**References with abstracts**

Abstracts not available or not entered for all citations

1\. Barbiroli, B., *et al.*, *Coenzyme Q10 improves mitochondrial
respiration in patients with mitochondrial cytopathies. An in vivo study
on brain and skeletal muscle by phosphorous magnetic resonance
spectroscopy.* Cellular and Molecular Biology, 1997. 43(5): p. 741-9.

2\. Barbiroli, B., S. Iotti, and R. Lodi, *Improved brain and muscle
mitochondrial respiration with CoQ. An in vivo study by 31P-MR
spectroscopy in patients with mitochondrial cytopathies.* Biofactors,
1999. 9(2-4): p. 253-60. We used in vivo phosphorus magnetic resonance
spectroscopy (31P-MRS) to study the effect of CoQ10 on the efficiency of
brain and skeletal muscle mitochondrial respiration in ten patients with
mitochondrial cytopathies. Before CoQ, brain \[PCr\] was remarkably
lower in patients than in controls, while \[Pi\] and \[ADP\] were
higher. Brain cytosolic free \[Mg2+\] and delta G of ATP hydrolysis were
also abnormal in all patients. MRS also revealed abnormal mitochondrial
function in the skeletal muscles of all patients, as shown by a
decreased rate of PCr recovery from exercise. After six-months of
treatment with CoQ (150 mg/day), all brain MRS-measurable variables as
well as the rate of muscle mitochondrial respiration were remarkably
improved in all patients. These in vivo findings show that treatment
with CoQ in patients with mitochondrial cytopathies improves
mitochondrial respiration in both brain and skeletal muscles, and are
consistent with Lenaz\'s view that increased CoQ concentration in the
mitochondrial membrane increases the efficiency of oxidative
phosphorylation independently of enzyme deficit.

3\. Boitier, E., *et al.*, *A case of mitochondrial encephalomyopathy
associated with a muscle coenzyme Q10 deficiency.* Journal of the
Neurological Sciences, 1998. 156(1): p. 41-6.

4\. Chen, R.S., C.C. Huang, and N.S. Chu, *Coenzyme Q10 treatment in
mitochondrial encephalomyopathies. Short-term double-blind, crossover
study.* European Neurology, 1997. 37(4): p. 212-8.

5\. Darley-Usmar, V.M., *et al.*, *Deficiency in ubiquinone cytochrome c
reductase in a patient with mitochondrial myopathy and lactic acidosis.*
Proc Natl Acad Sci U S A, 1983. 80(16): p. 5103-6. The skeletal muscle
of a patient with a mitochondrial myopathy was examined. A defect in the
electron transport chain was identified at the position of complex III
by activity measurements and the low levels of reducible cytochrome b.
The polypeptide composition of complex III in the patient\'s
mitochondria was determined by antibody binding experiments. The method
allowed detection of individual polypeptides at a lower limit of 10-40
ng of protein. Characterization of protein composition is thus possible
by using a biopsy sample of 1 g of tissue. The level of core proteins,
FeS protein, and subunit VI was greatly diminished in the patient\'s
mitochondria. Cytochrome c1 polypeptide was found at normal levels but
was sensitive to proteolysis by trypsin. These results show that complex
III is not assembled in the patient\'s mitochondria. The possible role
of cytochrome b as the site of the primary lesion is discussed.

6\. Di Giovanni, S., *et al.*, *Coenzyme Q10 reverses pathological
phenotype and reduces apoptosis in familial CoQ10 deficiency.*
Neurology, 2001. 57(3): p. 515-8. Two brothers with myopathic coenzyme
Q10 (CoQ10) deficiency responded dramatically to CoQ10 supplementation.
Muscle biopsies before therapy showed ragged-red fibers, lipid storage,
and complex I + III and II + III deficiency. Approximately 30% of
myofibers had multiple features of apoptosis. After 8 months of
treatment, excessive lipid storage resolved, CoQ10 level normalized,
mitochondrial enzymes increased, and proportion of fibers with
TUNEL-positive nuclei decreased to 10%. The authors conclude that muscle
CoQ10 deficiency can be corrected by supplementation of CoQ10, which
appears to stimulate mitochondrial proliferation and to prevent
apoptosis.

7\. England, J.D., *et al.*, *Mitochondrial myopathy developing on
treatment with the HMG CoA reductase inhibitors\--simvastatin and
pravastatin \[letter\].* Australian and New Zealand Journal of Medicine,
1995. 25(4): p. 374-5.

8\. Fosslien, E., *Mitochondrial medicine\--molecular pathology of
defective oxidative phosphorylation.* Ann Clin Lab Sci, 2001. 31(1): p.
25-67. Different tissues display distinct sensitivities to defective
mitochondrial oxidative phosphorylation (OXPHOS). Tissues highly
dependent on oxygen such as the cardiac muscle, skeletal and smooth
muscle, the central and peripheral nervous system, the kidney, and the
insulin-producing pancreatic beta-cell are especially susceptible to
defective OXPHOS. There is evidence that defective OXPHOS plays an
important role in atherogenesis, in the pathogenesis of Alzheimer\'s
disease, Parkinson\'s disease, diabetes, and aging. Defective OXPHOS may
be caused by abnormal mitochondrial biosynthesis due to inherited or
acquired mutations in the nuclear (n) or mitochondrial (mt)
deoxyribonucleic acid (DNA). For instance, the presence of a mutation of
the mtDNA in the pancreatic beta-cell impairs adenosine triphosphate
(ATP) generation and insulin synthesis. The nuclear genome controls
mitochondrial biosynthesis, but mtDNA has a much higher mutation rate
than nDNA because it lacks histones and is exposed to the radical oxygen
species (ROS) generated by the electron transport chain, and the mtDNA
repair system is limited. Defective OXPHOS may be caused by insufficient
fuel supply, by defective electron transport chain enzymes (Complexes
I - IV), lack of the electron carrier coenzyme Q10, lack of oxygen due
to ischemia or anemia, or excessive membrane leakage, resulting in
insufficient mitochondrial inner membrane potential for ATP synthesis by
the F0F1-ATPase. Human tissues can counteract OXPHOS defects by
stimulating mitochondrial biosynthesis; however, above a certain
threshold the lack of ATP causes cell death. Many agents affect OXPHOS.
Several nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit or
uncouple OXPHOS and induce the \'topical\' phase of gastrointestinal
ulcer formation. Uncoupled mitochondria reduce cell viability. The
Helicobacter pylori induces uncoupling. The uncoupling that opens the
membrane pores can activate apoptosis. Cholic acid in experimental
atherogenic diets inhibits Complex IV, cocaine inhibits Complex I, the
poliovirus inhibits Complex II, ceramide inhibits Complex III, azide,
cyanide, chloroform, and methamphetamine inhibit Complex IV. Ethanol
abuse and antiviral nucleoside analogue therapy inhibit mtDNA
replication. By contrast, melatonin stimulates Complexes I and IV and
Gingko biloba stimulates Complexes I and III. Oral Q10 supplementation
is effective in treating cardiomyopathies and in restoring plasma levels
reduced by the statin type of cholesterol-lowering drugs.

9\. Ogasahara, S., *et al.*, *Muscle coenzyme Q deficiency in familial
mitochondrial encephalomyopathy.* Proc Natl Acad Sci U S A, 1989. 86(7):
p. 2379-82. The electron transport system of muscle mitochondria was
examined in a familial syndrome of lactacidemia, mitochondrial myopathy,
and encephalopathy. The propositus, a 14-year-old female, and her
12-year-old sister had suffered from progressive muscle weakness,
abnormal fatigability, and central nervous system dysfunction since
early childhood. In the propositus, the state 3 respiratory rate of
muscle mitochondria with NADH-linked substrates and with succinate was
markedly reduced. The levels of cytochromes a + a3, b, and c + c1 were
normal. The activities of complexes I, II, III, and IV of the electron
transport chain were normal or increased. By contrast, the activities of
complex I-III and of complex II-III, both of which need coenzyme Q10
(CoQ10), were abnormally low. On direct measurement, the mitochondrial
CoQ10 content was 3.7% of the mean value observed in 10 controls. Serum
and cultured fibroblasts of the propositus had normal CoQ10 contents. In
the younger sister, the respiratory activities and CoQ10 level of muscle
mitochondria were similar to those observed in the propositus. The
findings establish CoQ10 deficiency as a cause of a familial
mitochondrial cytopathy and suggest that the disease results from a
tissue-specific defect of CoQ10 biosynthesis.

10\. Takusa, Y. and S. Yamaguchi, *\[Myopathies with miscellaneous
disorders related to mitochondrial fatty acid oxidation: defective
synthesis of ketone body, long-chain fatty acid transport defect, and
muscular coenzyme Q10 deficiency\].* Ryoikibetsu Shokogun Shirizu,
2001(36): p. 90-4.

11\. Tedeschi, D., *et al.*, *Potential involvement of ubiquinone in
myotonic dystrophy pathophysiology: new diagnostic approaches for new
rationale therapeutics.* Neurol Sci, 2000. 21(5 Suppl): p. S979-80. An
impairment of mitochondrial function may contribute to the
pathophysiology of myotonic dystrophy (MyD). Coenzyme Q10 (CoQ10)
deficiency has been previously observed, even if in a restricted sample
of patients. The aim of this investigation was to obtain more
information about coenzyme Q10 and its relationships to the aerobic
metabolism in a group of MyD patients. Serum CoQ10 appeared
significantly reduced with respect to normal controls: 0.93 +/- 0.22 vs.
1.58 +/- 0.28 micrograms/ml (p \< 0.05). Moreover, the results
demonstrated an inverse tendency between CoQ10 levels and the CTG
expansion degree. Basal blood lactate levels were significantly higher
than controls (p \< 0.05). A borderline inverse correlation between
CoQ10 and lactate, corresponding to lactate threshold, was found. These
data suggest a possible role of CoQ10 in the pathogenesis of MyD, which
may be mediated by mechanisms of cellular damage common to the oxidative
pathway. Therapeutic strategies may be devised by virtue of this
rationale.

12\. Fukuda, K., *et al.*, *Chronic multisymptom illness affecting Air
Force veterans of the Gulf War.* JAMA, 1998. 280: p. 981-988.

13\. Bowman, P.D., *et al.*, *Myopathic changes in diaphragm of rats fed
pyridostigmine bromide subchronically.* Fundam Appl Toxicol, 1989.
13(1): p. 110-7. To determine if alterations in muscle morphology occur
after subchronic oral administration of pyridostigmine bromide, rats
were fed 90 mg/kg continuously in meal and examined at 1, 2, 4, 7, and
15 days. Within the first day, cholinesterase activity was reduced by
87% and remained inhibited by 74-91% for the entire course of the
feeding. Light microscopy demonstrated that by the first day
approximately 1 in 100 myofibers was shrunken and contained centralized
nuclei. Electron microscopic examination showed that while presynaptic
areas of neuromuscular junctions were relatively unaffected by this
dose, postsynaptic areas invariably showed maximal changes.
Ultrastructural alterations included disruption of myofilaments,
mitochondrial changes consistent with accumulation of calcium, and
nuclear alterations. These effects appeared not to be cumulative and
were greatly diminished by 15 days even under constant drug
administration and inhibition of cholinesterase activity. We conclude
that subchronic feeding of pyridostigmine bromide induces primarily
myopathic rather than neurogenic changes in the diaphragm and that some
mechanism of accommodation may be activated that minimizes continued
muscle injury.

14\. Glass-Marmor, L., M. Chen-Zion, and R. Beitner, *Effects of
carbamylcholine and pyridostigmine on cytoskeleton-bound and cytosolic
phosphofructokinase and ATP levels in different rat tissues.* Gen
Pharmacol, 1996. 27(7): p. 1241-6. 1. The effects of carbamylcholine
(CaCh) (acetylcholine agonist) and pyridostigmine (Pyr)
(acetylcholinesterase inhibitor), on the activity of cytoskeleton-bound
and cytosolic phosphofructokinase (PFK), the rate-limiting enzyme in
glycolysis, and ATP levels, were studied in rat tibialis anterior (TA)
muscle, heart, and brain. 2. In the TA muscle, a marked (about
three-fold) increase in the allosteric activity of cytosolic (soluble)
PFK was found, 3-5 min following the injection of CaCh or Pyr. The
intracellular distribution of the enzyme was not affected by both drugs.
Stimulation of glycolysis in this muscle was also expressed by a
significant increase in the concentrations of glycolytic intermediates
and lactate. Glucose 1,6-bisphosphate (Glc-1,6-P2) levels were
unchanged, whereas fructose-2,6-bisphosphate (Fru-2,6-P2) was increased.
Glycogenolysis was also stimulated, as deduced from the decrease in
glycogen content. The stimulation of glycolysis, induced by both drugs,
was accompanied by an increase in ATP level in the TA muscle. 3. In
contrast to the stimulatory action of CaCh or Pyr on glycolysis in the
TA muscle, both drugs had no effect on cytosolic and cytoskeletal PFK in
heart and brain. However, ATP content in both heart and brain was
markedly reduced by these drugs, most probably due to their reported
harmful effects on mitochondrial function, leading to tissue damage. 4.
Electron microscopic studies of TA muscle and heart from rats treated
with CaCh or Pyr, revealed severe damage of heart but no harmful effects
on TA muscle, which is a muscle with high glycolytic and low oxidative
capacity. The present experiments suggest that the accelerated
glycolysis in this muscle induced by both drugs, supplies ATP, thus
preventing muscle damage.

15\. Glass-Marmor, L. and R. Beitner, *Effects of carbamylcholine and
pyridostigmine on mitochondrial-bound hexokinase in skeletal muscle and
heart.* Biochem Mol Med, 1996. 57(1): p. 67-70. We show here that
carbamylcholine (acetylcholine agonist) or pyridostigmine
(acetylcholinesterase inhibitor), drugs which are widely used in medical
treatments, exerted a rapid reduction in mitochondrial-bound hexokinase.
This reduction was inversely proportional to the changes in
glucose-6-phosphate levels in skeletal and heart muscle. Increased
concentration of acetylcholine, occurring in various diseases or induced
by acetylcholinesterase inhibitors, was reported to cause deterioration
of mitochondrial function, resulting in heart and muscle damage. The
present experiments suggest that the reduction in mitochondrial-bound
hexokinase, which is closely linked to intramitochondrial oxidative
metabolism, may play an important role in the mechanism which leads to
tissue damage.

