CITED . . . . . . . .
APPENDIX . . . . . . . . . . . .

Food Record Form . . . . . . .

Page

 

Nat-PH

22
29
32
32
35
38
39
40
40
44
47
51
53

55
61

62

LIST OF TABLES

Table Page

 

I. Comparison of the Analyzed Value of Vitamin A
Intake with Those Estimated From Three Standard
Tables 0 O O O O O O O O O O O O I O O O O O 0 O O 20

II. Comparison of the Mean values Obtained with the
Long and Short Methods of Dietary Analysis . . . . 23

III. Method of Administration and Estimation of
Ascorbic Acid and Thiamine . . . . . . . . . . . . 34

IV. The Average Nutritive Value of Food Consumed By
Two Subjects with Parkinson's Disease. . . . . . . 41

V. Serum Ascorbic Acid Concentrations . . . . . . . . 45

VI. Responses to Test Doses of Vitamins. . . . . . . . 48

INTRODUCTION

A nutritional survey of certain patients at the Ingham
County Hospital was conducted in 1953, by the Foods and
Nutrition Department of Michigan State University. This
study indicated that three patients with Parkinson‘s disease
.had serum total ascorbic acid concentrations from 0.15 to
0.22 mg. per 100 ml. (26). These values are considerably
lower than the 0.73 to 1.21 mg. per 100 ml. serum, that was
reported for healthy women who were receiving generous intakes
of vitamin C (48).

Investigators (8) have reported that blood serum ascorbic
acid values reflect previous dietary intake. The dietary
histories of these three patients with Parkinson's disease
suggested that their vitamin C intakes prior to admission to
the hospital had been reasonably satisfactory; and since they
had been hospitalized less than one month at the initiation
of the study, the low serum ascorbic acid concentrations
merited further investigation.

In the fall of 1954, the Foods and Nutrition Department
undertook a study of two of the patients with Parkinson's
.disease at the Ingham County Hospital. The urine of the
subjects was analyzed for excretion of ascorbic acid, thiamine,
and riboflavin, before and after the administration of an oral

test dose of each vitamin. The percentage excretion of the

test dose of ascorbic acid was 4.1 and 7.4, respectively (24).
The values of similar tests observed for healthy women on
self-selected diets ranged from 26 to 81 percent (5). After
eleven days of an oral supplement of 400 mg.. of ascorbic
acid per day, the patients' blood serum ascorbic acid concen-
trations were 1.70 and 1.72 mg. per 100 ml. These values
correspond favorably with those reported for healthy women
receiving a generous intake of this vitamin (48).

The percentage excretion of the test dose of thiamine
was 1.9 and 1.6. These values are low in comparison with
those obtained in healthy women, namely 12 to 38 percent (6).
The excretion of a test dose of riboflavin was calculated to
be 4.0 and 26.3 percent, respectively. The values observed
for healthy women on self-selected diets range from 26 to
38 percent (6). During the period of vitamin administration
the patients reported that they were "feeling better”.

The data that has been collected concerning the
nutritional status of Parkinson's patients was of two weeks
or less duration. Due to the observed feeling of well-being
of the two patients a more carefully controlled study of
longer duration seemed justified.'

The purpose of the present study has been to determine

the urinary excretions of ascorbic acid and thiamine and the

 

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concentration of blood serum total ascorbic acid of two
patients with Parkinson's disease preceding and following
the administration of test doses of the vitamins. Clinical

evaluations were made of the patients' general well-being.

REVIEW OF LITERATURE

Parkinson‘s Disease

 

Parkinson's disease is not a discrete entity. It has
been described as a slow, progressive degenerative disease
of the nervous system characterized by rigidity of the
skeletal musculature, fixidity in posture and a tremor of
regular, slow rhythm. It is not a disease in the sense of
being due to damage of a unitary physiologic system and
exhibiting a constant symptomatology. The disease is also
Iknown as paralysis agitans, shaking palsy, and Parkinsonism (16).

According to Christian (16) the classical clinical
description of the disease was reported by James Parkinson in
1817, and the disease was first described as a pathological
change at the base of the brain by Manchot in 1904.

The etiology of the disease is unknown. Usually the
disease occurs at ages past forty years, but it has been
reported in persons whose ages are in the twenties. A
history of the disease in the same family is found in
approximately 4 to 16 percent of the reported cases (16).

The disease causes cell degenerations in the basal ganglia.
'There is wide diversity of opinion concerning the precise

location within the basal ganglia of the pathological process.

 

There are several reports in the literature that present
a description of the progress of Parkinsonism (16, 55). The
disease begins gradually, usually with tremor in one or the
other hand. In the majority of cases, the disease sets in
abruptly after fright or trauma. Other early signs of
Parkinsonism are the loss of the normal swinging of an arm
while walking, and much fatigue.

The characteristic features of the disease are the
expressionless face, slow movement of the lips, the elevated
eyebrows and general facial immobility. The tremor is regular
in rhythm and appears in the small muscles of the hands and
feet, the lips, the tongue and mandible. Weakness is greatest
‘when the tremor is most developed.

Movements of a patient with Parkinson's disease are
characterized by great deliberation. These patients rise
from chairs slowly in a stooping attitude with the head
;projecting forward. Their steps are short and hurried.
Usually it is easier for these patients to run upstairs than
‘walk. Handwriting is tremulous and often micrographic.

Some patients with Parkinsonism complain of subjective
sensations of heat which may be present on one side only and
lassociated with an increase of surface temperature. In other
instances patients complain of cold. The mental condition

rarely shows any change until late in the disease.

There is no known cure for the disease. The treatment
is symptomatic, and is directed toward relief of the tremor
and rigidity. Death from Parkinsonism is usually due to
intercurrent infection, such as pneumonia.

Investigators have presented the data of studies that
have involved the use of vitamin therapy for the treatment
of Parkinsonism. Baker (1) treated 15 patients suffering
from paralysis agitans with pyridoxine hydrochloride,
supplemented in most cases by oral administration of brewers'
yeast. The patients received 50 to 100 mg. of the vitamin
intravenously at daily intervals over a period of two to
four weeks. The author stated that neither the age of the
patient nor the durationcf symptoms seemed to have an effect
on the treatment. The nature of improvement varied from
patient to patient. The indices of improvement were decreased
fatigability, better sleeping, and increased appetite. There
‘was "some" improvement noted in six subjects, and only in
one subject was there any objective benefit in the form of
observable decrease in tremor and rigidity.

It was Baker's opinion that the small number of patients
did not allow any definite conclusions to be reported.

Zeligs (66) reported the use of vitamin B5 in the
treatment of Parkinsonism. Daily doses of 50 to 100 mg.

repeated one to three times were administered intravenously

to 15 patients. Several days later there was no change in
the degree of muscular rigidity or tremor. On the basis of
the data, Zeligs concluded that vitamin B6 had no value for
the treatment of Parkinsonism.

Doshay (21) pointed out that loss of weight is especially
likely to occur among patients who are tense, excitable, and
tremulous. He reported that a number of Parkinsonism patients
have responded favorably to vitamin B12 injections of one mg.
twice a week supplemented by thiamine hydrochloride given

orally or parenterally.

Methods of Assessing Nutritional Status

 

Several techniques are available for use in the assess-
ment of the nutritive intakes for individuals or groups.

The main objective of a study of dietary habits, whether
sociological, physiological, or economic, usually determines
the techniques involved (38, 35, 61).

Stiebeling (61) has outlined the methods commonly used
in this country to determine food intake. They are as follows:
(a) food habit inquiry, (b) quantity of food commodity
available for family consumption, (0) family food records
with inventories, (d) family food list, and (e) nutrient

intake of an individual.

 

/\

Since this study is concerned with the dietary evaluation
of two subjects, the emphasis in this section of the review
of literature has been placed on method (e), the nutrient
intake of an individual.

Various techniques have been used to estimate the
nutrient intake of an individual. Some investigators have
determined the dietary intake from recall records, in which
the accuracy of the data has depended on the memory of the
subject. Other investigators have instructed the subjects
to record the amount of food eaten in household units, or
weights in grams. The nutritive value of the food thus
recorded has been calculated from tables of food values. A
third method of determining nutritive intake consists of the
chemical analysis of a representative sample of the food eaten.

Burke (9) has stated that although the most accurate
method of studying food intake was the controlled balance
study; the food intake as recorded by a trained interviewer
using the diet history, will result in a satisfactory method
of diet analysis. The author (9) pointed out that the
qualifications of the interviewer were most important for
obtaining valid data by use of the research dietary history.
The interviewer should be a "person with an alert and inquiring
mind, accuracy for detail, keen powers of observation, and

sound basic training in the allied sciences and nutrition!‘.

Burke and Stuart (10) suggested that the interviewer record
the subject's usual pattern of food eaten in household
measurements and list the food likes and dislikes. The
history should then be followed by a three-day food record.
Combined records are used to estimate a "typical" day's
dietary intake. The diets are evaluated by a rating scale
containing a range of values wide enough to take care of
most inaccuracies in the diet history.

