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LIBRARY
Michigan State
_ University

 

 

 

 

This is to certify that the

thesis entitled

EFFECTS OF DIETARY CARBOHYDRATE AND FAT ON
PANTOTHENIC ACID STATUS IN RATS

presented by

Debbie E. Goqu

has been accepted towards fulfillment
of the requirements for

M. S. degree in Human Nutrition

 

JMII%W/
Major éofessé

Date 8/10/88

 

 

0-7639 MS U is an Affirmative Action/Equal Opportunity Institution

 

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RETURNING MATERIALS:
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EFFECTS OF DIETARY CARBOHYDRATE AND FAT 0N
PANTOTHENIC ACID STATUS IN RATS

BY

Debbie E Gould

A THESIS

Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of

MASTER OF SCIENCE

Department of Food Science and Human Nutrition

1988

ABSTRACT

EFFECTS OF DIETARY CARBOHYDRATE AND FAT ON
PANTOTHENIC ACID STATUS IN RATS

BY
Debbie E. Gould

Effects of diet on pantothenic acid (PA) status in
weanling rats were examined to determine if a diet high
in fat results in higher PA requirements. In the first
of two studies, 24-hour urine samples were collected
daily from three groups of rats fed a high-fat (67.5%
kcal from fat) or a high-carbohydrate (67.5% kcal from
carbohydrate) diet deficient in PA, or a high-
carbohydrate diet supplemented with 312 ug PA/ 100 kcal
(12 mg PA/ kg diet) and analysed for PA content. The
second experiment examined PA content of tissues, whole
blood and plasma, and plasma triglycerides of rats fed

high-fat or high-carbohydrate diets deficient or

supplemented in PA. No differences were seen between high-

fat and high-carbohydrate PA deficient groups with the

exception of plasma triglycerides which were significantly

elevated in the high-fat PA deficient diet group. In this

study, a diet high in fat did not result in a higher PA

requirement then a high-carbohydrate diet.

ACKNOWLEDGEMENTS

To Dr Won Song, my deepest gratitude and respect, for her
guidance, support, and encouragement throughout this study.
My thanks to Dr Romsos, Dr Schemmel, and Dr Ullrey for their
valuable input. To Cheryl, Dana, Karen, Mary, Mary, and
Sherri, my appreciation for their friendship and support.
Special thanks to my family for their never ending support and
encouragement.

ii

TABLE OF CONTENTS

IntrOduction 00.00.000.00...00.00.0000. ....... 0.0.01
Literature Review
1) Dietary Effects of a High Fat Diet

vs a High Carbohydrate Diet ........ .......... 3
2) Pantothenic Acid Requirements and
Deficj-ency0000000000000000000.000000000 ...... .4

3) Dietary Effects of a High Fat vs a
High Carbohydrate Diet with or without
Pantothenic Acid Supplementation..... ......... 7
4) Synthesis and Degradation of Coenzyme A
and Acyl Carrier Protein......................9
5) Biochemical Role of Panothenic Acid
and Coenzyme A in Carbohydrate and
Fat Metabolism..... .......................... 12
6) Pantothenic Acid Status and Plasma
Triglyceride Levels..........................15
7) Tissue Pantothenic Acid and Coenzyme A
Levels.......................................17
8) Urinary Excretion of Pantothenic Acid........25
9) Reported Values for Blood Pantothenic
Acid.........................................28
10)Brief Review of Methods......................31
Methods...........................................32
Results ..... . ............. . ............... ........35
Discussion........................................47
Summary and Conclusion............................53
Recommendations...................................56
Appendix A: Diet Compositions......... ....... .....57
Appendix B: Experiment 1
1) Food Intake.................................59
2) Weight......................................60
3) Urine Excretion:raw data....................61
: Experiment 2
4) Blood and Tissues:raw data .. ............. ..63
List of References..................... ......... ..77

TABLE
TABLE
TABLE
TABLE
TABLE
TABLE

TABLE
TABLE

ooq

LIST OF TABLES

REPORTED LIVER PANTOTHENIC ACID
CONCENTRATIONS: TOTAL AND FREE..........19
REPORTED HEART PANTOTHENIC ACID
CONCENTRATIONS: TOTAL AND FREE .........20
REPORTED KIDNEY PANTOTHENIC ACID
CONCENTRATIONS: TOTAL AND FREE..........21
REPORTED LIVER PANTOTHENIC ACID

CONTENT: FASTED VS FED..................24
REPORTED PANTOTHENIC ACID VALUES OF

RAT URINE...............................27
REPORTED PANTOTHENIC ACID VALUES

OF RAT WHOLE BLOOD AND SERUM............30
WEEKLY WEIGHTS OF RATS: EXPERIMENT 2....39
WEEKLY FOOD INTAKE OF RATS:

EXPERIMENT 2............................40

iv

FIGURE

FIGURE

FIGURE

FIGURE

FIGURE

FIGURE

FIGURE

FIGURE

«more

LIST OF FIGURES

COENZYME A SYNTHESIS..................11
PANTOTHENATE DERIVATIVES MEDIATE
CARBOHYDRATE, LIPID AND AMINO ACID
METABOLISM............................15
DAILY PANTOTHENIC ACID
EXCRETION...................... ..... ..36
HEART PANTOTHENIC ACID CONTENT:

TOTAL AND FREE........................42
KIDNEY PANTOTHENIC ACID CONTENT:

TOTAL AND FREE........................43
LIVER PANTOTHENIC ACID CONTENT:

TOTAL AND FREE........................44
WHOLE BLOOD PANTOTHENIC ACID
CONTENT...................... ....... ..45
PLASMA PANTOTHENIC ACID
CONTENT...............................46

INTRODUCTION

Pantothenic acid (PA), one of the B-vitamins, is
the vitamin moiety of coenzyme A and the
phosphopantetheine of acyl carrier protein. As a part
of these coenzymes, PA is involved in carbohydrate,
lipid and amino acid metabolism. While distinct
deficiency signs are not as apparent in a PA deficiency
as they are in some of the other B-vitamins, it is known
that PA is essential in humans and some animals for
normal growth, reproduction, and physiological
functioning. Although it is generally assumed that PA
is adequate due to the wide variety of foods in which it
can be found, subclinical deficiencies may in fact exist
(Chipponi et al., 1982). These deficiencies and their
resultant signs may be difficult to relate to low levels
of PA in the diet because of the lack of overt clinical
signs as well as due to a lack of information on the
metabolic pathways affected by a PA deficiency.

Although PA deficiency has been induced in rats,

the clinical signs are far from specific. Deficiency
signs reported in the literature include growth
retardation, a rough hair coat, muscle weakness, and

eventual death (Reibel et al., 1982).

While no Recommended Dietary Allowance has yet been
established for PA, it is possible that as a result of
the extensive biochemical functions of the coenzyme form
of PA (coenzyme A and acyl carrier protein) in lipid
metabolism, a high fat diet might increase the PA
requirement compared to the amount needed when a high
carbohydrate diet is consumed. No studies to date have
looked at such indices as tissue or blood PA content
along with growth rate and clinical observations to
support or disprove the theory of a high fat diet
lacking in PA more rapidly inducing a vitamin deficiency
than a similar diet high in carbohydrate. It is this
theory upon which we based our hypothesis that weanling
rats fed a high fat diet deficient in PA would become
depleted of this vitamin (as measured in urine, tissue and
blood) more rapidly than would rats fed a high
carbohydrate diet lacking in PA The goal of this study
was to determine whether the PA requirement in rats is
altered by the composition of fat to carbohydrate in the
diet.

Literature Review
1) Dietary Effects of a High Fat Diet vs a High
Carbohydrate Diet

Examining the dietary effects of a high-fat diet
versus a high-carbohydrate diet on PA status in rats,
may help to determine some of the underlying metabolic
processes adversely affected during a PA deficiency. It
has been repeatedly shown that the energy from dietary
fat is utilized more efficiently than is energy from
dietary carbohydrate during growth (Leveille and
Cloutier, 1987; Donato, 1987). Regardless of sex or
strain, rats fed a high-fat ration (40% - 87% kcal from
fat) have been reported to gain weight more rapidly and
to reach a higher maximum weight than those on a high-
carbohydrate ration (Mickelsen et al., 1955; Schemmel et
al., 1969; Schemmel et al.,1970; Oscai et al., 1984;
Siedler et al., 1962; Jen et al., 1987;). Rats fed a
high-fat diet had a higher percentage of their total
body weight as fat than rats on the grain ration
(Schemmel et al., 1969; Schemmel et al., 1970; Donato
and Hegsted, 1985).

Sprague-Dawley rats fed a high-fat diet were
observed to consume 20% more calories as compared to
rats fed a high-carbohydrate ration (Schemmel et al.,
1970). Neither the high fat nor the high carbohydrate

diet had any effect on body protein through the 100th

day of life, after which time the animals fed the high-
fat ration had 10% more body protein. This increase may
be explained by the increased blood flow and organ size
of the rats on the high-fat diet (Schemmel et al.,
1969). Champigny and Hitier (1987), feeding a 55% fat
diet vs a 12% fat diet found that at equal consumption
(kJ/day) of both diets, the high-fat group gained more
weight than the high-carbohydrate group.
2) Pantothenic Acid Requirements and Deficiency

Early studies suggested that rats require 80 ug to
100 ug of calcium pantothenate per day for maximum
growth. The rate of growth paralleled the level of PA
intake up to 80 ug per day. Studies on the urinary
excretion of this vitamin indicated a marked increase in
the urinary excretion of PA as the level of intake was
raised from 80 to 150 ug. Up to 80 ug the excretion was
very low, and at 150 ug about 50 ug of PA was excreted
suggesting that 100 ug is approximately the requirement
for the growing rat (Henderson et al., 1941). Weanling
rats consuming 5.3 9 food daily required 80 ug to 100 ug
of PA for optimal growth as compared with a food
consumption of 15.5 g and a daily requirement of about
25 ug of PA in 10 week old rats. This suggests that the
pantothenate requirement is reduced with age (Unna and
Richards, 1942). It has therefore been suggested that
other parameters in addition to growth be used in

determining the PA status of adult rats (Henderson et

al., 1942). Brown and Sturtevant (1949) reviewed the
literature concerning PA requirements of the growing rat
and concluded that the optimal concentration lies
between 0.8 to 1.0 mg PA per 100 g of diet. A lower
level of vitamin supply tended to slow down the growth,
while higher vitamin concentrations did not result in
greater weight gains. Barboriak et al., (1956)
confirmed 0.8 mg to 1.0 mg PA per 100 g diet as the
requirement for growing and adult rats. The accepted PA
requirement for growth of young rats is based on a
purified diet containing 5% corn oil and 385 kcal/ 100 g
diet (Barboriak et al., 1956; NRC, Committee on Animal
Nutrition, 1962).