16\. Hudson, C.S., R.E. Foster, and M.W. Kahng, *Neuromuscular toxicity
of pyridostigmine bromide in the diaphragm, extensor digitorum longus,
and soleus muscles of the rat.* Fundam Appl Toxicol, 1985. 5(6 Pt 2): p.
S260-9. The neuromuscular junctions from diaphragm, soleus, and extensor
digitorum longus (EDL) muscles of male albino rats were assessed for
morphological alterations following acute (30-min) and subacute (2-day)
exposure to pyridostigmine bromide in Mestinon-equivalent buffer. These
muscles were selected to compare the effects of the drug on muscles of
different fiber type composition. The diaphragm has approximately equal
numbers of type I and type II fibers while the soleus and EDL possess
primarily type I and type II fibers, respectively. Pyridostigmine was
administered to each acute-exposure animal by a single subcutaneous
injection of 0.36 mg/kg pyridostigmine and to each subacute-exposure
animal by a subcutaneously implanted osmotic minipump containing 10
mg/ml pyridostigmine. Both treatments resulted in whole blood
cholinesterase (ChE) depression of approximately 60-70% as determined by
radiometric assay. Control animals received only Mestinon-equivalent
buffer. Both acute and subacute exposures resulted in morphological
alteration of the neuromuscular junctions (NMJs) of all three muscles,
although considerable variation in the extent of damage occurred even
within individual NMJs. The most frequently observed presynaptic
alterations were mitochondrial damage and partial withdrawal of nerve
terminal branches (partial denervation). Post-synaptic changes included
occasional rarefaction of mitochondrial matrices and disruption of the
myofibrillar organization in small numbers of subjunctional sarcomeres.
The data indicate that acute or subacute exposure to pyridostigmine
bromide at a whole blood ChE depression of 60-70% results in similar
alterations to the NMJs of three muscles with substantially different
fiber type compositions. Although the severity of the damage varies from
fiber to fiber, the variability appears random and not related to a
specific fiber type or dosage regimen.

17\. Kato, T., *et al.*, *Role of acetylcholine in
pyridostigmine-induced myocardial injury: possible involvement of
parasympathetic nervous system in the genesis of cardiomyopathy.* Arch
Toxicol, 1989. 63(2): p. 137-43. Although acetylcholine is known to be
involved in the genesis of skeletal muscle disturbance, its effect on
cardiac muscle has been scarcely studied. In the present paper, using
pyridostigmine, a cholinesterase inhibitor, the possible role of
acetylcholine in the genesis of cardiomyopathy was investigated. In a
mortality study, it was shown that pyridostigmine (100 mg/kg) caused
death of 9/10 rats within 8 h, and that the lethality of such a dose
could be significantly diminished by the subsequent administration of a
total dose of 4 mg/kg atropine. In all other experiments, rats were
divided into three groups; the control, untreated group; the
pyridostigmine + atropine group in which atropine (2 mg/kg) was
administered 5 min after pyridostigmine (60 mg/kg) administration; and
the pyridostigmine group in which pyridostigmine (60 mg/kg) was
administered orally. Rats were killed 3 h after pyridostigmine
administration, and hearts were isolated. Heart mitochondrial electron
transport activity (NADH-cytochrome c reductase, succinate-cytochrome c
reductase, and cytochrome c oxidase) were measured enzymatically, and
mitochondrial respiratory rates and control indices were measured
polarographically. Structural changes in cardiac muscles of each group
were observed by electron microscopy of cardiac sections. Acetylcholine
levels of left ventricle were measured by high performance liquid
chromatography. Activities of NADH-cytochrome c reductase and
succinate-cytochrome c reductase were not affected by pyridostigmine
administration; however, cytochrome c oxidase activity was significantly
reduced in the pyridostigmine group. Atropine markedly lessened this
reduction in activity. A protective effect of atropine was also observed
morphologically. A protective effect of atropine was also observed
morphologically. In the pyridostigmine group and the pyridostigmine +
atropine group, left ventricular acetylcholine levels were increased
significantly compared with the control.(ABSTRACT TRUNCATED AT 250
WORDS).

18\. Schuschereba, S.T., *et al.*, *Myopathic alterations in extraocular
muscle of rats subchronically fed pyridostigmine bromide.* Toxicol
Pathol, 1990. 18(3): p. 387-95. To determine if alterations in
extraocular muscle morphology occur after subchronic oral administration
of pyridostigmine bromide, rats were continuously fed 90 mg/kg in meal
and examined at 1, 2, 4, 7, and 15 days. Within the first day, blood
acetylcholinesterase activity was reduced by 87% and remained inhibited
by 74-91% during the study. Light microscopy demonstrated that by day 1
approximately 3% of the extraocular myofibers were shrunken and invaded
by inflammatory cells. The most severe degenerative changes consisting
of vacuoles and inflammatory cell infiltration occurred at day 1 with
progressively less severe changes at days 2 and 4. At days 7 and 15,
1.3-4.5% of the myofibers still exhibited damage. Ultrastructurally, all
presynaptic areas were normal but the postsynaptic areas of affected
myofibers at days 1, 2, and 4 showed myofilament and Z-band dissolution,
mitochondrial inclusions, subneural fold and T-tubule/sarcoplasmic
reticulum vacuolization and subneural fold depth reduction. By days 7
and 15, these changes were diminished in some cases and in others
alterations appeared similar to day 1. We conclude that subchronic
feeding of pyridostigmine bromide induces myopathic rather than
neurogenic changes in rat extraocular muscle and that the myopathy is
different in these muscles than in the diaphragm from the the same rats.

19\. Kawabuchi, M., *Neostigmine myopathy is a calcium ion-mediated
myopathy initially affecting the motor end-plate.* J Neuropathol Exp
Neurol, 1982. 41(3): p. 298-314. Morphological techniques were used to
determine the acute and chronic effects of neostigmine on rat muscles.
Transient calcium deposits, eliminated by prior treatment of sections
with ethyleneglycol bis (aminoethylether) tetracetate (EGTA), were
independent of fiber type and found at sites corresponding to
neostigmine-induced focal lesions. The dimension and number of focal
lesions and calcium deposits gradually decreased with chronic drug
treatment. Size, shape, and density of the calcium deposits varied.
Alterations in the motor nerve terminal, synaptic space, and junctional
fold persisted even when banding patterns at the motor end plates were
intact. Characteristic intermediate findings consisted of rod bodies and
ribosomal clusters. Such clusters were frequently mingled and clumped
sarcoplasmic reticulums, T-systems, or mitochondria. Despite continued
administration of neostigmine, focal myopathic changes, other than in
the synaptic region of the end plates, were reversible.

20\. Babu, S.R., *et al.*, *Effect of physostigmine on plasma lactate
and pyruvate in untrained/trained rats.* Pharmacol Biochem Behav, 1992.
42(1): p. 67-73. Physostigmine (Phy) is metabolized to eseroline, a
phenolic compound that appears to alter mitochondrial functions. The
effect of Phy on recovery from exercise and on time course of plasma
lactate and pyruvate levels following an acute bout of exercise (AE) was
examined in untrained and trained (ET) rats. Phy alone elicited
significantly higher plasma lactate and pyruvate levels than sedentary
control. AE + Phy had a significantly higher plasma lactate and pyruvate
levels compared to AE 2 min postadministration. From 5-30 min
postexercise, lactate and pyruvate levels did not differ between these
two acutely exercised groups. ET + Phy exhibited significantly lower
levels of plasma lactate and pyruvate from 5-60 min postexercise
compared to ET. The data show that the \"additive\" effect of Phy on
postexercise plasma lactate and pyruvate levels can be attenuated by an
enhanced fitness level in these rats.

21\. De Pinieux, G., *et al.*, *Lipid-lowering drugs and mitochondrial
function: effects of HMG-CoA reductase inhibitors on serum ubiquinone
and blood lactate/pyruvate ratio.* British Journal of Clinical
Pharmacology, 1996. 42(3): p. 333-7. 1. Statins inhibit synthesis of
mevalonate, a precursor of ubiquinone that is a central compound of the
mitochondrial respiratory chain. The main adverse effect of statins is a
toxic myopathy possibly related to mitochondrial dysfunction. 2. This
study was designed to evaluate the effect of lipid-lowering drugs on
ubiquinone (coenzyme Q10) serum level and on mitochondrial function
assessed by blood lactate/pyruvate ratio. 3. Eighty
hypercholesterolaemic patients (40 treated by statins, 20 treated by
fibrates, and 20 untreated patients, all 80 having total cholesterol
levels \> 6.0 mmol l-1) and 20 healthy controls were included.
Ubiquinone serum level and blood lactate/pyruvate ratio used as a test
for mitochondrial dysfunction were evaluated in all subjects. 4.
Lactate/pyruvate ratios were significantly higher in patients treated by
statins than in untreated hypercholesterolaemic patients or in healthy
controls (P \< 0.05 and P \< 0.001). The difference was not significant
between fibratetreated patients and untreated patients. 5. Ubiquinone
serum levels were lower in statin-treated patients (0.75 mg l-1 +/-
0.04) than in untreated hypercholesterolaemic patients (0.95 mg l-1 +/-
0.09; P \< 0.05). 6. We conclude that statin therapy can be associated
with high blood lactate/ pyruvate ratio suggestive of mitochondrial
dysfunction. It is uncertain to what extent low serum levels of
ubiquinone could explain the mitochondrial dysfunction.

22\. King, B.F. and S.M. Somani, *Distribution of physostigmine and
metabolites in brain subcellular fractions of the rat.* Life Sci, 1987.
41(17): p. 2007-15. The distribution of 3H-physostigmine (Phy) has been
studied in the rat brain subcellular fractions at various time intervals
following i.v. injection. 3H-Phy or its metabolites rapidly accumulate
into the cytoplasm of cells and penetrates the intracellular
compartments. Kinetic studies of the subcellular distribution of
radioactivity (RA) per gm of rat brain following i.v. injection of
3H-Phy show peak concentrations at 30 min in all subcellular fractions
with the exception of mitochondria. In the mitochondrial fraction the RA
levels continue to rise from 4682 +/- 875 DPM/gm at 5 min to 27474 +/-
2825 DPM/gm at 60 min (P less than .05). The cytosol contains the
highest RA: 223341 +/- 21044 DPM/gm at 30 min which declined to 53475
+/- 3756 DPM/gm at 60 min. RA in synaptosome, microsomes and myelin
increases from 5 to 30 min, and declines at 60 min. In vitro studies did
not show a greater uptake of RA by the mitochondrial or synaptosomal
fractions. The finding of relatively high concentrations of RA in the
mitochondrial fraction at 60 min increases the likelihood that Phy or
its metabolites could interfere with the physiological function of this
organelle.

23\. Li, L., *et al.*, *Pyridostigmine-induced acute and delayed
neuronal apoptosis \[abstract\].* Soc Toxicol Abstract, 1999.

24\. Li, L., *et al.*, *Muscarinic receptor-mediated
pyridostigmine-induced neuronal apoptosis.* Neurotoxicology, 2000.
21(4): p. 541-52. Pyridostigmine is a reversible cholinesterase (ChE)
inhibitor that is associated with neurologic dysfunction involving both
central and peripheral nervous systems. To determine the neurotoxic
potential of pyridostigmine, rats were sacrificed at intervals after
drug administration (0.5-1.85 mg/kg, i.p., twice daily for 4 days) and
brains examined histologically. ChE inhibition was used as a biomarker
of pyridostigmine activity. Using the in situ terminal deoxynucleotidyl
transferase nick-end labeling of DNA fragments (TUNEL) method and
electron microscopy, apoptotic brain cell death was noted in cerebral
cortex over a dose range of 0.5-1.85 mg/kg and at the higher dose (1.85
mg/kg), apoptosis was also noted in striatum and hippocampus. These
responses were blocked by pretreatment with atropine. Rat cortical cells
in culture also underwent apoptosis when exposed to pyridostigmine (250
microM for 24 hr), indicating that the pyridostigmine can initiate
apoptosis, independent of peripheral mechanisms. Pretreatment of cells
with atropine (10 microM) inhibited pyridostigmine-induced apoptosis,
confirming the response was mediated by muscarinic receptors. Short term
treatment of rats with pyridostigmine (1.85 mg/kg twice daily for 4
days) induced a prolonged apoptotic response, which was evident in rat
cortex up to 30 days after the last dose. Active apoptosis persisted,
despite recovery of serum ChE activity. These in vivo and in vitro
observations indicate that pyridostigmine can initiate a prolonged
neurodegeneration.

25\. Isom, G.E., *Pyridostigmine-induced neurodegeneration: role of
neuronal apoptosis*, in *Annual Report to Congress: Federally Sponsored
Research on Gulf War Veterans\' Illnesses for 1998. Appendices*, The
Research Working Group of the Persian Gulf Veterans Coordinating Board,
Editor. 1999. p. 100.

26\. Li, L., *et al.*, *Reactive oxygen species mediate
pyridostigmine-induced neuronal apoptosis: involvement of muscarinic and
NMDA receptors.* Toxicol Appl Pharmacol, 2001. 177(1): p. 17-25.
Pyridostigmine bromide (PB) is a reversible cholinesterase inhibitor
used for treatment of myasthenia gravis and for prophylactic protection
against organophosphate nerve agent. We previously showed PB can induce
apoptotic death in rat brain following systemic treatment. To study
mechanisms by which PB induces brain cell death, cultured rat cerebellar
granule cells were used. Cytotoxicity was determined after exposure to
PB (10-1000 microM) for 24 h; a high concentration of PB (\>500 microM)
significantly increased lactate dehydrogenase release, which was reduced
by pretreatment with the antioxidant, N-t-butyl-alpha-phenyl-nitrone
(PBN). Apoptosis, as determined by TUNEL staining, was concentration
dependent (10-250 microM) after a 24-h exposure and cytotoxicity was
confirmed by gel electrophoresis of DNA, release of cytochrome c from
mitochondria, elevation of caspase activity, and electron microscopy.
The oxidant-sensitive fluorescent dye 2\',7\'-dichlorofluorescin
diacetate was used to detect reactive oxidative species (ROS)
generation. Pretreatment with PBN, superoxide dismutase, catalase, or
the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester
(L-NAME) blocked PB-induced ROS generation and apoptotic cell death.
Pretreatment with atropine or MK-801 blocked ROS generation and the
subsequent neurotoxicity, showing that both muscarinic and NMDA
receptors mediate the response. DNA extracted from PB-treated cells
revealed oligonucleosomal fragmentation on gel electrophoresis and
antioxidants attenuated the DNA fragmentation, providing further
evidence for a link of ROS generation and apoptosis. These results
indicate that muscarinic receptor-mediated ROS generation is an
initiating factor in PB-induced apoptotic cell death and activation of
the NMDA glutamate receptor is directly linked to the response.