In 1954, Van Den Berg and Mayer (64) reported a study
which compared the dietary intake as recorded from the one-day
food record and the research dietary history. The subjects
were thirty-five obese pregnant women from the Research
Obesity Clinic of the Boston Lying In Hospital. The research
dietary history gave caloric intakes greater than those
given by one-day record, the difference being about 600
calories. Differences in estimates appeared to be due mainly
to inaccurate description of composition and size of dishes
on the one-day record, and to omission from the one-day
record of foods commonly consumed on the other days.

Several studies reported in the literature have compared
the data of a food record obtained by recall to similar data
obtained by the weighed food record. In 1950, Ohlson and

others (51) reported that different kinds of information were

-10-

obtained when more than one technique was applied in a given
situation. The estimated mean intake of calories, protein,
and calcium of 13 subjects as recorded by recall and weighed
diets was presented. The data revealed that the apparent
mean intake of all nutrients was greater when measured by
recall than from weighed diets.

Apparent reasons for the increase of nutritive value as
recorded by recall were: (a) there was less eating between
meals during periods on the weighed diets than during periods
of recall, (b) the size of the portion selected by many of
the aging women appeared smaller than that of the younger
women whose food habits formed the basis for the calculations
of the Leichsenring and Donnelson food composition table,
and (c) in the first weighed diet period the fear of social
rejection if mistakes were made resulted in emotional tension
which could have reduced the amount of food eaten.

Young and others (65) have reported a comparison of
calculated nutritive value of measured food intake against
the subject's estimation of intake from the data obtained in
the Northeastern Regional Research Nutritional Status Project
NE-4. The authors have pointed out that interpretations of
the data were difficult due to the fact that it was impossible
to follow a consistent or identical routine in collecting
data. The subjects were at one of six experimental stations

and varied in age and environment.

-11-

At the Maine Experiment Station, 25 eighth and ninth
grade children were observed at school lunch on three
consecutive days. They were aware that they were being
observed and would be asked to recall what they had eaten.

In New Jersey the noon meal of 56 male industrial
workers was the basis of comparison of measured and recorded
dietary intake. The meal was served in the plant cafeteria.
The authors stated that there was no means of checking plate
waste therefore the assumption was made that all food served
was eaten. Comparisons were made only on the basis of
measured food items and did not include items such as bread
and butter and cups of beverage. The subjects were unaware
that they were being observed.

At the New York Experiment Station 16 children twelve
to fourteen years of age were observed for one school lunch.
The amount of food served was estimated by three advisors from
the home economics department. The child's estimation was
obtained by questioning and recall directly after the meal.
The children were not aware that they had been observed.

At the Rhode Island Station, 14 college girls were
studied for one day during which they recorded their food
intake at the college cafeteria. Measured quantities Of food
were served and food not eaten was recorded. The subjects were

unaware that they were being observed.

-12-

In West Virginia, 21 college students in the Infirmary
were observed for one day. The food served was measured
and the unused portions were subtracted. The students
recorded their food intake. They were not aware that their
eating habits were being observed. The nutritive value of
the food was computed by Babcock's simplified method of
dietary calculation.

Considerable variation was found between the nutritive
values calculated from the estimated and from the measured
food intakes. In only one state, Maine, did the estimation
of half of the individuals yield calculated nutritive values
within ten percent of those of the measured values. Approxi-
mately two-thirds of the subjects estimated their food intake
in such a manner that the calculated nutritive value was within
twenty percent of that of the measured intake. It was noted
that the best estimations were made by subjects who were aware
they were being observed. The authors (65) pointed out that
"for only one group, Rhode Island college students, were there
pronounced under-estimations; for practically every nutrient,
the error of by far the greater majority of subjects was an
overestimation".

The greatest ranges of deviation of the nutritive value
from the measured intakes were found in the New York school

children's recalls and in the recorded estimations of the

-13-

New Jersey workers and the West Virginia college students.
There was no nutrient that could be singled out for all
states as being estimated most poorly.

From these surveys the authors (65) concluded that in
studying the nutrient intake of an individual, the subject's
ability to estimate the food portions could be a large source
of erroc. They recommended that in the absence of a weighed
record, measurement of intake in household units would be
the next most desirable procedure; and also "that it is wise
to check subjects' estimations periodically and to let them
know they are being checked".

Trulson (63) reported a study comparing dietary data
collected by the seven-day record, by dietary interview of
usual food practices, and three or more 24-hour recall diet
forms. No comparison was made with weighed diets. Data was
gathered from 47 subjects attending two Chicago clinics. The
differences in the mean values of the foods and nutrients
obtained by the three methods were not consistent. The
highest mean value for milk intake was obtained by the interview
method; the average of three or more 24—hour recalls provided
the lowest mean value. The highest mean value for intake of
foods high in carotene was the 7-day record method; the
interview method provided the lowest mean value. The author

stated that "it must be accepted that it is difficult, if not

ill-15‘ "lllllllllllllr.|

-14-

impossible, to determine which method gives correct food
intake". From the observations in the study, it was the
author's opinion that the long-range interview of usual
food practices was the method of choice, particularly in
clinical,studies.

In 1942, Huenemann and Turner (34) reported a study
comparing data obtained by weighing and by interview method.
Data was collected from 25 University of Chicago Clinic
patients whose ages ranged from 6 to 16 years. Each patient
was questioned regarding his usual food intake, his previous
day's intake and the frequency with which specific foods
were eaten. In addition, the family food purchases and
distribution of food among the members of the family were
determined. Three or four periods, each of 10 to 14 days
duration, of weighed food intake were obtained.

When estimations of food intake obtained from the
dietary history were compared with weighed dietary records,
the authors found that no dietary history agreed with the
weighed dietary record within twenty percent for all the
nutrients. Approximately one-half of the histories differed
from the weighed records in five or six dietary constituents
by a deviation of twenty percent. 0n the basis of this study
the author stated that data obtained by interview method

should be checked by a quantitative record of food intake.

-15-

Several studies have been reported concerning the
desirable length of the dietary study. In 1955, Trulson (62)
reported the finding of dietary studies based on records
gathered from one day, three days, and seven days. Informa-
tion was obtained from a random sample composed of 132
Chicago school children whose ages ranged from 10 to 12
years. The results of the study revealed that lengthening
the experimental period from one to three to seven days,
reduced the standard deviation respectively. She stated that
the length of the study should be determined by the nutrient
or food to be investigated. A nutrient, such as protein,
required shorter periods and smaller samples.

Chappell (15) stated that a short term survey should be
interpreted with caution. His statement was based on data
from a study of a 34-year—old woman observed for 70 weeks,
and a 66-year-old man observed for 13 weeks. The weekly
intake of calories and nutrients showed considerable
variation differing with the dietary component and with the
subjects. The woman's estimated average daily intake based
on one week sampled at random within the year had a
standard error of 8 to 15 percent for most of the dietary
components but a much larger percentage of error for some

of the vitamins.

The author stated that there was no reason to think the
woman's dietary habits were particularly variable. Therefore,
this suggested that the results of a short-term survey should
be interpreted with caution.

A comparison of the food intake of college women for
weekdays and for Saturday and Sunday was reported by
Leverton and Marsh (39). The data of the study consisted of
24 balance studies made of college women, 17 to 24 years of
age. The subjects were engaged in regular college work and
light housekeeping.

Results indicated a definite and significant variation
of food intakes for weekdays and for week ends when college
women were living on a self-chosen diet. The least difference
occurred between the period of five weekdays and the calendar
week. The author concluded that studies of food intake of
subjects on self-chosen diets would yield the most repre-
sentative data when conducted for a period of at least one
calendar week or in units of one week.

Church and others (17) have studied the ability of
different interviewers to obtain comparable dietary survey
data. The comparison of interviewers was made from material
gathered by three state experiment stations participating in
the Northeastern Regional Project NE-4. Data was collected

in Maine by three interviewers, working as a team in each of

-17-

two schools. Two interviewers, in each of two industrial
plants, collected the data obtained in New Jersey. In

New York, the dietary histories were taken by two interviewers
in one school.

The background of all the interviewers was similar. All
were nutritionists who had had some training in dietary
interviewing before collecting data for the project. The
same set of instructions was given to all interviewers. From
each subject, a diet history was taken and then verified by
crosschecking from a list of food groups. The comparison
was made between interviewers at the same station. The
author stated that with the available data, the difference
that may occur between interviewers at different stations
could not be tested statistically.

The differences that occurred among interviewers rarely
exceeded ten percent of the allowance for each nutrient,
the average being 5.4 percent. It appeared that these
differences were not likely to exceed sampling variations
inherent in dietary survey methods, therefore, it was
concluded that interviewers with similar background and
training for cooperative research could obtain comparable

data.

-18-

After food records have been obtained, their nutrient
content is assessed. Recommended dietary standards have been
proposed by the (a) National Research Council, United States
of America, 1953, (23); (b) the Canadian Council of Nutrition,
1950, (11); and (c) the British Medical Association, 1950,
(7). These three standards vary in philosophy and objective,
therefore in specific recommendations.