Mature rats, fed a diet deficient in PA, stop
growing in about one month (Reibel et al., 1982) and die
later, displaying typical PA deficiency signs prior to
their death. These signs include retarded growth rate
in young animals, achromotrichia, scaly dermatitis,
rusty fur coloration and alopecia (loss of hair)
(Henderson et al., 1942; Williams, 1943). Neuromuscular
disorders, gastrointestinal malfunction, adrenal
cortical failure and sudden death are also commonly seen
signs of PA deficiency (Nelson et al., 1947). Anorexia,
myelin nerve degeneration, sciatic nerve and spinal cord
damage are a few of the numerous other common signs

observed during a PA deficiency (Williams, 1943).

Voris et al. (1942) concluded that PA has a
specific growth promoting effect unrelated to appetite,
even though they observed appetite depression in rats
fed a diet deficient in PA. Pantothenic acid deficient
rats have a depletion of lipids and impaired
carbohydrate metabolism as demonstrated by decreased
glucose tolerance and increased insulin sensitivity
(Baker and Frank, 1968). Weanling rats, after 2-3 weeks
on a PA deficient diet had a marked drop in urinary PA
excretion, loss of weight and rusty coloration of the
fur. After 5 weeks on the same diet, marked weight loss
was observed, and urine levels of PA remained low,
similar to the values seen at weeks 2-3 (Hatano, 1962).
While PA deficiency signs are well documented, the
underlying metabolic processes which result in these
signs have yet to be elucidated.

There is evidence indicating that PA is synthesized
by intestinal microorganisms in rats and other animals
(Giovannetti, 1982: Mameesh et al., 1959; Henderson et
al., 1941). It is not known whether this PA is
available for absorption and use by the body, and if it
is, how much is actually available. By allowing
coprophagy in the rats, another variable is added to the
experiment, in that it is possible that this presents an
alternate or additional source of PA in the diet.
Treatment of PA deficient rats with oxytetracycline

caused a reduction in the deficiency signs (Mameesh et

al., 1959) in the presence and absence of coprophagy.
Oxytetracycline stimulates growth in animals fed diets
limiting in PA (1 mg PA/kg diet; 360 kcal/100 gm). As
growth was seen with and without copraphagy it is not
known whether the sparing effects of oxytetracycline are
as a result of increased production of PA in the
intestinal flora or due to improved absorption of the PA
produced.
3)Dietary Effects of a High-Fat vs a High-Carbohydrate
Diet with or without Pantothenic Acid Supplementation.
It has been suggested that a high-fat diet without
PA supplementation would result in the more rapid
development of a PA deficiency than would a high-
carbohydrate diet without PA supplementation. Hatano et
al. (1966) theorized that the increased dependence on
fat for energy may alter PA metabolism in rats.
Myszkowska (1964) determined that male rats fed a
diet containing 12% fat (by weight) without PA
supplementation displayed signs of PA avitaminosis more
rapidly than did rats consuming a similar diet
containing only 6% fat. (The protein content of both
diets was fairly constant.) The avitaminosis was
determined by changes in hair gloss, bald areas behind
the ears, on the neck and around the eyes, and the
average body weight gain which fell to zero. When
calcium pantothenate was orally administered in

therapeutic doses of 0.5 mg per day, improved conditions

of the rats were noted after two weeks time. No data
were provided in this study concerning the food intake
of the rats on the two different dietary regimens.
Without this information, it is difficult to ascertain
whether the resultant signs were in fact due solely to
different levels of fat in the diet or if level of
consumption may have been a factor. Similar effects of
a high-fat diet on pantothenate status have been
reported in pigs (Sewell et al., 1962) where a 12% fat
diet (corn oil, by weight) was compared with a 2% fat
diet.

Williams et al. (1968), comparing the effects of a
6% fat (cottonseed oil, by weight) diet with an 18% fat
semi-purified diet with or without PA supplementation,
found lower body weights in rats on the high-fat (18%)
PA deficient diet than in rats on the 6% fat diet
deficient in PA after 6 weeks. 0n diets supplemented
with graded levels of PA (200 ug/100 g diet to 3000
ug/100 g diet) these same reseachers found no evidence
that a high level of dietary fat reduced the growth
response to limiting intakes of PA or increased the PA
requirement for maximal growth of young male rats.
Again in this study, while the authors indicate that
food intake was measured two to three times per week,
this information was not reported. It is thus difficult
for readers to accurately interpret the data. In both of

the previous studies involving rats, the original weight

of the rats was similar ( approx. 60 g ). Differences
in results cannot therefore be dismissed for this
reason.

Carter and Hockaday (1962) found a significant
decrease in the concentration of total carcass lipids in
PA deficient animals as compared to supplemented
animals. They further found that this decrease was
uninfluenced by the level of fat in the diet (5% fat vs
24 % fat by weight), a theory also suggested elsewhere
(Williams et al., 1968). Differences in body weight
were also significant between PA deficient and
supplemented groups, but relatively unaffected by
whether the diet was high or low fat. No conclusive
evidence to support or disprove the theory of a high-fat
diet more rapidly inducing PA deficiency has yet been
produced.

4) Synthesis and Degradation of Coenzyme A and Acyl
Carrier Protein

Coenzyme A (CoA), synthesized from PA which must be
obtained from dietary sources (Robishaw and Neely,
1985), is an essential cofactor for numerous enzyme
reactions. Phosphopantetheine of acyl carrier protein
is another metabolically active form of PA. The
biosynthesis of CoA from PA also requires ATP and
cysteine. PA is phosphorylated to 4’-phosphopantothenic
acid by the action of pantothenate kinase. This is the

rate limiting step in CoA formation, as pantothenate

10

kinase is inhibited by 4’—phosphopantetheine, CoA and
acetyl CoA (Abiko, 1975; Halvorsen and Skrede, 1982)).
4'-phosphopantothenic acid and cysteine are then
converted to 4'-phosphopantothenoyl-cysteine. This
reaction is catalyzed by 4'-phosphopantothenoyl-cysteine
synthetase. Next, a dicarboxylase enzyme converts 4'-
phosphopantothenoyl-cysteine to 4'-phosphopantetheine.
These first three enzymes are located exclusively in the
cytosol (Skrede and Halvorsen, 1979). In the following
two steps in this pathway, 4'-phosphopantetheine is
adenylated to form dephospho-coenzyme A which is
phosphorylated at the 3' position of ribose to form CoA.
These last two reactions mediated by dephospho CoA
pyrophosphorylase and kinase respectively, are present
in both the cytosol and the mitochondria (Novelli et
al.,1949; Brown, 1958; Robishaw and Neely, 1985; Abiko,
1975).

The proposed route of degradation of CoA is the reverse
of the sythesis up to the point of 4’phosphopantetheine.
4'-phosphopantetheine is thought to breakdown to pantetheine,
and to PA and cysteamine. The cysteamine is further degraded
to hypotaurine and taurine. The fate of PA as a result of CoA
degradation is unknown. It may reenter the synthetic pathway
or be excreted (Abiko, 1975; Robishaw and Neely, 1985). There
are some problems with the proposed route of degradation of
CoA, one of which is that the majority of CoA is found in the

mitochondria, but the first degradative enzyme is lysosomal

II

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d3 As on
Pantothenic acid 4'-phosphopantothenic acid
coon ac-ou
uo-p-ocu’T-cucounca cu councucn'sa ' —— — =— —->
d cu . '

4'-phosphopantothenyloyl-l-cysteine

 

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HO-P-OCH C-CHCONHCH CH CONHCH CH SH —— — - — ->
| I ‘
OH CH

4'-phosphopantetheine

a $403

 

HO-P-OCH C-CHCONHCH CH CONHCH CH SH —-— —-—— -— >
O CH

HO-P-O

 

dephosphoCoA
H f-OH

AHO-P-OCH C-CHCONHCH CH CONHCH CH SH

CH

 

CoA
FIGURE 1: SYNTHETIC PATHWAY OF COENZYME A

(Brown 1958, Abiko,1975).

12

(Robishaw and Neely, 1985).

Experimentally, PA can be released from CoA by
incubation of the CoA with the enzymes pantetheinase and
alkaline phosphotase. (A step by step protocol can be
found in Wyse et al., 1985.) This enzymatic treatment
to release the bound form is essential for PA
determination by means of RIA as this procedure can only
measure free PA (Wyse et al., 1979). Most PA in
biological materials exist as CoA. In acyl carrier
protein, phosphopantetheine is linked to serine via a
phosphodiester bond (Majerus et al., 1965) and even with
enzymatic treatment, the pantothenate in acyl carrier
protein will not be released.

5) Biochemical Role of Pantothenic Acid and CoA in
Carbohydrate and Fat Metabolism

Dietary deficiency of PA in animals results in a
wide spectrum of biochemical defects that eventually
manifest themselves as signs described previously.

Abiko (1975) has listed more than 70 catalytic reactions
that involve CoA or phosphopantetheine as cofactors.

The physiological role of PA as a component of CoA, is
important in the release of energy from carbohydrates,
in gluconeogenesis, in the synthesis and degradation of
fatty acids, in the synthesis of ketone bodies, as well
as in the synthesis of sterols and steroid hormones,
porphyrins and other compounds (Goldman and Vagelos,

1964, Abiko 1975).

