# Acetylcholinesterase Inhibitor Brief: Reference Abstracts

1\. Fricker, R.D., *et al.*, *A Review of the Scientific Literature as
It Pertains to Gulf War Illnesses. Volume 12: Pesticide Use During the
Gulf War: A Survey of Gulf War Veterans*. 2000, Santa Monica, CA: RAND,
MR-1018/12-OSD.

2\. Golomb, B.A., *Illness in Persian Gulf War veterans: Evidence for a
causal role for acetylcholinesterase inhibitors (pyridostigmine bromide,
pesticides, and nerve agent).* Submitted.

3\. Nisenbaum, R., *et al.*, *Deployment stressors and a chronic
multisymptom illness among Gulf War veterans.* J Nerv Ment Dis, 2000.
188(5): p. 259-66. Unusual health problems have been reported by Gulf
War (GW) veterans, but no single etiology has been linked to these
illnesses. This study was conducted to determine the association between
self-reported GW deployment stressors and an illness defined by a
combination of fatigue, mood-cognition, and musculoskeletal symptoms. A
total of 1002 GW veterans from this cross-sectional survey of four Air
Force units completed a self-administered questionnaire that asked about
symptoms, demographic and military characteristics, and stressors during
deployment. Severe and mild-moderate illness was positively associated
with self-reports of pyridostigmine bromide use, insect repellent use
and belief in a threat from biological or chemical weapons. Injuries
requiring medical attention were only associated with severe illness.
These results suggest a link between self-reported chemical, emotional,
and physical exposures, and GW veterans\' illness. Further research is
needed to determine physiological and psychological mechanisms through
which such stressors could have contributed to this symptom complex.

4\. Unwin, C., *et al.*, *Health of UK servicemen who served in the
Persian Gulf war.* Lancet, 1999. 353: p. 169-178. BACKGROUND: Various
symptoms in military personnel in the Persian Gulf War 1990-91 have
caused international speculation and concern. We investigated UK
servicemen. METHODS: We did a cross-sectional postal survey on a random
sample of Gulf War veterans (Gulf War cohort, n=4248) and, stratified
for age and rank, servicemen deployed to the Bosnia conflict (Bosnia
cohort, n=4250) and those serving during the Gulf War but not deployed
there (Era cohort, n=4246). We asked about deployment, exposures,
symptoms, and illnesses. We analysed men only. Our outcome measures were
physical health, functional capacity (SF-36), the general health
questionnaire, the Centers for Disease Control and Prevention (CDC)
multisymptom criteria for Gulf War illness, and post-traumatic stress
reactions. FINDINGS: There were 8195 (65.1%) valid responses. The Gulf
War cohort reported symptoms and disorders significantly more frequently
than those in the Bosnia and Era cohorts, which were similar. Perception
of physical health and ability were significantly worse in the Gulf War
cohort than in the other cohorts, even after adjustment for confounders.
Gulf War veterans were more likely than the Bosnia cohort to have
substantial fatigue (odds ratio 2.2 \[95% CI 1.9-2.6\]), symptoms of
post-traumatic stress (2.6 \[1.9-3.4\]), and psychological distress (1.6
\[1.4-1.8\]), and were nearly twice as likely to reach the CDC case
definition (2.5 \[2.2-2.8\]). In the Gulf War, Bosnia, and Era cohorts,
respectively, 61.9%, 36.8%, and 36.4% met the CDC criteria, which fell
to 25.3%, 11.8%, and 12.2% for severe symptoms. Potentially harmful
exposures were reported most frequently by the Gulf War cohort. All
exposures showed associations with all of the outcome measures in the
three cohorts. Exposures specific to the Gulf were associated with all
outcomes. Vaccination against biological warfare and multiple routine
vaccinations were associated with the CDC multisymptom syndrome in the
Gulf War cohort. INTERPRETATION: Service in the Gulf War was associated
with various health problems over and above those associated with
deployment to an unfamiliar hostile environment. Since associations of
ill health with adverse events and exposures were found in all cohorts,
however, they may not be unique and causally implicated in
Gulf-War-related illness. A specific mechanism may link vaccination
against biological warfare agents and later ill health, but the risks of
illness must be considered against the protection of servicemen.

5\. Golomb, B.A., *A Review of the Scientific Literature as it Pertains
to Gulf War Illnesses, Vol 2: Pyridostigmine Bromide*. 1999, Santa
Monica, CA: RAND. 385.

6\. Cecchine, G., *et al.*, *A Review of the Scientific Literature as it
Pertains to Gulf War Illnesses, Vol 8: Pesticides*. 2000, Santa Monica,
CA: RAND. 182.

7\. 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.

8\. Li, L., *et al.*, *Muscarinic receptor-mediated
pyridostigmine-induced neuronal apoptosis.* Neurotoxicology, 2000.
21(4): p. 541-552. Abstract: Pyridostigmine is a reversible
cholinesterase (ChE) inhibitor that is associate 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.

9\. Pascuzzo, G.J., *et al.*, *The nature of the interactions of
pyridostigmine with the nicotinic acetylcholine receptor-ionic channel
complex I. Agonist, desensitizing, and binding properties.* Mol
Pharmacol, 1984. 25: p. 92-101.

10\. Tandon, P., *et al.*, *Fenthion produces a persistent decrease in
muscarinic receptor function in the adult rat retina.* Toxicol Appl
Pharmacol, 1994. 125: p. 271-280.

11\. Tandon, P., *et al.*, *Dose-response study of the tissue-specific
effects of fenthion on receptor function in rat CNS.* Toxicologist,
1994. 14: p. 257.

12\. Sherby, S.M., *et al.*, *Comparison of actions of carbamate
anticholinesterses on the nicotinic acetylcholine receptor.* Molecular
Pharmacology, 1985. 27: p. 343-348.

13\. Gupta, R.C., G.T. Patterson, and W.-D. Dettbarn, *Mechanisms of
toxicity and tolerance to diisopropylphosphorofluoridate at the
neuromuscular junction of the rat.* Toxicology and Applied Pharmacology,
1986. 84: p. 541-550.

14\. Eskenazi, B., A. Bradman, and R. Castorina, *Exposures of children
to organophosphate pesticides and their potential adverse health
effects.* Environ Health Perspect, 1999. 107 Suppl 3: p. 409-19. Recent
studies show that young children can be exposed to pesticides during
normal oral exploration of their environment and their level of dermal
contact with floors and other surfaces. Children living in agricultural
areas may be exposed to higher pesticide levels than other children
because of pesticides tracked into their homes by household members, by
pesticide drift, by breast milk from their farmworker mother, or by
playing in nearby fields. Nevertheless, few studies have assessed the
extent of children\'s pesticide exposure, and no studies have examined
whether there are adverse health effects of chronic exposure. There is
substantial toxicologic evidence that repeated low-level exposure to
organophosphate (OP) pesticides may affect neurodevelopment and growth
in developing animals. For example, animal studies have reported
neurobehavorial effects such as impairment on maze performance,
locomotion, and balance in neonates exposed (italic)in utero(/italic)
and during early postnatal life. Possible mechanisms for these effects
include inhibition of brain acetylcholinesterase, downregulation of
muscarinic receptors, decreased brain DNA synthesis, and reduced brain
weight in offspring. Research findings also suggest that it is
biologically plausible that OP exposure may be related to respiratory
disease in children through dysregulation of the autonomic nervous
system. The University of California Berkeley Center for Children\'s
Environmental Health Research is working to build a community-university
partnership to study the environmental health of rural children. This
Center for the Health Assessment of Mothers and Children of Salinas, or
CHAMACOS in Monterey County, California, will assess (italic)in
utero(/italic) and postnatal OP pesticide exposure and the relationship
of exposure to neurodevelopment, growth, and symptoms of respiratory
illness in children. The ultimate goal of the center is to translate
research findings into a reduction of children\'s exposure to pesticides
and other environmental agents, and thereby reduce the incidence of
environmentally related disease.

15\. Costa, L.G., B.W. Schwab, and S.D. Murphy, *Tolerance to
anticholinesterase compounds in mammals.* Toxicology, 1982. 25: p.
79-97.

16\. Buccafusco, J.J., *Chronic organophosphorus exposure and
cognition.* 1999 Annual Report to Congress on Gulf War Veterans\'
Illnesses, 2000: p. 101.

17\. Lev-Lehman, E., *et al.*, *Synaptogenesis and myopathy under
acetylcholinesterase overexpression.* Journal of Molecular Neuroscience,
2000. 14(1-2): p. 93-105. Abstract: Environmental, congenital, and
acquired immunological insults perturbing neuromuscular junction (NMJ)
activity may induce a variety of debilitating neuromuscular pathologies.
However, the molecular elements linking NMJ dysfunction to long-term
myopathies are unknown. Here, we report dramatically elevated levels of
mRNA encoding c-Fos and the \"readthrough\" (R) variant of
acetylcholinesterase (AChE) in muscles of transgenic mice overexpressing
synaptic (S) AChE in motoneurons and in control mice treated with the
irreversible cholinesterase inhibitor diisopropylfluorophosphonate
(DFP). Tongue muscles from DFP-treated and AChE-S transgenic mice
displayed exaggerated neurite branching and disorganized, wasting
fibers. Moreover, diaphragm muscles from both transgenic and DFP-treated
mice exhibited NMJ proliferation. 2\'-O-methyl-protected antisense
oligonucleotides targeted to AChE mRNA suppressed feedback upregulation
of AChE and ameliorated DFP-induced NMJ proliferation. Our findings
demonstrate common transcriptional responses to cholinergic NMJ stress
of diverse origin, and implicate deregulated AChE expression in
excessive neurite outgrowth, uncontrolled synaptogenesis, and
myopathology.

18\. Robinson, D. and R. McGee, *Agonist-induced regulation of neuronal
nicotinic acetylcholine receptor of PC12 cells.* Molecular Pharmacology,
1985. 27: p. 409-417.

19\. Sherby, S.M., *et al.*, *Comparison of actions of carbamate
anticholinesterses on the nicotinic acetylcholine receptor.* Molecular
Pharmacology, 1985. 27: p. 343-348.

20\. Shaw, K.-P., *et al.*, *The reversible cholinesterase inhibitor
physostgmine has channelblocking and agonist efects on the acetylcholine
receptor-ion channel complex.* Molecular Pharmacology, 1985. 28: p.
527-238.

21\. Anderson, R.J., *et al.*, *Decreased tetanic contracture of rat
skeletal muscle induced by pyridostigmine.* J Toxicol Environ Health,
1986. 18: p. 221-230.

22\. Haley, R.W., S. Billecke, and B.N. La Du, *Association of low PON1
type Q (type A) arylesterase activity with neurologic symptom complexes
in Gulf War veterans.* Toxicol Appl Pharmacol, 1999. 157(3): p. 227-33.
Previously Haley et al. described six possible syndromes identified by
factor analysis of symptoms in Gulf War veterans and demonstrated that
veterans with these symptom complexes were more neurologically impaired
than age-sex-education-matched well controls. They also uncovered strong
associations (relative risks 4-8) suggesting that these symptom
complexes were related to wartime exposure to combinations of
organophosphate pesticides, chemical nerve agents, high concentration
DEET insect repellant, and symptoms of advanced acute toxicity after
taking pyridostigmine. Here we have shown that compared to controls, ill
veterans with the neurologic symptom complexes were more likely to have
the R allele (heterozygous QR or homozygous R) than to be homozygous Q
for the paraoxonase/arylesterase 1 (PON1) gene. Moreover, low activity
of the PON1 type Q (Gln192, formerly designated type A) arylesterase
allozyme distinguished ill veterans from controls better than just the
PON1 genotype or the activity levels of the type R (Arg192, formerly
designated type B) arylesterase allozyme, total arylesterase, total
paraoxonase, or butyrylcholinesterase. A history of advanced acute
toxicity after taking pyridostigmine was also correlated with low PON1
type Q arylesterase activity. Type Q is the allozyme of
paraoxonase/arylesterase that most efficiently hydrolyzes several
organophosphates including sarin, soman, and diazinon. These findings
further support the proposal that neurologic symptoms in some Gulf War
veterans were caused by environmental chemical exposures.

23\. La Du, B.N., *et al.*, *Serum paraoxonase (PON1) isozymes: the
quantitative analysis of isozymes affecting individual sensitivity to
environmental chemicals.* Drug Metab Dispos, 2001. 29(4 Pt 2): p. 566-9.
In a recent study on Gulf War veterans who developed delayed
neurotoxicity symptoms, we found their levels of serum paraoxonase
(PON1) isozyme type Q to be significantly lower than in the control,
unaffected veteran group. These results were obtained in 25 ill veterans
and 20 well control subjects, of which 10 were deployed and 10 were
nondeployed battalion members who remained in the United States during
the Gulf War. The blood samples were also assayed for serum
butyrylcholinesterase in our laboratory, and more recently in Dr. C.
Broomfield\'s laboratory for somanase and sarinase activities. The
cholinesterase activities showed no significant correlation with the
PON1 isozyme levels or the severity of the clinical symptoms, but the
somanase and sarinase levels ran parallel to the PON1 type Q isozyme
concentrations. Although there is no direct evidence that these Gulf War
veterans were directly exposed to or encountered either of these nerve
gases, they may have been exposed to some environmental or chemical
toxin with a similar preference for hydrolysis by the PON1 type Q
isozyme. The number of subjects is relatively small, but the results
should encourage other investigators to examine both the individual
phenotypes and the levels of PON1 isozymes in other groups exhibiting
neurological symptoms.

24\. Mackness, B., P.N. Durrington, and M.I. Mackness, *Low paraoxonase
in Persian Gulf War Veterans self-reporting Gulf War Syndrome.* Biochem
Biophys Res Commun, 2000. 276(2): p. 729-33. Exposure to organophosphate
(OP\'s) insecticides and nerve gases during the Persian Gulf War has
been implicated in the development of Gulf War Syndrome. Paraoxonase
(PON1) present in human serum detoxifies OP\'s. We determined the levels
of PON1 in the serum of Gulf War Veterans and compared these to those
found in a control population. One hundred fifty-two Gulf War Veterans
from the UK who self-reported the presence of Gulf War Syndrome via a
questionnaire and 152 age and gender matched controls were studied. PON1
activity, concentration, and genotype were determined. In the Gulf War
Veterans, paraoxon hydrolysis was less than 50% of that found in the
controls (100.3 (14.8-233.8) vs 214.6 (50.3-516.2) nmol/min/ml, P \<
0.001). This low activity was independent of the effect of PON1
genotype. The serum PON1 concentration was also lower in the Gulf War
Veterans (75.7 (18.1-351.3) vs 88.2 (34.5-527.4) microg/ml, P \<
0.00025), which was again independent of PON1 genotype. There was no
difference in the rate of diazoxon hydrolysis between the groups (10. 2
+/- 4.1 micromol/min/ml vs 9.86 +/- 4.4, P = NS). A decreased capacity
to detoxify OP insecticides resulting from low serum PON1 activity may
have contributed to the development of Gulf War Syndrome.

25\. Cherry, N., *et al.*, *Paraoxonase (PON1) polymorphisms in farmers
attributing ill health to sheep dip.* Lancet, 2002. 359(9308): p. 763-4.
Human serum paraoxonase (PON1) hydrolyses diazinonoxon, the active
metabolite of diazinon, which is an organophosphate used in sheep dip.
In a case-referent study, 175 farmers with ill health that they
attributed to sheep dip nominated 234 referent farmers who also dipped
sheep and whom they believed to be in good health. We calculated odds
ratios for polymorphisms in PON1 at positions 192 and 55, and for PON1
activity with diazinonoxon as substrate. Cases were more likely than
referents to have at least one R allele at position 192 (glutamine to
arginine aminoacid substitution; odds ratio 1center dot93, 95% CI
1center dot24\--3center dot01), both alleles of type LL (1center dot70,
1center dot07\--2center dot68) at position 55, and to have diazoxonase
activity below normal median (1center dot77, 1center dot18\--2center
dot67). Our results support the hypothesis that organophosphates
contribute to the reported ill health of people who dip sheep.

26\. Cecchine, G., *et al.*, *A Review of the Scientific Literature as
it Pertains to Gulf War Illnesses, Vol 8: Pesticides*. 2000, Washington,
DC: RAND. 182.