27\. Quillet-Mary, A., *et al.*, *Implication of mitochondrial hydrogen
peroxide generation in ceramide-induced apoptosis.* J Biol Chem, 1997.
272(34): p. 21388-95. The key events implicated in ceramide-triggered
apoptosis remain unknown. In this study we show that 25 &mgr;M
C6-ceramide induced significant H2O2 production within 60 min, which
increased up to 180 min in human myeloid leukemia U937 cells. Inactive
analogue dihydro-C6-ceramide had no effect. Furthermore, no H2O2
production was observed in C6-ceramide-treated U937 rho degrees cells,
which are mitochondrial respiration-deficient. We also present evidence
that ceramide-induced activation of the transcription factors NF-kappaB
and AP-1 is mediated by mitochondrial derived reactive oxygen species.
Both H2O2 production, transcription factor activation as well as
apoptosis could be inhibited by rotenone and thenoyltrifluoroacetone
(specific mitochondrial complexes I and II inhibitors) and antioxidants,
N-acetylcysteine and pyrrolidine dithiocarbamate. These effects could be
potentiated by antimycin A (specific complex III mitochondrial
inhibitor). H2O2 production was also inhibitable by ruthenium red,
suggesting a role of mitochondrial calcium homeostasis alterations in
ceramide-induced oxidative stress. Finally, C6-ceramide had no influence
on mitochondrial membrane potential within the first 6 h. Altogether,
our study points to reactive oxygen species, generated at the ubiquinone
site of the mitochondrial respiratory chain, as an early major mediator
in ceramide-induced apoptosis.

28\. Haley, R.W., *et al.*, *Brain abnormalities in Gulf War syndrome:
Evaluation with 1H MR spectroscopy.* Radiology, 2000. 215: p. 807-817.
PURPOSE: To test for neuronal brain damage in the basal ganglia and
brainstem in Gulf War veterans by using magnetic resonance (MR)
spectroscopy. MATERIALS AND METHODS: Twenty-two Gulf War veterans with
one of three factor analysis-derived syndromes (case patients); 18 well
veterans matched for age, sex, and education level (control subjects);
and six Gulf War veterans with syndrome 2 from a different population
(replication sample) underwent long echo time (272 msec) proton
(hydrogen 1) MR spectroscopy on a 4 x 2 x 2-cm voxel in the basal
ganglia bilaterally and a 2 x 2 x 2-cm voxel in the pons. Syndromes 1-3
are described as \"impaired cognition,\" \"confusion-ataxia,\" and
\"central pain,\" respectively. RESULTS: The
N-acetylaspartate-to-creatine (NAA/Cr) ratio, which reflects functional
neuronal mass, was significantly lower in the basal ganglia and
brainstem of Gulf War veterans with the three syndromes than in those
structures of the control subjects (P =.007). The finding was
corroborated in the replication sample (P =.002). Veterans with syndrome
2 (the most severe clinically) had evidence of decreased NAA/Cr in both
the basal ganglia and the brainstem; those with syndrome 1, in the basal
ganglia only; and those with syndrome 3, in the brainstem only.
CONCLUSION: Veterans with different Gulf War syndromes have biochemical
evidence of neuronal damage in different distributions in the basal
ganglia and brainstem.

29\. Asa, P.B., Y. Cao, and R.F. Garry, *Antibodies to squalene in Gulf
War syndrome.* Exp Mol Pathol, 2000. 68(1): p. 55-64. Gulf War Syndrome
(GWS) is a multisystemic illness afflicting many Gulf War-era veterans.
The molecular pathological basis for GWS has not been established. We
sought to determine whether the presence of antibodies to squalene
correlates with the presence of signs and symptoms of GWS. Participants
in this blinded cohort study were individuals immunized for service in
Desert Shield/Desert Storm during 1990-1991. They included 144 Gulf
War-era veterans or military employees (58 in the blinded study), 48
blood donors, 40 systemic lupus erythematosus patients, 34 silicone
breast implant recipients, and 30 chronic fatigue syndrome patients.
Serum antibodies to squalene were measured. In our small cohort, the
substantial majority (95%) of overtly ill deployed GWS patients had
antibodies to squalene. All (100%) GWS patients immunized for service in
Desert Shield/Desert Storm who did not deploy, but had the same signs
and symptoms as those who did deploy, had antibodies to squalene. In
contrast, none (0%) of the deployed Persian Gulf veterans not showing
signs and symptoms of GWS have antibodies to squalene. Neither patients
with idiopathic autoimmune disease nor healthy controls had detectable
serum antibodies to squalene. The majority of symptomatic GWS patients
had serum antibodies to squalene.

30\. Powell, M.F. and M.J. Newman, eds. *Chapter 8. Adjuvant properties
of aluminum and calcium compounds.* Vaccine Design. The Subunit and
Adjuvant Approach. 1995, Plenum Press: New York.

31\. *Melinda Plaisier, FDA Associate Commissioner of Legislation,
Letter to Congressmand Jack Metcalf, 20 March 2000. Cited in:
\[Committee on Government Reform Hearings for the United States House of
Representatives, 2000 #1320\].* 2000. Cited in: Accountability of DoD,
FDA adn BioPort Officials for the Anthrax Vaccine Immunization Program
(AVIP\_. Committee on Government Reform Hearings for th United States
House of Representatives. Oct 3 and 11, 2000.

32\. Ivins, B., *et al.*, *Experimental anthrax vaccines: efficacy of
adjuvants combined with protective antigen against an aerosol Bacillus
anthracis spore challenge in guinea pigs.* Vaccine, 1995. 13(18): p.
1779-84. The efficacy of several human anthrax vaccine candidates
comprised of different adjuvants together with Bacillus anthracis
protective antigen (PA) was evaluated in guinea pigs challenged by an
aerosol of virulent B. anthracis spores. The most efficacious vaccines
tested were formulated with PA plus monophosphoryl lipid A (MPL) in a
squalene/lecithin/Tween 80 emulsion (SLT) and PA plus the saponin QS-21.
The PA+MPL in SLT vaccine, which was lyophilized and then reconstituted
before use, demonstrated strong protective immunogenicity, even after
storage for 2 years at 4 degrees C. The MPL component was required for
maximum efficacy of the vaccine. Eliminating lyophilization of the
vaccine did not diminish its protective efficacy. No significant
alteration in efficacy was observed when PA was dialyzed against
different buffers before preparation of vaccine. PA+MPL in SLT proved
superior in efficacy to the licensed United States human anthrax vaccine
in the guinea pig model.

33\. Ivins, B. and et al, *Adjuvant efficacy in experimental anthrax
vaccines: Protection studies in guinea pigs.* Abstracts of the 91st
General Meeting of the American Society for Microbiology, 1991: p. 121.

34\. Matsumoto, G., *The Pentagon\'s toxic secret*, in *Vanity Fair*.
1999. p. 83-96.

35\. Tomich, N., *Anthrax vaccine program scrutiny intensifies.* U.S.
Medicine, 2000. 36(11): p. 1,32,33,36.

36\. Carlson, B.C., *et al.*, *The endogenous adjuvant squalene can
induce a chronic T-cell-mediated arthritis in rats.* Am J Pathol, 2000.
156(6): p. 2057-65. Squalene is a cholesterol precursor, which
stimulates the immune system nonspecifically. We demonstrate that one
intradermal injection of this adjuvant lipid can induce joint-specific
inflammation in arthritis-prone DA rats. Histopathological and
immunohistochemical analyses revealed erosion of bone and cartilage, and
that development of polyarthritis coincided with infiltration of
alphabeta(+) T cells. Depletion of these cells with anti-alphabeta TcR
monoclonal antibody (R73) resulted in complete recovery, whereas
anti-CD8 and anti-gammadelta TcR injections were ineffective. The
apparent dependence on CD4(+) T cells suggested a role for genes within
the major histocompatibility complex (MHC), and this was concluded from
comparative studies of MHC congenic rat strains, in which DA.1H rats
were less susceptible than DA rats. Furthermore, LEW.1AV1 and PVG.1AV1
rats with MHC identical to DA rats were arthritis-resistant,
demonstrating that non-MHC genes also determine susceptibility. Some of
these genetic influences could be linked to previously described
arthritis susceptibility loci in an F2 intercross between DA and
LEW.1AV1 rats (ie, Cia3, Oia2 and Cia5). Interestingly, some F2 hybrid
rats developed chronic arthritis, a phenotype not apparent in the
parental inbred strains. Our demonstration that an autoadjuvant can
trigger chronic, immune-mediated joint-specific inflammation may give
clues to the pathogenesis of rheumatoid arthritis, and it raises new
questions concerning the role of endogenous molecules with adjuvant
properties in chronic inflammatory diseases.

37\. Gajkowska, B., *et al.*, *The experimental squalene
encephaloneuropathy in the rat.* Exp Toxicol Pathol, 1999. 51: p. 75-80.

38\. Lorentzen, J.C., *Identification of arthritogenic adjuvants of self
and foreign origin.* Scand J Immunol, 1999. 49(1): p. 45-50.

39\. Sverdrup, B., L. Klareskog, and S. Kleinau, *Common commercial
cosmetic products induce arthritis in the DA rat.* Environ Health
Perspect, 1998. 106(1): p. 27-32. squalene arthritis.

40\. Dijkstra, J., *et al.*, *Interaction of anti-cholesterol antibodies
with human lipoproteins.* J Immunol, 1996. 157(5): p. 2006-13.
Inoculation of mice with cholesterol-rich liposomes containing the
adjuvant monophosphoryl lipid A results in the production of antiserum
containing IgM Ab to cholesterol. The specificity of the Ab was to
cholesterol and structurally similar sterols containing a 3
beta-hydroxyl group. Anti-cholesterol binding activity was significantly
diminished if the 3 beta-hydroxyl was altered by either epimerization,
substitution, oxidation, or esterification. A similar specificity for 3
beta-hydroxy-sterols was observed for an anti-cholesterol IgM mAb. Both
hyperimmune serum and the mAb reacted with intact human
very-low-/intermediate-density lipoprotein (VLDL/IDL) and low-density
lipoproteins (LDL), but not high-density lipoproteins (HDL), in an
ELISA, but could react with total lipid extracts containing cholesterol
that were prepared from all three lipoprotein classes. Functionally,
immune serum or the mAb aggregated and induced a fusion-like reaction
with VLDL/IDL and LDL at low temperatures: these aggregates result in
spherical structures visible with light microscopy. Similarly, binding
of anti-cholesterol A to small cholesterol-rich liposomes resulted in
the appearance of vesicular structures with approximately 20- to
200-fold increased diameters. These data demonstrate that the
anti-cholesterol Ab recognize unesterified cholesterol in VLDL/IDL and
LDL; high-density lipoprotein cholesterol in the intact lipoprotein,
however, appears to be protected from reaction with these Ab.

41\. Ordovas, J.M., *Anticholesterol antibodies and plaque formation.*
Nutr Rev, 1996. 54(4 Pt 1): p. 124-7. Immunization of rabbits with a
protein-free formulation consisting of liposomes containing 71%
cholesterol and lipid A induced cholesterol antibodies in rabbits fed a
diet containing 0.5% and 1.0% cholesterol. Elevation in plasma
cholesterol levels was significantly less in immunized than in
nonimmunized rabbits. Immunization also resulted in a marked decrease in
the risk of developing atherosclerosis as determined by analysis of
aortic atherosclerosis by quantitative histological examination and
fatty streaks by automated morphometric probability-of-occurrence
mapping.

42\. Alving, C.R., *et al.*, *Immunization with cholesterol-rich
liposomes induces anti-cholesterol antibodies and reduces diet-induced
hypercholesterolemia and plaque formation.* J Lab Clin Med, 1996.
127(1): p. 40-9. Immunization of rabbits with a protein-free formulation
consisting of liposomes containing 71% cholesterol and lipid A as an
adjuvant induced anticholesterol antibodies that caused
complement-dependent lysis of liposomes lacking lipid A. The antibodies,
immunoglobulin G (IgG) and immunoglobulin M (IgM), also recognized
nonoxidized crystalline cholesterol as an antigen by enzyme-linked
immunosorbent assay (ELISA). The effects of immunization against
cholesterol on elevations in serum cholesterol and development of
atherosclerosis were examined in rabbits fed a diet containing 0.5% to
1.0% cholesterol. Although the mean serum cholesterol level, mainly in
the form of very-low-density lipoprotein cholesterol, rose as much as
60-fold in the nonimmunized rabbits, the elevation was significantly
less\--as much as 35% lower\--in the immunized rabbits. Elevation of
serum cholesterol was accompanied by an apparent drop in the level of
antibodies on initiating the diet, followed by a rebound on stopping the
diet, thus suggesting that the antibodies were adsorbed to cholesterol
that was present in circulating lipoproteins. When lipoprotein
fractions\--composed of either very-low-density and intermediate-density
lipoproteins derived from cholesterol-fed nonimmunized rabbits or human
low-density lipoproteins\--were tested as capture antigens by
solid-phase ELISA, reactivity was observed with IgG and IgM antibodies
present in the serum of immunized rabbits. Immunization also resulted in
a marked decrease in the risk of developing atherosclerosis. Analysis of
aortic atherosclerosis by quantitative histologic examination and fatty
streaks by automated morphometric probability-of-occurrence mapping
showed diminished atherosclerosis in most areas of the aorta in vaccine
recipients. It is proposed that immunization with liposomes containing
71% cholesterol and lipid A can reduce diet-induced hypercholesterolemia
and atherosclerosis.