13

The role of PA in carbohydrate metabolism comes
from the breakdown of glucose to ATP via glycolysis and
the citric acid cycle. At the end of glycolysis,
pyruvate combines with CoA to form acetyl CoA which can
then be incorporated into the citric acid cycle. In the
tricarboxylic acid cycle, CoA functions as an acyl
acceptor for the pyruvate and alpha-ketoglutarate
dehydrogenase complexes forming acetyl CoA and succinyl
CoA respectively. In gluconeogenesis, PA again as a
component of CoA, is necessary for the activity of the
pyruvate carboxylase enzyme which converts pyruvate to
oxaloacetate (0AA), as the first step in synthesizing
glucose from pyruvate. Biotin, a needed cofactor in the
conversion of pyruvate to 0AA, is not carboxylated (a
necessary step in the conversion) unless acetyl CoA (or
acyl CoA) is bound to the enzyme pyruvate carboxylase.
Pyruvate carboxylase also plays a critical (anapleurotic) role
in maintaining the level of citric acid cycle intermediates
at appropriate levels (Stryer, 1975).

Fatty acid synthesis, which takes place in the
cytosol, requires PA as a component of acyl carrier
protein (ACP). The fatty acid chain is elongated
(starting with malonyl ACP) by the sequential addition
of two carbon units derived from acetyl CoA (Stryer,
1975).

Fatty acids are oxidized in the mitochondria.

Prior to entry into the mitochondria, the fatty acids

14

are activated in a reaction involving CoA. These

activated acyl CoAs are then transported inside the
mitochondria by means of a special transport mechanism
involving carnitine. Once inside the mitochondria, the

acyl CoA compound is degraded by a recurring sequence of
B-oxidation involving CoA. The final product of the
degradative pathway is acetyl CoA for even numbered

fatty acids or propionyl CoA for odd numbered fatty

acids. Both these products can be incorporated in the
citric acid cycle and used for energy (Stryer, 1975).

If excess fatty acids are broken down to acetyl

CoA, ketone bodies will be formed. These ketone bodies

also involve CoA in their synthesis. Acetyl CoAs are
converted to acetoacetate which is further converted to
hydroxybutyrate and acetone. Acetyl CoA is the carbon

source for the biosynthesis of prostaglandins, cholesterol,
steroid hormones and. other compounds. Succinyl CoA, a
tricarboxlic acid cycle intermediate, is an essential
precursor for porphyrins and hence for hemoglobin and

cytochromes.

15

CARBOHYDRATE

FATTY ACIDS J]

PYRUVATE

1] yo ACIDS
ACETYL CoA

TCA STEROIDS
CHOLESTEROL
FIGURE 2: PANTOTHENATE DERIVATIVES MEDIATE

CARBOHYDRATE, LIPID AND AMINO ACID METABOLISM

6) Pantothenic Acid Status and Plasma Triglyceride Levels
Triglycerides are highly concentrated stores of
metabolic energy. For triglycerides to be utilized as
energy they must first be broken down into glycerol and
free fatty acids. Glycerol can be converted into
pyruvate or glucose in the liver, while free fatty acids
are degraded by beta-oxidation (which requires CoA) to
acetyl CoA which can then be incorporated into the
citric acid cycle (Stryer, 1975). Triglycerides from
the diet are transported by chylomicra from the
intestine to adipose tissue where they are stored until
needed, while triglycerides synthesized endogenously are
carried by very low density lipoproteins (VLDL) to the
adipose tissue. The fate of exogenous fatty acids may in
part be regulated by the cytosolic carnitine to CoA

ratio (Idell-Wenger et al., 1978; Idell-Wenger and

16

Neely, 1978; McGarry et al., 1975). Changes in the CoA
levels result in a concomitant change in PA metabolism.
An inadequate supply of PA has been shown to raise

blood triglyceride and free fatty acid levels in rats
fed a high fat diet. After two weeks on a high fat diet
(30% of kcal) with graded amounts of PA (0 mg/kg diet to
1600 mg/kg diet), Peterson et al. (1987) observed
significant differences in rat serum triglycerides and
free fatty acids (FFA). Those animals receiving less PA
in their diets had the higher serum triglycerides and
FFAs. The greatest differences in triglyceride levels
were observed to be between the deficient (0 mg/kg) and
the normally supplemented (16 mg/kg) groups suggesting
a vitamin related effect rather than a pharmacologic
one. Triglyceride differences were found to be
significant long before weight loss and other
conventional signs of deficiency appeared in this study.
In the high fat PA deficient group, triglyceride levels
were subsequently reduced with supplemental pantothenate
(amount not indicated) or pantethine.

Pantethine is a derivative of PA which contains PA
and cysteamine. It can be converted biochemically into
CoA and is used in some countries as a natural hypolipemic
drug to decrease total serum cholesterol and
triglycerides. The cholesterol lowering effect has been
shown to be due to the cysteamine fragment of pantethine

(Wittwer et al., 1985). Although still unproven, it is

17

widely assumed that the hypotriglyceridemic effect of
pantethine is as a result of increased levels of
pantothenate derivatives (i.e. as a result of the PA
portion of pantethine). Proposed mechanisms for the
hypotriglyceridemic effect include fatty acid oxidation
stimulation (Kameda and Abiko, 1980), activation of the
citric acid cycle (Prisco et al., 1984) and lipoprotein
lipase activation (Noma et al., 1984).

Hypertriglyceridemia can be caused by increased
secretion of VLDL by the liver, decreased degradation by
lipoprotein lipase or both. As PA is incorporated into
CoA which is extensively involved in lipid metabolism,
certain reactions may be suppressed by an inadequate
supply of CoA as a result of insufficient PA in the
diet. This could result in elevated serum
triglycerides.
7) Tissue Pantothenic Acid and CoA Levels

Levels of PA acid and CoA have been measured
extensively in the liver, heart, and kidney of rats for
a variety of reasons, using a number of different
experimental conditions. In general, it has been
concluded that when animals are fed on a diet deficient
in PA, tissue levels of the vitamin will drOp 70% to 90%
(Reibel et al., 1982) while CoA levels in the same
tissue remain relatively unchanged from the norm (liver
250-450 nmol/g wet tissue; heart 130-180 nmol/g; kidney

120-300 nmol/g) as seen in tables 1-3 below (Hatano,

I8

1962; Reibel et al., 1981, 1982; Moiseenok et al.,
1986).

Although tissue levels of CoA remain fairly
constant in a PA deficient state, the distribution of
CoA between free and acyl ester forms may vary over a
wide range. The ratio of free CoA to acyl CoA is
important in determining the rate of a number of key
metabolic reactions including those catalyzed by acyl
CoA synthetase (activates fatty acids prior to entering
the mitochondria) (Oram et al., 1973) pyruvate
dehydrogenase (pyruvate to acetyl CoA) and
alphaketogluterate dehydrogenase (alphaketogluterate to
succinyl CoA) (Rabinowitz and Swift, 1970). One
suggested explanation for the observed lack of change in
CoA levels in a PA deficiency is that the organs of the
body normally contain a large excess of PA (Reibel et
al., 1982).

In tables 1-3 below, it can be seen that there is
alot of variation in reported values of CoA and PA in
the tissues. Explanations for this variability include
differences in age and strain of rats, as well as
individual differences among animals and differences in
methodology between different laboratories.

Livers of rats fed PA deficient diets for 4 weeks,
contain about 30% of normal levels of free PA (Reibel et
al., 1982). Hatano (1962) measuring 9 different tissues

found that liver had the highest value of PA in the

19

TABLE 1: REPORTED LIVER PANTOTHENIC ACID
CONCENTRATIONS: TOTAL AND FREE
(nmol PA/g wet tissue)

Treatment Total Free Reference
Control 388 i 54 11.3 + 4.4 Smith et al
’ 1978
Control 251 t 46 27.2 t 3.3 Smith
1978
Control 264 + 12 Israel and
T Smith 1987
Control 445 (97.5 ug/g) 183 (40 ug/g) Hatano
Deficient 217 (47.5 ug/g) 137 (30 ug/g) 1962
Control 477 t 15 * 26 i 3 * Reibel et
Deficient 400 t 21 * 7 i 1 * al.,1982

all numbers represent mean i std dev
* values presented as nmol PA/ 9 dry tissue

20

TABLE 2: REPORTED HEART PANTOTHENIC ACID
CONCENTRATIONS: TOTAL AND FREE
(nmol PA/g wet tissue)

Treatment Total Free Reference
Control 130 i 15 94 t 21 Smith 1978
Control 138 i 36 36 t 6 Smith et a1
1978
Control 141 i 10 58 :_10 Israel and
Smith 1987
Control 180 (39.5ug/g) 73 (16 ug/g) Hatano
Deficient 85 (18.5 ug/g) 48 (10.5ug/g 1962
Control 617 i 26 * 284 :14 * Reibel et
Deficient 630 i 15 * 32 t 2 * al., 1982

all numbers represent mean i std dev
* values presented as nmol PA/ g dry tissue

21

TABLE 3: REPORTED KIDNEY PANTOTHENIC ACID
CONCENTRATIONS: TOTAL AND FREE
(nmol PA/g wet tissue)

Treatment Total Free Reference
Control 128 i 4 31 t 9 Israel and
Smith 1987
Control 324 (71 ug/g) 173 (38 ug/g) Hatano
Deficient 176 (38.5 ug/g) 130 (28.5ug/g) 1962
Control 474 t 49 * 322 t 35 * Reibel et
Deficient 489 t 52 * 27 1,11 * al., 1982

all numbers represent mean t_std dev
* values presented as nmol PA/ 9 dry tissue

22

animals on control diet (457 nmol/g). In more recent
studies, heart has been found to maintain higher pools
of PA and lower total CoA content than liver (Smith,
1978). Further study has quantified this observation,
reporting that heart (total organ) contains about ten
times as much free PA as liver (Reibel et al., 1981).
This relates in part to the different functions of the
two organs with respect to fat metabolism. In heart,
the major fate of fatty acids is oxidation. This
correlates well with the fact that cytosolic total CoA
levels are low compared with those of carnitine
(Lopaschuk et al., 1986). In liver, cytosolic CoA
levels are much higher and fatty acid conversion to
complex lipids is a more prominent function of the
tissue (McGarry et al., 1975).