The recommended dietary standards of the National
Research Council (23) were designed

"for the maintenance of good nutrition of healthy

persons in the United States under present condi-

tions. The recommendations are not requirements,

since they represent not merely minimal needs of

the average person but nutrient levels selected

to cover individual variations in a substantial

majority of the population".

The recommended dietary allowances were formulated by
the Food and Nutrition Board of the National Research Council.
The allowances were based on data published in the literature
that related to the requirements for the various dietary
essentials, and the combined judgment of a large number of
inutrition authorities, particularly those actively engaged in
research.

The dietary standards proposed by the British Medical
Association (7) differ from the National Research Council

Recommended Allowances. The British standard provides "values

which apply to the average person rather than values which will

-19-

cover essentially all individuals, recognizing, however,
that some members in every group may need more than the
average".

The Canadian standards differ considerably from the
American and British recommendation. The Canadian standard
represents a "nutritional floor" beneath which maintenance
of the health of the people cannot be assumed (11, 12, 13).

A possible source of error of the evaluation of dietary
intake results from the fact that food composition tables
differ. Hunscher and Macy (35) compared the analyzed value
of vitamin A with those estimated from three standard
tables. (Table I)

Differences between values obtained by analyses of a
dietary intake and those estimated from standard tables
result from several factors: (a) the numerous methods of
chemical analyses of foods, (27, 40, 62), (b) losses of
various nutrients during storage, preparation, and cooking,
(14, 43) and (c) the different types of soil, mode of
cultivation, amount of water available, geographic origin,
and marketing conditions involved in food production
(3. 45. 60).

Clear concepts are also lacking with regard to the
methods used to calculate the amount of energy derived by

the body from a given amount of food eaten.

 

 

 

-20..

TABLE I

COMPARISON OF THE ANALYZED VALUE OF VITAMIN A INTAKE
WITH THOSE ESTIMATED FROM THREE STANDARD TABLES

 

Method Intake of vitamin A

 

per day
Analyzed 341 I.U.
Estimated
Table l 1238 IOU.
Table 2 852 I.U.

 

-21_

The majority of studies have calculated energy values in
which either the Rubner or Atwater factors were used. In
general, all the studies done in the United States since
1900, particularly those using the Rose or Sherman tables
are based on the Atwater factors.

Maynard (44) has reviewed the Atwater system of
calculating the calorie value of diets. He pointed out that
the factor of 4-9-4 was meant to apply to diets in which kinds
and pr0portions of food were similar to the "average" mixed
diet. The Atwater figures should not be applied to diets
markedly different in make-up. "The figures 4-9-4 were never
intended to be applied without modification to all diets"
(44).

Throughout the study records of the food consumed by
the patients were obtained in the following manner: a food
record form listing the amount of a serving eaten was filled
out daily by a member of the nursing staff of the hospital.
This form was kept at the patient's bedside. This method
reduced extra work for the nursing staff. Both patients
were served their meals on trays in the same room.

Data gathered from other diet surveys indicated that this
method is adequate for recording information required in this

study.

-22-

The Leichsenring and Wilson (37) food composition table
was used to estimate the nutritive value of the dietary intake
of the two subjects. The reliability of this method had been
demonstrated previously in this and other laboratories.

Leichsenring and Wilson evaluated the accuracy of the
short methods table by the use of statistical tests. The
observed differences between the short method and long method
are presented in Table II. The data were based on 21 records
gathered over a period of three days. From the use of
statistical tests the observed differences were shown to be
due to errors in random sampling and not to a real difference
between the two methods.

Ohlson and others (51) in reporting the nutrition and
dietary habits of 13 women subjects, 51 to 77 years of age,
stated that the subjects' diets as computed by the short
method of Donelson and Leichsenring ”accurately reflected the
results of laboratory analyses for nitrogen, calcium, and
phosphorus except in cases of limited food intakes or where
predominance of bread, processed meats or intricate cookery

mixtures were found".

Ascorbic Acid Analysis

 

Early ascorbic acid research utilized the biological

technique that was dependent on the prevention of scurvy in

-23-

TABLE II

COMPARISON OF THE MEAN VALUES OBTAINED WITH
THE LONG AND SHORT METHODS OF DIETARY ANALYSIS*

 

Protein ‘IVitamin C Thiamine

 

Method Calories (gm.) (mg.) 1mg!)
Long 1984 65.8 75.9 1.13
Short 2004 65.1 74.4 1.12
Difference 20 0.7 1.5 0.01

 

*Leichsenring and Wilson (37)

-24-

guinea pigs (59). This method was followed by a similar
growth test (29). In 1947, Crampton (20) proposed a bioassay
for ascorbic acid that involved the growth of the odontoblast
cells of the incisor teeth of young guinea pigs. The
applicability of this method is limited if the potency of
the test material is too low for it to be measured accurately
by feeding trials, or if the sample is poorly absorbed by the
experimental animal.

In recent years biological methods of assay for ascorbic
acid have been replaced largely by chemical methods (28, 49, 58).
The technique most commonly used for the chemical determination
of the vitamin consists of the measurement of the reducing
capacity of ascorbic acid with 2, 6—dichlorophenolindophenol,
carried out in an acid medium (4, 36, 47). A second technique
involves the use of the osazone reaction, the estimation of
total ascorbic acid using 2, 4-dinitrophenylhydrazine (42, 57).
Micromethods of the latter have been adapted by Lowry and
others (42), thus subjects have not needed to experience the
trauma of venous punctures for blood ascorbic acid determinations.

Other methods include the photosensitized reaction with
methylene blue, and the reaction with iodine which has been
mainly of value in testing pure ascorbic acid or pharmaceutical

preparations free from interfering substances (58).

'11

-25-

According to Gyorgy (27) early reports of the indophenol
method appeared in 1927, from Tillman and his group. They
published the results of the estimation of oxidation-reduction
potential and its application to food chemistry. This group
devised the method of distinguishing natural from imitation
fruit juices by observation of the strong reducing power of
fresh lemon and other fruit juices against 2, 6~dichlor0pheno-
lindophenol.

Gyorgy (27) stated that Tillman showed that this reduction
of the dye to the leuco form was a measure of the antiscorbutic
activity of the lemon juice. From the investigations of
other workers there resulted numerous modifications of
Tillman's visual procedure. Harris and Ray (30) developed a
specific quantitative test for the determination of ascorbic
acid. Their method included the use of a preliminary extraction
process with trichloroacetic acid (later replaced by meta-
phosphoric acid) and the subsequent titration of the ascorbic
acid solution in a relatively strong acid solution. The
titrant was the indophenol dye solution. At this time it
became apparent that substances other than ascorbic acid
reduced the dye to its leuco form. It was shown that the
reaction of ascorbic acid was almost instantaneous while the
reaction of the interfering substances proceeded at a slower

rate.

-26-

Evelyn and others (22) believed that non—ascorbic acid
reducing substances could account for more than ninety
percent of the total indophenol reducing capacity of normal
urine. They presented a method of differentiating between
ascorbic acid and other reducing substances which was based
on the differences in the rate at which the dye was decolorized.
"It has been shown that the reaction of ascorbic acid is
almost instantaneous, while that of other known interfering
substances proceeds at a slower rate" (22). On the basis
of their experience, they suggested a method in which an
excess of the dye was mixed with a measured aliquot of urine,
and the amount of dye decolorized at five second intervals
was measured in a photoelectric colorimeter until the reaction
was complete. These results were used to plot a reaction
velocity curve from which the amount of decolorization due
to ascorbic acid was determined by extrapolation to zero time.
The authors stated that the method was empirical and could
only be expected to give approximate results; however, they
believed there were far less errors in this method than those
encountered from the use of visual titration.

King (36) pointed out two crucial points to consider when
using the indOphenol method: (a) the avoidance of oxidative
changes previous to the titration, and (b) awareness of

circumstances under which other reducing substances will or

-27-

will not interfere with the color reaction. The indophenol
dye is reduced by thiosulfate, cysteine, and other substances
containing sulfhydryl groups.

Mindlin and Butler (47) have stated that in using the
indophenol method the pH of the metaphosphoric acid extract
must not be low enough to cause significant fading of the
dye, and yet not so high as to result in immediate reduction
of the dye by reducing substances other than ascorbic acid
that are present in the extract.

Roe and Kuether (57) developed the method utilizing
the osazone reaction which is based on a direct colorimetric
determination of dehydroascorbic acid. The oxidized form
of the vitamin in metaphosphoric acid extract is coupled
with 2, 4-dinitrophenylhydrazine, and the resulting derivative
is treated with sulfuric acid to produce the red-colored
osazone which is measured photometrically. For the determina-
tion of total ascorbic acid the reduced ascorbic acid is first
oxidized to dehydroascorbic acid.

Lowry and others (42) adapted the dinitrophenylhydrazine
method for the determination of ascorbic acid in a small
amount of serum.