27\. Cherry, N., *et al.*, *Health and exposures of United Kingdom Gulf
war veterans. Part II: The relation of health to exposure.* Occup
Environ Med, 2001. 58(5): p. 299-306. OBJECTIVES: To investigate
whether, in personnel who served with the United Kingdom forces in the
Gulf war, self reported exposures were related to symptoms in a way that
was consistent, specific, and credible. METHODS: Responses to symptom
and exposure questionnaires, completed 7 or more years after the war,
were collected from 7971 subjects deployed in the Gulf, from two exposed
cohorts, in a study with an overall response rate of 85.5%. Exposures
were considered in three groups, those outside the control of the
subjects, the use of prophylaxis, and indicators of susceptibility.
Health indices derived from symptom questionnaires were related to
reports of 14 exposures in these three groups in a series of multiple
regression analyses to allow for confounding. The relation of exposure
to complaints of widespread pain and to symptoms suggesting peripheral
neuropathy were examined by logistic regression. RESULTS: Consistent but
weak correlations between exposures and with health effects were found
in independent analyses of the two (main and validation) cohorts. Three
exposures outside the control of the subject, the number of
inoculations, the number of days handling pesticides, and the days
exposed to smoke from oil fires, were consistently and independently
related to severity. The number of inoculations was also associated with
higher scores on a factor weighted on symptoms associated with skin and
musculoskeletal complaints. The number of days handling pesticides
related particularly to scores on a neurological factor and to symptoms
consistent with toxic neuropathy. CONCLUSION: The relations between
exposures and ill health were generally weak. Consistent, specific, and
credible relations, warranting further investigation, were found between
health indices and two exposures, the reported number of inoculations
and days handling pesticides.

28\. Gray, G.C., *et al.*, *Self-reported symptoms and medical
conditions among 11,868 Gulf War-era veterans: the Seabee Health Study.*
Am J Epidemiol, 2002. 155(11): p. 1033-44. US Navy Seabees have been
among the most symptomatic Gulf War veterans. Beginning in May 1997, the
authors mailed Gulf War-era Seabees a health survey in serial mailings.
As of July 1, 1999, 68.6% of 17,559 Seabees contacted had returned the
questionnaire. Compared with other Seabees, Gulf War Seabees reported
poorer general health, a higher prevalence of all 33 medical problems
assessed, more cognition difficulties, and a higher prevalence of four
physician-diagnosed multisymptom conditions: chronic fatigue syndrome,
posttraumatic stress disorder, multiple chemical sensitivity, and
irritable bowel syndrome. Because the four multisymptom conditions were
highly associated with one another, the authors aggregated them into a
working case definition of Gulf War illness. Among the 3,831 (22% cases)
Gulf War Seabee participants, multivariable modeling revealed that
female, Reserve, and enlisted personnel and participants belonging to
either of two particular Seabee units were most likely to meet the case
definition. Twelve of 34 self-reported Gulf War exposures were mildly
associated with meeting the definition of Gulf War illness, with
exposure to fumes from munitions having the highest odds ratio (odds
ratio = 1.9, 95% confidence interval: 1.5, 2.4). While these data do not
implicate a specific etiologic exposure, they demonstrate a strong
association and a high prevalence of self-reported multisymptom
conditions in a large group of symptomatic Gulf War veterans.

29\. Haley, R.W. and T.L. Kurt, *Self-reported exposure to neurotoxic
chemical combinations in the Gulf War. A cross-sectional epidemiologic
study.* Jama, 1997. 277(3): p. 231-7. OBJECTIVE: To identify risk
factors of factor analysis-derived Gulf War-related syndromes. DESIGN: A
cross-sectional survey. PARTICIPANTS: A total of 249 Gulf War veterans
from the Twenty-fourth Reserve Naval Mobile Construction Battalion. DATA
COLLECTION: Participants completed standardized booklets measuring
self-reported wartime exposures and present symptoms. MAIN OUTCOME
MEASURES: Associations of factor analysis-derived syndromes with risk
factors for chemical interactions that inhibit butyrylcholinesterase and
neuropathy target esterase. RESULTS: Risk of syndrome 1 (\"impaired
cognition\") was greater in veterans who reported wearing flea collars
during the war (5 of 20, 25%) than in those who never wore them (7 of
229, 3%; relative risk \[RR\], 8.7; 95% confidence interval \[CI\],
3.0-24.7; P\<.001). Risk of syndrome 2 (\"confusion-ataxia\") increased
with a scale of advanced adverse effects from pyridostigmine bromide
(chi2 for trend, P\<.001), was greater among veterans who believed they
had been involved in chemical weapons exposure (18 of 108, 17%) than in
those who did not (3 of 141, 2%; RR, 7.8; 95% CI, 2.3-25.9; P\<.001),
and was increased in veterans who had been in a sector of far
northeastern Saudi Arabia on the fourth day of the air war (6 of 21,
29%) than in those who had not been (15 of 228, 7%; RR, 4.3; 95% CI,
1.9-10.0; P=.004). Effects of perceived chemical weapons exposure and
advanced adverse effects from pyridostigmine were synergistic (Rothman
S, 5.3; 95% CI, 1.04-26.7). Risk of syndrome 3
(\"arthro-myo-neuropathy\") increased with an index of frequency and
amount of government-issued insect repellent containing 75% DEET
(N,N-diethyl-m-toluamide) in ethanol applied during the war (chi2 for
trend, P\<.001) and with advanced adverse effects from pyridostigmine
(chi2 for trend, P\<.001). CONCLUSION: Some Gulf War veterans may have
delayed, chronic neurotoxic syndromes from wartime exposure to
combinations of chemicals that inhibit butyrylcholinesterase and
neuropathy target esterase.

30\. Kang, H.K., *et al.*, *Illnesses among United States veterans of
the Gulf War: a population-based survey of 30,000 veterans.* J Occup
Environ Med, 2000. 42(5): p. 491-501. Despite numerous studies on
veterans of the 1990 to 1991 Gulf War, the fundamental questions of how
healthy they are and how their health compares with that of their
military peers who were not deployed to the Gulf have not been fully
answered. We conducted a health survey in which the health outcomes of a
population-based sample of 15,000 Gulf veterans representing various
military branches and unit components (regular, reserve, National Guard)
were compared with those of 15,000 non-Gulf veterans who were randomly
sampled to mirror the number in the same military strata in the Gulf
veteran group. In comparison with their peers, Gulf veterans had a
higher prevalence of functional impairment, health care utilization,
symptoms, and medical conditions and a higher rate of low general health
perceptions. A longitudinal follow-up of the health of these veterans
will be needed to detect changes in health status and to detect diseases
with a long latency period.

31\. Kang, H., *et al.*, *Evidence for a deployment-related Gulf War
syndrome by factor analysis*, . 2002, Arch Environ Health. p. 58.

32\. Kroenke, K., P. Koslowe, and M. Roy, *Symptoms in 18,495 Persian
Gulf War veterans. Latency of onset and lack of association with
self-reported exposures.* Journal of Occupational and Environmental
Medicine, 1998. 40(6): p. 520-8. Toxic or environmental exposures have
been suggested as a possible cause of symptoms reported by Gulf War
veterans. To further explore this hypothesis, we analyzed findings in
18,495 military personnel evaluated in the Department of Defense
Comprehensive Clinical Evaluation Program. The program was established
in 1994 to evaluate Persian Gulf veterans eligible for Department of
Defense medical care who had health concerns after service in the
Persian Gulf during Operation Desert Shield/Desert Storm. The evaluation
included a structured clinical assessment, a physician-administered
symptom checklist, and a patient questionnaire addressing self-reported
exposures, combat experiences, and work loss. Among 18,495 patients
examined, the most common symptoms were joint pain, fatigue, headache,
memory or concentration difficulties, sleep disturbances, and rash.
Symptom onset was often delayed, with two-thirds of symptoms not
developing until after individuals returned from the Gulf War and 40% of
symptoms having a latency period exceeding one year. There was no
association between individual symptoms and patient demographics,
specific self-reported exposures, or types of combat experience.
Increased symptom counts were associated with work loss, the number of
self-reported exposures, the number of types of combat experience, and
certain ICD-9 diagnostic categories, particularly psychological
disorders. Prolonged latency of symptom onset and the lack of
association with any self-reported exposures makes illness related to
toxic exposure less likely.

33\. Proctor, S.P., *et al.*, *Health-related quality of life in Persian
Gulf War Veterans.* Mil Med, 2001. 166(6): p. 510-9. OBJECTIVE: The
objective of this investigation is to describe the health-related
quality of life of Persian Gulf War (GW) veterans and to examine the
effects of current chronic medical conditions and psychiatric status on
physical functioning. METHODS: To measure health-related quality of
life, the Medical Outcomes Short Form Survey (SF36) was administered
approximately 4 years after the GW to a stratified, random sample of New
England-area GW-deployed veterans and a group of military personnel
deployed to Germany during the GW. The SF36 scores for the GW-deployed
study population (N = 141) were compared with those for the
Germany-deployed group (N = 46) and with published U.S. population
norms. Multiple linear regression analyses were performed to identify
risk factors associated with lower physical health functioning in the
GW-deployed study group. RESULTS: Functional health status was
significantly lower in the GW-deployed group compared with the
Germany-deployed group for each of the SF36 subscales and the two
summary scores (Physical Component Summary \[PCS\] and Mental Component
Summary). Compared with the general U.S. population, the GW-deployed
group median was between the 25th and 50th percentile for the Physical
Functioning subscale and the PCS score. Within the GW-deployed group,
lower education, psychological symptomatology, and a higher number of
chronic self-reported medical conditions were significant predictors of
the PCS score. CONCLUSION: GW-deployed veterans report lower functional
health status compared with a group of Germany-deployed veterans and
published general U.S. population norms. Within the group of GW-deployed
veterans, several current medical and psychological conditions
predictive of lower physical functioning levels were identified.

34\. Reid, S., *et al.*, *Multiple chemical sensitivity and chronic
fatigue syndrome in British Gulf War veterans.* Am J Epidemiol, 2001.
153(6): p. 604-9. The objective of this study was to measure the
prevalence of multiple chemical sensitivity (MCS) and chronic fatigue
syndrome (CFS) in British Gulf War veterans and to investigate their
association with reported exposures and psychologic morbidity. In
1997\--1998, the authors undertook a cross-sectional survey of three
cohorts of British military personnel comprising Gulf veterans (n =
3,531), those who had served in Bosnia (n = 2,050), and those serving
during the Gulf War but not deployed there (Era cohort, n = 2,614). MCS
and CFS were defined according to operational criteria. The prevalence
of MCS in the Gulf, Bosnia, and Era cohorts was 1.3%, 0.3%, and 0.2%,
respectively. For CFS, the prevalence was 2.1% (Gulf cohort), 0.7%
(Bosnia cohort), and 1.8% (Era cohort). In Gulf veterans, MCS was
strongly associated with exposure to pesticides (adjusted odds ratio =
12.3, 95% confidence interval: 5.1, 30.0). Both syndromes were
associated with high levels of psychologic morbidity. These findings
suggest that CFS and MCS account for some of the medically unexplained
illnesses reported by veterans after deployment to the Gulf. MCS was
particularly associated with Gulf deployment and self-reported exposure
to pesticides, findings that merit further exploration given the
controversial status of this diagnosis and the potential for recall bias
in a questionnaire survey.

35\. Schumm, W.R., *et al.*, *Pyridostigmine bromide and the long-term
subjective health status of a sample of female reserve component Gulf
War veterans: a brief report.* Psychol Rep, 2001. 88(1): p. 306-8. The
role of pyridostigmine bromide (PB) pills in explaining the long-term
subjective health status of a sample of over 100 female Reserve
Component Gulf War veterans was examined through regression analysis.
Results fell just short of significance (p \< .06) for the prediction of
subjective health approximately six years after the war and were clearly
not significant for the prediction of subjective health at previous
times. Results parallel Golomb\'s 1999 RAND report, which found
suggestive but not conclusive evidence for the possible adverse effects
of Gulf War veterans\' consumption of pyridostigmine bromide pills. Our
data suggest that use of more than 10 pills may have been especially
risky with respect to long-term subjective health.

36\. Schumm, W.R., *et al.*, *Pyridostigmine bromide and the long-term
subjective health status of a sample of over 700 male Reserve Component
Gulf War era veterans.* Psychol Rep, 2002. 90(3 Pt 1): p. 707-21. Data
from a 1996-1997 survey of approximately 700 Reserve Component male
veterans indicate that the consumption of pyridostigmine bromide pills,
used as a pretreatment for potential exposure to the nerve agent Soman,
was a significant predictor of declines in reported subjective health
status after the war, even after controlling for a number of other
possible factors. Reported reactions to vaccines and other medications
also predicted declines in subjective health. While higher military rank
generally predicted better health during and after the war, educational
attainment, minority status, number of days in theater, and age
generally did not predict changes in subjective health. Although
servicemembers were directed to take three pills a day, veterans
reported a range of compliance\--less than a fourth (24%) followed the
medical instructions compared to 61% who took fewer than three pills
daily and 6% who took six or more pills a day. Implications for use of
pyridostigmine bromide are discussed.

37\. White, R.F., *et al.*, *Neuropsychological function in Gulf War
veterans: relationships to self-reported toxicant exposures.* Am J Ind
Med, 2001. 40(1): p. 42-54. BACKGROUND: The present study was aimed at
(1) exploring evidence of central nervous system (CNS) dysfunction among
Gulf War (GW) veterans on neuropsychological tests and (2) examining
whether performance on neuropsychological tests was related to specific
neurotoxicant exposures experienced in the Gulf. METHODS: The
GW-deployed groups were selected using stratified random sampling
methods from two distinct cohorts of GW veterans. A comparison group
that had been called up for GW service but deployed to Germany rather
than the Gulf also was examined. Neuropsychological function was
assessed using a pre-determined battery chosen to include tests known to
be highly sensitive to the behavioral effects of the neurotoxicants
thought to have been present in the Gulf. RESULTS: Self-reported
exposures were related to neuropsychological test performance
controlling for post-traumatic stress disorder, major depression, and
other known covariates of neuropsychological test performance. Results
showed that GW-deployed veterans performed more poorly than the
Germany-deployed veterans on several specific neuropsychological tests,
but after adjustment for multiple comparisons, only the differences in
mood complaints remained significant. Within the GW-deployed group,
self-reported exposure to chemical warfare agents was associated with
poorer performance on cognitive tests involving specific functional
domains. CONCLUSIONS: Results provide evidence that there are subtle
differences in CNS function among GW-deployed veterans who report
chemical warfare agent exposure while in the GW theater.

38\. McCauley, L.A., *et al.*, *Symptoms of Gulf War veterans possibly
exposed to organophosphate chemical warfare agents at Khamisiyah, Iraq.*
Int J Occup Environ Health, 2001. 7(2): p. 79-89. During the 1991 Gulf
War, some Allied troops were potentially exposed to sarin/cyclosarin as
the result of the destruction of Iraqi munitions at Khamisiyah. To
evaluate the prevalence of past and current symptoms known to be
associated with exposure to these chemical warfare agents, the authors
conducted a computer-assisted telephone survey of 2,918 U.S. Gulf War
veterans. Veterans who had participated in or witnessed the demolition
in 1991 were more likely to report historical or extant symptoms than
were veterans from other military units. These results should be viewed
cautiously because they are based on symptoms recalled nine years after
the event without precise characterization of exposure. Nonetheless, the
findings suggest that symptoms consistent with low-level sarin exposure
may have initially occurred, and health effects may have persisted in
the veterans who were nearest to the demolition activity. Further
research is warranted.

39\. McCauley, L., *et al.*, *Illness experience of Gulf War veterans
possibly exposed to chemical warfare agents.* Am J Prev Med, 2002.
23(3): p. 200. BACKGROUND: During the 1991 Gulf War, some Allied troops
were potentially exposed to chemical warfare agents as the result of the
detonation of Iraqi munitions at Khamisiyah.METHODS: In 1999, we
conducted a computer-assisted telephone survey of 2918 Gulf War veterans
from Oregon, Washington, California, North Carolina, and Georgia to
evaluate the prevalence of self-reported medical diagnoses and
hospitalizations among this potentially exposed population and among
comparison groups of veterans deployed and nondeployed to the Southwest
Asia theater of operations.RESULTS: Troops reported to be within 50
kilometers of the Khamisiyah site did not differ from other deployed
troops on reports of any medical conditions or hospitalizations in the 9
years following the Gulf War. Hospitalization rates among deployed and
nondeployed troops did not differ. Deployed troops were significantly
more likely to report diagnoses of high blood pressure (odds ratio
\[OR\]=1.7); heart disease (OR=2.5); slipped disk or pinched nerve
(OR=1.5); post-traumatic stress disorder (OR=14.9); hospitalization for
depression (OR=5.1); and periodontal disease (OR=1.8) when compared to
nondeployed troops. There was a trend for deployed veterans to report
more diagnoses of any cancer (OR=3.0).CONCLUSIONS: These findings do not
provide evidence of any long-term health effect associated with exposure
to the detonation of chemical warfare agents, but support findings from
other investigations of increased morbidity among deployed troops. The
prevalence of cancer among this population of deployed troops merits
ongoing attention.