The kidney conserves whole body PA through
increased reabsorption (Reibel et al., 1981) when
dietary intake of PA is low. It is further believed
that this reabsorption is an active process at
physiological plasma concentrations and, that at higher
concentrations of PA, tubular secretion of plasma PA
occurs (Karnitz et al., 1984). When rats were fed PA
deficient diets for 4 weeks, CoA levels were maintained
in the kidney tissue, while free PA content was
reportedly reduced from 35-40% (Carter and Hockaday,
1962) to 80% (Reibel et al., 1981). Possible

explanations for this phenomenon are binding of PA to

23

the plasma proteins and/or tubular reabsorption of PA by
the kidney. Since no specific plasma protein binding of
PA was found, it appears that conservation mechanism for
free PA in the rat is through tubular reabsorption
(Karnitz et al., 1984). Tubular reabsorption of PA has
also been observed in dogs (Taylor et al., 1974). It
thus appears that two processs regulate excretion of PA
in the kidney, tubular reabsorption and tubular
secretion.

Differences were also noted in the PA content of
the organs between animals fasted prior to sacrifice
versus those fed right up until sacrifice (Table 4).
Fasting PA deficient animals resulted in increased PA
levels in heart, liver and kidney as compared with non-
fasted deficient animals (Reibel et al., 1981, 1982;
Smith et al., 1978). These researchers similarly found
the levels of pantothenate to rise in fasted versus fed
control rats. Smith et al. (1978) postulated that the
higher PA acid content in the liver of fasted versus fed
rats may reflect an influx of PA from other tissues not
being measured. Another possible explanation is that
the increase in fasting CoA levels arises from the
drastic decrease seen in the level of fatty acid
synthase in the liver during fasting. Since the enzyme
contains 4'-phosphopantetheine as a prosthetic group,
degradation of the enzyme could release 4'-

phosphopantetheine which could subsequently be converted

24

TABLE 4:REPORTED LIVER PANTOTHENIC ACID
CONTENT- FASTED VS FED
nmol PA/ 9 wet weight tissue

Group PA Supplemented PA Deficient Reference
Fed (26 i 3) * (7t 1 )* Reibel et al.
Fasted (46 1 4) * (18: 3) * 1981, 1982
(48h)

Fed 4.8 1.5 Israel and Smith
Fasted 3.6 t.6 1987
(24h)

Fed 6.3 1.6 "
Fasted 4.4 t.6

(24h)

Fed 3.3 t1.1 Smith et al.
Fasted 5.2 t 2.9 meal fed 1978
(21h)

Fasted 11.3 t4.4 ad lib "
(21h)
* (nmol/g dry wt)

numbers represent mean i std dev

25

to PA (Reibel et al., 1982). (The authors (Reibel et
al.) did not measure the activity of fatty acid
synthtase). Israel and Smith (1987) have shown higher
liver PA levels in fed versus fasted control rats. One
explanation for this discrepancy is the differences seen
in the length of time the animals were fasted prior to
sacrifice (24 vs 48 hrs) and in the type of diet they
were consuming during this time (meal fed vs ad
libitum). Treatment of the tissue samples was similar
in all studies.
8) Urinary Excretion of Pantothenic Acid

PA excretion in the urine is generally accepted
as an indication of nutritional status with regard to
this vitamin (Tao and Fox, 1976). A number of studies
(Srinivasan et al., 1981, Fox and Linkswiler, 1961,
Oldham et al., 1946; Cohenour and Calloway, 1972; Song
et al., 1984) looking at both intake and excretion of PA
in humans, suggest that urinary excretion of PA is
related to dietary intake. It has been shown that a
certain amount of excretion represents minimal daily
requirements of PA for tissue maintenance (Fox and
Linkswiler, 1961) although no specific quantity has been
identified at this time. These studies also suggest
that little or no bound PA appears in the urine.

Similar results have been found in rats with
respect to urinary excretion correlating well with

dietary intake (Hatano, 1962). It has been observed as

26

well, that urinary excretion of PA increases directly
with the age of the rat fed ad libitum (Nelson et al.,
1947; Tao and Fox, 1976) PA deficient or supplemented
diets. This is consistent with the theory that the PA
requirement of the rat decreases with age (Henderson et
al., 1942; Unna and Richards, 1942). In these studies,
all experimental diets were deficient in PA, therefore
increased urine excretion reported with age could not be
as a result of increased food intake and thus increased
PA intake. Many studies have measured the urinary PA
excretion in rats (Hatano et al., 1967; Hatano, 1962;
Nelson et al., 1947; Tao and Fox, 1976; Reibel et al.,
1981). A marked decrease in urinary PA was observed 11
days after initiation of a PA deficiency experiment in
young weanling rats. The excretion rate levelled off at
1 ug/day after 3 weeks on the deficient diet (Hatano,
1962). A summary is found in Table 5.

Although different methods were used to determine
urine PA levels (microbiological assay or RIA), good
correlation between these techniques has been documented
(Wyse et al., 1979; Srinivasan et al., 1981). Urinary
PA levels seem to be by far the most indicative
parameter when determining short term dietary
sufficiency of PA although further information is still
needed to relate urinary PA values to blood and tissue

PA concentrations.

27

TABLE 5 :REPORTED PANTOTHENIC ACID VALUES OF

RAT URINE
DIET PA IN DIET FOOD INTAKE URINARY PA REFERENCE
weight ug/100g g/day ug/24 hrs
68% sucrose Hatano
17% casein 0 N/A 1.0
10% fat 1962
basal N/A N/A 32.1:6.1 Hatano
et al.
1967
24% casein
64% sucrose 0 N/A 1.6 Nelson
8% fat et al.
1947
68.8% sucrose
20% casein 0 8.5 2.4 Tao
5% fat and Fox
1976
" 200 12.2 5.0 "
" 2000 15.6 69.6 "
N/A N/A N/A 250 i 37 Reibel
et al.
1981
20 .67 wkl
1.0 wk2
. 60 wk3 Henderson
1.4 wk4 et al.
2.2 wk5 1942
1.7 wk6
1.3 ug/24 hr average

N/A = not available

28

9) Reported Values for Blood Pantothenic Acid

Generally, the vitamin content of blood is
influenced by the amount in the diet. In the case of
some vitamins, level in the blood is used as a criterion
of the adequacy of intake. Many human studies have been
done to determine the PA levels of whole blood and serum
(or plasma ) in hopes of defining such a criterion to
distinguish adequate from inadequate PA intake in the
diet or PA status in the person in general. The results
of these studies have produced a wide range of numbers.
In humans, serum is known to contain only small amounts
of PA all in the form of free PA (Wyse et al., 1985).
Sauberlich et al., (1974) suggested that a total level
of PA less than 5 nmol/ml of whole blood may be
indicative of low levels of PA in the diet in human
subjects. Baker and Frank (1968) also with humans,
suggested a level of less than 0.7 nmol/ml whole blood
as a hypovitaminosis. This large variation may be due in
part to variations in technique between laboratories.
Cohenour and Calloway (1972) failed to relate
dietary intakes with blood levels of PA in their study
of pregnant teenagers. Srinivasan et al., (1981) also
found no correlation between PA intake and blood levels
of PA in an elderly population. Even when PA
supplementation was used, no correlation was seen
between PA intake and blood PA concentrations

(Srinivasan et al., 1981). This might be the result of

29

excretion of all excess PA being consumed. Fry et al.,
(1976) found that although blood PA levels decreased in
unsupplemented subjects and remained constant in
supplemented subjects, in general, blood PA levels
responded less readily to intake than did urinary PA
levels.

In animal studies, (rats and chickens) Pearson et
al., (1946) reported that the level of PA in the whole
blood, the plasma, and the cells is influenced by the
amount of PA in the diet. Pearson (1941) reported that
from 44 - 62% of the PA in blood occurs in the plasma.
Published data on the concentrations of PA in the serum
show large variations, even within the same species
(Robinson, 1966). These variations may be due to the
methods used to assay PA, differences in sample
preparation or due to genuine large variations existing
within and among species. RIA determination of PA in
blood seems to give more consistent results than
previously obtained with other methods as shown by the
results of Israel and Smith (1987), and Reibel et al.
(1981, 1982) in Table 6. Whole blood values range from
2.28 nmol/ml to about 4.57 nmol/ml whereas serum values
vary modestly from 1.5 nmol/ml to 1.75 nmol/ml (Table
6). Many of the studies reporting PA levels in the
whole blood and serum fail to provide information on PA
intake, making interpretation of their findings

difficult and creating problems when values from

30

TABLE 6:REPORTED PANTOTHENIC ACID VALUES OF

RAT WHOLE BLOOD AND SERUM

PA in SERUM
nmol/ml
(PLASMA)

REFERENCE

PA/DAY PA in WHOLE BLOOD
INTAKE nmol/ml

N/A 2.57+.32 *
390ug/day 4.84:1.64 *

" 2.461.32 *

" 4.15: 1.00 *

(fasted 24 hr)
" 4.75:.55 *

N/A

50 mg PA/
kg diet

(1.441: .26)*

1.75: 0.17

1.54:0.30

undetected

Hatano et
al.,1967

Israel and
Smith 1986

Reibel et
al. 1981

Reibel et
al. 1982

* units converted from ug/ml

numbers represent mean i std dev

31

different studies are compared.
10) Brief Review of Methods

PA concentrations of tissue, blood, or foodstuff
can be determined by means of radioimmunoassay (RIA).
D-pantothenic acid is conjugated with bovine serum
albumin by use of a bromoacetyl derivative of PA, and
antibody to this antigen is raised by injecting it into
footpads of rabbits. A 1:100 dilution of this resulting
antiserum is incubated with radiolabelled PA. The
antibodies are precipitated and dissolved, and the
radioactivity of the solution measured in a liquid
scintillation counter (Wyse et al., 1979). By using
known amounts of labelled and unlabelled PA in the RIA,
a standard curve can be derived based on the percent
binding of the unlabelled PA. This curve can be used in
determining the amount of PA in biological samples being

analyzed.