Objections to the osazone method were reported by Pijoan
and Gerjovich (53) who found that this method sometimes gave

unreliable results for ascorbic acid content of certain biological

-28-

preparations if loss in lactone structure of dehydroascorbic
acid had taken place. Penney and Zilva (52) showed that a
2,3-diketo-l-gulonic acid compound could be made from
dehydroascorbic acid by mutarotation and loss of lactone
structure. The d-glucoascorbic acid and d-araboascorbic
acid could not be distinguished from l-ascorbic acid by the
2,-4-dinitrophenylhydrazine method.

Roe and Kuether (57) pointed out that oxidizing agents
such as Fe+++ ions and H202 could produce interfering color
substances with the 2,4-dinitrophenylhydrazine.

In 1951, Hewston and coworkers (33) compared three methods
of determining ascorbic acid: the 2,6-dichlorophenolindo-
phenol, 2,4-dinitrophenylhydrazine, and the bioassay method
involving growth of the odontoblast cells of the incisor
teeth of young guinea pigs. The authors stated that when
extraneous reducing substances were present neither chemical
procedure could be relied upon to give an accurate measure of
vitamin C.

Total ascorbic acid in the blood serum was estimated
according to the method of Bessey and Lowry as listed in the
Northeast Regional Publication Number Five (19) except that
the standard was prepared daily. This procedure is one of
the most acceptable ones, since the amount of serum necessary

for the determination is obtained from a finger prick blood

-29-

sample. Other advantages of this method were that laboratory
facilities were readily available for the determination,
and the data obtained in this study could be compared readily

with data from similar studies on normal, healthy women.

Thiamine Analysis

 

Methods most often used to estimate thiamine are
(a) chemical procedures involving either the oxidation of
thiamine to thiochrome which can be measured by its fluorescence,
or a colorimetric procedure based upon a coupling reaction with
a diazotized amine, (b) the yeast fermentation procedure,

(c) microbiological assays, and (d) biological assays using
rats, chicks, or pigeons as test animals. Many modifications
of these various procedures have been published.

Two of the early reports using thiochrome method appeared
in 1935. Barger and coworkers (2) found that thiamine on
oxidation with ferricyanide in alkaline solution was transformed
into thiochrome. Robinson (56) reported that Jansen converted
thiamine into thiochrome, extracted the thiochrome with
isobutanol, and measured the fluorescence of the extract in
a fluorometer calibrated against standard solutions of quinine.

Many biological fluids, such as urine, contain interfering
substances that prevent accurate estimations of thiamine.
Hennessy and Cerecedo (32) reported that interfering substances

could be eliminated by adsorption of the thimaine on zeolite

-30-

and subsequent eluting with hot potassium chloride solution.

Mickelsen and others (46) pointed out that there is ‘
little information concerning the interfering fluorescent
substances in urine, and that there may be other substances
present which contribute to the total fluorescence and
therefore, are read as thiochrome. Mickelsen and co-workers
also made a statistical evaluation of the thiochrome method.
They found that in 33 pairs of duplicate samples the standard
deviation of the thiamine content was 4.6 percent of the mean.

Hennessy (31) reported that the oxidation of thiamine in
alkaline ferricyanide solution did not result in the production
of an equivalent amount of thiochrome. The amount of conversion
was not reported. He suggested the incomplete conversion was
due either to establishment of an equilibrium in the oxidation
process or to a side reaction.

Conner and Straub (18) suggested the isobutyl alcohol
be distilled between 106 and 107°C. which prevents any
fluorescent material from being carried over into the distillate.
A glass apparatus should be used.

In the present study the thiamine content of the urine
of two Parkinson's patients was determined by the thiochrome
technique of Conner and Straub (18). This method was chosen
since it is one'of the most accurate methods. Other

advantages of using the thiochrome technique were that laboratory

-31-

facilities were readily available, and the previous data
collected concerning the thiamine status of these patients

had been estimated by the thiochrome technique.

-32-

EXPERIMENTAL PROCEDURE

Experimental Plan

 

Since this study was undertaken to determine the possible
beneficial effects of ascorbic acid and/or thiamine, the
following experimental plan was designed. The three-month
study (Table III) was divided into four successive periods
during which the subjects were given daily oral administrations
of (a) a placebo, (b) 400 mg. of ascorbic acid only, (0)

400 mg. of ascorbic acid and 3 mg. of thiamine, (d) 3 mg. of
thiamine only. Prior to.Period.(a; the subjects had received
200 mg. of ascorbic acid per day for one month or longer.

During Period a, a sugar placebo was given daily. Twenty-
four-hour urine specimens were collected on three successive
days per week for Period a and analyzed for the vitamin C
content. The urine was collected in brown bottles which
contained metaphosphoric acid, sulfuric acid, and 8—hydrox-
yquinoline. The serum ascorbic acid concentration of each
subject was determined from fasting blood samples collected
on the last day of Period a.

On the first day of Period b, the 24—hour test dose
response of ascorbic acid was calculated. The formula used
to calculate percentage response of a test dose of the vitamin

was as follows:

-33

Excretion after tes’fl asal excretion
% test dose — lose (24 hr.) Avg. for 24 hr._ x 100

Test dose

The administration of 400 mg. of this vitamin was
continued throughout the 28-day period in an attempt to
saturate the patients. Analyses of the urinary excretion of
the vitamin were made from three 24—hour urine collections per
week. The serum ascorbic acid level was determined from a
fasting blood sample collected at two-week intervals throughout
the study. In the latter part of Period b, the basal urinary
thiamine excretion was measured from 24—hour urine specimens
collected during three successive days per week for a
two-week period.

On the first day of Period 0, the percentage response
to a 3-mg. test dose of thiamine was determined. Throughout
this period, 400 mg. of ascorbic acid and 3 mg. of thiamine
were given daily. The urinary concentrations of thiamine
and ascorbic acid were determined from 24—hour collections.

A maximum of three collections were made each week.

During Period d, the administration of ascorbic acid
Was discontinued, but the daily dose of 3 mg. of thiamine
continued. Urinary concentrations of thiamine were
determined from 24-hour collections twice per week. Serum

ascorbic acid was measured periodically.

TABLE III

METHOD OF ADMINISTRATION OF ESTIMATION OF
ASCORBIC ACID AND THIAMINE

 

Length ofCtime

 

Period (days) Vitamin Supplement
a 18 None
b 28 400 mg. ascorbic

acid per day

c 30 400 mg. ascorbic
’ acid, and 3 mg.
thiamine per day

d 28 3 mg. thiamine
per day

 

-35-

Clinical evaluations by a physician, occupational

tmerapist, and psychologist were scheduled for each period.

Analytical Procedures

 

The 2,6-dichlor0phenolindophenol method was used to
determine the concentration of ascorbic acid in the urine.
The method of Evelyn and others (22) was followed with some
modification. A calibration curve was used to calculate the
ascorbic acid content of the urine. The curve was prepared
from eight standard ascorbic acid solutions; the concentrations
of which ranged from 5 to 40 mcg. per 5 ml. The concentration
of urine was adjusted with one precent metaphosphoric acid
solution to the degree that permitted the galvanometer
reading of a 5 ml. sample to be within the linear portion of
the calibration curve. Frequent calibration curves using
standard ascorbic acid solutions were prepared to assure
stability of the indicator solution. To remove any error
which is due to a possible decomposition of the dye, fresh
solutions of dye were prepared daily.

The dye solution was standardized as follows: Five m1.
of one percent metaphosphOric acid were added from a blow
pipette to a colorimeter tube containing 5 ml. of dye solution.
Galvanometer readings were taken at five second intervals.
The dye solution was diluted so that the reading on the

galvanometer scale was approximately fifty.

a )1. lilll‘lut 1

 

 

Q r..‘.l..llbofi¢|'v.l; .I a u

Five m1. of diluted urine were added to a colorimeter
tube containing 5 m1. of standard dye solution, mixed, and
quickly placed in the cuvette holder of the instrument.
Galvanometer readings were taken at five-second intervals.
Three urine dilutions were made from each 24-hour urine
collection. Duplicate measurements of each urine solution
were made.

The galvanometer values of the urine samples were
plotted against time on linear graph paper and extrapolated
to zero time. The micrograms of ascorbic acid in a 5-ml.
sample were read directly from the calibration curve.

Fasting blood samples of each subject were collected
on September 15, and every two weeks thereafter until
December 19.9 The 2,4—dinitrophenylhydrazine procedure based
on that of Lowry and others (41) was used for the deter-
mination of total ascorbic acid in the serum samples. The
method as described in the Northeast Regional Publication
Number 5 (19) was modified in that the standards were prepared
fresh daily rather than frozen simultaneously with the serum
samples. The determination is dependent upon the separation
of dehydro-ascorbic acid as an osazone derivative of
2,4-dinitrophenlhydrazine. When the osazone is treated with
sulfuric acid a color is produced, the transmittance of

which shows good agreement with Beer's law.

-37...