43\. Alving, C.R., N.M. Wassef, and M. Potter, *Antibodies to
cholesterol: biological implications of antibodies to lipids.* Curr Top
Microbiol Immunol, 1996. 210: p. 181-6. Injection of silicone gel or
silicone oil intraperitoneally into BALB/c mice induced the formation of
antibodies that reacted by ELISA with highly purified crystalline
cholesterol and, to a much lesser extent, antibodies that reacted with a
phospholipid (dimyristoyl phosphatidylglycerol). Although IgM and IgG
antibodies to cholesterol were detected, the titers of IgG antibodies
were low when compared with IgM. The titers of IgM antibodies to
cholesterol in certain sera exhibited activities that reached baseline
values at dilutions as high as 1:5000, thus making them equivalent to
titers that have been previously published for ascites fluid containing
murine monoclonal antibodies to cholesterol. The antibodies to
cholesterol induced by silicone compounds are indistinguishable in their
binding to crystalline cholesterol from naturally-occurring antibodies
to cholesterol in normal human serum. They are also indistinguishable
from antibodies induced by a proposed vaccine to cholesterol that is
currently in late preclinical development for prevention of
hypercholesterolemia in humans. The anti-cholesterol vaccine, which
consists of liposomes heavily laden with cholesterol as an antigen and
lipid A as an adjuvant, induces antibodies that react with low density
lipoproteins (LDL) and opsonize them for removal by liver macrophages.
It appears that silicone gel or silicone oil causes recruitment and
adsorption of cholesterol at the injection site, and also serves as an
adjuvant that may have immunostimulant properties similar to lipid A for
inducing antibodies to lipids. Antibodies to lipids such as cholesterol
or phospholipids are not harmful to intact cell membranes because of
steric hindrance from surrounding lipids and larger macromolecules that
block binding of the antibodies.

44\. Lahita, R.G., *et al.*, *Low levels of total cholesterol,
high-density lipoprotein, and apolipoprotein A1 in association with
anticardiolipin antibodies in patients with systemic lupus
erythematosus.* Arthritis Rheum, 1993. 36(11): p. 1566-74. OBJECTIVE. To
determine if there is an association between low levels of high-density
lipoprotein cholesterol (HDL), apolipoprotein A1 (Apo A1), total
cholesterol, and anticardiolipin antibody (aCL) in patients with
systemic lupus erythematosus (SLE) who are not taking corticosteroids.
METHODS. We studied 75 outpatients with documented SLE who were
attending our hospital clinics: 57 were aCL positive and 18 were aCL
negative. Both IgG and IgM aCL levels were determined by enzyme-linked
immunosorbent assay. Lipid fractions (total cholesterol, HDL,
low-density lipoprotein, very-low-density lipoprotein, and
triglycerides) were determined by standard enzymatic techniques. Apo A1
and Apo B levels were determined by nephelometry. RESULTS. Patients with
SLE who were IgG aCL+ had low levels of serum cholesterol (mean +/- SD
173.6 +/- 34.6 mg/dl) and HDL (43.9 +/- 16.3 mg/dl) compared with aCL-
SLE patients, normal donors, and patients with other diseases. Apo A1
levels were also low in the aCL+ group (95.5 +/- 50.9 mg/dl) compared
with the aCL- group (152.7 +/- 32.6 mg/dl). There was no association of
total cholesterol level or aCL titer with clinical activity. CONCLUSION.
These data indicate that in SLE patients, there is an association
between antibody against the phospholipid cardiolipin and low levels of
cholesterol, HDL, and Apo A1.

45\. Alving, C.R. and G.M. Swartz, Jr., *Antibodies to cholesterol,
cholesterol conjugates and liposomes: implications for atherosclerosis
and autoimmunity.* Crit Rev Immunol, 1991. 10(5): p. 441-53. Polyclonal
and monoclonal antibodies to cholesterol are readily induced by
injecting cholesterol-loaded liposomes containing lipid A as an
adjuvant. Analysis of the literature reveals that conjugates of
cholesterol, and conjugates of analogues of cholesterol, with
heterologous proteins or lipids have been used as antigens in various
studies since 1925, and this has led to successful development of
immunoassays for steroid hormones. It is concluded that cholesterol is a
highly immunogenic molecule. The ability of monoclonal antibodies to
cholesterol to react with liposomes containing cholesterol to cause
complement-dependent immune damage to the liposomes is strongly
influenced by the lipid composition of the liposomes, the amount of
cholesterol in the liposomes, and the reaction temperature. The
antibodies also react with crystalline cholesterol in a solid-phase
ELISA and, depending on the particular monoclonal antibody, immune
reactivity may or may not be observed with cholesterol esters,
cholesterol analogues, or steroid hormones. Analysis by ELISA has
revealed that virtually all normal human sera contain varying levels of
naturally occurring IgG and IgM autoantibodies to cholesterol. Naturally
occurring autoantibodies to cholesterol are also observed in pigs, but
not in guinea pigs. Possible implications of these investigations for
theories of immune mechanisms that may have beneficial or detrimental
roles in processes of aging, atherosclerosis, and vascular diseases are
discussed.

46\. Alving, C.R., *Antibodies to liposomes, phospholipids, and
cholesterol: implications for autoimmunity, atherosclerosis, and aging.*
Prog Clin Biol Res, 1990. 343: p. 41-51.

47\. Swartz, G.M., Jr., *et al.*, *Antibodies to cholesterol.* Proc Natl
Acad Sci U S A, 1988. 85(6): p. 1902-6. Cholesterol-dependent complement
activation has been proposed as a factor that might influence the
pathogenesis of atherosclerosis. Although antibodies to cholesterol
conjugates have been reported, cholesterol is widely regarded as a
poorly immunogenic substance. Monoclonal IgM complement-fixing
antibodies to cholesterol were obtained in the present study after
immunizing mice with liposomes containing high amounts of cholesterol
(71 mol % relative to phosphatidylcholine) and lipid A as an adjuvant.
Clones were selected for the ability of secreted antibodies to react
with liposomes containing 71% cholesterol but not with liposomes
containing 43% cholesterol. The antibodies also reacted with crystalline
cholesterol in a solid-phase enzyme-linked immunosorbent assay. Binding
of monoclonal antibodies to the surface of crystalline cholesterol was
demonstrated by electron microscopy by utilizing a second antibody
(anti-IgM) labeled with colloidal gold. The immunization period required
to induce monoclonal antibodies was very short (3 days) and a high
fraction of the hybrid cells (at least 70%) were secreting detectable
antibodies to cholesterol. The results demonstrate that cholesterol can
be a highly immunogenic molecule and that complement-fixing antibodies
to cholesterol can be readily obtained.

48\. Folkers, K. and J.L. Nilsson, *\[Coenzyme Q regarded as a
vitamin\].* Nord Med, 1970. 83(16): p. 489-93.

49\. Estornell, E., *et al.*, *Effects of vitamin A deficiency on
mitochondrial function in rat liver and heart.* Br J Nutr, 2000. 84(6):
p. 927-34. The aim of this study was to investigate comparative effects
of vitamin A deficiency on respiratory activity and structural integrity
in liver and heart mitochondria. Male rats were fed a liquid control
diet (control rats) or a liquid vitamin A-deficient diet (vitamin
A-deficient rats) for 50 days. One group of vitamin-A deficient rats was
refed a control diet for 15 days (vitamin A-recovered rats). To assess
the respiratory function of mitochondria the contents of coenzyme Q
(ubiquinone, CoQ), cytochrome c and the activities of the whole electron
transport chain and of each of its respiratory complexes were evaluated.
Chronic vitamin A deficiency promoted a significant increase in the
endogenous coenzyme Q content in liver and heart mitochondria when
compared with control values. Vitamin A deficiency induced a decrease in
the activity of complex I (NADH-CoQ reductase) and complex II
(succinate-CoQ reductase) and in the levels of complex I and cytochrome
c in heart mitochondria. However, NADH and succinate oxidation rates
were maintained at the control levels due to an increase in the CoQ
content in accordance with the kinetic behaviour of CoQ as an
homogeneous pool. On the contrary, the high CoQ content did not affect
the electron-transfer rate in liver mitochondria, whose integrity was
preserved from the deleterious effects of the vitamin A deficiency.
Ultrastructural assessment of liver and heart showed that vitamin A
deficiency did not induce appreciable alterations in the morphology of
their mitochondria. After refeeding the control diet, serum retinol,
liver and heart CoQ content and the activity of complex I and complex II
in heart mitochondria returned to normality. However, the activities of
both whole electron transfer chain and complex I in liver were increased
over the control values. The interrelationships between physiological
antioxidants in biological membranes and the beneficial effects of their
administration in mitochondrial diseases are discussed.

50\. Anderson, S.S., I.G. Lyle, and R. Paterson, *Electron transfer
across membranes using vitamin K1 and coenzyme Q10 as carrier
molecules.* Nature, 1976. 259(5539): p. 147-8.

51\. Sobreira, C., *et al.*, *Mitochondrial encephalomyopathy with
coenzyme Q10 deficiency.* Neurology, 1997. 48(5): p. 1238-43.

52\. DiMauro, S., *et al.*, *Mitochondria in neuromuscular disorders.*
Biochimica et Biophysica Acta, 1998. 1366(1-2): p. 199-210.

53\. Suzuki, Y., *et al.*, *A case of diabetic amyotrophy associated
with 3243 mitochondrial tRNA(leu; UUR) mutation and successful therapy
with coenzyme Q10.* Endocrine Journal, 1995. 42(2): p. 141-5.

54\. Hansen, I.L., *et al.*, *Bioenergetics in clinical medicine. IX.
Gingival and leucocytic deficiencies of coenzyme Q10 in patients with
periodontal disease.* Res Commun Chem Pathol Pharmacol, 1976. 14(4): p.
729-38. The specific activities of the succinate dehydrogenase-coenzyme
Q10 reductase in mitochondria were determined for patients from a normal
periodontal practice. The criteria for selection were patients having a
bone score of 1.0-4.0 and a pocket depth of 2.5-5.2 mm. All 29 patients
showed a deficiency of 20-63% of CoQ10-enzyme activity in gingival
biopsies. The mean value was elevated (P less than 0.001) over that of
controls. For corresponding blood samples, 24/28 (86%) showed
deficiencies of 20-66% and a higher (P less than 0.001) mean value than
that of controls. Periodontal patients frequently have significant
gingival and leucocytic deficiencies of CoQ10. The leucocytic deficiency
indicates a systemic nutritional imbalance and is not likely caused by
neglected oral hygiene. A gingival deficiency could predispose this
tissue to periodontitis and this disease could even augment the
deficiency. These results support previously suggested adjunctive use of
CoQ10 with oral hygiene for improved treatment presumably through
bioenergetics.

55\. Wilkinson, E.G., *et al.*, *Bioenergetics in clinical medicine. II.
Adjunctive treatment with coenzyme Q in periodontal therapy.* Res Commun
Chem Pathol Pharmacol, 1975. 12(1): p. 111-23. Eight patients under
routine care for periodontitis received oral treatment with a form of
coenzyme Q (7 / CoQ10 and 1 / hexahydrocoenzyme Q4). An unchanged plaque
score showed the patients cooperated and were under plaque control. The
periodontal score decreased (p less than 0.01) on CoQ treatment.
Unexpectedly, the periodontal pocket depth decreased (P less than 0.05)
on CoQ treatment since all patients were considered candidates for
surgical intervention. Healing was so excellent 5-7 days post-biopsy
that the biopsy sites were difficult to locate. The healing was viewed
as extraordinarily effective. The mean value of the specific activities
of the succinate dehydrogenase-coenzyme Q10 reductase of gingival
biopsies increased (P less than 0.05) during treatment which could
correlate with the extraordinarily healing. Treatment of periodontitis
with coenzyme Q should be considered as adjunctive treatment with
current dental practice.

56\. Folkers, K. and R. Simonsen, *Two successful double-blind trials
with coenzyme Q10 (vitamin Q10) on muscular dystrophies and neurogenic
atrophies.* Biochimica et Biophysica Acta, 1995. 1271(1): p. 281-6.

57\. Kuz\'menko, I.V., *et al.*, *\[Efficiency of ubiquinone and
p-oxybenzoic acid in prevention of E-hypovitaminosis-induced development
of muscular dystrophy\].* Ukr Biokhim Zh, 1981. 53(5): p. 73-9. It is
shown that E-hypovitaminosis-induced muscular dystrophy in rabbits is
accompanied by a sharp decrease in the body mass, an increase in the
urine creatine-index, a decrease in the vitamin E and ubiquinone
contents in the liver and skeletal muscle tissues. In the myocardium
mitochondria a decrease in the vitamin E content and an increase in the
ubiquinone content are observed. The activity of NADH-cytochrome c-,
NADH-ubiquinone- and succinate-ubiquinone-reductase also varies in
mitochondria of the studied tissues. In myocardium organellas a direct
dependence is found between the content of ubiquinone, NADH- and
succinate-ubiquinone-reductase activity and an inverse one-between its
content and the activity of the NADH-cytochrome c-reductase system. It
is established that p-oxybenzoic acid as well as vitamin E prevents
development of muscular dystrophy and causes changes analogous in
direction in the activity of the ubiquinone-dependent enzymic systems of
mitochondria. Ubiquinone-9 is less efficient in preventing the
development of muscular dystrophy.

58\. Shimomura, Y., *et al.*, *Protective effect of coenzyme Q10 on
exercise-induced muscular injury.* Biochem Biophys Res Commun, 1991.
176(1): p. 349-55. The effect of coenzyme Q10 administration on
exercise-induced muscular injury was examined in rats. Coenzyme
Q10-treated and control rats were exercised by 90 min of downhill
treadmill running. A part of the animals in both groups were killed
immediately after exercise and the others were 40 h postexercise. After
the exercise bout, serum creatine kinase and lactate dehydrogenase
activities were elevated in the control rats, but not in the coenzyme
Q10-treated rats. These enzyme activities in the latter increased to the
similar level of the former 40 h postexercise. The muscle coenzyme Q10
content increased by the coenzyme Q10 treatment. These results suggest
that the coenzyme Q10 treatment protected skeletal muscles against
injury caused during exercise, but not against damage related with
inflammatory processes after exercise.