METHODS

This study included 2 experiments:

W

This experiment was undertaken to measure
daily PA excretion in urine of rats fed one of three
diets: high-fat PA deficient (67.5% kcal from fat);
high-CHO PA deficient (AIN-76 diet 67.5% kcal from
CHO);high-CHO PA supplemented (312 ug PA/ 100
kcal;Appendix A). Fifteen weanling Sprague-Dawley rats
(Harlan Sprague-Dawley Inc. Indianapolis, Indiana) were
randomly divided into equal groups (5 rats per group),
housed individually in metabolism cages, and fed one of
the three experimental diets described above.
Twenty-four hour urine samples were collected daily in
the mid morning hours. Each urine sample was diluted to
50 ml with distilled water and 100 ul of this fresh
solution was then used to determine PA content of the
urine daily by means of radioimmunoassay (Wyse et a1,
1979). Food intake was measured daily and body weights
were recorded weekly (Appendix B).

After a one week depletion period, all groups were
fed a stock diet for repletion (Wayne Research Animal
Diet, Chicago, Illinois). This was done to observe if
differences could be detected between fat and CH0 PA
deficient groups with respect to repletion of PA. Nine

days after the beginning of the repletion period, the

32

 

33

rats were again randomly divided into three groups and
fed the three experimental diets as above. The
redepletion phase of the experiment was planned for a
week because the rapid decline in PA in the urine
observed in the first depletion period had not
previously been reported in the literature.

W

In this experiment, eighty-seven male weanling
Sprague-Dawley rats were randomly divided into four
treatment groups: high-fat PA deficient; high-CHO PA
deficient; high-fat PA supplemented pair fed to high-fat
PA deficient; high-fat PA supplemented ad libitum (diet
composition as in the first experiment). PA
supplemented groups received 312 ug PA/ 100 kcal (17 mg
PA/kg high-fat diet; 12 mg PA/kg high-CHO diet). Rats
were housed individually in wire bottom cages.

At weeks 0.5, 1.5, 2.5, and 4.0 five rats per group
were killed by exsanguination via cardiac puncture and
liver, kidneys and heart were removed at this time.
Organs were rinsed in 0.9% saline and weighed. Heart
was divided into two equal portions approximately 0.3 g
each, to measure total and free PA content. One kidney
was used for measuring total PA and the other kidney was
used to measure free PA. Two liver pieces, weighing
approximately 0.5 g each, were used for these two
determinations. These tissues were then homogenized

using the Brinkmann polytron at speed setting 3 for one

34

minute. Kidney and heart were homogenized with 0.5 ml
of distilled water, while 1.0 ml of distilled water was
used for liver.

Tissue samples homogenized for free PA measurements
were immediately deproteinized with equimolar saturated
Ba(OH)2 and 10% ZnSO4 (approximately 300 ul total volume,
the ratio being determined by titration using
phenolphthalein as an indicator) and centrifuged at
4,000 x g for 10 minutes in a Sorvall Superspeed RC2-B
centrifuge. Supernatants were stored at -200 C. Tissue
samples to measure total PA were treated with an enzyme
mixture containing 30 units alkaline phosphatase and 15
units pantetheinase in phosphate buffered saline (PBS).
The optimum amount of enzyme needed for each sample to
release PA bound as CoA had previously been determined
in this laboratory. These samples were then incubated
overnight (8-10 h) at 370 C.

Whole blood was separated into two portions. One
portion was divided into packed cells and plasma by
centrifugation at 4,000 x g for 10 minutes in a Sorvall
Superspeed RC2-B centrifuge and the other portion was
saved as whole blood. Whole blood was hemolyzed by
rapidly freezing and thawing the samples three
consecutive times. Two 50 ul aliquots of the hemolyzed
blood was used for measurement of total and free PA.

The aliquot for free PA was immediately deproteinized

with equimolar saturated Ba(OH)2 and 10% ZnSO4

35

(approximately 200 ul total volume). The aliquot for total
PA measurement was treated with 10 units of alkaline
phosphatase and 20 units of pantetheinase in PBS prior to
deproteinization. Plasma was immediately deproteinized and
all the supernatants were stored at -200 C.

Fifty ul of tissue sample supernatant and 100 ul of
blood supernatant were analyzed for PA content by means of
RIA. Plasma triglyceride concentrations were measured by
means of colorimetric determination at 410 nm (Fletcher,
1968). Statistical analysis was performed in both studies
using ANOVA followed by the Bonferroni t-test (Miller, 1977)
which allow any number of contrasts to be compared using the
type I error rate based on all the contrasts.

RESULTS

In experiment 1, it was found that PA content of the
urine decreased significantly after one day on the
experimental diets (Figure 3). At day 8, when all
groups were fed stock diet for PA repletion, differences
between control and previously deficient groups became
significant for the following 2 days. No significant
differences were found at any time between the group
receiving the high-fat PA deficient diet and the high-

CHO PA deficient diet. After switching the two groups
on deficient diets onto the stock diet, increased

excretion of PA was apparent by the following day. No
significant differences were shown between the high-fat

and high-CHO deficient groups although excretion was

36

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37

significantly lower in the groups which had been on
deficient diets as compared to the group which had been
on the control (PA supplemented) diet. Nine days after
beginning to consume a stock diet, no significant
differences were seen among PA excretion of the

three groups (268145; 234119; 24717 ug PA/

24 hr for high-fat PA deficient, high-CHO PA deficient
and high-CHO PA supplemented groups respectively). At
day 9, rats were randomly redivided into three treatment
groups of 5 rats each, and immediately (the following
morning) after receiving a PA deficient high-fat or
high-CHO diet, PA excretion began to decrease
significantly (p <.01) as compared to control values
(15815; 184127; 30717 ug PA/ 24 hr). PA excretion
stayed significantly lower in the PA deficient groups
for the remainder of the study (6 days). Differences in
excretion over time were only significant between days 8
and 9 where rats were being switched from experimental
diets to stock diet and between days 17 and 18 when rats
were being switched from stock diet to experimental
diet. In this experiment, weight differed significantly
over time, but not among the groups (Appendix B).

As PA excretion in the urine dropped so
dramatically in one day in the first experiment, a
second experiment was undertaken to measure tissue and
blood PA levels and plasma triglyceride concentrations.

A dietary deficiency of PA would likely take longer to

38

affect changes in blood and tissue PA levels. By
examining these parameters at frequent intervals
beginning 3 days after the initiation of the experiment,
any existing differences in PA concentrations as a
result of a high-fat or a high-CHO diet would be
observed.

Food intake of the high-fat PA supplemented ad
libitum fed group was significantly greater (p<.01)
than all other groups by the third week of the
study (Table 7). A difference in food intake between
the high-CHO and the high-fat diets was not seen in the
PA deficient groups. Body weight increased significantly
over time in all four treatment groups: high-fat PA
deficient, high-CHO PA deficient, high-fat PA
supplemented pair fed to high-fat PA deficient, or high-
fat PA supplemented fed ad libiutm (Table 8).

Differences in weight were significant beginning at
week 2 in the high-fat PA supplemented ad lib group as
compared with high-fat PA deficient group (12118 vs
11119 g) and beginning at week 3 in PA supplemented pair
fed group as compared with the high-fat PA deficient
group (145112 vs 131112 9). No significant difference,
however, was observed between the two PA deficient
groups at any time point measured.

Wet tissue weight of individual organs at time of
sacrifice increased significantly over time but did not

differ among the groups at any one time (Appendix C).

39

Table 7 1
Weekly Food Intake (kcal) of Rats
Treatment Week
1 2 3 4
HF PA- 305 347 360 249
+26 +38 134 148
3
HC PA- 283 322 334 278
+57 155 +25 +20
4
HF PA+ 305 347 360 249
PR FED 126 138 134 148
5 2 2
HF PA+ 304 373 471 473
AD LIB +22 150 +36 149
1

mean std dev; n=20 at wk 0, 15 at wk 1, 10 at wk 2,
5 at wk 3 & 4

p < .01 compared to high-fat PA deficient group
(HF PA-)

HC PA- = high-carbohydrate diet deficient in PA

HF PA+ pr fed = high-fat PA supplemented diet
(312 ug / 100 kcal) pair fed to HF PA- gp

HF PA+ ad lib = high-fat PA supplemented diet
(312 ug / 100 kcal) fed ad libitum

40

Table 8 1
Weekly Weights (g) of Rats

Treatment Week
0 1 2 3 4
HF PA- 40 80 111 131 131
13 16 +9 112 +11
3
HC PA- 39 76 103 127 131
14 16 19 111 +10
4 2 2
HF PA+ 40 78 116 145 160
PR FED +2 16 18 113 +23
5 * 2 2
HF PA+ 39 77 121 164 212
AD LIB +3 16 18 +8 +13
1

mean std dev; n=20 at wk 0, 15 at wk 1, 10 at wk 2, 5
at wk 3 8 4

p < .01 compared to high-fat PA deficient group
(HF PA-)

* p <.05 compared to high-fat PA deficient group
(HF PA-)

HC PA- = high-carbohydrate diet deficient in PA

HF PA+ pr fed = high-fat PA supplemented diet
(312 ug / 100 kcal) pair fed to HF PA- group.