The blood samples were obtained in the following manner:
the subject's hand was placed in a basin of warm water for
approximately five minutes in order to stimulate circulation
of the blood and then a finger was pricked with the point
of a surgical blade. A few drops of blood were collected in
a small glass tube and immediately centrifuged. Using a
Levy Lang pipette three lO-mm3 samples of serum were placed
into serological tubes, and 4O-mm3 of five percent tri-
chloroacetic acid were added. The samples were mixed,
capped, and placed in ice, and after approximately thirty
minutes placed in storage in the deep freeze.

The thiamine content of the urine was determined by
the thiochrome technique. The procedure followed was that
reported by Mickelsen and others (46). The urine was passed
through a column of Decalso.* Modifications of the oxidation
step were made according to recommendations reported by
Conner and Straub (18).

The oxidation of thiamine to thiochrome was accomplished
as follows: (a) 3 ml. of 15 percent sodium hydroxide and
0.1 ml. of 1 percent potassium ferricyanide were placed in
an oxidizing chamber, (b) 5 ml. of eluate were transferred

to the oxidizing chamber and 15 ml. of isobutyl alcohol were

 

*The Permutit Co., 330 West 42nd Street, N. Y.

 

o

J“. ‘5“;

.\

 

 

. I flu-Hui”

.....uu. .‘. a

 

.J
ilxlvl“

-38-

added, (c) the solution was shaken for 1.5 minutes, and then
centrifuged for 1.5 minutes, (d) the aqueous layer was
removed with a pipette, and two grams of anhydrous sodium
sulfate were added, and the solution was centrifuged for

one minute. The isobutyl alcohol solution which contained
the thiochrome was transferred to a cuvette, and the amount - “TIT
of light transmission read in the Coleman fluorophotometer.
The instrument was standardized against a quinine sulfate 3

solution.

 

Estimation of Food Intake

Throughout the study, records of food consumed by the
two patients were obtained by a food record form (appendix)
for each patient, which was filled out daily by a member of
the nursing staff of the hospital. From these records, the
nutrient value of the food consumed was calculated with the
use of the short form of Leichsenring and.-YWilson (37).
The average daily intake of nutrients in the food consumed
during the first week of each month was considered to be

representative of that for the entire month.

nun...‘

 

 

 

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-39-

Administration of Vitamins

 

The vitamins were given orally. Gelatin capsules
were filled with a weighed amount of cerelose-vitamin
mixture so that the capsules contained either 400 mg. of
ascorbic acid, or 3 mg. of thiamine. To assure correct
dosage, quantitative determinations were made from capsules
which were selected by random sampling. The error in the
weight of ascorbic acid was found to be i 3.5 percent and of
thiamine, t 2.5 percent. Capsules were also filled with
cerelose which were administered as a placebo during the
first period of the study. Throughout the study the patients

were not aware of the content of the capsules.

[I ~..-' v m : ‘fl‘m int-l": mmvfig’vfi
‘ 5 u

-40-

RESULTS AND CONCLUSIONS

Evaluation of the Foods Records

 

The nutritive value of the food intakes of the two
patients was estimated according to the short form of
Leichsenring and Wilson (37). These data are presented in
Table IV. Both subjects knew that daily records were kept.
This awareness may have influenced their intakes since they
took considerable interest in the study, and as roommates,
usually compared foods eaten at each meal.

The diets of the two subjects furnished the National
Research Council recommended allowances of calcium, protein,
vitamin A, and riboflavin. Both subjects consumed a
low-iron diet. The mean daily intakes of iron were 8.5, 8.3,
8.0, and 8.4 mg. in September, October, November, and
December, respectively, for subject H.L.; and 7.3, 7.4, 7.7,
and 8.8 mg. respectively, for subject P.P. These mean daily
intakes of iron were 31 and 35 percent below the National
Research Council recommended daily allowance of 12 mg. for
a sixty-year-old woman. In future work with Parkinson patients
it would be desirable to include the determination of
hemoglobin as this provides a means of estimating the adequacy

of dietary iron.

 

;

Lh-

 

-41-

TABLE IV

THE AVERAGE NUTRITIVE VALUE OF FOOD CONSUMED BY TWO
SUBJECTS WITH PARKINSON’S DISEASE

Subject H.L.

 

 

Nutritive Month N.R.C.
Value of Sept. Oct. Nov. Dec. Recommended
Allowances
Protein gm 63.5 63.5 68.2 64.8 55.0
Calcium gm 1.12 1.10 1.11 1.13 0.80
Iron mg 8.5 8.3 8.0 8.4 12.0
Vitamin A (I.U.) 10,900 3,235 3,130 5,884 5,000
Thaimine mg 1.00 1.07 1.02 1.04 1.00
Riboflavin mg 2.20 1.80 1.86 1.96 1.40
Niacin mg 9.32 9.24 9.40 9.55 10.00
Ascorbic Acid mg 77 62 48 63 70
Calories 1863 1874 1856 1878 1800

 

Subject P.P.

 

 

Nutritive Month N.R.C.
Value of Sept. Oct. Nov. Dec. Recommended
Allowances
Protein gm 60.0 55.9 64.4 69.4 55.0
Calcium gm 1.03 1.03 1.10 1.17 0.80
Iron mg 7.3 7.4 7.7 8.8 12.0
Vitamin A (I.U.) 10,147 4,159 3,530 4,383 5,000
Thiamine mg 0.83 0.88 0.83 1.21 1.00
Riboflavin mg 2.18 1.85 1.86 2.25 1.40
Niacin mg 8.71 8.80 10.13 11.11 10.00
Ascorbic Acid mg 80 53 57 68 70
Calories 1644 1716 1831 1964 1800

 

*The National Research Council Recommended Dietary Allowances
-of the above constituents for a 60-year-old woman.

K. v...

-42-

The National Research Council has recommended an intake
of 10 mg. of niacin per day for women 65 years or older.

For subject H.L. the mean daily intakes of niacin were 9.32,
9.24, 9.40, and 9.55 mg. in September, October, November,
and December, respectively; and for subject P.P. 8.71, 8.80,
10.13, and 11.11 mg., respectively.

The greatest day—by-day variation was observed in the
intake of vitamin A. For subject H.L. the mean daily intake
varied from 3, 130 I.U. during November, to 10,900 I.U. during
September. Comparable values for subject P.P. ranged from
3,530 I.U. (November daily mean) to 10,147 I.U. (September
daily mean). The mean intakes during the four months study
were 5,787 I.U. and 5,555 I.U. for subjects H.L. and P.P.,
respectively. Similar findings of such variation have been
reported in other nutritional surveys. ,It is generally
accepted that the body has the capacity to store vitamin A;
therefore, the average intake of this vitamin was judged to
be satisfactory.

The mean daily intakes of thiamine for subject P.P.
during September, October, November, and December, were
0.83, 0.88, 0.83, and 1.21 mg., respectively. With the
exception of the final month of the study the daily intakes

f thiamine were lower than the one mg. per day recommended

by the National Research Council. The mean daily intakes of

-43-

thiamine for subject H.L. during the four months of the
study were 1.00, 1.07, 1.02, and 1.04 mg., respectively.

Both subject's intakes of ascorbic acid were the greatest
during the first month of the study when cantaloupe was
available. In September the mean daily intake of this vitamin
was 77 mg. for subject H.L., and 80 mg. for subject P.P.

For both subjects the mean daily intakes of ascorbic acid
the following three months of the study were below the 70 mg.
recommended by the National Research Council. For subject
H.L. the mean daily intakes of ascorbic acid were 62, 48,

and 63 mg. in October, November, and December, respectively.
The comparable values for subject P.P. were 53, 57, and

68 mg., respectively.

A Subject H.L. had a relatively constant caloric intake
throughout the four months of the study. Her mean daily
caloric intakes were 1863, 1874, 1856, and 1878 calories
during September, October, November, and December, respectively.
No significant weight change was observed. Subject H.L.
weighed 147 pounds in September, and 150 pounds in April.

The caloric intake of subject P.P. increased at the
rate of approximately one—hundred calories per month
throughout the study. Her mean daily caloric intakes were
1644, 1716, 1831, and 1964 during September, October,

November,and December, respectively. Although no weight

 

 

-44-

records were obtained for this bedfast patient, it can be
assumed that her caloric intake was in excess of her energy
expenditure since the nurses reported that she was gradually
becoming too heavy for nursing care. Ohlson and others (50)
studied the dietary habits of aging women and reported that
it was possible that the total food needs were lower after
seventy. It appeared that this reduction occurred rather
abruptly in response to decreased number of activities and
interests. These observations suggest that further study
is needed in order to fully understand the nutritional
requirements of patients who must decrease activity, such

as those confined to bed.

Serum Ascorbic Acid Concentrations

 

The serum ascorbic acid concentrations of the two patients
were determined on the various regimens. These values along
with similar data gathered in two previous studies in this
laboratory (24, 26) are presented in Table V.