40\. Gray, G., *et al.*, *The postwar hospitalization experience of US
veterans of the Persian Gulf War.* New Engl J Med, 1996. 335(20): p.
1505-1513.

41\. Gray, G.C., *et al.*, *Are Gulf War veterans suffering war-related
illnesses? Federal and civilian hospitalizations examined, June 1991 to
December 1994.* Am J Epidemiol, 2000. 151(1): p. 63-71. A previous
epidemiologic study demonstrated no unexplained increase in risk for
postwar hospitalization among Gulf War veterans who had remained on
active duty. The authors sought to expand this study to include Reserve
and separated military personnel. They examined hospitalization data
from the Department of Defense, the Department of Veterans Affairs (VA),
and the California Office of Statewide Health Planning and Development
hospital systems for the years 1991-1994. Since denominator data were
not available, the authors compared the proportional morbidity ratios
(PMRs) of hospitalization discharge diagnoses (both large categories and
specific diagnoses) between Gulf War veterans and other veterans of the
same era. There were no indications that Gulf War veterans were
suffering increased PMRs for infectious diseases; neoplasms; endocrine
diseases; blood diseases; skin conditions; or diseases of the nervous
system, circulatory system, or musculoskeletal system. However, these
veterans did experience proportionally more hospitalizations for various
specific diagnoses, namely, fractures and bone and soft-tissue injuries
(Department of Defense and California Office of Statewide Health
Planning and Development), various diseases of the respiratory
(including asthma) and digestive systems (VA), and diverse symptom
diagnoses (VA). While these findings may be influenced by chance or by a
number of potential confounders, including health registry
participation, they merit further examination using other study designs.

42\. Kaiser, K.S., *Pyridostigmine bromide intake during the Persian
Gulf War is not associated with postwar handgrip strength.* Mil Med,
2000. 165(3): p. 165-8. Many Persian Gulf War veterans took
pyridostigmine bromide (PB) during the Persian Gulf War. Previous
research suggests that PB intake and insecticide exposure may reduce
muscular strength. During 1994 and 1995, we examined the relationships
between self-reported PB intake, self-reported exposures, and handgrip
strength among 527 Gulf War veterans (GWVs) and 969 nondeployed veterans
of that era (NDVs). We found that 25.4% and 6.7% of the GWVs and NDVs,
respectively, reported generalized musucle weakness (for 1 month or
longer) since the Gulf War (July 1990). Many veterans also reported
exposure to insecticide during the war. Dominant handgrip strength was
measured three times with a hand-held dynamometer in subjects standing
with the elbow bent at a right angle. Multiple linear regression
revealed that handgrip strength was negatively associated with age (p =
0.001) and female gender (p \< 0.001). Handgrip strength was also found
to be positively associated with height (p \< 0.001), but it was not
associated with PB intake (p = 0.558). Exposure to insecticides had no
major effect on handgrip strength. These data suggest no association
between PB intake and postwar handgrip strength.

43\. Golomb, B.A., *Chapter 14. Chronic Effect. In: A Review of the
Scientific Literature as it Pertains to Gulf War Illnesses, Vol 2:
Pyridostigmine Bromide*. 1999, Santa Monica, CA: RAND. 385.

44\. Lotti, M., *Low-level exposures to organophosphorus esters and
peripheral nerve function.* Muscle Nerve, 2002. 25(4): p. 492-504. This
review evaluates the epidemiological and clinical evidence linking
low-level and prolonged exposures to organophosphorus esters, used as
insecticides or nerve agents, to peripheral nerve dysfunction. The
clinical effects of large doses of these chemicals-including the
cholinergic syndrome, the intermediate syndrome, and the delayed
polyneuropathy-are all well established and are summarized. Based on
these clinical observations and experimental studies, dose-effect
relationships indicate that peripheral neuropathy always develops after
cholinergic toxicity. However, several studies have suggested that this
relationship may be different after low-level prolonged exposures, as,
for instance, those experienced by Gulf War veterans and British sheep
farmers, thereby leading to the development of peripheral neuropathy
without preceding cholinergic toxicity. A critical assessment of these
studies, involving subjects with either current or past exposures,
indicates that changes in peripheral nerve function were mild,
inconsistent, and unexplained and that most studies lack exposure data.
Suggestions made about individual hypersusceptibility to delayed
polyneuropathy lack support. It is concluded that there is no evidence
of peripheral nerve dysfunction caused by low-level prolonged exposures
to organophosphate insecticides or nerve agents.

45\. Ray, D.E., *Chronic effects of low level exposure to
anticholinesterases.* Toxicology Letters, 1998. 95(1001): p. 26.

46\. 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.

47\. Francesconi, R., R. Hubbard, and M. Mager, *Effects of
pyridostigmine on ability of rats to work in the heat.* J Appl Physiol,
1984. 56(4): p. 891-5. Adult, male rats (300-325 g) were treated with
pyridostigmine bromide (n = 22) or saline (n = 22) to quantitate the
effects of cholinesterase inhibition (64%) on the ability to work (9.14
m/min, level treadmill) in the heat (35 degrees C).
Pyridostigmine-treated rats had a mean endurance of 23 min, whereas
saline-treated animals ran for nearly 35 min (P less than 0.001). Rates
of rectal and skin temperature increments were significantly higher (P
less than 0.001) in pyridostigmine-treated rats as were water losses (P
less than 0.001). Exercise in the heat to hyperthermic exhaustion
effected anticipated increments in circulating urea nitrogen,
creatinine, lactate dehydrogenase, and potassium levels, whereas
pyridostigmine pretreatment had additive effects on lactate and creatine
kinase concentrations. Additionally, pyridostigmine elicited a
significant (P less than 0.01) hyperglycemia before exercise, an effect
noted also with other organophosphate simulants. We concluded that
pyridostigmine-induced cholinesterase inhibition had a variety of
debilitating effects during work in the heat.

48\. 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.

49\. 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.

50\. 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.

51\. Chariot, P., *et al.*, *Determination of the blood lactate:pyruvate
ratio as a noninvasive test for the diagnosis of zidovudine myopathy.*
Arthritis Rheum, 1994. 37(4): p. 583-6. OBJECTIVE. To evaluate the use
of the lactate: pyruvate ratio as a test for the detection of zidovudine
myopathy. METHODS. Twenty consecutive human immunodeficiency
virus-infected patients with muscle involvement and 20 without muscle
involvement were studied prospectively. Blood lactate and pyruvate
levels and serum creatine kinase levels were tested, muscle involvement
was assessed both clinically and electrophysiologically, and muscle
biopsy was performed in patients with myopathy. RESULTS. Nine patients
had biopsy-proven zidovudine myopathy. All 9 had a high lactate:pyruvate
ratio, with elevations on 2 of 2 determinations in 6 patients and on 1
of 2 in 3 patients. Two of 11 patients with other myopathies and 2 of 20
patients without myopathy had a high lactate:pyruvate ratio on 1 of 2
determinations. CONCLUSION. The lactate:pyruvate ratio, when determined
repeatedly, is a sensitive test for detecting mitochondrial muscular
toxicity of zidovudine.

52\. 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.

53\. 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.

54\. 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.

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

56\. 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.

57\. 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.

58\. 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.

59\. 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.

60\. 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.

61\. 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.

62\. 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.

63\. 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.

64\. 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.

65\. 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.

66\. 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.

67\. Elble, R., E. Giacobini, and C. Higgins, *Choline levels are
increased in cerebrospinal fluid of Alzheimer patients.* Neurobiol
Aging, 1989. 10(1): p. 45-50. We measured choline (Ch),
acetylcholinesterase activity (AChE) and total protein in the
cerebrospinal fluid (CSF) of 66 Alzheimer patients (ages 54-89 years)
and 22 age-matched controls (ages 52-80 years), looking for markers of
the well-established cholinergic deficit and neuronal degeneration in
Alzheimer disease (AD). Three or more lumbar punctures were performed in
21 patients over a span of 24 months in order to study the changes in
these CSF components with disease progression. We found a statistically
significant reduction in AChE and an increase in Ch with advancing
dementia. These changes were not related to patient age. We suggest that
the rise in CSF choline is related to neuronal membrane breakdown and
reduced Ch uptake by cholinergic neurons. The reduction in CSF AChE is
consistent with the depletion of cholinergic neurons in AD.

68\. Berrettini, W.H., *et al.*, *Intravenous physostigmine increases
cerebrospinal fluid neuropeptide-Y.* Biol Psychiatry, 1989. 26(6): p.
623-30. The ability to measure directly central nervous system (CNS)
neurotransmitter changes after an acute pharmacological challenge would
be a useful clinical tool in psychiatric research. As one approach to
this possibility, we attempted to measure cerebrospinal fluid (CSF)
neuropeptide changes produced by an intravenous infusion of the indirect
cholinergic agonist physostigmine. Six rhesus monkeys, with indwelling
CSF catheters, had serial CSF samples removed before and after a 15
micrograms/kg physostigmine infusion. Five of six monkeys studied showed
at least a 50% increase in CSF neuropeptide-Y (NPY) levels. Normal human
subjects (n = 27) had CSF sampled before and 15, 30, and 45 min after an
acute intravenous infusion of physostigmine (either 0, 5, or 15
micrograms/kg). An Analysis of Variance revealed a significant (p =
0.04) dose-time interaction, suggesting that physostigmine increased CSF
NPY at the 15 micrograms/kg dose. CSF levels of seven other
neuropeptides remained unchanged. These results suggest that the
pharmacological challenge paradigm can be adapted to CSF neuropeptides,
providing new measures of CNS stimulus-induced response beyond the
peripheral plasma determinations usually employed.

69\. Davis, K.L., *et al.*, *Alterations in cerebrospinal fluid dopamine
metabolites following physostigmine infusion.* Psychopharmacology
(Berl), 1981. 72(2): p. 155-60. The effect of IV physostigmine
administration on the cerebrospinal fluid (CSF) levels of homovanillic
acid (HVA) and dihydroxyphenylacetic acid (DOPAC) in normal subjects was
determined. After an adjustment for differing CSF concentrations of
probenecid, physostigmine was found to elevate CSF HVA and DOPAC
concentrations. The authors discuss these changes in CSF HVA and DOPAC
and their possible relationship to the ability of physostigmine to
decrease the symptoms of some patients with tardive dyskinesia.

70\. Kaye, J.A., *et al.*, *Cerebrospinal fluid neurochemistry in the
myoclonic subtype of Alzheimer\'s disease.* Ann Neurol, 1988. 24(5): p.
647-50. Monoamine metabolites, biopterin, acetylcholinesterase (AChE)
activity, and somatostatin-like immunoreactivity (SLI) were determined
in the lumbar cerebrospinal fluid (CSF) of 24 patients with dementia of
the Alzheimer type (DAT) without myoclonus or extrapyramidal signs, in 8
patients with DAT and myoclonus, and in 14 age-matched healthy control
subjects. In patients with DAT with myoclonus as compared with both DAT
patients without myoclonus and control subjects, the concentrations of
homovanillic acid and biopterin were significantly decreased.
5-Hydroxyindoleacetic acid was significantly lower in patients with
myoclonic DAT as compared to patients with nonmyoclonic DAT, but not
significantly lower than in control subjects. CSF AChE and SLI were
significantly reduced in patients with DAT with or without myoclonus, as
compared with control subjects, but AChE and SLI were not significantly
different between dementia groups. These results suggest that DAT
patients with myoclonus represent a distinct clinical and neurochemical
DAT subtype.

71\. Jolkkonen, J., *et al.*, *Somatostatin-like immunoreactivity in the
cerebrospinal fluid of patients with Parkinson\'s disease and its
relation to dementia.* J Neurol Neurosurg Psychiatry, 1986. 49(12): p.
1374-7. Acetylcholinesterase, somatostatin-like immunoreactivity, and
homovanillic acid levels were measured in the cerebrospinal fluid of 36
patients with early stages of Parkinson\'s disease and in 19 control
patients. In patients with Parkinson\'s disease the levels of
somatostatin-like immunoreactivity were lower than in the controls (p
less than 0.01); these values were lowest in the demented Parkinsonian
patients. Concentrations of homovanillic acid were also significantly
lower in Parkinsonian patients (p less than 0.05). In contrast, no
changes were observed in the acetylcholinesterase activity of patients
with Parkinson\'s disease. The reduced somatostatin-like
immunoreactivity in CSF agrees with previous post-mortem studies and
indicates that Parkinson\'s disease and Alzheimer\'s disease may have
some neurochemical features in common.

72\. Gomez, S., *et al.*, *Somatostatin-like immunoreactivity and
acetylcholinesterase activities in cerebrospinal fluid of patients with
Alzheimer disease and senile dementia of the Alzheimer type.*
Psychoneuroendocrinology, 1986. 11(1): p. 69-73. Somatostatin-like
immunoreactivity (SLI) and acetylcholinesterase (AchE) activities were
measured in the cerebrospinal fluid of 25 patients with senile dementia
of the Alzheimer type (SDAT). Both SLI levels and AchE activities were
reduced in the CSF of SDAT patients. The SLI levels and AchE activities
were not correlated with the duration and the dementia score. However,
in two patients the CSF SLI concentration was in agreement with the SLI
levels in the frontal cortex obtained by biopsy. Our findings suggest
that CSF SLI may be a good index of cortical SLI activities.

73\. Hartikainen, P., *et al.*, *Neurochemical markers in the
cerebrospinal fluid of patients with Alzheimer\'s disease, Parkinson\'s
disease and amyotrophic lateral sclerosis and normal controls.* J Neural
Transm Park Dis Dement Sect, 1992. 4(1): p. 53-68. Several
neurotransmitter markers were investigated in the cerebrospinal fluid
(CSF) from patients with Alzheimer\'s disease (AD) (n = 27),
Parkinson\'s disease (PD) (n = 35) and ALS (n = 26) and from control
subjects (n = 34) to compare the possible alterations in the biochemical
profiles of these different neurodegenerative diseases. The main
proportion of the patients represented an early phase of the illness at
the time of the diagnosis. Correlations of the degree of dementia and
the stage of the disease with CSF measures were evaluated. The CSF
levels of somatostatin like-immunoreactivity (SLI) were significantly
reduced in AD patients when compared with those of normals and ALS
patients. The CSF concentrations of homovanillic acid (HVA) were
significantly decreased for PD patients and the decrease focused on the
non-demented patients. A trend of decreasing HVA values towards the most
advanced stage of Parkinson\'s disease assessed by Webster\'s scale was
also displayed. The content of 3-methoxy-4-hydroxyphenylglycol (MHPG) in
the CSF was higher for ALS patients than for other groups. The lowest
5-hydroxy-indoleacetic acid (5HIAA) levels were observed in the PD group
and the lowest acetylcholinesterase (AChE) activities were found in the
PD patients with the most severe disease. Changes in CSF measures were
too subtle to be beneficial for diagnostic purposes, but adequate for
reflecting the different neurochemical profiles of these three
degenerative neurological disorders.

74\. Davis, K.L., *et al.*, *Neurotransmitter metabolites in the
cerebrospinal fluid of man following physostigmine.* Life Sci, 1977.
21(7): p. 933-6.

75\. Popovic, M., *et al.*, *Effect of physostigmine and verapamil on
active avoidance in an experimental model of Alzheimer\'s disease.* Int
J Neurosci, 1997. 90(1-2): p. 87-97. The present study was performed to
investigate and compare the effect of acetylcholinesterase inhibitor,
physostigmine (0.045, 0.060 and 0.075 mg/kg sc, 30 min before the tests)
and Ca-antagonist, verapamil (1.0, 2.5, 5.0 and 10.0 mg/kg sc, 30 min
before the tests), on two-way active avoidance (AA) learning
(acquisition and performance) in nucleus basalis magnocellularis
(NBM)-lesioned rats. Bilateral electrolytic lesions of NBM induced
significant decrease of acquisition and performance of AA responses in
rats. Physostigmine (0.060 mg/kg) significantly improved only
acquisition of AA, while verapamil (2.5 and 5.0 mg/kg) significantly
improved both type of AA behavior in NBM-lesioned rats. These results
suggest that altered calcium homeostasis might play significant role in
pathogenesis of experimental induced Alzheimer\'s disease (AD) and that
administration of calcium antagonist such as verapamil might
successfully ameliorate disturbances of learning and memory appeared
after lesions of NBM.