HF PA+ ad lib = high-fat PA supplemented diet
(312 ug / 100 kcal) fed ad libitum

41

By week 1.5, heart and kidney free PA concentration
(Figure 4, 5) in both pair fed and ad libitum fed
groups consuming diets containing PA had significantly
(p <.01) greater amounts of free PA than did the high-
fat PA deficient group (7114, 87111 vs 412 nmol PA/ 9
heart tissue: 13518, 13118 vs 27120 nmol PA/ g kidney).
No significant difference was found in free PA content
of heart or kidney between high-fat and high-CHO PA
deficient groups at any time point measured. No
significant differences were seen between pair fed and
ad libitum fed PA supplemented groups in heart and
kidney free PA content. In liver, only the high-fat PA
supplemented ad lib group had significantly greater
amounts of free PA at weeks 2.5 (94120 vs 20110 nmol PA/
g liver) and 4.0 (49134 vs 1315 nmol PA/ g liver) as
compared to the high-fat PA deficient group (Figure
6).

Whole blood, total and free PA content (Figure 7),
and plasma free PA (Figure 8) showed similar trends to
one another. Pair fed and ad libitum fed rats receiving
PA in their diets had significantly greater (p<.01)
amounts of PA than the high-fat PA deficient treatment
group by 0.5 week (the first sacrifice time; 2.010.5,
1.610.9 vs 0.410.1 nmol PA/ml blood). No significant
differences were seen between the high-fat PA deficient
and the high-CHO PA deficient groups in blood or plasma

PA content at any time in the study.

42

 

 

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47

Previous studies (Beck et a1, 1988; Peterson et a1,
1987) have reported that plasma triglyceride
concentrations are correlated inversely with PA intake
at vitamin but not pharmacologic levels when rats were
fed a high fat diet. Plasma triglycerides were measured
in our study to further examine the relationship between
PA intake and its effects on lipid metabolism in rats
fed high-fat and high-CHO diets. By week 2.5, high-fat
PA supplemented pair fed, high-fat PA supplemented ad
lib and high-carbohydrate PA deficient groups all
exhibited significantly lower (p <.01) plasma
triglyceride levels than the high-fat PA deficient
treatment group (119123 in high-fat deficient group vs
59129, 2916 and 70133 mg/dl in the other three
respective groups). The plasma triglycerides of both of
the high-fat PA supplemented and high-CHO PA deficient
groups compared closely to normal plasma triglyceride
values in rats (Carlile et al., 1981).

DISCUSSION

Differences in PA status were not found between
high-fat and high-CHO PA deficient treatment groups in
weanling rats in any parameter measured except plasma
triglycerides. Significant differences were found
in free PA content of tissue and plasma, and in total
and free PA content of whole blood between PA
supplemented and PA deficient groups. This suggests

that the presence or absence of PA in the diet is more

48

important with respect to PA status than is the amount
of fat or carbohydrate present. These conclusions agree
with Williams et al. (1968) and Carter and Hockaday
(1962) who concluded that a high fat diet did not
increase the PA requirement in rats.

Urinary excretion of PA showed no significant
differences at any time in the study between high-fat
and high-CHO PA deficient diet groups. Within 1 day,
excretion of PA dropped significantly in PA deficient
treatment groups, which was a much more rapid response
than previously reported by Hatano (1962). This
suggests a mechanism in the body sensitive to PA levels.
Karnitz et al. (1984) concluded earlier that the only
mechanism for conservation of free PA in the rat is
tubular reabsorption in the kidney. PA excretion in
this study, however, never dropped to a zero level,
indicating that a small amount of PA is not reabsorbed.
Fox and Linkswiler (1961) observed a similar trend in
their humans studies and suggested this as the minimal
daily requirement of PA for tissue maintenance.

The lack of significant differences in urinary PA
excretion between the control and deficient groups for
the first week of the depletion in the study could be
the result of low food intake of the semipurified diet
by the control group and the rapid growth resulting in a
higher PA requirement than that added during this time.

As rats age, their requirement for PA decreases (Unna

49

and Richards, 1943). This can also partially explain
the PA excretion pattern of the control group in that as
they grow, their requirements decrease and therefore PA
excretion increases. While the National Research
Council recommends 8 - 10 mg PA /kg diet to be
sufficient for growth and maintenance in the rat, it has
been previously determined that weanling rats require 80
- 100 ug PA per day for optimal growth (Unna and
Richards, 1942). Although the diet supplemented in PA
contained 12 mg PA/ kg diet, it is possible that due to
the small quantity of food consumed by the weanling rat,
not enough PA was ingested for optimum growth. This
would result in lower than expected excretion of PA in
the urine. It would be desirable to repeat the first
part of this study to confirm the results obtained.
Hatano et al. (1968) reported a larger increase in

PA excretion in the control group versus the PA
deficient group when both groups were given a PA load of
4 mg Ca(PA)z subcutaneously. This pattern was similar
to the excretion patterns observed in this study when
all groups were fed stock diet. Since urinary excretion
of PA fell after 1 day on the experimental diets and
remained at low levels until a stock diet was fed, we
felt it was not necessary to measure urinary excretion
again in the second experiment as the diets fed in both

experiments were the same.

50

It is possible that blood PA concentrations which
were significantly lower in PA deficient treatment
groups by 0.5 week in experiment 2, had dropped prior to
this first measurement and in so doing had triggered the
reaborption of PA from the kidney tubules. Urine and
blood PA concentrations have previously been found to be
indicative of PA levels in the diet (Tao and Fox, 1976:
Hatano, 1962; Pearson et al., 1946) and our studies offer
further proof of this. We need, however, further
examination of the relationship between urine and blood
PA, with tissue PA concentrations to be able to
elaborate on the urine and blood PA concentrations
indicative of depleted stages.

Although total PA levels did not change among the
treatment groups in any of the tissues analysed, there
was a decrease of 80% to 90% in free PA in heart and of
50% to 60% in kidney in the deficient groups over the 4
weeks of the study (mostly occuring between 0.5 and 1.5
weeks). Reibel et al. (1981) reported decreases of 70%
to 90% in free PA in organs in adult rats fed PA
deficient diets for 4 weeks. The observed conservation
of total PA content might suggest that organs normally
contain a large excess of PA (Reibel et al., 1982).
Alternative explanations for this lack of change in
total PA per gram tissue include increased synthesis or
increased absorption of PA from the intestine, or

possibly increased copraphagy. Decreased PA content in

51

other unmeasured tissues is another possibility which is
presently being explored in this laboratory. As well,
while the diet was deficient in PA, it was not
completely devoid of the vitamin, offering still another
source of PA to maintain total PA concentrations per
gram tissue at constant levels. Tissue concentrations
of CoA which are normally maintained within a narrow
range have been shown to change under some pathological
conditions (e.g. diabetes) (Robishaw and Neely, 1985).
These changes could have a significant effect on fatty
acid metabolism or other pathways utilizing CoA. Apart
from measuring total PA it would be useful to have
information on the ratio of free CoA to acyl CoA as well
as on the distribution of CoA between the cytosol and
mitochondria as these factors can alter certain
metabolic functions.

' It appears from this study that PA has a
hypotriglyceridemic effect for elevated plasma lipids
induced by feeding a high fat diet. This effect has
been proposed to be as a result of a vitamin related
effect of PA versus a pharmacologic one (Peterson et
al., 1987). Proposed mechanisms for the
hypotriglyceridemic effect include fatty acid oxidation
stimulation (Kameda and Abiko, 1980), activation of the
citric acid cycle (Prisco et al., 1984) and lipoprotein

lipase activation (Noma et al., 1984).

52

Hypertriglyceridemia can be caused by increased
secretion of VLDL by the liver or decreased degradation
by lipoprotein lipase or both. As PA is incorporated
into CoA which is extensively involved in lipid
metabolism, certain reactions may be suppressed by an
inadequate supply of CoA as a result of insufficient PA
in the diet. This could result in elevated

triglycerides in rats not receiving PA in their diet.

53

SUMMARY

Urinary PA excretion drOpped significantly (p<.01)
by day 2 of experiment 1 in rats on diets deficient in
IPA” No differences were found between high-fat and
high-CHO PA deficient treatment groups.
Significant differences (p<.01) were found
Ibeginning at week 0.5 of experiment 2 in whole blood
(total and free) and in plasma (free) between PA
supplemented and PA deficient treatment groups. No
significant differences were seen between high-fat and
high-CHO PA deficient treatment groups.
Free PA concentrations in the liver, kidney, and
Iheart were significantly lower (p<.01) in the PA
(deficient groups beginning at week 1.5 of experiment 2.
'No significant differences were found among treatment
groups with respect to total PA in any organ analyzed.
Plasma triglycerides were significantly higher
(p<.01) in the high-fat PA deficient group than in the
other three treatment groups at weeks 2.5 and 4 in
experiment 2.
The high-fat PA deficient group had significantly
lower PA in tissue and blood and higher triglyceride
concentrations (p<.01) than the high-fat PA supplemented pair
fed or ad libitum fed groups. The underlying mechanisms are

unknown at this time.

54

Neither food intake nor weight gain differed
significantly between the high-fat and high-CHO
PA deficient groups. Food intake of the high-fat PA
supplemented ad libitum fed group was significantly
greater (p<.01) than all other treatment grous beginning
at week 3 of experiment 2. Weight of the ad libitum and
pair fed PA supplemented groups was significantly
greater (p<.01) than the PA deficient groups beginning

at week 2 of experiment 2.

CONCLUSIONS

The results of these studies do not demonstrate
significant differences in total and free PA in blood
and tissue, or urine PA levels between high fat PA
deficient and high carbohydrate PA deficient groups,
though the differences were significant between PA
supplemented and PA deficient groups beginning at week
0.5 of the second experiment. From these studies, it
was concluded that the PA requirement in rats as
determined by blood, tissue, and urinary PA levels is
not altered significantly by the quantity of fat and

carbohydrate in the diet.