None of the serum ascorbic acid values recorded in the
present study are below the range of 0.73 to 1.21 mg. per
100 ml. of serum observed in healthy women on self-selected
diets (48). On September 15, 1955, the serum ascorbic acid
concentrations were 2.10 mg. for subject H.L., and 1.86 mg.
for subject P.P. A factor to be considered when interpreting

 

-45-

TABLE V
SERUM ASCORBIC ACID CONCENTRATIONS

_~_~_-_—_-—u‘-——-_—_-——_——_—o_—_~_—_—_‘_~fl—
——‘___——_————_———-—-—.-_—.——-.————.——-n————-—n————-——¢——_——n——-—u—————-—-—‘

 

 

 

Daily Vitamin Serum Ascorbic Acid
Date _kSupplement(mg) (Mg./100 ml. of serum)
Ascorbic PThiamIne Sfibject
Acid H.L. P.P.
Spring of 1954+ none none 0.15 0.22
Fall of 1954* 400 none 1.72 1.70
Fall of 1955
September 15 **none none 2.10 1.86
September 28 400 none 2.52 2.64
October 13 400 3 2.64 2.43
October 27 400 3 2.98 3.43
November 10 none 3 2.39 2.54
November 23 none. 3 1.62 1.75
December 8 none none 1.12 0.87
December 19 none none 0.83 0.85
+

Food and Nutrition Department (24)

* A daily oral supplement of 400 mg. of ascorbic acid had
been given for the preceeding 11 days. Furnival (26).

** Prior to September 1, both subjects had received 200 mg.
of ascorbic acid per day for a period of one month or
more. From September 1 to September 15, no supplements
were given.

-45-

these data is that both subjects had received 200 mg. of
ascorbic acid per day during August. The 200 mg. dose was
discontinued on September 1, thus there was a two-week
period when no vitamin supplement was given. It is likely
that the serum ascorbic concentration of the Parkinson's
patients recorded on September 15, partly reflect the 200 mg.
supplement administered during August.

During the period of the oral supplementation of ascorbic
acid, the serum values for both subjects were above 2.4 mg.
per 100 m1. of serum. The serum values for subject H.L. were
2.52, 2.64, 2.98, and 2.39 mg. on September 28, October 1,
October 27, and November 10, respectively. For subject P.P.
the serum values were 2.64, 2.43, 3.43 and 2.54, respectively.

After the discontinuation of the ascorbic acid supplemen-
tation on November 10, a downward trend in the serum values
for both subjects was observed. The serum ascorbic acid
concentrations for subject H.L. decreased from 2.39 mg.
percent on November 10, down to 0.83, mg. percent on
December 19. Similarly the values for subject P.P. declined
from 2.54 mg. percent down to 0.85 mg. percent.

From October 1 to the 27, daily supplements of 3 mg. of
thiamine and 400 mg. of ascorbic acid were administered to
the Parkinson's subjects. The serum values recorded in this
study reflected no apparent synergistic effect of these two

vitamins.

 

5‘41

In previous studies (24, 26) of these subjects the serum
ascorbic acid concentrations were 1.72 mg. for subject H.L.,
and 1.70 mg. for subject P.P. during the fall of 1954; 0.15
mg. and 0.22 mg. respectively, during the spring of 1954.

As previously stated the serum ascorbic acid values
recorded in the present study, during and immediately after
the period of ascorbic acid supplementation, were above 2.4
mg. percent which is considerably above the values of 0.73
to 1.21 mg. percent reported for healthy subjects (48).

Lowry and others (42) reported that ascorbic acid plasma
levels may rise to as much as 1.4 mg. percent on high intakes
of the vitamin, and that, in general, the kidney excretes
most of the larger doses, consequently the blood level does
not rise farther. Pijoan and Lozner (54) stated that plasma
ascorbic acid levels indicate essentially an overflow of a
phase of positive balance, and that when this becomes high
enough, urinary excretion of the vitamin takes place. Friedman
and others (25) observed that plasma concentrations above
1.5 mg. percent resulted in an increase of urinary excretion
of vitamin C. They found that below this plasma level urinary

clearance of the vitamin was of small magnitude.

Responses to Test Doses of Vitamins

 

The percent responses to test dose of ascorbic acid and
thiamine are presented in Table VI. Data from the fall of

1954 are also included (24).

 

.J‘.

v‘.-....1ut,

'-

_48_

TABLE VI

RESPONSES TO TEST DOSES OF VITAMINS

 

 

 

*Percefit5

Vitamin Date Subject Response
Ascorbic Acid* 1954 P.P. 4.1
Ascorbic Acid“ 1955 P.P. 15.9
Ascorbic Acid* 1954 H.L. 7.4
Ascorbic Acid” 1955 H.L. 15.0
Thiamine* 1954 P.P. 1.9
Thiamine“ 1955 P.P. 21.9
Thiamine* 1954 H.L. 1.6
Thiamine‘ 1955 H.L. 12.5

 

*Furnival (26).

 

-49-

On September 15, 1955, the percent responses to test
doses of ascorbic acid were 15.0 and 15.9 for subjects H.L.
and P.P., respectively. These data are below the range of
26 .to, 81 percent response observed for healthy women on
self-selected diets (5). Values from the fall of 1954 were
7.4 and 4.1 percent for subjects H.L. and P.P., respectively.

A low basal urinary excretion of the vitamin was observed
at the time the test dose was given. The average 24-hour
urinary excretion of a three-day period, week of September
12, 1955, for subject H.L. was 5.9 mg., and for subject
P.P. 3.3 mg. Following the daily oral dose of 400 mg. of
ascorbic acid, the urinary excretion of the vitamin increased.
For subject P.P. the greatest excretion of ascorbic acid was
observed during the week of October 25, with an average of
317 mg. per day for two 24-hour urine collections. For
subject H.L. the maximum excretion occurred during the week
of September 27, with an average of 217 mg. for three 24-hour
urine collections.

The relatively high serum ascorbic acid values observed
for these subjects along with the relatively low urinary
excretion of ascorbic acid following a test dose might indicate
a disturbed metabolism of ascorbic acid, particularly with
relation to kidney function. However, there is a possibility
that the total ascorbic acid values determined by the Roe and

-50-

Kuether method (57) may have included substances other than
ascorbic acid which reacted with the 2,4-dinitropheny1hydrazine.
Such substances may have arisen from drugs or medication and
would not ordinarily appear in blood serum of healthy
individuals. The fact that the rise and fall of the blood
serum ascorbic acid values paralleled the period of supple-
mentation, however, would indicate that the substance measured
was ascorbic acid.

Further the fact that the test dose of ascorbic acid was
of the same magnitude as the daily supplement may have
accounted for the relatively small increase in urinary excretion
of ascorbic acid after the test dose.

Certainly these findings are only indicative of the need
for further exploration of the metabolism of ascorbic acid in
patients with Parkinson's disease.

During the fall of 1955, the percent responses to the
test doses of thiamine were within the 12 to 28 percent range
observed in healthy women on self—selected diets (6). The
values obtained were 12.5 and 21.9 percent for subjects H.L.'
and P.P., respectively. In the study (24) of the fall of 1954,
the percent responses to a similar test dose were 1.6 for the
subject H.L. and 1.9 for subject P.P. No explanation is

apparent for the differences in response obtained in 1954 and

 

 

.I. 1.1] i. I - 1

 

lflllfl1

 

-51-

1955, however, a contributing factor may have been the levels
of thiamine supplied by the daily diet. During the 1955 study,
the diet of subject H.L. furnished over 1 mg. of thiamine per
day, and that of subject P.P. averaged a little less than 1 mg.
daily. Since thiamine intakes were not obtained in 1954,

no conclusive statement can be made.

Clinical Evaluations

 

The physical well-being of the subjects was assessed from
clinical examinations* of a physician, a physical therapist,
and a psychologist. The physician saw both subjects on
October 10, November 7, and December 18, 1955. He reported
that there were no changes either in the neurological findings
or blood pressure or general well-being of the subjects. During
each visit a specimen of the signature of each subject was
obtained. No significant differences were observed.

The physical therapist examined the subjects on August 29,
September 6, October 2, and November 30, 1955, and reported no
significant changes in the physical strength of either subject.
Due to paralysis of the lower extremeties of subject P.P.,

testing was difficult.

 

*The author is indebted to Dr. Mansel Dunn, Lansing,
Michigan, for the general physical examinations; to Mr. Face,
Ingham County Hospital, for the physical therapist's reports;
and to Dr. Fink, Ingham County Hospital, for the psychologist's
report.

-52-

The psychologist observed no psychological changes
attributable to the therapy.

The data on these two subjects do not permit the
establishment of definite conclusions, but they appear to
justify additional investigation of the ascorbic acid

nutrition of Parkinson's subjects.

-53-

SUMMARY

Literature pertinent to the etiology, symptoms, and
treatment of Parkinson's disease was reviewed.

Methods of assessing nutritional status were described.

The nutritional status of two patients with Parkinson's
disease at the Ingham County Hospital was assessed. Daily
food records were obtained. Urinary excretions of ascorbic
acid were determined by the 2,6~dichlorophenolindophenol
method, and of thiamine by the thiochrome method. Serum
ascorbic acid concentrations were estimated by the 2,
4-dinitr0phenylhydrazine method.