59\. Muller, D.P., J.K. Lloyd, and O.H. Wolff, *Vitamin E and
neurological function: abetalipoproteinaemia and other disorders of fat
absorption.* Ciba Found Symp, 1983. 101: p. 106-21. Evidence that
vitamin E is important for normal neurological function in humans is
presented. First, in abetalipoproteinaemia early therapy with vitamin E
delays and may prevent the development of the neurological
complications, and in patients with established lesions treatment can
arrest or reverse the neuropathy. Second, in other chronic disorders of
fat absorption with severe vitamin E deficiency, neurological
manifestations which are very similar to those described in untreated
abetalipoproteinaemia can be improved by vitamin E. Vitamin E
supplementation is therefore advisable for all patients with chronic fat
malabsorption who have low serum vitamin E concentration. Serum vitamin
E concentrations should also be measured in patients with
spinocerebellar disorders, whatever the aetiology.

60\. Muller, D.P.R., J.K. Lloyd, and A. Bird, *Long-term management of
abetalipoproteinemia. Possible role for vitamin E.* Arch Dis Child,
1977. 52: p. 209-14.

61\. Azizi, E., *et al.*, *Abetalipoproteinemia treated with parenteral
and oral vitamin A and E and with medium chain triglycerides.* Acta
Pediatr Scand, 1978. **67**: p. 797-801.

62\. Hegele, R.A. and A. Angel, *Arrest of neuropathy and myopathy in
abetalipoproteinemia with high-dose vitamin E therapy.* Can Med Assoc J,
1985. 132(1): p. 41-4. A 16-year-old girl, one of dizygotic twins,
presented in 1976 complaining of a 1-year history of a lack of
coordination and an inability to run. The results of biochemical tests
confirmed the diagnosis of classic abetalipoproteinemia. In addition to
the recognized neurologic features of this disorder, she had a reduced
evoked motor unit potential and markedly elevated serum levels of muscle
enzymes, which suggested myositis. The serum vitamin E level was
markedly decreased. Oral therapy with vitamin E, 800 mg daily, was
begun, and in 1981 the dosage was increased to 3200 mg daily. Over the 7
years of follow-up she improved clinically, there was an increase in the
evoked motor unit potential, the serum levels of some of the muscle
enzymes decreased to normal, and the serum and tissue vitamin E levels
increased significantly. It was concluded that treatment with high doses
of vitamin E was responsible for the arrest of the usually progressive
neuropathy and myopathy.

63\. Iannaccone, S.T. and R.J. Sokol, *Vitamin E deficiency in
neuropathy of abetalipoproteinemia.* Neurology, 1986. 36(7): p. 1009.

64\. Alleva, R., *et al.*, *Coenzyme Q blocks biochemical but not
receptor-mediated apoptosis by increasing mitochondrial antioxidant
protection.* FEBS Lett, 2001. 503(1): p. 46-50. Generation of free
radicals is often associated with the induction and progression of
apoptosis. Therefore, antioxidants can prove anti-apoptotic, and can
help to elucidate specific apoptotic pathways. Here we studied whether
coenzyme Q, present in membranes in reduced (ubiquinol) or oxidised
(ubiquinone) forms, can affect apoptosis induced by various stimuli.
Exposure of Jurkat cells to alpha-tocopheryl succinate (alpha-TOS),
hydrogen peroxide, anti-Fas IgM or TRAIL led to induction of apoptosis.
Cell death due to the chemical agents was suppressed in cells enriched
with the reduced form of coenzyme Q. However, coenzyme Q did not block
cell death induced by the immunological agents. Ubiquinol-10 inhibited
reactive oxygen species (ROS) generation in cells exposed to alpha-TOS,
and a mitochondrially targeted coenzyme Q analogue also blocked
apoptosis triggered by alpha-TOS or hydrogen peroxide. Therefore, it is
plausible that ubiquinol-10 protects cells from chemically-induced
apoptosis by acting as an antioxidant in mitochondria. Our results also
indicate that generation of free radicals may not be a critical step in
induction of apoptosis by immunological agents.

65\. Fernandez-Ayala, D.J., *et al.*, *Coenzyme Q protects cells against
serum withdrawal-induced apoptosis by inhibition of ceramide release and
caspase-3 activation.* Antioxid Redox Signal, 2000. 2(2): p. 263-75.
Coenzyme Q10 (CoQ10) is a component of the antioxidant machinery that
protects cell membranes from oxidative damage and decreases apoptosis in
leukemic cells cultured in serum-depleted media. Serum deprivation
induced apoptosis in CEM-C7H2 (CEM) and to a lesser extent in CEM-9F3, a
subline overexpressing Bcl-2. Addition of CoQ10 to serum-free media
decreased apoptosis in both cell lines. Serum withdrawal induced an
early increase of neutral-sphingomyelinase activity, release of
ceramide, and activation of caspase-3 in both cell lines, but this
effect was more pronounced in CEM cells. CoQ10 prevented activation of
this cascade of events. Lipids extracted from serum-depleted cultures
activated caspase-3 independently of the presence of mitochondria in
cell-free in vitro assays. Activation of caspase-3 by lipid extracts or
ceramide was prevented by okadaic acid, indicating the implication of a
phosphatase in this process. Our results support the hypothesis that
plasma membrane CoQ10 regulate the initiation phase of serum
withdrawal-induced apoptosis by preventing oxidative damage and thus
avoiding activation of downstream effectors as neutral-sphingomyelinase
and subsequent ceramide release and caspase activation pathways.

66\. Kagan, T., *et al.*, *Coenzyme Q10 can in some circumstances block
apoptosis, and this effect is mediated through mitochondria.* Ann N Y
Acad Sci, 1999. 887: p. 31-47. The mitochondrial component coenzyme Q10
(CoQ10) has been used for many years as a dietary supplement intended to
promote good health by trapping free radicals, thus preventing lipid
peroxidation and DNA damage. We have tested its use as a generic
anti-apoptotic compound and have found that its ability to protect
against apoptosis varies depending on both cell type and mode of cell
death induction. We have further established that this protection may be
mediated by its effect on mitochondrial function and viability. We
provide additional evidence that CoQ10\'s protective effect on
mitochondrial membrane potential does not always result in altered
mitochondrial enzyme activity and neither does it guarantee survival.
These observations open the way for further investigations into the
mechanisms involved in mitochondrial control of apoptosis.

67\. Mosca, L., *et al.*, *Modulation of apoptosis and improved redox
metabolism with the use of a new antioxidant formula.* Biochem
Pharmacol, 2002. 63(7): p. 1305-14. Oxidative stress is involved in the
pathogenesis of a wide spectrum of diseases, implicating that strategies
directed at counterbalancing oxidative processes could have a role in
clinical medicine. There is also an evidence that oxidative stress acts
as a major determinant of apoptotic cell death. Many studies have
reported favourable effects of antioxidant formulas on several
parameters of the oxidant-antioxidant balance, but none of them has
focused whether antioxidant formulas could modulate apoptosis. We
investigated in 20 healthy individuals the effect of supplementation
with a formula containing alpha-tocopherol, alpha-lipoic acid, coenzyme
Q(10), carnitines, and selenomethionine, on plasma oxidant status and
peroxide levels, erythrocyte antioxidant enzymes, lymphocyte apoptosis,
and generation of ROS at the mitochondrial level. Control subjects
received only carnitines or an incomplete formula with alpha-tocopherol,
alpha-lipoic acid, coenzyme Q(10), and selenomethionine. Supplementation
with the complete formula resulted in a significant increase in the
plasma antioxidant status that was mirrored by a decrease in blood
peroxide levels and a reduced generation of ROS at the mitochondrial
level. This was associated with a significant decrease in the frequency
of peripheral blood lymphocytes, with either CD4 or CD8 phenotype,
undergoing apoptosis. Less consistent results were found when either
incomplete formula was used. Our study suggests that supplementation
with antioxidant formulas can modulate the process of apoptosis under in
vivo conditions. The clinical potential of this strategy in the
treatment of diseases with an elevated commitment to apoptosis should be
explored.

68\. Buckenmeyer, P.J., *et al.*, *Effect of concurrent exercise and
physostigmine on lactate and pyruvate in plasma, muscle, and brain
tissue of rats.* Pharmacol Biochem Behav, 1994. 47(4): p. 779-88. The
purpose of this investigation was to determine the effect of
physostigmine (Phy) and/or concurrent exercise on lactate, pyruvate, and
L/P ratio in plasma, skeletal muscle, and brain tissue in male
Sprague-Dawley rats. The Phy-dosed (Phy-D) and Phy-dosed + concurrent
acute exercise (Phy-D + CAE) groups elicited significantly higher L/P
ratios in plasma compared to the acutely exercised (AE) group at 30 min
postexercise. Physostigmine dosing, with or without exercise, resulted
in significantly lower muscle pyruvate levels, from 30 to 50 min
postdrug administration, in Phy-D and Phy-D + CAE groups compared to the
AE group. In the brain, lactate values were significantly elevated in
the acutely exercised groups at 5 min postexercise with or without Phy
dosing. However, at 15 to 30 min postexercise, lactate values were
significantly elevated in the Phy-D + CAE compared to the AE group.
These data suggest that when Phy is administered prior to a 20-min
moderately intensive exercise bout, there is an accumulation of lactate
for a prolonged period of time in recovery.

69\. Phillips, P. and et al, *Title Not Yet on Current Contents.* Annals
of Internal Medicine, 2002. Oct 1, 2002: p. Pages to be secured.

70\. Ackrell, B.A., *et al.*, *Effect of iron deficiency on succinate-
and NADH-ubiquinone oxidoreductases in skeletal muscle mitochondria.* J
Biol Chem, 1984. 259(16): p. 10053-9. The effects of iron deficiency on
the NADH- and succinate-oxidizing complexes of rat skeletal muscle
mitochondria have been investigated. Both systems were similarly
affected: activities were about 30% of normal in dehydrogenase,
ubiquinone reductase, and oxidase assays, and similar reductions in the
concentration of their respective flavin prosthetic groups were also
evident in the iron-deficient membranes. Thus, the turnover numbers of
the two enzymes were unchanged in iron deficiency. Sodium dodecyl
sulfate-polyacrylamide gel electrophoresis revealed similarly reduced
levels of those peptide components of Complexes I and II that could be
unequivocally distinguished. Soluble beef heart succinate dehydrogenase
added to alkaline-treated rat skeletal muscle mitochondrial membranes
attached to binding sites exposed by the treatment, forming a hybrid
complex indistinguishable from the original skeletal muscle complex,
with restoration of succinoxidase and succinate-ubiquinone reductase
activities to the levels observed in the original rat membranes.
Iron-deficient particles behaved like the normal in these tests. No
unfilled binding sites for the enzyme could be detected prior to
alkaline treatment. The data are interpreted as indicating that the
lower activities of these two respiratory complexes in iron deficiency
are due to lower content of the enzymes rather than to the presence of
impaired enzymes in the membrane, that only fully competent complexes
are present in these membranes, and that iron-deficient complexes are
either not assembled or are lost after assembly.

71\. Shingwekar, A.G., M. Mohanram, and V. Reddy, *Effect of zinc
supplementation on plasma levels of vitamin A and retinol-binding
protein in malnourished children.* Clin Chim Acta, 1979. 93(1): p.
97-100. The relationship between plasma levels of vitamin A and zinc was
studied in 45 children suffering from vitamin A deficiency and 20
children with protein-energy malnutrition. Thirty apparently normal
children of the same age group were also studied for comparison. The
mean levels of plasma vitamin A, retinol-binding protein and zinc were
significantly lower in vitamin A-deficient children and in children with
PEM, as compared to controls. Supplementation with 40 mg zinc daily for
5\--10 days resulted in a significant increase in plasma vitamin A and
RBP levels in children with PEM but not in the vitamin A-deficient
group. There was, however, no correlation between plasma levels of
vitamin A and zinc. The data suggest that in children with PEM, apart
from deficiencies of protein and vitamin A, zinc deficiency may also
contribute to the lowering of plasma vitamin A levels. They also suggest
that in vitamin A-deficient children, without protein-energy
malnutrition, zinc deficiency does not seem to have a role.

72\. Vadhanavikit, S. and H.E. Ganther, *Decreased ubiquinone levels in
tissues of rats deficient in selenium.* Biochem Biophys Res Commun,
1993. 190(3): p. 921-6. The effect of selenium deficiency (-Se) on the
levels of ubiquinones in liver, heart, kidney, and leg muscle was
studied in the rat. Levels of ubiquinone 9 and ubiquinone 10 in the
liver of -Se rats were about 50% of the levels in selenium adequate
animals. Both ubiquinones in the heart were about 15% lower in -Se rats.
Only ubiquinone 9 was significantly lower in the kidney of -Se rats.
There was no difference in ubiquinone levels in leg muscle. Glutathione
peroxidase activity in the tissues of -Se rats was \> 95% lower. It is
concluded that Se, as an integral part of the enzyme glutathione
peroxidase, may protect tissues from oxidative damage, thereby
preserving the ability of the cells to synthesize ubiquinone and
preventing ubiquinone from oxidative degradation.

73\. Pedersen, H.S., *et al.*, *High serum coenzyme Q10, positively
correlated with age, selenium and cholesterol, in Inuit of Greenland. A
pilot study.* Biofactors, 1999. 9(2-4): p. 319-23.

74\. Pelton, R., *et al.*, *Drug-Induced Nutrient Depletion Handbook
1999-2000*. 1999, Hudson, Ohio: Lexi-Comp, Inc. 485.

75\. *Pfizer reports 38% increase*, in *San Diego Union Tribune, January
24*. 2002: San Diego.

76\. Sinzinger, H., *Does vitamin E beneficially affect muscle pains
during HMG-Co-A-reductase inhibitors without CK-elevation.*
Atherosclerosis, 2000. 149: p. 225.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10799017.