55

RECOMMENDATIONS
The relationship between high-fat and high-

carbohydrate diets and PA status needs to be further
examined by using diets containing marginal PA, that is,
inadequate amounts to support growth and maintenance. In
this way, the presence and/or absence of PA would not be
such a strong factor and other more subtle effects could be
observed. As well, it might prove beneficial to examine
other tissues in rats fed experimental diets as in this
study. Since kidney, heart, and liver are all quite
important tissues metabolically, it is likely that these

are the same tissues which would be affected last in a
deficiency state in the rat. Muscle and fat tissue are

less active and thus might provide interesting information
about what happens when rats are fed high-fat or high-
carbohydrate PA deficient diets vs rats fed these same

diets supplemented with PA. These tissues could

potentially be acting as stores of PA from which liver,
kidney, and heart draw to maintain their CoA levels as was
observed. More generally, in order to more accurately
determine the correlation between dietary intake of PA and
blood levels of the vitamin, measuring blood PA content of
rats daily for the first three or four days they are fed
experimental diet is recommended. This information will be
useful in helping to determine a Recommended Dietary

Allowance for pantothenic acid.

56

APPENDICES

APPENDIX A

57

HIGH CARBOHYDRATE PANTOTHENIC ACID DEFICIENT
1,2,3
DIET COMPOSITION

 

INGREDIENT WEIGHT KCAL
Vitamin Free Casein 20 21%
DL-Methionine 0.3 _

Cornstarch 15 1 67.5%
Sucrose 50 _1

Fiber - Celufil 5

Corn oil 5 12%
AIN-76 Mineral mix 3.5

AIN-76 Vitamin mix 1.0

without calcium pantothenate

Choline bitartrate 0.2

1

PA supplemented diet mixed as above with the addition of
12 ug PA/g diet (312 ug PA/ 100 kcal) D-calcium
pantothenate

2
this diet contains 385 kcal ME/100 g

3
all ingredients purchased from United States
Biochemical Corporation

58

HIGH FAT PANTOTHENIC ACID DEFICIENT
1,2,3
DIET COMPOSITION

INGREDIENT WEIGHT KCAL
Vitamin Free Casein 20 21%
DL-methionine _ 0.3
Cornstarch l 11.25 12%
Sucrose _1
Fiber - Celufil 5 _

Corn oil 5 l

Crisco * 23.89 _1 67.5%
AIN-76 Mineral mix 3.5

AIN-76 Vitamin mix 1.0

without calcium pantothenate
Choline bitartrate 0.2

 

pantothenic acid supplemented diet mixed as above
with the addition of 17 ug PA/ 9 diet (312 ug PA/ 100
kcal) D-calcium pantothenate

this diet contains 385 kcal ME/ 70 g

all ingredients purchased from United States
Biochemical Corporation

* Crisco is made of partially hydrogenated soybean,
palm, and sunflower oils

 

APPENDIX B

59

Food Intake - Experiment 1

1
Day Treatment
High Fat PA- High CHO PA- * High CHO PA+
1 g 51 1 51 1 41 1
2 kcal 331 9 271 4 271 1
3 " 281 9 341 3 251 3
4 " 311 3 391 2 331 2
5 " 321 4 391 2 381 4
6 " 351 2 421 2 401 5
7 " 371 3 391 1 371 2
8 g 81 1 81 1 81 1
9 " 111 1 111 2 121 2
10 " 131 1 141 1 141 1
11 " 151 1 161 1 171 1
12 " 151 1 151 1 151 1
13 " 131 2 141 2 151 1
14 " 161 1 161 2 171 1
15 " 181 3 181 2 181 1
16 " 191 1 191 1 191 1
17 kcal 791 6 621 5 611 6
18 " 761 5 731 6 631 1
19 " 651 8 74: 6 691 4
20 " 651 4 671 5 661 3
21 " 681 6 681 5 631 4
22 " 711 3 701 6 681 4
23 " 671 6 731 6 691 5

* food spilt into urine not accounted for, therefore
reported values are higher than actual intake.

day 1- 7 and 17- 23 rats fed experimental diets
day 8- 16 rats fed stock diet

60

Weight - Experiment 1

Week Treatment

0 37+ 2 381 2 381 3
1 681 7 711 5 701 5
2 1111 9 1201 10 1181 7
2'* 1161 10 1171 12 1171 6
3 1571 12 1561 12 1521 7

* rats randomly divided into three new equal groups
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76

KIWI AI SACRIFICE

IAKLIK 11111
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KAI 105.55 105.55 101.35 .114.”
575 IV 5.42 5.75 4.14 5.3
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7 114.55 121.” 105.40 144.55
5 113.” 147.” 122.“ 155.3
5 122.25 140.75 100.50 13.”
10 121.10 131.55 104.05 145.25
KAI 121.35 13.“ 1N.52 145.42
575 IV 5.55 5.” 5.25 11.53
KB 4.0
15 125.0 171.“ 145.20 204.10
17 125.55 150.15 133.00 152.10
15 13.55 135.45 13.” 225.00
15 115.55 18.45 124.” 215.20
25 144.” 150.” 115.“ 217.50
KAI 11.55 155.5 130.74 211.50

575 IV 11.02 22.57 10.27 13.32

LIST OF REFERENCES

LIST OF REFERENCES

ABIKO Y., Metabolism of Coenzyme A pp 1-25 in D.M.
Greenberg (ed.) Metabolism of sulfur compounds. Vol 7
Metabolic pathways. Academic Press, New York.

American Institute of Nutrition. (1977) Report of AIN Ad
Hoc Committee on Standards for Nutritional Studies. J.
Nutr. 107:1340-1348.

BAKER, H. & FRANK, 0. (1968) Clinical Vitaminology:
Methods and Interpretations New York: Interscience.

BARBORIAK, J.J., KREHL, W.A., & COWGILL, G.R. (1956)
Pantothenic Acid Requirement of the Growing Adult Rat.

BROWN, G.M. (1958) The Metabolism of Pantothenic Acid.
J. Biol. Chem. 234: 370- 378.

BROWN, R.A. & STURTEVANT, M. (1949) The Vitamin
Requirements of the Growing Rat Vitamins and Hormones,
71181-182, Academic Press, New York.

CARLILE, S.I. & LACKO, A.G. (1981) Strain Difference in
the Age Related Changes of Rat Lipoprotein Metabolism.
Comp. Biochem. Physiol. 70B1753-758.

CARTER, C.W. & HOCKADAY, T.D.R. (1962) Liver Lipids
and Ketone-Body Formation in Rats Deficient in
Pantothenate. Biochem. J. 84:275-280.

CHAMPIGNY, o. & HITIER, Y. (1987) Lipoprotein Lipase
Activity in Skeletal Muscle and Brown Adipose Tissue of
Pregnant and Lactating Rats. J. Nutr. 1171349-354.

CHIPPONI, J.K., BLEIER, J.C., SANTI, M.T., & RUDMAN, D.
(1982) Deficiencies of Essential Nutrients. Am J. Clin.
Nutr. 35:1112-1116.

COHENOUR, S.H. & CALLOWAY, D.H. (1972) Blood, Urine and

Dietary Pantothenic Acid Levels of Pregnant Teenagers.
Am. J. Clin. Nutr. 25:512-517.

77

78

DONATO, K.A. (1987) Efficiency and Utilization of
Various Energy Sources for Growth. Am. J. Clin. Nutr.
45:164-167.

FLETCHER, M.J. (1968) A Colorimetric Method for
Estimating Serum Triglycerides. Clin. Chim. Acta
22:393-397.

FRY, P.C., Fox, H.M., & TAO, H.G. (1976) Metabolic
Response to a Pantothenic Acid Deficient Diet in Humans.
J. Nutr. Sci. Vitaminol. 22:339-346.

FOX, H.M. & LINKSWILER, H. (1961) Pantothenic Acid
Excretion on Three Levels of Intake. J. Nutr. 75:451-
454.

GIOVANNETTI, P.M. (1982) Effect of Coprophagy on
Nutrition. Nutr. Res. 2:335-349.

GOLDMAN, P. & VAGELOS, P.R. (1964) Acyl-transfer
reactions pp 71-92 in M. Florkin and E.H. Stotz (eds.)
Comprehensive biochemistry. Vol 15 Elsevier Pub. Co.,
New York.

GUEHRING, R.R., HURLEY, L.S. 1 MORGAN, A.F. (1952)
Cholesterol Metabolism in Pantothenic Acid Deficiency.
J. Biol. Chem. 197:485-493.

HALVORSEN, o. 1 SKREDE, s. (1982) Regulation of the
Biosynthesis of CoA at the Level of Pantothenate Kinase.
Eur. J. Biochem. 124:211-215.

HATANO, M. (1962) Pantothenic Acid Deficiency in Rats.
J. Vitaminol. 81143-159.

HATANO, M., HODGES, R.E., EVANS, T.C., HAGEMANN, R.F.,
LEEPER, 0.3., BEAN, W.B., & KREHL, W.A. (1966) Urinary
Excretion and Organ Distribution of Pantothenic Acid in
Normal, Diuretic Treated and Diabetic Rats. Fed Proc.
25:721.

HATANO, 11., HODGES, 12.3., EVANS, T.C., HAGEMANN, R.F.,
LEEPER, D., 8 KREHL, W.A. (1967) Urinary Excretion of
Pantothenic Acid by Diabetic Patients and by Alloxan-
Diabetic Rats. Am. J. Clin. Nutr. 20:960-967.

HENDERSON, L.M., MCINTIRE, J.R., WAISMAN, H.A., &
ELVEHJEM, C.A. (1942) Pantothenic Acid in the
Nutrition of the Rat. J. Nutr. 23:47-58.

IDELL-WENGER, J.A., GROTYOHANN, L.W., & NEELY, J.R.
(1978) Coenzyme A and Carnitine Distribution in Normal
and Ischemic Hearts. J.Biol. Chem. 253:4310-4318.

79

IDELL-WENGER, J.A. & NEELY, J.R. (1978) Regulation Of
Uptake and Metabolism of Fatty Acids by Muscle.
In:Disturbances in Lipid and Lipoprotein Metabolism.
Bethesda, MD: Am. Physiol. Soc. p.269-284.

ISRAEL, B. C., & SMITH, C.M. (1987) Effects of Acute
and Chronic Ethanol Ingestion on Pantothenate and CoA
Status of Rats. J. Nutr. 117: 443-451.