76\. Peskind, E.R., *et al.*, *Oral physostigmine in Alzheimer\'s
disease: effects on norepinephrine and vasopressin in cerebrospinal
fluid and plasma.* Biol Psychiatry, 1995. 38(8): p. 532-8. Physostigmine
is a cholinesterase inhibitor which enhances central and peripheral
cholinergic activity. In this study, we explored in persons with
Alzheimer\'s disease (AD) the effects of an acute dose of physostigmine
in patients receiving chronic physostigmine treatment on the activity of
the cholinergically regulated noradrenergic and arginine vasopressin
(AVP) systems. Specifically, we estimated the effects of sustained
release oral physostigmine on central and peripheral noradrenergic and
AVP systems by measuring norepinephrine (NE) and AVP in cerebrospinal
fluid (CSF) and plasma. Lumbar punctures were performed in both
physostigmine and no drug treatment conditions. In some subjects the
effects of physostigmine on the plasma AVP response to the osmolar
stimulus of a hypertonic saline infusion also were measured. NE
concentrations in both CSF and plasma were significantly lower in the
physostigmine than in the no drug condition. AVP concentrations did not
differ between conditions in either compartment, nor did physostigmine
affect the AVP response to hypertonic saline. Physostigmine appears to
decrease both central and peripheral noradrenergic activity in AD.

77\. degli Uberti, E.C., *et al.*, *Acute administration of human
galanin in normal subjects reduces the potentiating effect of
pyridostigmine-induced cholinergic enhancement on release of
norepinephrine and pancreatic polypeptide.* Neuroendocrinology, 1996.
64(5): p. 398-404. The neuropeptide galanin (GAL) is widely distributed
in the central and peripheral nervous systems where it often coexists
with catecholamines and acetylcholine. Recently we have reported that
human GAL (hGAL) in man depresses the release of norepinephrine (NE) and
the responses to both assumption of upright posture and insulin-induced
hypoglycemia. To gain an insight into the action of hGAL on sympathetic
nervous system activity in man, we investigated the effects of a 60-min
infusion (80 pmol/kg/min) of hGAL or saline on the release of NE,
epinephrine (E) and pancreatic polypeptide (PP) induced by an
acetylcholinesterase inhibitor, pyridostigmine bromide (PD), in nine
healthy volunteers. PD (120 mg orally) induced a significant rise in
plasma concentrations of NE (1.6 +/- 0.04 vs. 1.08 +/- 0.06 nmol/l), E
(0.34 +/ 0.05 vs. 0.12 +/- 0.04 nmol/l) and PP (178.06 +/- 33 vs. 37.57
+/- 7.35 pmol/l), whilst it significantly reduced heart rate (HR; 61 +/-
2 vs. 71 +/- 4 beats/min). Changes in plasma levels of PP were
determined as an indirect measure of amplification of endogenous
cholinergic activity produced by PD. Administration of hGAL blunted the
release of NE and PP evoked by PD. The mean (+/- SEM) area under the
curve produced by PD of NE (50.05 +/- 3.97 nmol/l.90 min) and PP
(8,692.87 +/- 1,724 pmol/l.90 min) was significantly (p \< 0.001)
reduced by hGAL infusion (2.65 +/- 1.57 nmol/l.90 min and 248.1 +/- 148
pmol/l.90 min, for NE and PP, respectively). hGAL failed to affect
significantly the E release evoked by PD. hGAL was able to enhance HR
significantly (104 +/- 5 vs. 69 +/- 3 beats/min), and completely
prevented the PD-induced slowing of HR. Both PD and hGAL did not alter
supine systolic and diastolic blood pressure. We conclude that hGAL
significantly reduces the release of NE and PP stimulated by PD-induced
enhancement of cholinergic activity. These findings are consistent with
a functional interrelationship between GAL and the cholinergic system in
man, and may suggest the participation of a cholinergic pathway in the
galaninergic modulation of the autonomic nervous system.

78\. Araki, S. and H. Shibasaki, *\[Variations in serum amylase
responding to the neostigmine test in amyotrophic lateral sclerosis\].*
Igaku To Seibutsugaku, 1966. 73(5): p. 233-9.

79\. Gowen, G.F., *A second look at the neostigmine morphine test.* Am
Surg, 1989. 55(10): p. 640-4. In a prospective five-year study of 65
consecutive patients with upper abdominal pain the Neostigmine Morphine
Test (NMT) was applied as a screen for biliary, ampullary, and
pancreatic disease. Three facts emerged from this study: 1) the amylase
and lipase were overly sensitive, but not specific and had only a 10 per
cent predictive value for ampullary obstruction; 2) the bilirubin,
alkaline phosphatase, and serum glutamic oxaloacetic transaminase (SGOT)
were positive in patients with ampullary obstruction if they were
postcholecystectomy but not so in patients with an intact gallbladder,
which may be explained by the third observation; 3) the gallbladder can
compensate for partial ampullary obstruction by dilatation five to ten
times and by its ability to absorb 90 per cent of the water content of
the bile. The Neostigmine Morphine Test is not a reliable screen for
ampullary obstruction and positive findings must be confirmed by other
studies.

80\. Bell, I.R., *et al.*, *Serum neopterin and somatization in women
with chemical intolerance, depressives, and normals.*
Neuropsychobiology, 1998. 38(1): p. 13-8. The symptom of intolerance to
low levels of environmental chemicals (CI, chemical intolerance) is a
feature of several controversial polysymptomatic conditions that overlap
symptomatically with depression and somatization, i.e., chronic fatigue
syndrome, fibromyalgia, multiple chemical sensitivity, and Persian Gulf
syndrome. These syndromes can involve many somatic symptoms consistent
with possible inflammation. Immunological or neurogenic triggering might
account for such inflammation. Serum neopterin, which has an inverse
relationship with l-tryptophan availability, may offer a marker of
inflammation and macrophage/monocyte activation. This study compared
middle-aged women with CI (who had high levels of affective distress; n
= 14), depressives without CI (n = 10), and normals (n = 11). Groups did
not differ in 4 p.m. resting levels of serum neopterin. However, the CI
alone had strong positive correlations between neopterin and all of the
scales measuring somatization. These preliminary findings suggest the
need for additional research on biological correlates of \'unexplained\'
multiple somatic symptoms in subtypes of apparent somatizing disorders.

81\. Amici, S., *et al.*, *Cerebrospinal fluid acetylcholinesterase
activity after long-term treatment with donepezil and rivastigmina.*
Mech Ageing Dev, 2001. 122(16): p. 2057-62. At present
acetylcholinesterase (AChE) inhibitors (AChEIs) represent the only
reliable therapeutic resource for symptomatic treatment of Alzheimer
Disease (AD). This study was designed to assess the effects of 6-12
month treatment with AChEIs donepezil and rivastigmine on cerebrospinal
fluid (CSF) AChE and butyrylcholinesterase (BuChE) activity in AD
patients. The pattern of AChE isoforms (G4, G1, G2) before and after
treatment was investigated as well. In AD patients treated with
donepezil a significant increase of CFS AChE activity was observed,
whereas treatment with rivastigmine induced a significant decrease of
AChE activity. Both drugs did not change BuChE activity and tended to
restore the physiological pattern of AChE isoform. The possible
significance of the influence of AChEIs on CSF AChE activity and
isoforms is discussed.

82\. Appleyard, M.E., *et al.*, *Cholinesterase activities in
cerebrospinal fluid of patients with senile dementia of Alzheimer type.*
Brain, 1987. 110 ( Pt 5): p. 1309-22. Acetylcholinesterase (AChE) and
nonspecific cholinesterase (nsChE) activities of lumbar ceresbrospinal
fluid (CSF) from patients with a clinical or histological diagnosis of
Alzheimer\'s disease have been compared with those of normal age-matched
control patients and patients with dementia of non-Alzheimer aetiology.
No significant differences in the AChE activity of lumbar CSF from
histologically and clinically diagnosed Alzheimer\'s disease patients
and normal age-matched controls were found, although they could be
distinguished from controls and other dements by their lower lumbar CSF
levels of nsChE activity and by their elevated ratio of AChE/nsChE. A
lower level of AChE activity was observed in the lumbar CSF of patients
with dementia of non-Alzheimer aetiology. The AChE and nsChE activities
of ventricular CSF obtained at postmortem have also been examined. The
AChE activity of the ventricular CSF of patients with histologically
confirmed Alzheimer\'s disease was 66% lower than that of age-matched
controls; these patients could also be distinguished from normals by
their lower levels of nsChE and by the elevated ratio of AChE/nsChE
activities. A molecular defect in the AChE in the ventricular CSF of
Alzheimer patients is indicated by the finding that the enzyme failed to
show inhibition by high concentrations of substrate. The lower level of
AChE in ventricular CSF may reflect the changes in this enzyme in
forebrain regions of Alzheimer patients. Although it is at present not
possible to correlate the lower levels of nsChE found in CSF with any
known brain pathology, the significantly altered ratio of AChE/nsChE
activities in lumbar CSF may possibly form the basis for a diagnostic
test of Alzheimer type dementia.

83\. Appleyard, M.E. and B. McDonald, *Acetylcholinesterase and
butyrylcholinesterase activities in cerebrospinal fluid from different
levels of the neuraxis of patients with dementia of the Alzheimer type.*
J Neurol Neurosurg Psychiatry, 1992. 55(11): p. 1074-8.
Acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities
of cerebrospinal fluid (CSF) collected post mortem from the lateral
ventricles, cisterna magna, and lumbar regions of the spinal cord of
patients with a histologically confirmed diagnosis of Alzheimer\'s
disease were compared with those of normal, age matched control
patients, patients with dementia of non-Alzheimer aetiology, and
patients with non-dementing neurological disorders. The AChE activity of
the ventricular CSF of patients with Alzheimer\'s disease was 48% lower
(p \< 0.005) than that of age matched controls or patients with other
types of dementia, and the AChE activity of CSF sampled from the basal
cistern was 40% lower (p \< 0.005) in patients with Alzheimer\'s
disease. There were no significant differences between the AChE activity
in Alzheimer\'s disease and control patients in CSF collected from the
lumbar cistern. AChE activity was lower in CSF sampled from the basal
and lumbar cistern of patients with dementia of non-Alzheimer aetiology,
while ventricular activity was in the normal range. BuChE activity in
ventricular CSF of Alzheimer\'s disease patients was 41% lower than
normal (p \< 0.05) and in the normal range in all other samples. The
secretion of AChE from forebrain and hindbrain regions is reduced in
Alzheimer\'s disease patients, leading to decreased ventricular and
cisternal levels of the enzyme. Secretion from more caudal regions of
the central nervous system seems to be unaffected by the disease,
resulting in AChE in the lumbar CSF of patients with Alzheimer\'s
disease being in the control range.(ABSTRACT TRUNCATED AT 250 WORDS).

84\. Abeyta, B.J., R.M. Albrecht, and C.R. Schermer, *Retrospective
study of neostigmine for the treatment of acute colonic
pseudo-obstruction.* Am Surg, 2001. 67(3): p. 265-8; discussion 268-9.
Acute colonic pseudo-obstruction (ACPO) typically develops
postoperatively or after severe illness. Studies suggest that
pharmacologic manipulation with intravenous (i.v.) neostigmine (NSM) may
be an effective and less invasive treatment modality for ACPO with
minimal side effects. The purpose of this study was to retrospectively
assess the efficacy and incidence of complications of an i.v. NSM bolus
in patients with ACPO. Eight patients with ten episodes of ACPO were
treated with a bolus dose of NSM. Rapid and effective decompression of
the colon was achieved in six episodes after a single dose of NSM at a
mean of 22.8 +/- 13.5 minutes. In three episodes decompression occurred
after a second dose of NSM at a mean of 44.7 +/- 37.7 minutes. One
patient failed NSM treatment but responded to a Cystografin enema. One
patient experienced significant bradycardia. NSM is a simple, safe, and
effective treatment for ACPO and based on result comparison of this
study and previous studies both bolus and slow infusion dosing practices
of NSM are effective. The NSM bolus dosing side effect profile has been
shown to include significant bradycardia, whereas when NSM was infused
over one hour significant bradycardic episodes requiring treatment have
not been encountered. We propose that a prospective study evaluating NSM
dosing as an i.v. bolus versus an i.v. infusion would be useful in
determining whether NSM infusion can be proven safer than bolus dosing
for the treatment of ACPO.

85\. Adams, S.B. and M.A. MacHarg, *Neostigmine methylsulfate delays
gastric emptying of particulate markers in horses.* Am J Vet Res, 1985.
46(12): p. 2498-9. Eight horses were allotted to 2 groups, each of 4
horses. All horses were given 100 plastic markers intragastrically via a
nasogastric tube. One group of animals (control group) was not given
medication after marker administration. The other group (test group) was
given neostigmine methylsulfate (0.022 mg/kg of body weight) in the
subcutaneous tissue at the time of marker administration and 30, 60, and
90 minutes later. All horses were killed 135 minutes after marker
administration to locate the beads in the gastrointestinal tract.
Gastric emptying of the markers was significantly delayed (P less than
0.05) in horses given neostigmine methylsulfate.

86\. Amaro, R. and A.I. Rogers, *Neostigmine infusion: new standard of
care for acute colonic pseudo-obstruction?* Am J Gastroenterol, 2000.
95(1): p. 304-5. This study was designed to assess the efficacy of i.v.
infusion of neostigmine in patients with acute colonic
pseudo-obstruction, which was defined as colonic distention with a cecal
diameter of at least 10 cm on plain radiographs and no radiographic
evidence of mechanical obstruction. Patients who failed to respond to
conventional management (nothing by mouth, nasogastric suction, postural
changes, i.v. fluids, electrolyte replacement, and discontinuation of
any drugs that affect colonic motility) for 24 h were included in the
study. Those with bradycardia (heart rate \<60/min), hypotension
(systolic blood pressure \<90 mm Hg), active bronchospasm, clinical or
radiographic evidence of perforation, history of partial colonic
resection, active gastrointestinal bleeding, pregnancy, or serum
creatinine \>3 mg/dL were excluded. Twenty patients were included in
this prospective, randomized, double-blind, placebo-controlled study.
Eleven patients received neostigmine 2.0 mg i.v. over 3-5 min with
electrocardiographic monitoring, and 10 received placebo. Patients were
evaluated for immediate clinical response (passage of flatus or stools
associated with decreased abdominal distention within 30 min) and
sustained response with decreased abdominal girth and reduced colonic
dilation on radiographs 3 h after infusion. Ten patients in the
neostigmine group had an immediate clinical response (median time, 4
min) compared to none in the placebo group (p\<0.001). Three patients in
the neostigmine group (27%) and eight in the placebo group (80%) failed
to show sustained improvement 3 h after infusion (p = 0.04). Eight
patients (one-neostigmine; seven-placebo) who failed to respond received
open-label treatment with neostigmine. Seven patients responded; one
patient from the placebo group failed and eventually required colonic
resection. In conclusion, from a total of 18 patients treated with
neostigmine, 17 (94%) had immediate clinical response, and 16 (89%) did
not have recurrent colonic dilation. The most common side effect was
crampy abdominal pain reported in 13 patients, although usually mild
(nine). Symptomatic bradycardia requiring atropine occurred in two
patients. Two patients in the neostigmine group died, but death was felt
not to be related to acute colonic pseudo-obstruction or its treatment.

87\. Alitalo, I. and A. Hietala, *Effects of atropine, isoprenaline,
neostigmine and practocol on cardiac arrhythmias induced by potassium in
anesthetized dogs.* Nord Vet Med, 1977. 29(11): p. 498-501. Either
atropine, isoprenaline, neostigmine or practolol was given intravenously
in therapeutic doses to dogs anesthetized with pentobarbitone sodium.
Cardiac arrhythmias were induced by potassium chloride intravenously as
a bolus injection in a dose of 0.3 mmol/kg. The standard
electrocardiographic lead II was recorded continuously. pH, Po2 and Pco2
were checked from arterial blood samples before and after the injection.
The isoprenaline treated dogs showed the smallest amount of arrhythmias
and all the dogs survived. More severe arrhythmias were seen in the
neostigmine and practolol treated dogs and the most severe occurrences
were encountered in the atropine treated dogs.

88\. Anzueto, A., *et al.*, *Acute inhalation toxicity of soman and
sarin in baboons.* Fundam Appl Toxicol, 1990. 14(4): p. 676-87. Adult
baboons (Papio sp.; 8-12 kg) were anesthesized with sodium pentobarbital
(20 mg/kg iv). The animals were instrumented for measurement of mean
blood pressure (MBP), pulmonary artery pressure (PAP), ECG, arterial and
mixed venous blood gases, lung volumes, lung pressures, and efferent
phrenic nerve activity. Bronchoalveolar lavage (BAL) was performed.
Studies were done prior to exposure, at intervals during the first 4 hr
postexposure, and at 4 and 28 days after exposure. Control animals
received a sham exposure to 2-propanol (N = 5). Soman (pinacolyl
methylphosphonofluoridate) at 13.14 micrograms/kg (2 X LD50) was
vaporized into the upper airway in a second group of animals (N = 5),
and sarin (isopropyl methylphosphonofluoride) 30 micrograms/kg (2 X
LD50) was vaporized into a third group of animals (N = 4). Controls
showed no change in any parameter either immediately after diluent
exposure or during the monitoring period. Soman and sarin produced a
decline in MBP and bradyarrhythmias that were reversed with atropine.
Apnea occurred in all soman- and sarin-exposed animals within 5 min
postexposure, and was associated with absence of phrenic nerve signal.
Ventilation was mechanically supported until the animal could maintain
normal arterial blood gases during spontaneous breathing. BAL studies
revealed an increase in total white cell population and neutrophils at 4
hr in all three groups. There were signs of impaired hemodynamics and
persistent lung injury for 4 days that resolved by 28 days after
exposure. In conclusion, inhalation of soman and sarin in the baboon is
associated with cardiac arrhythmias, development of apnea, and a
significant decrease in MBP. Inhalation exposure also resulted in a
persistent influx of neutrophils and hypoxemia.