77\. Sinzinger, H., F. Chehne, and G. Lupattelli, *Oxidation Injury in
Patients Receiving HMG-CoA Reductase Inhibitors: Occurrence in Patients
Without Enzyme Elevation or Myopathy.* Drug Saf, 2002. 25(12): p.
877-83. BACKGROUND: Myopathy in its severe forms including
rhabdomyolysis is a very rare adverse effect occurring during
monotherapy with the HMG-CoA reductase inhibitors (\'statins\') and is
associated with pronounced signs of oxidation injury. This has been
found at a local (muscle) as well as at a systemic level (blood).
Several lines of evidence indicate that even mild forms of myopathy
during statin treatment may be associated with in vivo oxidation injury.
In contrast, statin therapy has been shown to be associated with a
decrease in oxidation injury. OBJECTIVE: The aim of this study was to
investigate whether patients with heterozygous familial
hypercholesterolaemia who did not exhibit any symptoms or abnormalities
in safety parameters during 6 months of treatment with various statins
(atorvastatin, fluvastatin, lovastatin, pravastatin, simvastatin) did
exhibit a change in oxidation injury as assessed by the isoprostane
levels. METHODS: Blood (plasma and serum) as well as urine was tested
before and 1, 3 and 6 months after starting statin therapy. Results: Out
of 111 treated patients (63 males, 48 females; aged 19 to 58 years) who
did not experience any adverse effects during statin treatment, 11
(seven males, four females; aged 24 to 51 years) showed a pronounced
increase in 8-epi-prostaglandin (PG) F(2alpha) in all the compartments
examined. In the remaining 100 patients (56 males, 44 females; aged 19
to 58 years) there was either no change in or even an apparent decrease
in 8-epi-PGF(2alpha). This increase was monitored with all the statins
administered. If elevated, the increase in 8-epi-PGF(2alpha) remained
without change throughout the entire follow-up period. No sex difference
or differential response between smokers and nonsmokers was observed.
DISCUSSION: These findings indicate that in the absence of other
clinically observable adverse effects, in some of the patients, for an
as yet unknown reason, statin therapy may be associated with increased
oxidation injury. The fact that changing to another statin is apparently
not necessarily associated with an identical response raises the
question of a specific predisposition for certain compounds in a given
patient. These data add a further piece of evidence that mild adverse
effects of statins that are difficult to assess might be much more
prevalent than widely considered. The clinical relevance and consequence
of these findings still remains to be assessed.

78\. Sinzinger, H., P. Schmid, and J. O\'Grady, *Two different types of
exercise-induced muscle pain without myopathy and CK -elevation during
HMG-Co-enzyme-A-reductase inhibitor treatment.* Atherosclerosis, 1999.
143(11): p. 459-460.

79\. Orsi, A., O. Sherman, and Z. Woldeselassie, *Simvastatin-associated
memory loss.* Pharmacotherapy, 2001. 21(6): p. 767-9. The statins are
widely used to treat dyslipidemias. They are generally associated with
mild adverse effects, but rarely, more serious reactions may occur. A
51-year-old man experienced delayed-onset, progressive memory loss while
receiving simvastatin for hypercholesterolemia. His therapy was switched
to pravastatin, and memory loss resolved gradually over the next month,
with no recurrence of the adverse effect.

80\. Muldoon, M.F., *et al.*, *Effects of lovastatin on cognitive
function and psychological well-being.* Am J Med, 2000. 108(7): p.
538-46.

81\. Bruckert, E., C. Simonetta, and P. Giral, *Compliance with
fluvastatin treatment characterization of the noncompliant population
within a population of 3845 patients with hyperlipidemia. CREOLE Study
Team.* J Clin Epidemiol, 1999. 52(6): p. 589-94. The objectives of the
study were to analyze 1) compliance with an HMG-CoA reductase inhibitor,
2) the relationship between treatment compliance and sociodemographic,
clinical, and psychological criteria, and 3) the effect of raising
patients\' awareness through distribution of an information notice. The
results analyzed in this article compare the noncompliant and compliant
population independently of awareness. This open-label study was
conducted in two randomized parallel groups: a control group that
received the information normally given by the practitioners, and an
awareness group that received specific informational brochures on diet
and cardiovascular risk factors and the reasons for the treatment. Male
and female patients (n = 3845) aged 18 to 75 years with primary
hypercholesterolemia (type IIa and IIb), not taking a
cholesterol-lowering drug or for whom an ongoing treatment was poorly
tolerated or ineffective, were to be included. Cholesterol levels had to
be greater than 250 mg/dL (or 200 mg/dL if previous coronary history)
and triglyceride levels less than 350 mg/dL. A total of 2888 subjects
(75%) were defined as compliant (taking more than 90% of the
prescription) and 957 (25%) noncompliant. Both populations are identical
for age, sex ratio, and different risk factors, with the exception of
diabetes. The adverse effects in noncompliant subjects were very clearly
different, with an overrepresentation of gastrointestinal and neurologic
effects and the noncompliant patients more frequently having more than
one adverse effect. Noncompliant patients had an identical duration of
follow-up, and the number of patients claiming to have a symptom related
to hypercholesterolemia, self-evaluation of cardiovascular risk level,
and source of knowledge about cholesterol and diet was similar in both
groups. In contrast, in the noncompliant group, there were a larger
number of symptomatic patients who thought the drug did not improve the
symptoms. In practice, these results show that physicians should
systematically evaluate compliance by looking for and analyzing adverse
effects and by reassuring the patient when these effects are minor or
probably unrelated to the treatment. Diabetics and polymedicated
patients deserve special attention in this regard.

82\. Gaist, D., *et al.*, *Statins and risk of polyneuropathy: A
case-control study.* Neurology, 2002. 58: p. 1333-1337.
***Background:*** Several case reports and a single epidemiologic study
indicate that use of statins occasionally may have a deleterious effect
on the peripheral nervous system. The authors therefore performed a
population-based study to estimate the relative risk of idiopathic
polyneuropathy in users of statins. ***Method:*** The authors used a
population-based patient registry to identify first-time-ever cases of
idiopathic polyneuropathy registered in the 5-year period 1994 to 1998.
For each case, validated according to predefined criteria, 25 control
subjects were randomly selected among subjects from the background
population matched for age, sex, and calendar time. The authors used a
prescription register to assess exposure to drugs and estimated the odds
ratio of use of statins (ever and current use) in cases of idiopathic
polyneuropathy compared with control subjects. ***Results:*** The
authors verified a diagnosis of idiopathic polyneuropathy in 166 cases.
The cases were classified as definite (35), probable (54), or possible
(77). The odds ratio linking idiopathic polyneuropathy with statin use
was 3.7 (95% CI 1.8 to 7.6) for all cases and 14.2 (5.3 to 38.0) for
definite cases. The corresponding odds ratios in current users were 4.6
(2.1 to 10.0) for all cases and 16.1 (5.7 to 45.4) for definite cases.
For patients treated with statins for 2 or more years the odds ratio of
definite idiopathic polyneuropathy was 26.4 (7.8 to 45.4).
***Conclusions:*** Long-term exposure to statins may substantially
increase the risk of polyneuropathy.

83\. Blasiak, J. and Z. Walter, *Protective action of cholesterol
against changes in membrane fluidity induced by malathion.* Acta
Biochimica Polonica, 1992. 39(1): p. 49-52. no abstract.

84\. Nevin, D.N., *et al.*, *Paraoxonase genotypes, lipoprotein lipase
activity, and HDL.* Arterioscler Thromb Vasc Biol, 1996. 16(10): p.
1243-9. Paraxonase, an enzyme associated with the high density
lipoprotein (HDL) particle, hydrolyzes paraoxon, the active metabolite
of the insecticide parathion. Several studies have shown that paraxonase
levels in humans have a distribution characteristic of two alleles, one
with low activity and the other with high activity. Paraoxonase also has
arylesterase activity, which does not exhibit activity polymorphism and
can therefore serve as an estimate of enzyme protein. Although the
ability of paraoxon to irreversibly inhibit lipoprotein lipase (LPL) has
been exploited experimentally for many years, the role of plasma
paraoxonase in lipoprotein metabolism is unknown. Seventy-two normal
individuals were examined for paraoxonase genotypes, plasma paraoxonase
and arylesterase activities, postheparin LPL and hepatic lipase (HL)
activities, and lipoprotein levels to determine whether (1) paraoxonase
activity or genotype determines lipoprotein levels via an effect on LPL
or HL activity or (2) variation in LPL and HL activities determines HDL
levels and indirectly affects paraoxonase activity and protein levels in
plasma. In the entire group, paraoxonase activity was related to
arylesterase activity and genotype. Whereas arylesterase activity was
correlated with HDL cholesterol (HDL-C) and apolipoproteinA-I (apoA-I)
levels, neither arylesterase nor paraoxonase was correlated with LPL or
HL activity. Furthermore, LPL activity was positively correlated and HL
inversely correlated with HDL cholesterol and apoA-I levels, whereas LPL
was inversely correlated with triglyceride levels. The paraoxonase
genotypes of the study group were 30 individuals homozygous for the
low-activity allele, 38 heterozygotes, and 4 individuals homozygous for
the high-activity allele. Paraoxonase genotype accounted for
approximately .75 of the variation in paraoxonase activity. Paraoxonase
activity was linearly related to arylesterase activity within each
subgroup. No difference in either LPL or HL activity was seen as a
function of paraoxonase genotype, nor were differences seen in plasma
triglyceride or HDL-C by genotype by ANOVA. The relation between LPL and
HL and components of HDL in the paraoxonase genotypic subgroups in
general reflected the associations seen in the group as a whole.
Multivariate analysis showed that LPL, HL, and arylesterase, a measure
of paraoxonase mass, were independent predictors of HDL cholesterol,
while paraoxonase genotype or activity was not. Thus, variation in LPL
and HL appears to be significantly related to HDL cholesterol and apoA-I
levels. The levels of HDL are a major correlate of paraoxonase protein
levels, while paraoxonase genotype is the major predictor of plasma
paraoxonase activity.

85\. Kishi, H., T. Kishi, and K. Folkers, *Bioenergetics in clinical
medicine. III. Inhibition of coenzyme Q10-enzymes by clinically used
anti-hypertensive drugs.* Res Commun Chem Pathol Pharmacol, 1975. 12(3):
p. 533-40. Background data revealed that some American and Japanese
patients with essential hypertension, including many who were not being
treated with any anti-hypertensive drug, had a deficiency of coenzyme
Q10. Eight clinically used anti-hypertensive drugs have now been tested
for inhibition of two mitochondrial coenzyme Q10-enzymes of heart
tissue, succinoxidase and NADH-oxidase. Diazoxide and propranolol
significantly inhibited the CoQ10-succinoxidase and CoQ10-NADH-oxidase,
respectively. Metoprolol did not inhibit succinoxidase, and was
one-fourth as active as propranolol for inhibition of NADH-oxidase.
Hydrochlorothiazide, hydralazine, ans clonidine also inhibited
CoQ10-NADH-oxidase. Reserpine did not inhibit either CoQ10-enzyme, and
methyldopa was a very eak inhibitor of succinoxidase. The
internationally recognized clinical side-effects of propranolol may be
due, in part, to inhibition of CoQ10-enzymes which are indispensable in
the bioenergetics of cardiac function. A pre-existing deficiency of
coenzyme Q10 in the myocardium of hypertensive patients could be
augmented by subsequent treatment with propranolol, possibly to the
\"life-threatening\" state described by others.

86\. Kishi, T., T. Watanabe, and K. Folkers, *Bioenergetics in clinical
medicine XV. Inhibition of coenzyme Q10-enzymes by clinically used
adrenergic blockers of beta-receptors.* Res Commun Chem Pathol
Pharmacol, 1977. 17(1): p. 157-64. Adrenergic blockers for
beta-receptors were studied for inhibition of mitochrondrial
CoQ10-enzymes. These enzymes are indispensable for the bioenegetics of
the myocardium. Propranolol is frequently used to treat hypertension; in
some patients, it depresses myocardial function as an adverse reaction.
This side effect may be related to the inhibition by propranolol of
CoQ10-enzymes of the myocardium. Timolol showed negligible inhibition of
the CoQ10-enzyme, NADH-oxidase. Metoprolol was less inhibitory than
propranolol. Five alprenolols showed inhibition which approached that of
propranolol. The 1-isomer of alprenolol showed weak inhibition of
another CoQ10-enzyme, succinoxidase, but the other beta-blockers were
essentially non-inhibitory to this enzyme. The drug of choice is
timolol, based on negligible inhibition of these bioenergetic enzymes of
the heart, which correlates with its pharmacologically low cardiac
depressant effects.

87\. Reyes, A.J., *et al.*, *Diuretics and zinc.* S Afr Med J, 1982.
62(11): p. 373-5. Distal tubule diuretics (DTDs), including
chlorothiazide, hydrochlorothiazide, bendroflumethiazide, chlorthalidone
and xipamide, have been found to increase urinary zinc output through a
poorly understood mechanism which could involve both direct and
hormone-mediated processes. Significant zinc depletion may occur during
long-term administration of DTDs, principally in conditions associated
with diminished total body zinc levels such as hepatic cirrhosis,
diabetes mellitus, gastro-intestinal disorders and several renal
diseases. Attention to the early symptoms of zinc deficiency such as
hypogeusia, hyposmia, abnormal dark adaptation and impotence and the
monitoring of serum zinc levels are advisable during long-term treatment
with common DTDs.

88\. Cohanim, M. and E.R. Yendt, *The effects of thiazides on serum and
urinary zinc in patients with renal calculi.* Johns Hopkins Med J, 1975.
136(3): p. 137-41. Urine zinc (Zn) excretion was significantly higher in
338 male patients with renal calculi than in 21 normal male subjects,
but there was no significant difference between 102 female patients with
renal calculi and 15 normal female subjects. In 33 patients with renal
calculi urinary Zn was less than 50% of the mean normal value; in these
patients there is a possibility that lower urine Zn excretion
contributes to the formation of renal calculi. Hydrochlorothiazide
produced a striking and sustained increase in urinary Zn excretion,
which might contribute to the efficacy of this drug in the prevention of
calcific renal stones. Serum Zn levels remained normal in the majority
of subjects on long term thiazide therapy, but the occasional finding of
subnormal levels suggests that long term thiazide therapy carries with
it the hazard of Zn deficiency.