JEN C., PIRO, G., & LIN, p. (1987) Effects of
Infantile Overnutrition and Adulthood High Fat Feeding
on Adult Body Weight Regulation in Rats. Nutr. Research
7:421-431.

KAMEDA, K. & ABIKO, Y. (1980) Stimulation of Fatty Acid
Metabolism by Pantethine Natural Sulfur Compounds. Novel
Biochemical and Structural Aspects ed.D. Cavallini, G.E.
Gaull, V. Zappia. Plenum Pub. Corp. N.Y. pp443-452.

KARNITZ, L.M., GROSS, C.J., & HENDERSON, L.M. (1984)
Transport and Metabolism of Pantothenic Acid by Rat
Kidney. Biochim Biophy. Acta. 769:486-492.

LEVEILLE, G.A., & CLOUTIER, P.F. (1987) Isocaloric
Diets: Effects of Dietary Changes. Am. J. Clin. Nutr.
45:158-163.

LOPASCHUK, G.D., Hansen, C.A., & NEELY, J.R. (1986)
Fatty Acid Metabolism in Hearts Containing Elevated
Levels of Coenzyme A. Am. J. Physiol. 250:H351-H359.

MAJERUS, P.W., ALBERTS, A.W., & VAGELOS P.R. (1965)
Acyl Carrier Protein VII. The Primary Structure of the
Substrate Binding Site. J.Biol. Chem. 240:4723-4726.

MAMEESH, M.S., WEBB, R.E., NORTON, H.W.,& CONNOR
JOHNSON, B. (1959) The Role of Coprophagy in the
Availability of Vitamins Synthesized in the Intestinal
Tract with Antibiotic Feeding. J. Nutr. 69:81-84.

McGARRY, J.D., RABLES-VALDES, C. & FOSTER, D.W. (1975)
Role of Carnitine in Hepatic Ketogenesis. Proc. Natl
Acad Sci USA 72:4385-4388.

MICKELSEN, O., TAKAHASHI, S., & CRAIG, C. (1955)
Experimental Obesity I: Production of Obesity in Rats by
Feeding High Fat Diets. J. Nutr. 57:541-554.

MILLER, R.G. (1977) Development in Multiple
Comparisons, 1966-1976. J Am Stat Assoc 72:779-788.

80

MOISEENOK, A.G., SHEIBAK, V.M., & GURINOVICH, V.A.
(1986) Hepatic CoA,S-Acyl-CoA,Biosynthetic Precursors
of the Coenzyme and Pantothenate-Protein Complexes in
Dietary Pantothenic Acid Deficiency. Internat. J. Vit.
Nutr. Res. 57:71-77.

MYSZKOWSKA, K. (1964) Effect of Pantothenic Acid and
Phenyl Pantothenate on Certain Metabolic ProceSses.
Acta Physiol. Pol. 15:236-247.

NOMA, A., KITO, M., & OKAMIYA, T. (1984) Effect Of
Pantethine on Post-Heparin Plasma Lipolytic Activities
and Adipose Tissue Lipoprotein Lipase in Rats. Horm.
Metabol. Res. 16:233-236.

NELSON, M.M., NOUHUYS, F.V., 8 EVANS, E.H. (1947) The
Sparing Action of Protein on the Pantothenic Acid
Requirement of the Rat. J. Nutr. 34:189-203.

NOVELLI, G.D. (1953) Metabolic Functions of
Pantothenic Acid. Physiol. Rev. 33:525-543.

OLDHAM, H.G., DAVIS, M.V., & ROBERTS, L.J. (1946)
Thiamine Excretion and Blood Levels of Young Women on
Diets Containing Varying Levels Of the B-Vitamins with
Observations on Niacin and Pantothenic Acid. J. Nutr.
32:163-180.

ORAM, J.E., BENNETCH, S. & NEELY, J.R. (1973)
Regulation of Fatty Acid Utilization in Isolated
Perfused Rat Hearts. J. Biol. Chem. 248:5299-5309.

OSCAI, L.B., BROWN, M.M., & MILLER, W.C. (1984) Effect
of Dietary Fat on Food Intake, Growth and Body
Composition in Rats. Growth 48:415-424.

PEARSON, P. (1941) The Pantothenic Acid Content of the
Blood Of Mammalia. J. Biol. Chem. 140:423-426.

PEARSON, P., MELASS, v., & SHERWOOD, R. (1946) The
Pantothenic Acid Content of the Blood and Tissues of
Chicken as Influenced by the Level in the Diet. J.
Nutr. 32:187-193.

PETERSON, M.A., WITTWER, C.T., JORGENSEN, T., WINDHAM,
C.T., & WYSE, B.W. (1987) The Effects of Pantothenate
Status on Serum Triglycerides and Free Fatty Acids in
Rats Fed a High Fat Diet. Fed. Proc. 46:1489.

PRISCO, D., ROGASI, P.G., MATUCCI, M., COSTANZO, G., &
GENSINI, G.F. (1984) Effect of Pantethine Treatment
on Platelet Aggregation and Thromboxane A Production.
Curr. Ther. Res. 35:700-706.

81

RABINOWITZ, M. & SWIFT, H. (1970) Mitochondrial
Nucleic Acids and Their Relation to the Biogenesis of
Mitochondria Physiol. Rev. 50:376-427.

REIBEL, D.K., WYSE, B.W., BeRKICH, D.A., PALKO, W.M., &.
NEELY, J.R. (1981) Effects of Diabetes and Fasting on
Pantothenic Acid Metabolism in Rats. Am. J. Physiol.
240:E597-601. ’

REIBEL, D.K., WYSE, B.W., BERKICH, D.A., & NEELY, J.R.
(1982) Coenzyme A Metabolism in Pantothenic Acid-
Deficient Rats. J. Nutr. 112:1144-1150.

ROBINSON, F. (1966) Pantothenic Acid pp406-484. In:
The Vitamin Co-Factors of Enzyme Systems. New York.
Pergamon.

ROBISHAW, J. & NEELY, J.R. (1985) Coenzyme A
Metabolism Am. J. Physiol. 248:E1-9.

SAUBERLICH, H.E., DOWDY, R.P.,& SKALA, J.H. (1974)
Laboratory Tests for the Assessment of Nutrition Status.
CRC Press, Cleveland, Ohio pp 88-91.

SCHEMMEL, R., MICKELSEN, 0., & TOLGAY, Z. (1969)
Dietary Obesity in Rats: Influence of Diet, Weight,Age
and Sex on Body Composition. Am. J. Physiol. 216:373-
379.

SCHEMMEL, R., MICKELSEN, O. & GILL, J.L. (1970)
Dietary Obesity in Rats: Body Weight and Body Fat
Accretion in Seven Strains of Rats. J. Nutr. 100:1041-
1048.

SEWELL, R.F., PRICE,D.G., & THOMAS, M.C. (1962)
Pantothenic Acid Requirement of the Pig as Influenced by
Dietary Fat. Fed. Proc. 21:468.

SIEDLER, A.J., RICE, M.S., MALONEY, P.A., LUSHBOUGH,
C.M., & SCHWEIGERT, 3.8. (1962) The Influence of
Varying Levels of Dietary Protein, Carbohydrate, and
Fats in the Nutrition of the Rat. J. Nutr. 77:149-154.

SKREDE, S. & HALVORSEN, O. (1979) Mitochondrial
Biosynthesis of Coenzyme A. Biochem. Biophys. Res.
Comm. 91:1536-1542.

SMITH, C.M., LINN CANO, M., & POTYRAJ, J. (1978) The
Relationship Between Metabolic State and Total CoA
Content of rat Liver and Heart. J. Nutr. 108:854-862.

SMITH, C.M. (1978) he Effect of Metabolic State on
Incorporation of [14-C]Pantothenate into CoA in Rat
Liver and Heart. J. Nutr. 108:863873.

82

SRINIVASAN, V., CHRISTENSEN, N., WYSE, B., & HANSEN,
R.G. (1981) Pantthenic Acid Nutritional Status in
the Elderly-Institutionalized and Noninstitutionalized
Am. J. Clin. Nutr. 34:1736-172.

STRYER, L. (1975) Biochemistry 2nd ed. W.H. Freeman
and Co. San Francsco.

TAO, H. & FOX, H.M. (1976) Protein-Pantothenic Acid
Interrelationships in Growing Rats. Nutr. Reports Int'l
1497-106.

UNNA, K., & RICHARDS, G.V. (1942) Relationship Between
Pantothenic Acid Requirement and Age in the at. J.
Nutr. 23:545-553.

VORIS, L., BLACK, A., SWIFT, R.W. & FRENCH, C.E.
(1942) Thiamine, Riboflavin, Pyridoxine and
Pantothenate Deficiencies as Affecting the Apetite and
Growth of the Albino Rat. J. Nutr. 23:555-566.

WILLIAMS, R.J. (1943) The Chemistry and Biochemistry
of Pantothenic Acid. Advances in Enzymol. 3:253-287.

WILLIAMS, M.A., CHU, L.-C., MCINTOSH, D.J., &
HINCENBERGS, I. (1968) Effects of Dietary Fat Level on
Pantothenate Depletion and Liver Fatty Acid Composition
in the Rat. J. Nutr. 94:377-382.

WITTWER, C., WYSE, B.W., & HANSEN, R.G. (1982) Assay
of the Enzymatic Hydrolysis of Pantetheine. Anal.
Biochem. 122:312-315.

WYSE, B.W., WITTWER, C.W., & HANSEN, R.G. (1979)
Radioimmunoassay for Pantothenic Acid in Blood and Other
Tissues. Clin. Chem. 25:108-111.

WYSE, B.W., SONG, W.O., WALSH, J.R., & HANSEN, R.G.
(1985) Pantothenic Acid pp 399-416 In Augustin J.,
Klein B.P., & Becker D. eds. Methods of Vitamin Assay.
John Wiley and Sons. N.Y.

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