89\. Haley, R.W., *et al.*, *Abnormal control of heart rate variability
in veterans with Gulf War Syndrome.* Submitted for publication.

90\. Ally, A., *et al.*, *Cardiovascular effects elicited by central
administration of physostigmine via M2 muscarinic receptors in conscious
cats.* Brain Res, 1995. 677(2): p. 268-76. The cardiovascular effects of
an intracerebroventricular (i.c.v.) injection of physostigmine were
studied using conscious cats. Physostigmine (5-25 micrograms: 5
microliters) caused a dose-dependent increase in mean arterial pressure
(MAP) and heart rate (HR). The highest dose (25 micrograms) increased
MAP and HR by 32 +/- 3 mmHg and 45 +/- 5 beats/min, respectively (n =
5). Pre-administration of the muscarinic receptor antagonist, atropine
(25 micrograms; i.c.v.) blocked the effects of physostigmine (25
micrograms; i.c.v.). Also, the pre-administration of the M2 muscarinic
antagonist, methoctramine (25 micrograms; i.c.v.), antagonized the
cardiovascular effects of physostigmine without altering the baseline
variables. However, the M1 muscarinic antagonist, pirenzepine (100
micrograms; i.c.v.) did not alter baseline MAP or HR, and also failed to
inhibit the cardiovascular responses to physostigmine. Similarly, the M3
muscarinic blocker, 4-diphenyl-acetoxy-N-methylpiperidine methiodide (50
micrograms; i.c.v.), neither changed baseline cardiovascular variables
nor blocked the effects of physostigmine. When the same cats were
anesthetized with intravenous injection of sodium pentobarbital (25-30
mg/kg), physostigmine (25 micrograms; i.c.v.) evoked a decrease in MAP
and HR of 13 +/- 6 mmHg and 15 +/- 6 bpm, respectively (n = 5). These
results demonstrate that the increases in MAP and HR to the i.c.v.
administration of physostigmine in conscious cats are possibly mediated
through stimulation of central M2 muscarinic receptors. In addition,
anesthesia reverses the effects elicited by the central administration
of physostigmine to a decrease in MAP and HR.

91\. Ally, A., G.A. Hand, and J.H. Mitchell, *Central injection of
physostigmine attenuates exercise-induced pressor response in conscious
cats.* Am J Physiol, 1997. 273(1 Pt 2): p. R393-9. The effects of
intracerebroventricular administration of physostigmine, a
cholinesterase inhibitor, on the cardiovascular responses evoked by
static voluntary exercise were investigated using conscious cats. Four
cats were trained to press a bar (200-650 g) with one forelimb for at
least 20 s. The changes in mean arterial pressure (MAP), heart rate
(HR), and developed force during the first five trials in 30 min by each
individual cat were averaged, and a mean of the four values was then
calculated. After the cats exercised for 30 min, either artificial
cerebrospinal fluid (CSF) or physostigmine (5 micrograms) was
administered intracerebroventricularly. Before physostigmine, exercise
trials by the cats increased MAP and HR by 17 +/- 3 mmHg and 42 +/- 4
beats/min, respectively. Administration of physostigmine did not alter
the resting MAP and HR but attenuated the MAP and HR responses to
exercise (5-30 min postphysostigmine: MAP = 8 +/- 3 mmHg, HR = 25 +/- 7
beats/min; 30-60 min postphysostigmine: MAP = 4 +/- 3 mmHg, HR = 19 +/-
8 beats/min). Intracerebroventricular administration of CSF had no
effect on the cardiovascular responses to static exercise. Pretreatment
with the muscarinic antagonist, atropine (25 micrograms icv), blocked
the attenuating effects of subsequent intracerebroventricular
administration of physostigmine. These results demonstrate that
stimulation of central muscarinic receptors attenuates the
cardiovascular responses to static exercise by conscious cats. In
addition, the present study suggests that there is no tonic effect of
central muscarinic receptors on the cardiovascular responses to
voluntary exercise.

92\. Jancin, B., *Pyridostigmine reduced orthostatic intolerance.*
Internal Medicine News, 2002. June 15: p. 32. Cites Dr. Wolfgang Singer
at annnual meeting of Amer Acad of neurology.

18 pts with orthostatic intolerance with HR increase of =30 w standing,
w sx cerebral hypoperfusion or symp overactivity. All underwent
tilt-table testing before and at 1h after taking 60mg of oral PB. Mean
HR increas of 45 dropped to 38 with rx (significant). Also supine HR
fell 79 to 73; upright fell 124 to 111. Sx severity score 0-10 scale
improved 5.6 on baseline tilt table to 4.2 after PB (on 0-10 scale)

Plasma NE increased predrug 214 and 491pg/ml supine and upright to 263
and 5552pg/ml after drug. \"No side effects were noted in assoc with PB
therapy\".

Current most widely used drugs are beta blockers, vasoconstrictors, and
volume expansion agents, but resp\'s are highly variable.

93\. Park, K.H., J.P. Long, and J.G. Cannon, *Reversal of
antihypertensive agent-induced postural hypotension with physostigmine.*
Arch Int Pharmacodyn Ther, 1991. 311: p. 155-65. Physostigmine,
administered intravenously, reversed postural hypotension induced by
hypotensive agents, guanethidine, clonidine and dopamine2-receptor
agonists. Postural hypotension, induced by pentobarbital sodium, was
also reversed by physostigmine. Neostigmine reversed postural
hypotension induced by clonidine following intracerebroventricular, but
not intravenous administration. It is proposed that centrally acting
cholinomimetic agents may be used to manage postural hypotension
resulting from suppression of sympathetic nervous system activity.

94\. 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.

95\. Aoyagi, M., *et al.*, *Central cholinergic control of cerebral
blood flow in the baboon. Effect of cholinesterase inhibition with
neostigmine on autoregulation and CO2 responsiveness.* J Neurosurg,
1975. 43(6): p. 689-705. Cerebral autoregulation and vastomotor
responsiveness to carbon dioxide (CO2) were measured quantitatively by
the use of the autoregulation index and chemical index, respectively, in
normal baboons before and after intravertebral and intracarotid infusion
of the anticholinesterase agent, neostigmine methylsufate (Prostigmin).
Continuous measurements were made of cerebral blood flow (measured as
bilateral internal jugular venous outflow), arterial and cerebral venous
pO2 and pCO2, cerebral arteriovenous oxygen differences, and
endotracheal CO2. The effect of intravertebral infusion of neostigmine
(12.5 mug/kg body weight) was compared to intravertebral infusion of
neostigmine (25 mug/kg body weight) for assessment of any specific
action of the drug on a hypothetical cholinergic vasomotor center,
presumed to be located in the territory of the vertebrobasilar supply.
No significant or persistent changes in cerebral blood flow (CBF) and
cerebral metabolic rate for oxygen (CMRO2) followed either
intravertebral or intracarotid infusion of neostigmine. Cerebral
vascular resistance (CVR) and cerebral perfusion pressure (CPP),
however, decreased significantly after intravertebral infusion. Cerebral
autoregulatory vasoconstriction during increases of CCP was
significantly reduced following both intravertebral and intracarotid
infusion. Cerebral autoregulatory vasodilatation was not altered as CPP
was lowered. Cerebral vasodilatory reactivity to CO2 inhalation was
significantly enhanced following intravertebral neostigime but not
following intracarotid neostigmine. Cerebral vasoconstrictive response
to hyperventilation was not influenced by neostigmine. These results
support the view that central cholinergic cerebrovascular influences
exist, and are vasodilatory in nature.

96\. Eneroth-Grimfors, E., *et al.*, *Noninvasive test of microvascular
endothelial function in normal and hypertensive pregnancies.* Br J
Obstet Gynaecol, 1993. 100(5): p. 469-71. OBJECTIVE: To examine
microvascular endothelial cell function in vivo in pre-eclampsia.
DESIGN: Iontophoresis of acetylcholine (ACh), which gives rise to
endothelial cell dependent vasodilatation, and of sodium nitroprusside
(SNP), which elicits vasodilatation independently of functioning
vascular endothelium. SETTING: Sodersjukhuset, Stockholm, Sweden.
SUBJECTS: Ten pre-eclamptic patients, ten healthy pregnant women and ten
healthy nonpregnant women were examined. MAIN OUTCOME MEASURES: The
degree of vasodilatation following iontophoretic administration of ACh
compared with SNP was recorded with a laser Doppler technique, the data
being analysed on a personal computer. RESULTS: Both ACh and SNP
administration resulted in marked vasodilatation; the magnitude of the
vasodilatation was similar in the three groups of women. CONCLUSION:
Following iontophoretic administration of endothelial cell dependent or
independent vasodilatators, laser Doppler measurement of blood flow
demonstrated no microvascular endothelial cell dysfunction in
pre-eclamptic women.

97\. Arora, S., *et al.*, *Estrogen improves endothelial function.* J
Vasc Surg, 1998. 27(6): p. 1141-6; discussion 1147. PURPOSE: To
determine the effect of estrogen on endothelium-dependent relaxation in
the cutaneous microcirculation of women. METHODS: Three groups of women
participated in the study. Group 1 (n = 20) was premenopausal and had a
mean age of 39 years (range 24-50 years). Group 2 (n = 9) was
postmenopausal and had a mean age of 58 years (range 53-65 years). Group
3 (n = 11) was postmenopausal and taking estrogen replacement therapy;
the mean age was 53 years (range 43-58 years). Eleven women in group 1
underwent testing twice, once during menstruation (mean serum estradiol
level 73 +/- 30 pg/ml) and once during midcycle (mean serum estradiol
level 268 +/- 193 pg/ml; p = 0.003). Single-point laser Doppler
ultrasound and laser Doppler imaging with a scanner were used to measure
vasodilatation in the forearm skin in response to iontophoresis of 1%
acetylcholine (endothelium dependent) and 1% sodium nitroprusside
(endothelium-independent smooth muscle relaxant). RESULTS: All three
groups were matched for body mass index and fasting glucose, total,
high-density lipoprotein, and low-density lipoprotein cholesterol and
triglyceride levels. All women had normal blood pressure, and none
smoked. Mean serum estradiol levels were 196 +/- 170 pg/ml (group 1), 35
+/- 12 pg/ml (group 2), and 107 +/- 78 pg/ml (group 3) (p = 0.004).
Maximum microvascular vasodilatation (percentage increase over baseline)
in response to acetylcholine was reduced in group 2 (93% +/- 43%)
compared with group 1 (187% +/- 63%) and group 3 (142% +/- 56%) (p =
0.001). The response to sodium nitroprusside also was diminished in
group 2 (73% +/- 27%) compared with group 1 (126% +/- 45%) and group 3
(100% +/- 32%) (p = 0.02). Within group 1 the acetylcholine response was
higher during the midcycle phase (186% +/- 31%) compared with the
menstrual phase (147% +/- 57%) (p \< 0.05). The sodium nitroprusside
response also was higher during the midcycle phase (144% +/- 31%)
compared with the menstrual phase (94% +/- 41%) (p \< 0.05) CONCLUSION:
The results indicate that estrogens might enhance endothelium-dependent
and endothelium-independent vasodilatation in the microcirculation of
women.

98\. Kubli, S., *et al.*, *Reproducibility of laser Doppler imaging of
skin blood flow as a tool to assess endothelial function.* J Cardiovasc
Pharmacol, 2000. 36(5): p. 640-8. Endothelial dysfunction might be an
important and early event in the pathogenesis of major cardiovascular
diseases. Therefore, the evaluation of endothelial function in humans
may be of great clinical relevance. Usual methods for that purpose are
either invasive and/or technically demanding. In the dermal
microcirculation, endothelial function may be assessed noninvasively
from the laser Doppler measurement of increases in blood flow after
either the transdermal application of acetylcholine by iontophoresis, or
the release of transient arterial occlusion (reactive hyperemia). An
endothelium-independent response may be provided by the iontophoresis of
sodium nitroprusside. This approach is notable for technical simplicity,
but of uncertain reproducibility. Sixteen young, healthy, nonsmoking
males were examined in the fasting state. Changes in skin blood flow
were measured with a laser Doppler imager during the iontophoresis of
acetylcholine and sodium nitroprusside, as well as during reactive
hyperemia, on two different days, at each of two different sites on the
volar face of the forearm. Nonspecific effects related to the
stimulation of terminal nerve fibers by the iontophoretic current were
suppressed by prior surface anesthesia. The iontophoresis of
acetylcholine and sodium nitroprusside induced a seven- to eightfold
increase in dermal blood flow. The corresponding figure for peak
reactive hyperemia was approximately fourfold. The mean coefficients of
variation of responses recorded on different days, on the same site, in
the same individual were \<10% for iontophoresis of acetylcholine and
for peak reactive hyperemia, and between 10 and 20% for iontophoresis of
sodium nitroprusside. This day-to-day variation was significantly
smaller than the site-to-site variation (p \< 0.01 for all three
responses). Endothelium-dependent and -independent responses of dermal
blood flow evaluated with laser Doppler imaging are highly reproducible
from day to day, at least in healthy nonsmoking young male subjects, and
provided some simple precautions are observed, foremost among which is
the strict standardization of the recording site. These observations may
have implications for the testing of endothelial function in clinical
studies.

99\. Williams, M.R., *et al.*, *Variations in endothelial function and
arterial compliance during the menstrual cycle.* J Clin Endocrinol
Metab, 2001. 86(11): p. 5389-95. Female sex hormones have been
implicated in the cardioprotection of premenopausal women. However, the
cardiovascular actions of these hormones and the effects of their
natural fluctuations during the menstrual cycle are not fully
understood. We studied changes in vascular function during the menstrual
cycle in 15 healthy premenopausal women. Four noninvasive procedures
were performed during the early follicular (EF), late follicular (LF),
early luteal (EL), and late luteal (LL) phases: flow-mediated dilatation
(FMD) of the brachial artery during reactive hyperemia, laser Doppler
velocimetry (LDV) with direct current iontophoresis of acetylcholine
(ACh) and nitroprusside, whole body arterial compliance (WBAC), and
pulse wave velocity. Hormone levels were consistent with predicted cycle
phase and showed that all subjects ovulated during the cycle studied.
FMD, LDV with ACh, and WBAC varied cyclically, with significant
increases from the F to LF phase, sharp falls in the EL phase, and
significant recoveries in the LL phase. These changes were most marked
for FMD \[EF, 8.8 +/- 0.6% (mean +/- SEM); LF, 10.0 +/- 0.7; EL, 4.2 +/-
0.6; LL, 8.6 +/- 0.9\] and the LDV response to ACh (EF, 2.7 +/- 0.2
V/min; LF, 3.3 +/- 0.4; EL, 1.8 +/- 0.3; LL, 2.7 +/- 0.4). WBAC changed
similarly (EF, 0.58 +/- 0.08 arbitrary units; LF, 0.84 +/- 0.06; EL,
0.65 +/- 0.05; LL, 0.68 +/- 0.06). Sodium nitroprusside-induced
vasodilatation decreased significantly from EF to EL, with no other
significant difference, and pulse wave velocity did not vary
significantly over the four time points. Conductance and resistance
artery endothelial reactivity and smooth muscle sensitivity to nitric
oxide and arterial compliance are modulated significantly in response to
the changing hormonal patterns of the menstrual cycle. These findings
emphasize the importance of menstrual phase in the interpretation of
data on endothelial function and may provide insights into the
mechanisms underlying sex differences in cardiovascular risk and other
disease processes in premenopausal women.