89\. Mountokalakis, T., *et al.*, *Zinc deficiency in mild hypertensive
patients treated with diuretics.* J Hypertens Suppl, 1984. 2(3): p.
S571-2. Mean hair zinc was found to be significantly lower in 20 mild
hypertensive patients treated with thiazides for 6-36 months than in 19
mild hypertensive patients who had not been given diuretics for at least
six months before study. Mean serum zinc did not differ significantly
between the two groups. Chronic diuretic treatment can result in zinc
deficiency through enhanced urinary excretion of zinc.

90\. Lockett, C.J., *et al.*, *Zinc, angiotensin I-converting enzyme and
hypertension.* S Afr Med J, 1983. 64(26): p. 1022-4.
Angiotensin-converting enzyme, or kininase II, is a zinc metallo-enzyme;
the cation forms part of its active site. The enzyme transforms
angiotensin I into angiotensin II and hydrolyses bradykinin into
inactive peptides. Antihypertensive diuretics may decrease the activity
of the angiotensin-converting enzyme by increasing urinary zinc
excretion. Captopril inhibits the angiotensin-converting enzyme, but
also affects electrolyte excretion and should be reassessed as a
possible first-choice antihypertensive agent in the treatment of
essential hypertension.

91\. Bunning, P. and J.F. Riordan, *The functional role of zinc in
angiotensin converting enzyme: implications for the enzyme mechanism.* J
Inorg Biochem, 1985. 24(3): p. 183-98. Zinc is essential to the
catalytic activity of angiotensin converting enzyme. The enzyme contains
one g-atom of zinc per mole of protein. Chelating agents abolish
activity by removing the metal ion to yield the inactive, metal-free
apoenzyme. Zinc does not stabilize protein structure since the native
and apoenzymes are equally susceptible to heat denaturation. Addition of
either Zn2+, Co2+, or Mn2+ to the apoenzyme generates an active
metalloenzyme; Fe2+, Ni2+, Cu2+, Cd2+, and Hg2+ fail to restore
activity. The activities of the metalloenzymes follow the order Zn
greater than Co greater than Mn. The protein binds Zn2+ more firmly than
it does Co2+ or Mn2+. Hydrolysis of the chromophoric substrate,
furanacryloyl-Phe-Gly-Gly, by the active metalloenzymes is subject to
chloride activation; the activation constant is not metal dependent.
Metal replacement mainly affects Kcat with very little change in Km,
indicating that the role of zinc is to catalyze peptide hydrolysis.

92\. Reeves, P.G. and B.L. O\'Dell, *An experimental study of the effect
of zinc on the activity of angiotensin converting enzyme in serum.* Clin
Chem, 1985. 31(4): p. 581-4. The activity in serum of zinc-dependent
angiotensin converting enzyme (ACE), is measured to aid in diagnosis and
monitor treatment of certain diseases. This report shows the effect of
dietary zinc deprivation on ACE activity in the serum of rats. The mean
(and SE) of the zinc concentration (mumol/L) in serum was 3.5 (0.3) in
rats deprived of dietary zinc for four days, 16.3 (0.2) in control rats,
and 19.8 (0.9) in rats deprived of zinc for four days, then repleted
with zinc for 12 h. The respective mean (and SE) of ACE activities
(nmol/mL per min) in serum were 390 (15), 543 (13), and 545 (20). Serum
ACE activity was restored also by adding zinc to the assay mixture in
vitro. The Vmax for ACE was 1.4 times greater when serum was diluted
40-fold as compared with twofold dilution. There was a small effect on
the Km for the substrate, but the Km for zinc was decreased by 22-fold
when serum was diluted 40-fold. The Vmax under these conditions was
decreased by only 9%.

93\. Reeves, P.G. and B.L. O\'Dell, *Effects of dietary zinc deprivation
on the activity of angiotensin-converting enzyme in serum of rats and
guinea pigs.* J Nutr, 1986. 116(1): p. 128-34. Angiotensin-converting
enzyme (ACE) is a zinc-dependent peptidyldipeptide hydrolase found in
blood and in the endothelium of many tissues. This study was designed to
determine the effects of dietary zinc deprivation in rats and guinea
pigs on ACE activity in serum. Rats were fed zinc-deficient (less than 1
mg/kg) or zinc-adequate (25 mg/kg) diets for 3 d. Guinea pigs were fed
zinc-deficient (less than 1 mg/kg) or zinc-adequate (100 mg/kg) diets
for 3 wk. The plasma zinc concentration in the zinc-deprived rats was
26% and that of the deprived guinea pigs 35% of the control values. When
serum was diluted by one-half in the assay medium, the respective ACE
activities were 42 and 59% of controls. In spite of decreased ACE
activity there were no differences in plasma angiotensin II levels in
either species or in bradykinin levels in the rat. To determine the
effect of zinc on the kinetics of serum ACE, rat serum was treated to
remove low-molecular-weight components and diluted 30-fold in an assay
medium that contained a final zinc concentration of 0.68 microM.
Supplemental zinc (total, 24 microM) increased both Vmax and Km. EDTA
inhibited the enzyme activity, and the inhibition was totally reversible
by zinc. Copper at 30 microM had no effect on ACE activity. The results
of this study show that ACE activity in serum of rats and guinea pigs is
highly sensitive to the dietary intake of zinc and suggest that
metabolism of the vasoactive hormones, angiotensin II and bradykinin,
might be affected in vivo.

94\. White, C.L., *et al.*, *The effect of zinc in vivo and in vitro on
the activities of angiotensin converting enzyme and kininase-I in the
plasma of rats.* Biochem Pharmacol, 1986. 35(15): p. 2489-93. Two groups
of rats were pair fed, for 18 days, diets containing either 2.6 (Zn
deficient) or 100 mg Zn/kg (control diet). Plasma was assayed
spectrophotometrically for the activity of kininase-I and angiotensin
converting enzyme (ACE) in the presence of varying concentrations of
added Zn2+. Zinc deficient rats had only 76% of the activity of both
kininase-I and ACE compared with zinc supplemented control rats. There
was a significant linear relationship between enzyme activity and
concentration of zinc in plasma for both enzymes. When zinc was added to
the enzyme incubation mixture for zinc deficient rats, the activity of
ACE increased by 73% and that of kininase-I by 33%. This Zn2+-stimulated
increase in enzyme activity was negatively correlated with the in vivo
concentration of zinc in plasma, and a plateau in enzyme activity was
seen at concentrations of plasma zinc that were commensurate with normal
zinc status (over 14 mumol/1). The results demonstrate that the
activities of both kininase-I and ACE are dependent on the concentration
of zinc in vivo and in vitro, and suggest that information concerning
the concentration of zinc in plasma and assay solutions be a
prerequisite solutions be a prerequisite for the use of these enzymes in
the clinical diagnosis of disease states. The results also showed that
the activity of ACE and kininase-I in plasma could be used for the
biochemical diagnosis of a suboptimal zinc status.

95\. Schullek, J.R. and I.B. Wilson, *The binding of zinc to
angiotensin-converting enzyme.* Arch Biochem Biophys, 1988. 265(2): p.
346-50. The equilibrium constant for the dissociation of zinc ion from
angiotensin-converting enzyme (ACE) was measured using zinc ion buffers
of zinc chloride and nitrilotriacetic acid (NTA). The dissociation
constant is 6.4 X 10(-10) M. The fraction of active enzyme at
equilibrium is independent of the presence of substrate which indicates
that hippuryl-histidylleucine binds equally well to the holoenzyme and
apoenzyme. The rate constant for the dissociation of zinc from ACE was
measured as 0.68 min-1 for the free enzyme; the rate constant for the
enzyme substrate complex was roughly 0.18 min-1. The association of zinc
ion and ACE is very fast; the rate constant is 1.06 X 10(9) M-1 min-1.
Ethylenediaminetetraacetic acid (EDTA) and NTA rapidly remove zinc from
ACE with rate constants of 1.27 X 10(3) and 2.2 X 10(3) M-1 min-1. The
equilibrium constant for the reaction of NTA with ACE was measured as
4.6 X 10(-2) and was calculated for EDTA as 3.8 X 10(3).

96\. Dahlheim, H., *et al.*, *Effect of zinc depletion on angiotensin
I-converting enzyme in arterial walls and plasma of the rat.* Miner
Electrolyte Metab, 1989. 15(3): p. 125-9. The effect of zinc depletion
and of additional angiotensin I-converting enzyme (ACE) inhibitor
treatment was studied on ACE in aortic and other tissues, in plasma and
on systolic blood pressure of the rat. Zinc deprivation significantly
reduced plasma zinc concentration, plasma and testicular ACE activities
and blood pressure, but stimulated aortic ACE while lung values remained
constant. Zinc deficiency combined with ACE inhibition further
suppressed plasma ACE and stimulated the aortic enzyme earlier. Zinc
repletion experiments (in vitro) suggest the existence of a feedback
mechanism controlling ACE synthesis depending on plasma ACE activity.

97\. Kagan, B.L., *et al.*, *Elevated lipid levels in Vietnam veterans
with chronic posttraumatic stress disorder.* Biological Psychiatry,
1999. 45(3): p. 374-377. Elevated cholesterol levels have been reported
in panic disorder and anger attacks, but not major depression. No data
have been reported in posttraumatic stress disorder (PTSD). In this
study, 73 male Vietnam veterans (mean age 43.8 yrs) with chronic PTSD
had serum lipid screening upon entry to a 90-day inpatient program.
Results showed that elevated cholesterol, low-density lipoprotein,
triglycerides, and reduced high-density lipoprotein were frequent in
Vietnam veterans with chronic PTSD and are significant risk factors for
coronary artery disease. It is concluded that routine lipid screening
may be warranted in this at-risk population. Altered lipid levels may
result from activation of the noradrenergic system.

98\. Peter, H., S. Tabrizian, and I. Hand, *Serum cholesterol in
patients with obsessive compulsive disorder during treatment with
behavior therapy and SSRI or placebo.* International Journal of
Psychiatry in Medicine, 2000. 30(1): p. 27-39. 33 patients with
obsessive compulsive disorder (OCD) participated in the study. Serum
cholesterol was measured as pretreatment and at the end of a 10-wk
treatment-period. All patients received behavior therapy and, in a
double-blind fashion, fluvoxamine or placebo. Severity of OCD was
assessed by the Yale-Brown Obsessive Compulsive Scale (Y-BOCS).
Pretreatment cholesterol values of OCD patients were compared with
cholesterol levels of 30 panic disorder patients and 30 normal controls.
**OCD patients had elevated cholesterol levels comparable with those of
panic disorder patients**. Cholesterol levels decreased significantly
from pre- to posttreatment. OCD patients with high cholesterol levels
could make best use of the treatment whereas patients with desirable
cholesterol levels did not change their cholesterol during treatment.
Data support the assumption that not only panic disorder but also other
anxiety disorders, e.g., obsessive compulsive disorders, may be
associated with serum cholesterol elevations.

99\. Kuczmierczyk, A.R., *et al.*, *Serum cholesterol levels in patients
with generalized anxiety disorder (GAD) and with GAD and comorbid major
depression.* Canadian Journal of Psychiatry, 1996. 41(7): p. 465-8.
Investigated risk for cardiovascular disease in 38 patients (mean age
40.2 yrs) with general anxiety disorder (GAD), as well as the effects of
comorbid major depression (MD). Predrug-trial serum cholesterol and
triglyceride levels were assessed in Subjects with pure GAD and compared
with those of 21 patients (mean age 36.7 yrs) with mixed GAD and
comorbid MD. **Significantly higher cholesterol and triglyceride levels
were found in the GAD group.** It is concluded that increased
noradrenergic activity may be responsible for elevations in lipid levels
in patients with pure GAD.

100\. Bajwa, W.K., *et al.*, *High cholesterol levels in patients with
panic disorder.* American Journal of Psychiatry, 1992. 149(3): p.
376-378. 30 outpatients (mean age 37.7 yrs) with panic disorder were
found to have higher serum cholesterol levels than 30 outpatients with
major depression and 30 matched controls. Current or past histories of
anxiety disorders in Subjects with major depression were associated with
higher cholesterol levels. **It is hypothesized that higher levels of
cholesterol in panic disorder are a result of increased noradrenergic
activity,** which may be the underlying mechanism for symptoms of panic
disorder.

OBJECTIVE: This study was undertaken to help clarify whether the higher
cholesterol levels found in patients with panic disorder are a
complication of panic disorder only or are associated with any
psychiatric disorder. METHOD: The subjects of the study were 30 patients
with panic disorder and 30 patients with major depression, diagnosed
according to the Structured Interview for DSM-III-R, and 30 normal
control subjects. The three groups were matched for sex and age, and
none of the subjects had alcohol/drug abuse, abnormal ECGs, or unstable
medical conditions. Blood samples were drawn at random times, and serum
cholesterol levels were determined. RESULTS: The patients with panic
disorder had significantly higher serum cholesterol levels than did the
patients with major depression and the normal control subjects. Among
the patients with major depression, histories (current or past) of
anxiety disorders were associated with significant elevation of serum
cholesterol levels. The presence of stable medical conditions was not
associated with higher cholesterol levels in any of the three groups of
subjects. CONCLUSIONS: Higher cholesterol levels were particularly
associated with panic disorder in comparison with major depression.
Higher levels of cholesterol in panic disorder are hypothesized to be a
result of increased noradrenergic activity, which may be the underlying
biological/neurochemical mechanism for symptoms of panic disorder,
including anticipatory anxiety.

101\. Bajwa, W.K., *et al.*, *High cholesterol levels in patients with
panic disorder.* Am J Psychiatry, 1992. 149(3): p. 376-8. OBJECTIVE:
This study was undertaken to help clarify whether the higher cholesterol
levels found in patients with panic disorder are a complication of panic
disorder only or are associated with any psychiatric disorder. METHOD:
The subjects of the study were 30 patients with panic disorder and 30
patients with major depression, diagnosed according to the Structured
Interview for DSM-III-R, and 30 normal control subjects. The three
groups were matched for sex and age, and none of the subjects had
alcohol/drug abuse, abnormal ECGs, or unstable medical conditions. Blood
samples were drawn at random times, and serum cholesterol levels were
determined. RESULTS: The patients with panic disorder had significantly
higher serum cholesterol levels than did the patients with major
depression and the normal control subjects. Among the patients with
major depression, histories (current or past) of anxiety disorders were
associated with significant elevation of serum cholesterol levels. The
presence of stable medical conditions was not associated with higher
cholesterol levels in any of the three groups of subjects. CONCLUSIONS:
Higher cholesterol levels were particularly associated with panic
disorder in comparison with major depression. Higher levels of
cholesterol in panic disorder are hypothesized to be a result of
increased noradrenergic activity, which may be the underlying
biological/neurochemical mechanism for symptoms of panic disorder,
including anticipatory anxiety.