The three-month study was divided into four successive
periods during which the subjects were given daily oral
administrations of (a) a placebo, (b) 400 mg. of ascorbic
acid, (0) 400 mg. of ascorbic acid and 3 mg. of thiamine,
and (d) 3 mg. of thiamine.

Evaluation of the daily food records suggested that both
subjects consumed a low-iron diet. Their mean daily intakes
of iron were 31 and 35 percent below the 12 mg. per day
recommended by the National Research Council. The diets of
both subjects furnished adequate amounts of protein, calcium,
vitamin A, and riboflavin, but on the average supplied less
ascorbic acid and niacin than the amounts recommended by the

National Research Council. For one subject the mean daily

l

intake of thiamine was below one mg. except during the final
month of the study. The caloric intake of one subject
increased at the rate of approximately one-hundred calories
per month throughout the study, while the other subject had
a relatively constant intake.

Blood serum ascorbic acid values were 2.10 and 1.86 mg.
per 100 ml. for the two subjects during period (a). An
increase in blood serum ascorbic acid occurred during the
periods when 400 mg. ascorbic acid were given daily. Values
were decreased to 0.83 and 0.85 mg. per 100 ml. six weeks
after termination of the ascorbic acid supplement.

The percentage responses to a test dose of ascorbic
acid were 15.0 and 15.9. These values are below the range of
values observed for healthy women on self-selected diets.

The percentage responses to a test dose of thiamine were
12.5 and 21.9. This is within the range of values observed
in healthy women.

There were no significant alterations in the subjects
well-being or physical strength, or psychological status
attributable to the therapy as assessed from the clinical
examinations of a physician, a physical therapist, and a
psychologist.

The data on these two subjects do not permit the establish-
ment of definite conclusions, but they appear to justify
additional investigation of the ascorbic acid nutrition of

Parkinson's subjects.

(l)

(2)

(3)

(4)

(5)

(6)

(7)

(3)

(9)
(10)

(ll)
(12)

-55-

LITERATURE CITED

BAKER, A. B.: Treatment of paralysis a itans with
vitamin B6 (pyridoxine hydrochloride . J.A.M.A.
116: 2484, 1941.

BARGER, G., BERGEL, F., AND TODD, A.R.: A crystalline
Fluorescent dehydrogenation product from Vitamin B1'
Nature 136: 259, 1935.

BEESON, K.C.: The soil factor in human nutritional
problems. Nutr. Reviews 7: 353, 1949.

BESSEY, O.A.: A method 1231' determination of small
quantities of ascorbic acid and dehydroascorbic acid
in turbid and colored solution in the presence of
other reducing substances. J. Biol. Chem. 126: 771, 1938.

BREWER, W .D., CEDERQUIST, D.C ., STRINGER, C .J ., AND
OHLSON, M.A.: Studies of food intake and requirements
of women with active and arrested tuberculosis. Am.
Rev. Tuberc. 60: 455, 1949.

BREWER, W.D., TOBEY, H.L., DA HWEI PENG KAN, OHLSON, M.A.,
AND STRINGER, C.J.: Riboflavin, nitrogen, and thiamine
metabolism of women with active tuberculosis. J. Am.
Dietet. A. 26: 861, 1950.

BRITISH MEDICAL ASSOCIATION: Committee On Nutrition.
London, 1950.

BRYAN, A.H., TURNER, D.F., HUENEMANN, R.L., AND LOTWIN, G.:
The relation between plasma and dietary ascorbic acid.
Am. J. Med. Science 202: 77, 1941.

BURKE, B.S.: The dietary history as a tool in research.
J. Am. Dietet. A. 23: 1041, 1947.

BURKE, B.S., AND STUART, H.C.: A method of diet analysis.
J. Pediatrics 12: 493, 1938.

THE CANADIAN BULLETIN ON NUTRITION: 2: l, 1950.

CANADIAN COUNCIL ON NUTRITION: A new dietary standard
for Canada, 1949. Canadian J. Publ. Health 40: 420, 1949.

(l3)

(l4) ,

(15)

(16)
(17)
(18)
(19)
(20)
(‘21)
(22)

(23)

(24)

'CRAMPTON, E.W.:

CANADIAN COUNCIL ON NUTRITION: The construction and
use of dietary standards. Canadian J. Publ. Health
36: 272, 1945.

CAUSEY, K., HAUSRATH, M.E., RAMSTAD, D.F., FENTON, F.:
Effect of thawing and cooking methods on palatability
and nutritive value of frozen ground meat. II. Beef.
Food Research 15: 249, 1950.

CHAPPELL, 0.: Long-term individual dietary surveys.
Brit. J. Nutrition 9: 323, 1955.

CHRISTIAN, H.A.:
16th ed. New York:

p. 1358.

CHURCH, H.N., CLAYTON, M.M., YOUNG, C.M., AND FOSTER, W.D.:
Can different interviewers obtain comparable dietary
survey data? J. Am. Dietet. A. 30: 777, 1954.

Principles and Practice of Medicine.
Appleton-Century Co., 1947,

 

r LMQKIV—l
I .

CONNER, R.T., AND STRAUB, G.J.: Determination of thiamine
by thiochrome reaction. Ind. & Eng. Chem. Anal. Ed.
13: 380, 1941.

Cooperative Nutritional Status Studies in the Northeast
Region. Cornell Univ. Agri. Expt. Stn. Memoir No. 307,
(Northeast Regional Publication No. 5) March 1951.

The growth of the odontoblasts of the
incisor tooth as a criterion of the vitamin C intake
of the guinea pig. J. Nutrition 33: 491, 1947.

Problem situations in the treatment of

DOSHAY, L.J.:
J.A.M.A. 156: 680, 1954.

paralysis agitans.

EVELYN, K., MALLOY, H., AND ROSEN, C.: The determination
of ascorbic acid in urine with the photoelectric
colorimeter. J. Biol. Chem. 126: 645, 1938.

FOOD AND NUTRITION BOARD: Recommended Dietary Allowances,
Revised 1953. Natl. Acad. Sci. Natl. Research Council

Pub. 302, 1953, p. 9.

FOOD AND NUTRITION DEPARTMENT: Michigan State University,
E. Lansing, Mich.: Unpublished data.

 

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9
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(25)

(29)

(27)

(28).

(29)

(30)

(31)

(32)

(33)

(34)

(35)

-57-

FRIEDMAN, G.J., SHERRY, 8., AND RALLI, E.P.: The
.mechanism of excretion of vitamin C by the human
kidney at low and normal plasma levels of ascorbic
acid. J. Clin. Invest. 19: 685, 1940.

FURNIVALL, M.E.: Evaluation of The Nutritional Status
of Patients at a Rehabilitation Center, M.S. thesis.
Mich. State Univ. Library, E. Lansing, Mich., 1954.

GYORGY, P.: Vitamin Methods, N.Y. 10, N.Y.: Academic
Press, 1950, Vol. I, p.201.

HARRIS, L.J.: Brit. Med. Bulletin 12:
57 1 9 1956-

HARRIS, L.J., AND OLIVER, M.: Vitamin methods. The
reliability of the method for estimating vitamin C
by titration against 2; 6-dichlorophenolindophenol. ;
1. Control tests with plant tissues. Biochem. J. é
36: 155. 1942. 5

Vitamin C.

 

HARRIS, L.J., AND RAY, S.N.: Vitamin C and the supra-
renal cortex. II. Loss of potency of guinea-pig
suprarenals in scurvey. With notes on a method for
determining antiscorbutic activity (hexuronic acid)
by chemical means.' Biochem. J. 27: 303, 1933.

Chemical methods for determination of

HENNESSY, D.J.:
Ind. & Eng. Chem. Anal. Ed. 13: 216, 1941.

vitamin B1.

HENNESSY, D.J., AND CERECEDO, L.R.: The determination
of free and phosphorylated thiamine by a modified
thiochrome assay. J. Amer. Chem. Soc. 61: 179, 1939.

.HEWSTON, E.M., FISHER, M., AND ORENT-KEILES, E.:

Comparison of the 2,6-dichlorophenolindophenol and
2,4-dinitrophenylhydrazine methods with Crampton
bio-assay for determination of vitamin C values in
foods. U.S.D.A. Tech. Bulletin NO. 1023, 1951.

HUENEMANN, H.L., AND TURNER, D.: Methods of dietary
investigation. J. Am. Dietet. A. 18: 562, 1942.7

HUNSCHER, H.A., MACY, I.G.: Dietary study methods.
I. Uses and abuses of dietary study methods. J.
Dietet. A. 27: 558, 1951.

Am.

 

 

 

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(36)

(37)

(38)

(39)

(40)

(41)

(42)

(43)

(44)
(45)
(46)
(47)

(48)

-58-

KING, C.G.:
Vitamin 0.

Chemical methods for determination of
Ind. & Eng. Chem. Anal. Ed. 13: 225, 1941.