100\. Newton, D.J., F. Khan, and J.J. Belch, *Assessment of
microvascular endothelial function in human skin.* Clin Sci (Lond),
2001. 101(6): p. 567-72. Endothelial dysfunction is an important factor
in many cardiovascular diseases, and is commonly associated with
impaired endothelium-mediated vasodilatation. Information about the
mechanisms behind this dysfunction has come largely from animal studies
or, in humans, through invasive techniques that are not specific to one
vascular bed. We have developed protocols to assess endothelial function
non-invasively in the cutaneous microcirculation by measuring blood flow
responses to four receptor-specific vasoactive compounds. Cumulative
doses of acetylcholine, methacholine, bradykinin and substance P were
administered iontophoretically to the forearm skin of healthy volunteers
on two to three occasions. Dose-dependent increases in skin
microvascular blood flow in response to these drugs were measured with
laser Doppler imaging. Vascular responses to acetylcholine and
methacholine were reasonably consistent, with coefficients of variation
of approx. 17%. The coefficients of variation for bradykinin and
substance P were much poorer, as high as 70% for some doses. This might
partly be a consequence of the more unpredictable effects of histamine
release in the vasoactive behaviour of these two agonists. Although it
might be advantageous to find other agonists with which to test the
function of different receptor pathways, we have shown that just
acetylcholine and methacholine can currently be used with iontophoresis
to allow sensitive and reproducible assessment of endothelial function.

101\. Elherik, K., *et al.*, *Circadian variation in vascular tone and
endothelial cell function in normal males.* Clin Sci (Lond), 2002.
102(5): p. 547-52. The existence of circadian rhythms in the time of
onset of acute cardiovascular events has been described previously. This
report describes the circadian variation in endothelial cell products,
such as nitric oxide (NO) and endothelin-1 (ET-1) levels, and
endothelium-dependent and -independent vasodilation in normal males.
Plasma ET-1 and NO were measured every 4 h in nine subjects (20-41 years
old) over a 24 h period. Endothelium-dependent and -independent vascular
responses were measured in the forearm skin every 4 h using laser
Doppler imaging after iontophoresis of increasing doses of acetylcholine
(ACh) and sodium nitroprusside respectively. A statistically significant
circadian variation was demonstrated for the mean ACh response
(P=0.0001, ANOVA). The peak response \[in arbitrary perfusion units
(AU)\] occurred at 16.00 hours (8.90+/-1.91 AU) and the lowest response
at 08.00 hours (4.57+/-0.66 AU). A significant circadian variation was
also seen for the highest dose of sodium nitroprusside (P=0.036, ANOVA),
the peak occurred at 16.00 hours (3.97+/-1.80 AU), and the lowest at
04.00 hours (2.62+/-0.58 AU) and 08.00 hours (2.58+/-1.16 AU). There was
a significant circadian variation in the ET-1 levels (P=0.04) with two
peaks, one at 20.00 hours (0.80+/-0.28 pg/ml) and the other at 08.00
hours (0.84+/-0.15 pg/ml). The lowest value occurred at 16.00 hours
(0.61+/-0.24 pg/ml). There was also a borderline trend for a circadian
variation in NO levels (P=0.06), with higher levels at 20.00 hours
(15.53+/-8.42 micromol/l), and low levels at 04.00 hours (10.87+/-4.70
micromol/l) and 08.00 hours (9.82+/-3.15 micromol/l). ACh responses were
significantly correlated with ET-1 (r=-0.3, P=0.02) and NO (r=0.30,
P=0.02) levels. Our findings suggest that endothelial activity has a
circadian variation with attenuation in the morning. These circadian
variations in endothelial activity might play an important role in the
occurrence of acute cardiovascular events at this time, which are
precipitated through the interplay between ET-1, NO and vascular
function.

102\. Lim, S.C., *et al.*, *The effect of hormonal replacement therapy
on the vascular reactivity and endothelial function of healthy
individuals and individuals with type 2 diabetes.* J Clin Endocrinol
Metab, 1999. 84(11): p. 4159-64. Estrogens protect healthy women from
cardiovascular disease. However, epidemiological data suggest that women
with diabetes are denied the cardioprotection associated with estrogens.
Whether or not hormonal replacement therapy (HRT) confers cardiovascular
benefits in postmenopausal women with diabetes is not known. The aim of
this study was to examine the effects of HRT on the microvascular
reactivity and endothelial function of individuals with and without
diabetes. We studied the following groups of individuals: premenopausal
healthy women \[n = 28, age 41 +/- 8 yr (mean +/- SD)\], premenopausal
women with type 2 diabetes (n = 16, age 43 +/- 6 yr); postmenopausal
healthy women (n = 12, age 57 +/- 4 yr), postmenopausal women with
diabetes (n = 17, age 62 +/- 5 yr); postmenopausal healthy women on HRT
(n = 13, age 51 +/- 5 yr), postmenopausal women with diabetes on HRT (n
= 11, age 57 +/- 7 yr). We used laser Doppler flowmetry to measure
forearm cutaneous vasodilatation in response to iontophoresis of 1%
acetylcholine (endothelium dependent) and 1% sodium nitroprusside
(endothelium independent). The endothelium-dependent vasodilation was
significantly higher in premenopausal healthy women (180 +/- 67%;
increase over baseline) compared to premenopausal diabetic women (87 +/-
41%; P \< 0.001). endothelium-dependent vasodilation was also higher in
postmenopausal healthy women on HRT (143 +/- 52) compared with
postmenopausal diabetic women on HRT (86 +/- 61), postmenopausal healthy
women without HRT (104 +/- 43), and postmenopausal diabetic women
without HRT (74 +/- 28; P \< 0.001). A similar pattern of responses was
observed in the endothelium-independent vasodilation (premenopausal
healthy women, 126 +/- 56; premenopausal diabetic women, 88 +/- 26;
postmenopausal healthy women on HRT, 121 +/- 37; postmenopausal diabetic
women on HRT, 88 +/- 41; postmenopausal healthy women without HRT, 84
+/- 36; and postmenopausal diabetic women without HRT, 73 +/- 36; P \<
0.001). Soluble intercellular adhesion molecule (sICAM) was also
measured among all the women with diabetes. Premenopausal women with
diabetes (248.9 +/- 56 ng/ml) and postmenopausal women with diabetes on
HRT (257.7 +/- 49 ng/ml) had lower sICAM levels compared with the
postmenopausal diabetic women without HRT (346.4 +/- 149 ng/ml; P \<
0.05). We conclude that menopausal status and type 2 diabetes are
associated with impaired microvascular reactivity. HRT substantially
improves microvascular reactivity in postmenopausal healthy women. In
contrast, the effect of HRT on the microvascular reactivity of
postmenopausal diabetic women is less apparent. However, the use of HRT
among women with diabetes is associated with lower sICAM levels,
suggesting an attenuation in endothelial activation.

103\. Wirtschafter, J.D., C.R. Volk, and R.J. Sawchuk, *Transaqueous
diffusion of acetylcholine to denervated iris sphincter muscle: a
mechanism for the tonic pupil syndrome (Adie syndrome).* Ann Neurol,
1978. 4(1): p. 1-5. The accepted hypothesis for the pathophysiology of
tonic pupil syndrome (Adie syndrome) was reexamined in light of recent
developments concerning denervation supersensitivity of cholinergically
innervated smooth muscle. Kinetic analysis suggests that enzymatic
hydrolysis is unimportant relative to convective diffusion in the
turnover of acetylcholine in the aqueous humor. We postulate that the
greater response to near stimuli than to light stimuli and the delay in
iris sphincter contraction and relaxation can be explained by release of
acetylcholine from the neuromuscular junction of the ciliary muscle
followed by transaqueous diffusion to receptor sites on denervated,
supersensitive iris sphincter muscle.

104\. Fountoulakis, K., *et al.*, *Changes in pupil reaction to light in
melancholic patients.* Int J Psychophysiol, 1999. 31(2): p. 121-8. The
aim of the current study was to compare the pupil reaction to light in
depressed patients and normal control subjects. Seven depressed patients
with melancholic features according to DSM-IV criteria and 14 age- and
gender-matched control subjects took part in the study. All were free of
any medication for at least 2 weeks. All were aged between 25 and 50
years. An optical method was used to assess the pupil reaction to a
single flash. Depressed patients manifested shorter latency for
constriction than control subjects, and a marginal difference in the
total work produced by acetylcholine. The results of the current study
support the theory that there is a norepinephrine hypoactivity in
melancholic depression, with less affected acetylcholine activity.

105\. Elliott, A. and C. Carter, *Pupil size after extracapsular
cataract extraction and posterior chamber lens implantation: a
prospective randomized trial of epinephrine and acetylcholine.*
Ophthalmic Surg, 1989. 20(8): p. 591-4. The effects of using epinephrine
in the irrigating fluid and intracameral acetylcholine were studied by
measuring changes in pupil size in the 48 hours following extracapsular
cataract extraction and intraocular lens implantation in 39 eyes.
Epinephrine reduced peroperative pupil constriction, but its effect was
insignificant thereafter. The pupil constriction following acetylcholine
was maximal at 2 hours and was still significant at 4 hours, but pupils
redilated by 6 hours. Neither drug had any effect after this time. The
edge of most lens implants was visible at 6 hours, after which pupils
steadily constricted.

106\. Suzuki, R., S. Kobayashi, and H. Yoshino, *\[The inhibitory
cholinergic innervation of the dilator on the movement of the pupil\].*
Nippon Ganka Gakkai Zasshi, 1984. 88(12): p. 1555-7.

107\. Naito, M., *\[The autonomic nervous system and eyes: the role of
the sympathetic nerve on extra-ocular muscle and pupil\].* Nippon Ganka
Gakkai Zasshi, 1971. 75: p. 988-95.

108\. Pohjanpelto, P., *\[Carbachol in cataract surgery used for
constriction of the pupil\].* Duodecim, 1969. 85(24): p. 1504-7.

109\. Cook, J.E., M.A. Kolka, and C.B. Wenger, *Chronic pyridostigmine
bromide administration: side effects among soldiers working in a desert
environment.* Mil Med, 1992. 157(5): p. 250-4. The side effects of
chronic pyridostigmine bromide administration were studied in seven male
soldiers performing moderate-intensity exercise in a desert environment.
A 2-week, double-blind, placebo-controlled crossover design was employed
in which pyridostigmine was administered for 7 consecutive days (30 mg
orally, t.i.d.). Four hours each day were spent in the heat (42 degrees
C, 20% relative humidity); 2 hours rest followed by 2 hours moderate
exercise (40% maximal aerobic power). Each day, subjects completed four
symptom questionnaires and received three focused physical examinations.
Symptoms reported did not differ between treatment groups except for
fewer headaches during pyridostigmine treatment. Soldiers were unable to
distinguish the effects of pyridostigmine from placebo. Pyridostigmine
was associated with lower resting diastolic blood pressure
(approximately 4 mmHg, p less than 0.05), smaller pupil diameter
(approximately 0.5 mm, p less than 0.01), decreased handgrip strength
(approximately 3%, p less than 0.05), and higher final rectal
temperature (approximately 0.1 degree C, p less than 0.01). Effects of
this magnitude are not likely to appreciably limit performance. We
conclude that chronic pyridostigmine administration does not negatively
impact on soldiers\' ability to perform physical work over repeated days
in a desert environment.

110\. Borland, R.G., *et al.*, *Studies on the possible central and
peripheral effects in man of a cholinesterase inhibitor
(pyridostigmine).* Hum Toxicol, 1985. 4(3): p. 293-300. The effect of a
reversible cholinesterase inhibitor (pyridostigmine: 30 mg, 8-hourly for
3 days) on psychomotor performance and visual function and on the
electrical activity of the brain was studied in healthy man. It was not
possible to detect any change during the six individual experimental
sessions over the 3 days, but with pooling of the data there was a drug
effect. The threshold for detection of a flickering light was increased
and less responses were missed on a dynamic visual-acuity task.
Visuo-motor coordination was impaired. The observations were consistent
with the known activity of the drug and suggest an increase in central
arousal and minimal alteration in motor coordination.

111\. Goel, A., S. Joseph, and T.K. Dutta, *Organophosphate poisoning:
predicting the need for ventilatory support.* J Assoc Physicians India,
1998. 46(9): p. 786-90. The present study evaluated 103 consecutive
patients of organophosphate poisoning with special reference to the need
for ventilatory support. Of 103 patients, 36(34.95%) required assisted
ventilation. The need for ventilatory support was significantly more
with greater time duration for institution of specific treatment, low
level of sensorium at admission, pin-point pupils and generalized
fasciculations, presence of convulsions, presence of respiratory
insufficiency at admission, and higher initial atropine requirement for
atropinization. The study also proposes a system of grading of severity,
modified from the existing systems proposed.

112\. Kay, C.D. and J.D. Morrison, *The effects of ingestion of 60 mg
pyridostigmine bromide on contrast sensitivity in man.* Hum Toxicol,
1988. 7(4): p. 347-52. The effects on vision of ingestion of the
anticholinesterase pyridostigmine bromide (60 mg), assessed from
pharmacokinetic data to provide at least 20% inhibition of blood
cholinesterase over the 1 1/2-4 1/2 h experimental period, was compared
with 60 mg lactose in a double-blind crossover protocol. Contrast
sensitivity to stationary oscilloscope-generated gratings of 3-40 c/deg
showed a small but significant increase of 7% which was consistent with
a small reduction in pupil diameter, surmised to cause a small
improvement in optical quality. This reduction in pupil diameter was,
however, overshadowed by a larger though still non-significant reduction
on the second visit to the laboratory compared with the first. Contrast
sensitivities to laser interference fringes observed in the Maxwellian
view, by which the effects of the optical media are essentially
bypassed, thus providing an entirely neural assessment, were unchanged
after pyridostigmine. It is concluded that pyridostigmine may be given
as a prophylactic in anticipation of exposure to an organophosphorus
anticholinesterase without a deleterious effect on stationary visual
function.

113\. Loupe, D.N., *et al.*, *Pupillary response to tropicamide in
patients with Alzheimer disease.* Ophthalmology, 1996. 103(3): p.
495-503. PURPOSE: To determine whether pupillary responses to dilute
tropicamide could be used as a diagnostic test for Alzheimer disease
(AD). The authors also investigated whether concurrent use of an oral
acetylcholinesterase inhibitor (tacrine) alters the pupillary response
to dilute tropicamide in patients with AD, and whether pupillary
responses to dilute tropicamide differ in young versus older control
subjects. METHODS: Pupillary diameter and area of both eyes were
measured in light and darkness, at 10-minute intervals for 40 minutes
after random instillation of 0.01% tropicamide to one eye. Four groups
of subjects were studied: 9 patients with AD, 10 who were treated with
tacrine, 11 older control subjects, and 10 young control subjects.
RESULTS: Mean change in anisocoria was not significantly different among
groups at any of the measurement time points. Mean percent change in
diameter of the treated eyes showed a trend toward faster maximum
dilatation in the AD groups, but change in pupillary measurements did
not identify individuals with AD. CONCLUSION: Pupillary response to
dilute tropicamide did not effectively distinguish individual patients
with AD from young or older control subjects.

114\. Macic, I., *et al.*, *\[Antagonism of miosis induced by sarin and
VX in rabbits\].* Vojnosanit Pregl, 1986. 43(2): p. 103-8.

115\. Nozaki, H., *et al.*, *Secondary exposure of medical staff to
sarin vapor in the emergency room.* Intensive Care Med, 1995. 21(12): p.
1032-5. OBJECTIVE: To clarify the risk of secondary exposure of medical
staff to sarin vapor in the emergency room, and to warn emergency room
staffs of the hazard. DESIGN: Retrospective observational survey.
SETTING: Emergency department of a university hospital in a metropolitan
area of Japan. PARTICIPANTS: Fifteen doctors treating victims of a
terrorist attack with sarin in the Tokyo subways on the day of the
attack. MEASUREMENTS AND RESULTS: Of the 15 doctors who worked in the
emergency room treating the victims, 13 became simultaneously aware of
symptoms during the resuscitation of two victims who were exposed to
sarin. Among 11 doctors (73%) who complained of dim vision, the pupils
were severely miotic (\<2 mm) in 8 (73%). Other symptoms included
rhinorrhea in eight (53%), dyspnea or tightness of the chest in four
(27%), and cough in two (13%). Atropine sulfate was given to six, and
pralidoxime was given to one of these six doctors. To decontaminate the
emergency room of sarin vapor, ventilation was facilitated and all
belongings of the patients were sealed up. None of the doctors noticed
worsening of their symptoms thereafter. CONCLUSIONS: Careful attention
to the risks of secondary exposure to toxic gas in the emergency room
and prompt decontamination if such exposure should occur are necessary
in the case of large-scale disasters caused by sarin.