102\. Hayward, C., *et al.*, *Plasma lipid levels in patients with panic
disorder or agoraphobia.* Am J Psychiatry, 1989. 146(7): p. 917-9.

103\. Patterson, S., *et al.*, *Effects of acute mental stress on serum
lipids: mediating effects of plasma volume.* Psychosomatic Med, 1993.
55: p. 525-32.

104\. Peterson, J., R. Keith, and A. Wilcox, *Hourly changes in serum
cholesterol concentration: Effects of the anticipation of stress.*
Circulation, 1962. 25: p. 798-803. Over 100mg/dl variability in
cholesterol over a period of hours in some people \-- in response to
stress or concern about stress, putatively \-- though not well demo\'d.

105\. Thomas, C. and E. Murphy, *Further studies on cholesterol levels
in the Johns Hopkins medical students: the effect of stress at
examinations.* J Chron Dis, 1958. 8(6): p. 661-8. chol high at anatomy
exam, falls thereafter;

No good preexam unstressed measure: done on adm to med school, within 3
wks, also high.

106\. Wertlake, P., *et al.*, *Relationship of mental and emotional
stress to serum cholesterol levels.* Proc Soc Exper Biol & Med, 1958.
97: p. 163.

107\. Muldoon, M., *et al.*, *Effects of acute psychological stress on
serum lipid levels, hemoconcentration, and blood viscosity.* Arch Intern
Med, 1995. 155: p. 615-20.

108\. Muldoon, M., *et al.*, *Acute cholesterol responses to mental
stress and change in posture.* Arch Intern Med, 1992. 152: p. 775-80.

109\. McCabe, W., *et al.*, *Elevations of serum cholesterol in man in
association with life stress and independent of diet and exercise.*
Abstract, J. Lab & Clin Med, 1959. 54: p. 922.

110\. Dreyfuss, F. and J. Czaczkes, *Blood cholesterol and uric acid of
healthy medical students under the stress of an examination.* Ann Intern
Med, 1959. 103: p. 708-11.

111\. Muldoon, M., *et al.*, *Effects of acute psychological stress on
serum lipid levels, hemoconcentration, and blood viscosity.* Arch Intern
Med, 1995. 155: p. 615-20.

112\. Patterson, S., *et al.*, *Effects of acute mental stress on serum
lipids: mediating effects of plasma volume.* Psychosomatic Med, 1993.
55: p. 525-32.

113\. Hata, S., H. Kunida, and Y. Oyama, *\[Antihypertensive effects of
coenzyme Q10 in essential hypertension\--in relation to the
renin-aldosterone system\].* Horumon To Rinsho, 1977. 25(9): p. 1019-23.

114\. Hamdy, B.H., *et al.*, *Effectiveness of periodic oral vitamin A
dosage on hypovitaminosis in Egypt.* Int J Vitam Nutr Res, 1982. 52(3):
p. 235-40. Seventy boys, aged 6-13 years were subjected to periodic oral
vitamin A program. Non-parasitized boys (n = 63) responded to the oral
treatment with significant increase in plasma retinol level, even five
weeks after the end of oral treatment. On the other hand, boys infested
with parasites, none of whom had originally acceptable plasma-retinol
level, did not respond significantly to the oral treatment. Vitamin A
was found also to have an effect on growth, and there was highly
significant correlation between retinol level and percentile weight for
age.

115\. Marinho, H.A., *et al.*, *Influence of enteral parasites on the
blood vitamin A levels in preschool children orally supplemented with
retinol and/or zinc.* Eur J Clin Nutr, 1991. 45(11): p. 539-44. A sample
of 471 pre-school children who frequented schools and creches in a poor
district of Manaus (Amazonas), Brazil, were randomly submitted to faecal
parasitological tests. Two-hundred-and-forty children from both sexes
between the ages of 3 and 7 years with Ascaris lumbricoides and/or
Giardia lamblia were selected. The objective of the study was to
determine the possible influence of these two intestinal parasites and
vitamin A and/or zinc supplementation on the serum retinol levels of
primary school children. The children were submitted to clinical and
anthropometric examinations, dietary interviews and biochemical
examinations of retinol and carotene in the serum and of zinc in the
hair. The parasitic incidence was 85.0% and about 54% of the children
were polyparasitic. During the pretreatment phase, the retinol and
carotene serum levels were 36% and 57%, respectively, below the normal
levels. Using the Waterlow classification, the anthropometric analyses
revealed that 88% of the children showed normal growth. A significant
effect was observed of the anti-parasitic medicine on the serum retinol
levels.

116\. Morris, M.C., Archives of Neurology, 2002. 59: p. 1125-32.

117\. *Vitamin E slows cognitive decline in the elderly.* Internal
Medicine World Report, 2002: p. 4.

118\. Sato, Y., *et al.*, *Hypovitaminosis D and decreased bone mineral
density in amyotrophic lateral sclerosis.* Eur Neurol, 1997. 37(4): p.
225-9. To assess the bone health of patients with amyotrophic lateral
sclerosis (ALS), we evaluated the bone density and serum biochemical
indices of bone metabolism in 11 ALS patients. The serum concentration
of 25-hydroxyvitamin D (25-OHD) was significantly lower in patients
(14.0 +/- 3.7 ng/ml) than in controls (25.2 +/- 4.0 ng/ml), at deficient
levels (\< 10 ng/ml) in 2, and at insufficient levels (10-20 ng/ml) in 9
patients. Serum levels of parathyroid hormone (PTH) and ionized calcium
were elevated in 8 and 6 patients, respectively. Dietary intake of
vitamin D was below the recommended level (100 IU) in 10 patients, and
10 patients were in a sunlight-deprived state. The metacarpal bone
density (MBD) and the metacarpal index (MCI) of the second metacarpal
bone were measured by computed X-ray densitometry. Z scores of the MBD
and the MCI were negative in 7 and 6 patients, respectively. The serum
concentration of 25-OHD was positively correlated with the Z score of
the MBD (p \< 0.05, r = 0.727) and negatively with the PTH level (p \<
0.05, r = -0.410). The degree of dysfunction of hand grip also
correlated with the Z score of the MBD (p \< 0.05, r = 0.749). These
data underscore the importance of hypovitaminosis D and compensatory
hyperparathyroidism in the development of osteopenia in patients with
ALS.

119\. Prabhala, A., R. Garg, and P. Dandona, *Severe myopathy associated
with vitamin D deficiency in western New York.* Arch Intern Med, 2000.
160(8): p. 1199-203. Five cases of severe myopathy associated with
vitamin D deficiency are described. Each patient was confined to a
wheelchair because of weakness and immobility. Two were elderly, 1 was a
37-year-old African American with type 1 diabetes mellitus, 1 was being
treated for carcinoid syndrome, and 1 was severely malnourished due to
poor oral intake. In each, weakness had previously been attributed to
other causes, including old age, concomitant diabetic neuropathy, or
general debility. Correct diagnosis was made initially by a high index
of suspicion, following the demonstration of clinical proximal myopathy;
confirmation was made by the demonstration of low 25-hydroxyvitamin D
and elevated parathyroid hormone concentrations. Treatment with vitamin
D caused a resolution of body aches and pains and a restoration of
normal muscle strength in 4 to 6 weeks. Four patients became fully
mobile and had normal 25-hydroxyvitamin D concentrations, and the fifth
also became mobile. In the 4 fully recovered cases, parathyroid hormone
levels on follow-up were lower but still elevated. This finding suggests
a degree of autonomy of parathyroid secretion known to occur in cases of
long-standing vitamin D deficiency. Myopathy, due to chronic vitamin D
deficiency, probably contributes to immobility and ill health in a
significant number of patients in the northern United States. An
awareness of this condition may significantly improve mobility and
quality of life in patient populations vulnerable to vitamin D
deficiency.

120\. Gennari, A., *et al.*, *Organotins induce apoptosis by disturbance
of \[Ca(2+)\](i) and mitochondrial activity, causing oxidative stress
and activation of caspases in rat thymocytes.* Toxicol Appl Pharmacol,
2000. 169(2): p. 185-90. Di-n-butyltin dichloride (DBTC) and
tri-n-butyltin chloride (TBTC) cause thymus atrophy in rodents. At low
doses, antiproliferative modes of action have been shown to be involved,
whereas at higher doses apoptosis seems to be the mechanism of
thymotoxicity by these chemicals. In vitro, a similar
concentration-dependency has been observed. The purpose of the present
research was to investigate the mechanisms underlying DNA fragmentation
induced by these organotin compounds in freshly isolated rat thymocytes.
As previously shown for TBTC, DBTC is also able to significantly
increase intracellular Ca(2+) level (\[Ca(2+)\](i)). The rise in
\[Ca(2+)\](i), already evident 5 min after treatment, was followed by a
dose- and time-dependent generation of reactive oxygen species (ROS) at
the mitochondrial level. Simultaneously, organotins induced the release
of cytochrome c from the mitochondrial membrane into the cytosol. ROS
production and the release of cytochrome c were reduced by BAPTA, an
intracellular Ca(2+) chelator, or rotenone, an inhibitor of the electron
entry from complex I to ubiquinone, indicating the important role of
Ca(2+) and mitochondria during these early intracellular events.
Furthermore, we demonstrated that rotenone prevents apoptosis induced by
3 microM DBTC or TBTC and, in addition, that both BAPTA and Z-DEVD FMK
(mainly a caspase-3 inhibitor) decreased apoptosis by DBTC (already
shown for TBTC). Taken together these data show the apoptotic pathway
followed by organotin compounds starts with an increase of
\[Ca(2+)\](i), then continues with release of ROS and cytochrome c from
mitochondria, activation of caspases, and finally results in DNA
fragmentation.

121\. Ravikumar, A., *et al.*, *Isoprenoid pathway and free radical
generation and damage in neuropsychiatric disorders.* Indian J Exp Biol,
2000. 38(5): p. 438-46. Two substances which are products of the
isoprenoid pathway, can participate in lipid peroxidation. One is
digoxin, which by inhibiting membrane Na(+)-K+ ATPase, causes increase
in intracellular Ca2+ and depletion of intracellular Mg2+, both effects
contributing to increase in lipid peroxidation. Ubiquinone, another
products of the pathway is a powerful membrane antioxidant and its
deficiency can also result in defective electron transport and
generation of reactive oxygen species. In view of this and also in the
light of some preliminary reports on alteration in lipid peroxidation in
neuropsychiatric disorders, a study was undertaken on the following
aspects in some of these disorders (primary generalised epilepsy,
schizophrenia, multiple sclerosis, Parkinson\'s disease and CNS
glioma)\--1) concentration of digoxin, ubiquinone, activity of HMG CoA
reductase and RBC membrane Na(+)-K+ ATPase 2) activity of enzymes
involved in free radical scavenging 3) parameters of lipid peroxidation
and 4) antioxidant status. The result obtained indicates an increase in
the concentration of digoxin and activity of HMG CoA reductase, decrease
in ubiquinone levels and in the activity of membrane Na(+)-K+ ATPase.
There is increased lipid peroxidation as evidenced from the increase in
the concentration of MDA, conjugated dienes, hydroperoxides and NO with
decreased antioxidant protection as indicated by decrease in ubiquinone,
vit E and reduced glutathione in schizophrenia, Parkinson\'s disease and
CNS glioma. The activity of enzymes involved in free radical scavenging
like SOD, catalase, glutathione peroxidase and glutathione reductase is
decreased in the above diseases. However, there is no evidence of any
increase in lipid peroxidation in epilepsy or MS. The role of increased
operation of the isoprenoid pathway as evidenced by alteration in the
concentration of digoxin and ubiquinone in the generation of free
radicals and protection against them in these disorders is discussed.

122\. Schults, C.W., *et al.*, *Effects of coenzyme Q10 in early
Parkinson disease. Evidence of slowing of the functional decline.*
Archives of Neurology, *2002*. 59(10): p. 1541-50. Background: Parkinson
disease (PD) is a degenerative neurological disorder for which no
treatment has been shown to slow the progression. Objective: To
determine whether a range of dosages of coenzyme Q10 is safe and well
tolerated and could slow the functional decline in PD.
Design**  **Multicenter, randomized, parallel-group, placebo-controlled,
double-blind, dosage-ranging trial. Setting:**  **Academic movement
disorders clinics. Patients:**  **Eighty subjects with early PD who did
not require treatment for their disability. Interventions**:  **Random
assignment to placebo or coenzyme Q10 at dosages of 300, 600, or 1200
mg/d.Main Outcome Measure**: **The subjects underwent evaluation with
the Unified Parkinson Disease Rating Scale (UPDRS) at the screening,
baseline, and 1-, 4-, 8-, 12-, and 16-month visits. They were followed
up for 16 months or until disability requiring treatment with levodopa
had developed. The primary response variable was the change in the total
score on the UPDRS from baseline to the last visit. Results:** **The
adjusted mean total UPDRS changes were +11.99 for the placebo group,
+8.81 for the 300-mg/d group, +10.82 for the 600-mg/d group, and +6.69
for the 1200-mg/d group. The *P* value for the primary analysis, a test
for a linear trend between the dosage and the mean change in the total
UPDRS score, was .09, which met our prespecified criteria for a positive
trend for the trial. A prespecified, secondary analysis was the
comparison of each treatment group with the placebo group, and the
difference between the 1200-mg/d and placebo groups was significant (*P*
= .04). Conclusions:** **Coenzyme Q10 was safe and well tolerated at
dosages of up to 1200 mg/d. Less disability developed in subjects
assigned to coenzyme Q10 than in those assigned to placebo, and the
benefit was greatest in subjects receiving the highest dosage. Coenzyme
Q10 appears to slow the progressive deterioration of function in PD, but
these results need to be confirmed in a larger study.