LEICHSENRING, J.M. AND WILSON, E.D.: Food composition
table for Short Method of dietary analysis. J. Am.
Dietet. A. 27: 386, 1951.

Technique and interpre-
Nutrition Abst. & Reviews.

LEITCH, I., AND AITKEN, F.C.:
tation of dietary surveys.

19: 507, 1950.

LEVERTON, R.M., AND MARSH, A.G.: Comparison of food
intakes for weekdays and for Saturday and Sunday.
J. Home Econ. 31: 111, 1939.

,n---1
3

LOCKHART, E.E., HARRIS, R.S., TAPIA, E.W., 11. LOCKHART,
H.S.; NUTTER, M.K., TIFFANY, V., AND NAGEL, A.H.: Study
of nutritional quality of dietaries by chemical analysis.
J. Am. Dietet. A. 20: 742, 1944.

"I- m. hi“

Effects of
Proc. Soc.

LOWRY, O.H., BESSY, 0.A., AND BURCH, H.B.:
prolonged high dosage with ascorbic acid.-
Exp. Biol. Med. 80: 361, 1952.

LOWRY, O.H., LOPEZ, J.A., AND BESSY, O.A.: The determina-
tion of ascorbic acid in small amounts of blood serum.
J. Biol. Chem. 160: 609, 1945.

MAYFIELD, H.L., AND HEDRICK, M.T.: Thiamine and ribo-
flavin retention in beef during roasting, canning,
and corning. J. Am. Dietet. A. 25: 1024, 1949.

mmmmLma
caloric value of diets.

The Atwater system of calculating the

J. Nutrition 28: 443, 1944.

MAYNARD, L.A.: Soils and Health. J.A.M.A. 143: 807, 1950.

MICKELSEN, 0., CONDIFF, H., AND KEYS, A.: The determin-
ation of thiamine in urine by means of the thiochrome
technique. J. Biol. Chem. 160: 361, 1945.

MINDLIN, H.L., AND BUTLER, A.M.: The determination Of
ascorbic acid in plasma. J. Biol. Chem. 122: 673, 1937.

MORGAN, A.F., GILLUM, H.L.,
status of the aging. III.
intake. J. Nutrition 55:

AND WILLIAMS, R.I.: Nutritional
Serum ascorbic acid and

“'31) 1955 o

 

 

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-59..

/
NUTRITION FOUNDATION, INC.: Present Knowledge In
Nutrition. 2nd ed.:N.Y. 16, N.Y., 1956, p. 76.

OHLSON, M.A., JACKSON, L., BEEGLE, R.M., DUNSING, D.,
AND BROWN, E.G.: Utilization of an improved diet
by older women. J. Am. Dietet. A. 28: 1138, 1952.

OHLSON, M.A., JACKSON, L., BOEK,
BREWER, W.D., AND BROWN, E.G.:
habits of aging women. Am. J.
1950. ‘

PENNEY, J.R., AND ZILVA, 3.3.: The determination of ‘ .
2:3 dikeo-l-gulonic acid: Biochem. J. 37: 39, 1943. i

J., CEDERQUIST, D.C.,
Nutrition and dietary
Pub. Health, 40: 1101,

PIJOAN, M., AND GERJOVICH, H.J.: Use of 2,4edinitro-
phenylhydrazine for the determination of ascorbic
acid. Science 103: 202, 1946.

PIJOAN, M., AND LOZNER, E.L.:
ficance of vitamin C in man.
14, 1944.

REIMANN, H.A.: Treatment in General Medicine, 4th ed.
Philadelphia: F.A. Davis, Co., 1948, p. 218.

The physiologic signi- _
New Eng. J. Med. 231: '“

 

ROBINSON, F.A.: The Vitamin B Complex. New York:

John.Wiley and Sons, Inc., 1951, p. 38.

ROE, J.H., AND'KUETHER, O.A.: The determination of
ascorbic acid in whole blood and urine through the
2,4-dinitropheny1hydrazine derivative of dehydro-
ascorbic acid. J. Biol. Chem. 147: 399, 1943.

SEBRELL, W.H., AND HARRIS, B.S.: The Vitamins. New York:
Academic Press Inc., 1954, Vol. I, p. 242.
.SHERMAN, H.C., LAMER, V.K., AND CAMPBELL, H.L.: The

Quantitative Determination of the Antiscorbutic
vitamin (Vitamin 0). Am. Chem. Soc. 44: 165, 1922.

SOMERS, G.F., AND BEESON, K.C.: The influence of climate
and fertilizer practices upon the vitamin and mineral
content of vegetables. Advances in Food Research 1:
291, 1948.

 

 

 

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(61)

(62)

(63)

(64)

(65)

(66)

-60-

STIEBELING, H.K.: Techniques of finding out what people
eat. Fed. Proc. 4: 253, 1945.

TRULSON, M.F.: Assessment of dietary study methods. I.‘
Comparison of methods for obtaining data for clinical
work. J. Am. Dietet. A. 30: 991, 1954.

TRULSON, M.F.: Assessment of dietary study methods II.
Variability of eating practices and determination of
sample size and duration of dietary surveys. J. Am.
Dietet. A. 31: 797, 1955.

VAN DEN BERG, A.S., AND MAYER, J.: Comparison of one-
day food record and research dietary history on a
group of obese pregnant women. J. Am. Dietet. A.

YOUNG, C.M., CHALMERS, F.W., CHURCH, H.N., CLAYTON, M.M.,
MURPHY, G.C., AND TUCKER, R.E.: Subjects' Estimation
of food Intake and Calculated Nutritive Value of the
Diet. J. Am. Dietet. A. 29: 1216, 1953.

I“: m'.-‘_m-IJWV 1‘0.”ij

ZELIGS, M.A.: Use of pyridoxine hydrochloride (vitamin B5)
in Parkinsonism. J.A.M.A. 116: 2148, 1941. ’

films «(z—’1‘}: an

 

 

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APPENDIX

 

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.1. I"...-

-62-

 

 

 

 

 

Food Record Form . RESEARCH
Name
Date
Day or Week
Amount Subjects'
FOOd Eaten Serving Eaten Comments
Breakfast

 

 

 

_." ‘.'

 

' What foods were on the tray that subject did not eat?
What did subject eat between breakfast and lunch?

 

Lunch

 

 

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".~.-’...I -". .M - «vs...»

 

 

What foods were on the tray that subject did not eat?
What food did subject eat between lunch and dinner?

 

Dinner

 

 

 

 

What foods were on the tray that subject did not eat?
What did subject eat before going to bed?

Form recorded by (initials)

 

A.

Ash-van».

I.

 

., r4.“ M 1,.

. .4” . ...i

1|. .1‘

ABSTRACT

The nutritional status of two patients with Parkinson's
disease at the Ingham County Hospital was assessed. Daily
food records were obtained. Urinary excretions of ascorbic
acid and thiamine, and blood serum ascorbic acid concentrations
were determined by chemical analyses. FT“

The three-month study was divided into four successive 1

periods during which the subjects were given daily oral

 

 

administrations of (a) a placebo, (b) 400 mg. of ascorbic

flu-BLUbgfi “ __,__

acid only, (c) 400 mg. of ascorbic acid and 3 mg. of thiamine,
(d) 3 mg. of thiamine only. Prior to period (a) the subjects

had received 200 mg. of ascorbic acid per day for one month or
longer.

Evaluation of the daily food records suggested that both
subjects consumed a low-iron diet. Their mean daily intakes
of iron were 31 to 35 percent below the 12 mg. per day
recommended by the National Research Council. The diets of
both subjects furnished adequate amounts of protein, calcium,
vitamin A, and riboflavin, but on the average supplied less
ascorbic acid and niacin than the amounts recommended by the
National Research Council. For one subject the mean daily
intake of thiamine was below 1 mg. except during the final
month of the study. The caloric intake of one subject increased
at the rate of approximately one-hundred calories per month

throughout the study, while the other subject had a relatively

constant intake.

 

I I I1. 1 .. IF?

‘M‘Gfllfl: an,

 

Blood serum ascorbic acid values were 2.10 and 1.86 mg.
per 100 ml. for the two subjects during period (a). An
increase in blood serum ascorbic acid occurred during the
periods when 400 mg. ascorbic acid were given daily. Values
were decreased to 0.83 and 0.85 mg. per 100 ml. six weeks
after termination of the ascorbic acid supplement.

The percentage responses to a test dose of ascorbic acid 8“
were 15.0 and 15.9. These values are below the range of
values observed for healthy women on self-selected diets.

The percentage responses to a test dose of thiamine

 

tfixd ". “.-.-2

were 12.5 and 21.9, which is within the range of values
observed in healthy women.

There were no significant alterations in the subjects
well-being or physical strength, or psychological status
attributable to the therapy as assessed from the clinical
examinations of a physician, a physical therapist, and a
psychologist.

The data on these two subjects do not permit the
establishment of definite conclusions, but they appear to
justify additional investigation of the ascorbic acid

nutrition of Parkinson's subjects.

 

 

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