116\. Nozaki, H., *et al.*, *Relationship between pupil size and
acetylcholinesterase activity in patients exposed to sarin vapor.*
Intensive Care Med, 1997. 23(9): p. 1005-7. OBJECTIVE: To elucidate the
effect of sarin vapor on pupil size and erythrocyte acetylcholinesterase
activity (AchE). DESIGN: Retrospective observational survey. SETTING:
Emergency department of an urban teaching hospital. PATIENTS: 80
patients who were exposed to sarin in a terrorist attack in Tokyo
subways. MEASUREMENTS AND RESULTS: Pupil size and AchE activity on the
day of exposure were measured. Among the 80 patients, the pupils were
miotic (\< 3 mm) in 50 patients (62.5%), while AchE activity was below
the normal range (\< 1.2 U) in 34 patients (42.5%). AchE was
significantly lower in the miotic group than in the group with normal
pupils (1.0 +/- 0.5 U vs 1.5 +/- 0.3 U, p \< 0.01). In the miotic group,
AchE activity was lower than normal in 32 patients (64.0%) but was
decreased in only 2 patients in the normal pupil group (6.7%) (p \<
0.01). CONCLUSIONS: Miosis is a more sensitive index of exposure to
sarin vapor than erythrocyte AchE. Systemic poisoning is apparently less
likely to develop if the patient\'s pupil size is normal on arrival at
the hospital.

117\. Rengstorff, R.H., *Accidental exposure to sarin: vision effects.*
Arch Toxicol, 1985. 56(3): p. 201-3. Two men were accidentally exposed
to vapors of sarin, a cholinesterase inhibitor and extremely toxic nerve
gas. Diagnosis was confirmed by depressed cholinesterase activity, and
fixed extremely miotic pupils. No other signs or symptoms developed and
neither man required treatment. Recovery to normal cholinesterase
activity was gradual over a 90-day period. Pupillary reflexes were not
detectable until 11 days after exposure; the miotic pupils dilated
slowly over a 30-45 day-period. Eye pain and blurred vision did not
occur; visual acuity and amplitude of accommodation were improved for
several weeks. Other functions not affected significantly were
intraocular pressure, visual fields, color vision, heterophorias, and
vergences.

118\. Sidell, F.R. and W.A. Groff, *The reactivatibility of
cholinesterase inhibited by VX and sarin in man.* Toxicol Appl
Pharmacol, 1974. 27(2): p. 241-52.

119\. Wiley, R.W., J.C. Kotulak, and I. Behar, *The effects of
pyridostigmine bromide on visual performance.* Aviat Space Environ Med,
1992. 63(12): p. 1054-9. The effects of pyridostigmine bromide (PB) on
selected visual functions were measured on four healthy aviator
candidates. Following a pretreatment day during which baseline
measurements were completed, subjects were administered currently
recommended doses (30 mg, t.i.d.) of PB for 3 d during which their
visual functions were assessed using a repeated measures design. Spatial
resolution ability was evaluated with high and low contrast visual
acuity charts and contrast sensitivity charts at three luminance levels.
Dark adaptation was evaluated by measuring visual thresholds for 40 min
after a standardized retinal photopigment bleach. Also, refractive error
and several oculomotor functions (lateral phoria, fusional vergence,
accommodative amplitude, and pupil size) were measured. On days that the
subjects ingested PB, only refractive error and pupil diameter were
significantly different, and these only minimally. We conclude that the
use of PB at doctrinal doses will not significantly compromise an
aviator\'s visual ability.

120\. Schneider, R., *A test for the assessment of the duration of the
action of neostigmine and of the relative potencies of various
anticholinesterases using the pupil of the mouse.* J Pharm Pharmacol,
1970. 22(4): p. 298-301.

121\. Beaver, K.M. and T.J. Gavin, *Treatment of acute anticholinergic
poisoning with physostigmine.* Am J Emerg Med, 1998. 16(5): p. 505-7.
Five cases of acute anticholinergic poisoning presenting to an
inner-city emergency department (ED) are discussed. All five patients
presented with classic signs and symptoms of anticholinergic toxicity,
which included tachycardia, hot, dry and flushed skin, markedly dilated
and fixed pupils, and pronounced delirium. The patients were violently
agitated, and physical restraint was required. Initial treatment with
benzodiazepines did not diminish their combative behavior. Treatment
with intravenous physostigmine salicylate resulted in a decrease in
agitation within 15 to 20 minutes of therapy. No untoward effects
occurred as a result of treatment with physostigmine.

122\. Crowell, E.B., Jr. and J.S. Ketchum, *The treatment of
scopolamine-induced delirium with physostigmine.* Clin Pharmacol Ther,
1967. 8(3): p. 409-14.

123\. Wilhelm, B., *et al.*, *\[Pupillography as an objective attention
test\].* Wien Med Wochenschr, 1996. 146(13-14): p. 387-9. Infrared video
pupillography (IVP) allows continuous recording of spontaneous pupillary
oscillations in darkness which change characteristically with fatigue.
Pupil size in darkness is age-related, has its maximum in the second
decade and subsequently decreases during life time. Normally, the pupil
oscillates in darkness with a frequency of about 1 Hz and amplitudes of
less than 0.3 mm. Excessive daytime sleepiness causes instability of
this pupillary behaviour which is constant in alert normals, and so
called fatigue waves appear with an amplitude reaching several
millimeters. The frequency profile is dominated by slow frequencies
below 0.5 Hz, while average pupil size decreases continuously with time.
As IVP is an objective and time-saving method it could become an
important supplement to test procedures used in sleep medicine and sleep
research to measure daytime sleepiness.

124\. Wilhelm, B., *et al.*, *\[Pupillography for objective vigilance
assessment. Methodological problems and possible solutions\].*
Ophthalmologe, 1996. 93(4): p. 446-50. To measure vigilance disorders in
healthy normals or in patients (narcolepsy, sleep apnea syndrome) is
difficult, time-consuming and hardly objective with present methods.
Recording and analysis of spontaneous pupillary behaviour in darkness by
infrared video pupillography is an objective and time-saving method to
measure daytime sleepiness. However, certain external conditions must be
satisfied (avoid light, noise, stress) to get reliable results.
Spontaneous pupillary oscillations are recorded in darkness over 10 min
and data are analyzed by fast Fourier transformation, with additional
calculation of the mean pupillary diameter for each time segment
(approx. 1 min). While in the alert normal, pupil remains dilated during
the measurement in darkness and oscillates with an amplitude below 0.3
mm and a frequency about 1 Hz, there are characteristic changes in
fatigue: (1) low-frequency components dominate the spontaneous pupillary
oscillations, with an amplitude reaching several millimeters, and (2)
pupil diameter decreases with time. Infrared video pupillography could
play a role as a screening method and therapy control for hypersonic
patients (most frequent: sleep apnea syndrome) with excessive daytime
sleepiness. An objective, time-saving method like infrared video
pupillography would be useful in sleep medicine and psychiatry when
testing the level of vigilance, and in psychology or industrial medicine
as well, providing informations about acute vigilance problems in
healthy normals.

125\. Morad, Y., *et al.*, *Pupillography as an objective indicator of
fatigue.* Curr Eye Res, 2000. 21(1): p. 535-42. PURPOSE: To determine
whether parameters calculated from pupillary activity can identify
subjects with sleep deprivation, and whether the objective values
correlate with a subjective feeling of fatigue. METHODS: pupil size in
the dark was recorded continuously for 10 minutes in 12 healthy
volunteers using an infrared video camera. Two recordings were made for
each subject: after a full night\'s sleep, and after 24 hours of sleep
deprivation. Several parameters calculated from pupil size and activity
were analyzed and compared with a subjective rating of the state of
alertness provided by the participants in each test. RESULTS: All
pupillary parameters differed significantly between alertness and
fatigue (p = 0.0076-0. 0186). Changes in one of the parameters - average
pupillary diameter - correlated with changes in the subjective level of
sleepiness (r = -0.51, p = 0.028). Although the values of most
parameters differed among subjects, an absolute value of more than 25 in
one parameter, cumulative pupillary variability ratio, was always
associated with sleep deprivation. CONCLUSION: On-line analysis of the
pupillogram using the suggested parameters can be performed easily to
produce a real-time assessment of an individual\'s state of alertness or
fatigue that correlates with his/her subjective assessment of this
state.

126\. Danker-Hopfe, H., *et al.*, *Time-of-day variations in different
measures of sleepiness (MSLT, pupillography, and SSS) and their
interrelations.* Psychophysiology, 2001. 38(5): p. 828-35. The aim of
the present study is to analyze how well physiological measures of
sleepiness derived from pupillography and the Multiple Sleep Latency
Test correlate with a subjective measure, the Stanford Sleepiness Scale
(SSS) score. The results are based on data from 12 healthy participants,
who underwent these tests every 2 hr from 7:00 a.m. until 11:00 p.m.
Sleep latencies were correlated with four different variables derived
from pupillography and the SSS score. The results indicate that the
physiologically based variables correspond very well. This is reflected
by similar patterns of time-of-day variations, a good agreement at the
group level, and correlations at the individual level, whereas the SSS
shows a quite different pattern of variation. The two physiological
measures of sleepiness seem to reflect the same aspect of the level of
tonic central nervous activation, which is not correlated with the
subjective feeling of sleepiness.

127\. Coelho, F. and J. Birks, *Physostigmine for Alzheimer\'s disease.*
Cochrane Database Syst Rev, 2001(2): p. CD001499. BACKGROUND: The main
pharmacological approach for the treatment of Alzheimer\'s disease (AD)
has been based on the use of agents potentiating cholinergic
transmission, particularly by inhibiting acetylcholinesterase (AChE),
the enzyme that destroys acetylcholine after it has been secreted into
the synaptic clefts. Physostigmine is an AChE inhibitor originally
extracted from calabar beans. It is licensed in many countries as an
agent for reversing the effect of drugs and poisons causing the
anticholinergic syndrome. Studies conducted more than 20 years ago
suggested that physostigmine could improve memory in people with or
without dementia. Investigation of this property has been limited by the
very short half-life of physostigmine. Various forms of administering
the drug have been tried to overcome this problem, most recently a
controlled-release (CR) oral formulation, and a skin patch. It has been
proposed as a potential drug for the symptomatic treatment of AD.
OBJECTIVES: To determine whether there is evidence of beneficial effects
for the use of physostigmine in Alzheimer\'s disease. To assess the
incidence and severity of adverse effects. SEARCH STRATEGY: The Cochrane
Controlled Trials Register was searched using the following terms:
\'physostigmine\', \'physostigmine salicylate\', \'Synapton\' and
\'Antilirium\' in accordance with the Cochrane Dementia and Cognitive
Improvement Group\'s search strategy. The pharmaceutical company was
contacted. SELECTION CRITERIA: All relevant unconfounded, double-blind,
randomized, placebo-controlled trials in which physostigmine was
administered for more than one day to patients with dementia of
Alzheimer type. Trials in which the allocation to the treatment was not
randomized, or in which the allocation to the treatment was not
concealed were excluded. DATA COLLECTION AND ANALYSIS: Data were
extracted independently by two reviewers (JMC & JB), pooled where
appropriate and possible, and the weighted or standardized mean
differences or Peto odds ratios (95% CI) were estimated. Where possible,
intention-to-treat analysis was used. MAIN RESULTS: Fifteen studies were
included using four different methods of administration of
physostigmine. Four studies, involving 29 people in total, used
intravenous infusion; seven, involving 131 people, used a conventional
oral form; four, involving 1456 participants, used a controlled-release
oral form, and one study of 181 people used a verum skin patch. There
are no usable results from the intravenous infusion trials, and the few
results from the conventional oral form showed no benefit of
physostigmine compared with placebo. The results from two of the four
studies of the controlled-release physostigmine apply only to a group of
patients identified as responders in a pre-randomization titration
period. The best dose physostigmine (mean 25mg/day) was associated with
a 1.75 point improvement on ADAS-Cog score (mean difference -1.75, 95%
confidence interval -2.90, -0.60 on an intention-to-treat basis) and a
0.26 point improvement on the CGIC score (treated as a continuous scale)
(mean difference -0.26, 95% confidence interval 0.06, 0.46 on an
intention-to-treat basis) compared with placebo at 6 weeks. There were
statistically significantly higher numbers of patients from the
physostigmine group withdrawing from the trial (22/183 vs 2/183)(OR
5.92, 95% confidence limits 2.59, 13.54) and suffering at least one
event of nausea, vomiting, diarhoea, anorexia, dizziness, stomach pain,
flatulence or sweating compared with placebo at 6 weeks. The best dose
physostigmine (mean 27mg/day) was associated with a 2.0 point
improvement on ADAS-Cog score (mean difference -2.02, 95% confidence
interval -3.59, -0.45 on an intention to treat basis) compared with
placebo at 12 weeks. There were statistically significantly higher
numbers of patients from the physostigmine group withdrawing from the
trial due to adverse events (13/83 vs 5/93)(OR 3.05, 95% confidence
limits 1.15, 8.07) and suffering at least one event of nausea, vomiting,
diarhoea, anorexia, dizziness, stomach pain, tremor, asthenia or
sweating compared with placebo at 12 weeks.

When no attempt was made to identify responders and all relevant
patients with Alzheimer\'s disease were randomized, fixed dose
physostigmine (mean 33 mg/day) was associated with a statistically
significantly higher number withdrawing (234/358 vs 31/117)(OR 4.82, 95%
confidence limits 3.17, 7.33), withdrawing due to adverse events
(196/358 vs 10/117) (OR 6.54, 95%confidence limits 4.29, 9.95) and
suffering at least one event of nausea, vomiting, diarhoea, anorexia,
dizziness, stomach pain, dyspepsia, sweating, asthenia, dyspnoea or
abnormal dreaming compared with placebo at 24 weeks. The results from
the study of the verum patch physostigmine show that the double dose
(delivering mean dose 12mg/day) was associated with statistically
significantly higher numbers suffering at least one adverse event of
vomiting, nausea or abdominal cramps compared with placebo at 24 weeks,
but placebo was associated with statistically significantly greater
numbers of gastrointestinal complaints at 24 weeks compared with
single-dose physostigmine. \[BG: Does not specify what the placebo was\]
REVIEWERS\' CONCLUSIONS: The evidence of effectiveness of physostigmine
for the symptomatic treatment of Alzheimer\'s disease is limited. Even
in a controlled release formulation designed to overcome the short
half-life, physostigmine showed no convincing benefit and adverse
effects remained common leading to a high rate of withdrawal.

128\. Thal, L.J., *et al.*, *A 24-week randomized trial of
controlled-release physostigmine in patients with Alzheimer\'s disease.*
Neurology, 1999. 52(6): p. 1146-52. OBJECTIVE: To evaluate the safety
and efficacy of controlled-release physostigmine, an
acetylcholinesterase inhibitor, in patients with probable AD of mild to
moderate severity. METHODS: A prospective, 24-week, randomized,
multicenter, double-blind, parallel group study of patients was
conducted. The study enrolled 475 patients at 24 sites. Patients met
criteria for probable AD and were randomized to one of three arms:
placebo, controlled-release (CR) physostigmine 30 mg daily, or CR
physostigmine 36 mg daily. Dosage was escalated by a forced upward
titration during the first 6 to 9 weeks of the trial, then maintained at
a constant dose to 24 weeks. Primary outcome measures were the
Alzheimer\'s Disease Assessment Scale-Cognitive subscale (ADAS-Cog) and
the Clinician\'s Interview-Based Impression of Change-Plus with
caregiver input (CIBIC+). Secondary outcome measures included the
Clinical Global Impression of Change (CGIC), the Geriatric Evaluation by
Relatives Rating Instrument, and an Instrumental Activities of Daily
Living Scale. RESULTS: In an intent-to-treat population, the last
observation carried forward analysis revealed a 2.9-point ADAS-Cog (p =
0.002) difference between physostigmine and placebo-treated patients for
both dosages, and a 0.26 to 0.31-point difference on the CIBIC+ (p =
0.048). There were no significant differences on the secondary outcome
measures except for a difference on the CGIC when analyzed by use of the
Cochran-Mantel-Haenszel statistic (p = 0.014). There were significant
increases in gastrointestinal side effects including nausea, vomiting,
diarrhea, anorexia, dyspepsia, and abdominal pain for patients on either
dose of physostigmine, resulting in a high dropout rate. Agitation was
decreased significantly. There was no evidence of cardiac rhythm
disturbance or liver function abnormalities. CONCLUSION: CR
physostigmine enhanced cognitive and global function. It is relatively
safe for the treatment of cognitive dysfunction secondary to AD.
However, in light of the gastrointestinal side effects, a lower starting
dose and a flexible titration schedule might lead to a more favorable
adverse event profile in the clinical arena.

129\. van Dyck, C.H., *et al.*, *Extended-release physostigmine in
Alzheimer disease: a multicenter, double-blind, 12-week study with dose
enrichment. Physostigmine Study Group.* Arch Gen Psychiatry, 2000.
57(2): p. 157-64. BACKGROUND: The efficacy of extended-release
physostigmine salicylate, an acetylcholinesterase inhibitor, was
evaluated in 850 subjects with mild-to-moderate Alzheimer disease (AD)
in a multicenter trial. METHODS: Subjects initially entered a
dose-enrichment phase in which they received 1 week each of
physostigmine salicylate, 24 mg/d and 30 mg/d, and daily placebo. Among
the subjects who completed this phase, 35.9% responded to physostigmine
treatment, whereas 62.4% were considered nonresponders, and 1.6% could
not be evaluated because of missing data. After a 4-week placebo-washout
phase, 176 responder subjects were randomized to receive their best dose
of physostigmine or placebo in a 12-week double-blind phase. Primary
efficacy measures included the cognitive subscale of the Alzheimer\'s
Disease Assessment Scale (ADAS-Cog), the Clinician\'s Interview-Based
Impression of Change With Caregiver Input (CIBIC+), and the Clinical
Global Impression of Change (CGIC). RESULTS: In the intent-to-treat
analysis of the double-blind phase, physostigmine-treated subjects
scored -2.02 points better than placebo-treated subjects on the ADAS-Cog
(F1,167 = 6.42 \[P = .01\]) and 0.33 points higher on the CIBIC+ (F1,150
= 5.68 \[P = .02\]). No significant improvement was observed on the CGIC
or the secondary outcome measures. Nausea and vomiting were experienced
by 47.0% of all physostigmine-treated subjects during the double-blind
phase. CONCLUSIONS: Physostigmine demonstrated a statistically
significant benefit compared with placebo on a clinical global rating of
change and an objective test of cognitive function. Given the frequency
of gastrointestinal side effects, the role of this agent in clinical use
remains to be determined.

130\. *Memory enhancement with oral physostigmine in Alzheimer\'s
disease.* N Engl J Med, 1983. 308(12): p. 720-1.

131\. Aigner, T.G. and M. Mishkin, *The effects of physostigmine and
scopolamine on recognition memory in monkeys.* Behav Neural Biol, 1986.
45(1): p. 81-7. The visual recognition of rhesus monkeys was evaluated
by means of a delayed nonmatching-to-sample task with trial-unique
objects. Each daily session consisted of two lists of 20 objects each,
which untreated animals were able to recognize at approximately 75%
accuracy. When they were performing at this level reliably, doses of
physostigmine (0.32, 1.0, 3.2, 10.0, 32.0, 56.0 micrograms/kg),
scopolamine (1.0, 3.2, 5.6, 10.0, 17.8, 32.0 micrograms/kg), or saline
were administered 20 min prior to the session. Physostigmine and
scopolamine produced dose-related increases and decreases, respectively,
in the number of objects correctly remembered. The systematic changes in
performance support the view that cholinergic mechanisms contribute to
recognition memory and suggest that tasks with trial-unique objects may
be particularly useful for studying the mnemonic effects of cholinergic
drugs.

132\. Aigner, T.G., *et al.*, *Effects of scopolamine and physostigmine
on recognition memory in monkeys with ibotenic-acid lesions of the
nucleus basalis of Meynert.* Psychopharmacology (Berl), 1987. 92(3): p.
292-300. Monkeys with bilateral ibotenic-acid lesions of the nucleus
basalis of Meynert, an area rich in cholinergic neurons that innervate
the cerebral cortex, were compared with unoperated control monkeys on a
recognition memory task. Although animals with large lesions had
substantial reductions of cortical choline acetyltransferase activity,
none showed impairment in the task. Lesion effects were observed,
however, when performance was assessed following administration of a
muscarinic receptor blocker (scopolamine) or a cholinesterase inhibitor
(physostigmine). Although scopolamine produced dose-related impairments
in both groups, this effect was greater in the experimental animals.
Conversely, whereas physostigmine produced modest improvement in
performance in the control group, no such improvement was observed in
the experimental animals. The altered sensitivity to the mnemonic
effects of cholinergic agents in the experimental group suggests that
the cholinergic neurons of the nucleus basalis of Meynert contribute to
recognition memory.

133\. Alagiakrishnan, K., W. Wong, and P.L. Blanchette, *Use of
donepezil in elderly patients with Alzheimer\'s disease\--a Hawaii based
study.* Hawaii Med J, 2000. 59(2): p. 57-9. Donepezil (Aricept) is a
reversible acetylcholinesterase inhibitor which is indicated for the
treatment of mild to moderate dementia of the Alzheimer\'s type. We did
a retrospective analysis of 41 elderly Alzheimer\'s subjects of
different ethnic groups including a large number of Asian and Hawaiian
patients. Donepezil appears to be clinically effective in patients of
different ethnicities with mild to moderate Alzheimer\'s disease, even
at advanced age.

134\. Almkvist, O., *et al.*, *Responder characteristics to a single
oral dose of cholinesterase inhibitor: a double-blind placebo-controlled
study with tacrine in Alzheimer patients.* Dement Geriatr Cogn Disord,
2001. 12(1): p. 22-32. A proportion of Alzheimer\'s disease (AD)
patients treated for several months with cholinesterase (ChE) inhibitors
have shown some favorable response on cognition, but the characteristics
of the responders are still unclear. This study attempts to identify the
characteristics of individuals with a positive behavioral response after
a double-blind randomized administration of a single oral dose of
tacrine (40 mg) and placebo to AD patients. Furthermore, the
relationship between single-dose and long-term responders are examined.
Twenty-four mildly to very mildly demented AD patients participated in
the study. They all fulfilled the diagnosis of probable AD according to
NINCDS-ADRDA criteria. Active treatment (tacrine 40 mg) and placebo was
administered in random order on 2 consecutive days, and the effects were
evaluated within 2 h using neuropsychological tests (assessing
visuospatial ability, episodic memory and attention), registration of
EEG activity and measurement of red blood cells (RBC)
acetylcholinesterase (AChE), ChE activity and concentrations of tacrine
and its metabolites in plasma. Results demonstrated significant
improvement, tacrine compared to placebo, in measures of attention, but
not in episodic memory or visuospatial ability. A single-dose response
was therefore defined in terms of improvement in attention. The tacrine
plasma concentration (pcTHA) showed a positively skewed distribution
(mean +/- SD: 10.5 +/- 11.8, range: 1.0-51.8 ng/ml). There were no
significant differences between single-dose responders compared to
nonresponders in pcTHA, metabolites of tacrine, inhibition of AChE in
RBC, tau levels in CSF, AChE activity in CSF or plasma and demographic
variables. However, single-dose responders showed a higher right frontal
alpha/theta ratio on EEG and had lower glucose metabolism in the
parietal-temporal association cortex at baseline. In addition, the
frequency of apolipoprotein E (APOE) epsilon 4 alleles was higher in
responders. Interestingly, the single-dose response was related to the
long-term response, although not significantly, which probably was due
to lack of power. To conclude, the present study identified single-dose
responders in terms of improved attentional performance associated with
a relatively higher alpha/theta activity in the right frontal regions of
the brain measured on EEG and predominance of APOE epsilon 4 allele.

135\. Arsland, D. and K. Laake, *\[Should Alzheimer disease be treated
with tacrine? Review of the literature\].* Tidsskr Nor Laegeforen, 1996.
116(23): p. 2791-4. The authors describe the rationale for use of the
acetylcholinesterase inhibitor, tacrine, in Alzheimer\'s disease and
critically review the controlled clinical trials using this drug. The
data shows that tacrine improves cognition and function, and that 20-40%
of the patients may benefit from such treatment. A large proportion of
the patients do not tolerate the highest dose, mainly because of an
increase in serum liver-enzymes, which is reversible and asymptomatic.
The authors recommend that tacrine should be considered for patients
with Alzheimer\'s disease of mild and moderate severity, and present
guidelines for clinical use.

136\. Arendt, T., M.M. Schugens, and V. Bigl, *The cholinergic system
and memory: amelioration of ethanol-induced memory deficiency by
physostigmine in rat.* Acta Neurobiol Exp (Warsz), 1990. 50(4-5): p.
251-61. Male Sprague-Dawley rats were treated with a solution of ethanol
(20% v/v) as the only source of fluid for 28 weeks while controls
received tap water. Profound reductions of acetylcholine content, the
activities of choline acetyltransferase and acetylcholinesterase and
choline uptake were observed in the cortex, hippocampus, substantia
innominata and striatum four weeks after withdrawal from ethanol. These
changes in cholinergic markers were accompanied by an impaired
performance in a spontaneous alternation task tested at long
acquisition-retention intervals. This ethanol induced behavioural
deficiency, which most likely represents a memory impairment, could be
reversed by application of physostigmine (0.45 g/kg; i.p) whereas
neostigmine did not show any behavioural effect. The results suggest
that ethanol-induced memory impairment can be ameliorated by
pharmacological manipulation of central cholinergic function.

137\. Ashford, J.W., *et al.*, *Physostigmine and its effect on six
patients with dementia.* Am J Psychiatry, 1981. 138(6): p. 829-30.

138\. Asthana, S., *et al.*, *Clinical pharmacokinetics of physostigmine
in patients with Alzheimer\'s disease.* Clin Pharmacol Ther, 1995.
58(3): p. 299-309. OBJECTIVE: To study the pharmacokinetic and
pharmacodynamic properties of physostigmine in subjects with
Alzheimer\'s disease. METHODS: Plasma physostigmine concentration and
butyrylcholinesterase inhibition were measured in blood samples
collected during and after a single high-dose (1 to 1.5 mg for 45 to 60
minutes) and a sustained low-dose steady-state intravenous infusion in
nine subjects with Alzheimer\'s disease. Escalating doses (0.5 to 25
mg/day) were administered during a 2-week period. A dose (2 to 12
mg/day) that optimized cognition in each subject was identified and then
administered in a randomized, double-blind, placebo-controlled crossover
design for 1 week. RESULTS: The elimination half-life of physostigmine
was 16.4 +/- 3.2 (SE) minutes. Clearance and volume of distribution were
7.7 +/- 0.9 (SE) L/min and 2.4 +/- 0.6 (SE) L/kg, respectively.
Butyrylcholinesterase inhibition half-life was 83.7 +/- 5.2 (SE)
minutes. During sustained steady-state infusion, plasma physostigmine
concentration (r = 0.95) and butyrylcholinesterase inhibition (r = 0.99)
were linearly correlated with the dose. In five cognitive responders,
the memory enhancement was significantly correlated (r = 0.86; p \<
0.05) with butyrylcholinesterase inhibition. CONCLUSIONS: These results
showed that, in cognitive responders, memory enhancement by
physostigmine in Alzheimer\'s disease is correlated directly to the
magnitude of plasma cholinesterase inhibition. Furthermore, during
single-dose conditions, the dynamic half-life is five-fold longer than
the kinetic half-life.

139\. Lacey, S.C., *et al.*, *Evaluation of working memory in Persian
Gulf War veterans with chronic fatigue \[abstract\].* Proceedings of the
Conference on Federally Sponsored Gulf War Veteransí Illnesses Research,
Pentagon City, Jun 1999, 133.134, 1999. thus, for instance, in a set of
PGW veterans with chronic fatigue, slower reaction times were found only
during a dual-task condition (p \< 0.007), suggesting to the authors
that there may be ìinefficient allocation of attentional resources
necessary to process simultaneous tasks adequately.

140\. Leclercq, M., *et al.*, *Dual task performance after severe
diffuse traumatic brain injury or vascular prefrontal damage.* J Clin
Exp Neuropsychol, 2000. 22(3): p. 339-50. The ability to perform two
tasks simultaneously is a key function of the central executive of
working memory (Baddeley, 1986). This study addressed dual-task
performance after diffuse very severe traumatic brain injury (TBI) (mean
coma duration = 21 days, mean post-traumatic amnesia = 70 days) or
prefrontal damage due to a ruptured aneurysm of the anterior
communicating artery (AACA). Mean time since injury was 8 and 16 months
in the TBI and the AACA group respectively. A simple visual reaction
time and random number generation were used as single and dual tasks.
Randomization was self-paced, to control for individual differences in
speed. Both patient groups had greater reaction time decrements than
controls under the dual-task condition, suggesting a divided attention
deficit. In addition, patients with AACA performed significantly poorer
in random generation. These results suggest that patients with AACA and
with severe TBI suffer from an impairment of the central executive
system.

141\. Overstreet, D.H., *et al.*, *Potential animal model of multiple
chemical sensitivity with cholinergic supersensitivity.* Toxicology,
1996. 111(1-3): p. 119-34. Multiple Chemical Sensitivity (MCS) is a
clinical phenomenon in which individuals, after acute or intermittent
exposure to one or more chemicals, commonly organophosphate pesticides
(OPs), become overly sensitive to a wide variety of chemically-unrelated
compounds, which can include ethanol, caffeine and other psychotropic
drugs. The Flinders Sensitive Line (FSL) rats were selectively bred to
be more sensitive to the OP diisopropylfluorophosphate (DFP) compared to
their control counterparts, the Flinders Resistant Line (FRL) rats. The
present paper will summarize evidence which indicates that the FSL rats
exhibit certain similarities to individuals with MCS. In addition to
their greater sensitivity to DFP, the FSL rats are more sensitive to
nicotine and the muscarinic agonists arecoline and oxotremorine,
suggesting that the number of cholinergic receptors may be increased, a
conclusion now supported by biochemical evidence. The FSL rats have also
been found to exhibit enhanced responses to a variety of other drugs,
including the serotonin agonists m-chlorophenylpiperazine and 8-OH-DPAT,
the dopamine antagonist raclopride, the benzodiazepine diazepam, and
ethanol. MCS patients report enhanced responses to many of these drugs,
indicating some parallels between FSL rats and MCS patients. The FSL
rats also exhibit reduced activity and appetite and increased REM sleep
relative to their FRL controls. Because these behavioral features and
the enhanced cholinergic responses are also observed in human
depressives, the FSL rats have been proposed as a genetic animal model
of depression. It has also been reported that MCS patients have a
greater incidence of depression, both before and after onset of their
chemical sensitivities, so cholinergic supersensitivity may be a state
predisposing individuals to depressive disorders and/or MCS. Further
exploration of the commonalities and differences between MCS patients,
human depressives, and FSL rats will help to elucidate the mechanisms
underlying MCS and could lead to diagnostic approaches and treatments
beneficial to MCS patients.

142\. Servatius, R.J., *et al.*, *Persistently exaggerated startle
responses in rats treated with pyridostigmine bromide.* J Pharmacol Exp
Ther, 1998. 287(3): p. 1020-8. Troops in the Persian Gulf War have
registered complaints consistent with CNS dysfunction that emerged after
returning from the Gulf. A common experience among Persian Gulf War
veterans was exposure to pyridostigmine bromide (PB) for prophylaxis
against nerve gas exposure. To determine whether PB causes emergent CNS
dysfunction, Wistar-Kyoto (WKY) and Sprague-Dawley (SD) rats were given
PB for 7 consecutive days in their drinking water. The WKY, but not the
SD, rats exhibited a delayed-onset, persistently exaggerated startle
response. The WKY rats exhibited exaggerated startle responses that
appeared 15 days after the end of PB treatment and were still evident 22
days after the end of treatment. Both the duration and the magnitude of
the exaggerated startle responses were related to the dosage of PB. The
PB-treated rats exhibited normal short-term and long-term habituation.
However, exaggerated startle responses were related to the development
of enhanced short-term sensitization. Treating the rats for a second
time, 7 weeks after the end of the first PB treatment, induced an
exaggerated startle response that appeared sooner and dissipated faster
than was evident after the first PB treatment. Inasmuch as the WKY rat
has inherently low butyrylcholinesterase activity, a scavenger for PB,
these results suggest that prophylactic PB may influence CNS function in
individuals with low butyrylcholinesterase activity. Elaboration of the
factors that mediate enhanced sensitization in the WKY rat may provide
insight into some of the complaints registered by veterans of the
Persian Gulf War.

143\. Janowsky, D.S., *et al.*, *Parasympathetic suppression of manic
symptoms by physostigmine.* Arch Gen Psychiatry, 1973. 28(4): p. 542-7.

144\. Janowsky, D., *et al.*, *Increased vulnerability to cholinergic
stimulation in affective disorder patients.* Psychopharmacol Bull, 1980.
16: p. 29-31.

145\. Janowsky, D.S., *et al.*, *Cholinergic supersensitivity in affect
disorder patients: Behavioral and neuroendocrine observations.*
Psychopharmacol Bull, 1981. 17: p. 129-132.

146\. Janowsky, D., C. Risch, and J. Gillin, *Adrenergic-cholinergic
balance and the treatment of affective disorders.* Prog
Neuropsychopharmacol Biol Psychiatry, 1983. 7: p. 297-307.

147\. Janowsky, D. and C. Risch, *Cholinomimetic and anti cholinergic
drugs used to investigate an acetylcholine hypothesis of affective
disorders adn stress.* Drug Dev Res, 1984.

148\. Bannon, A.W., K.L. Gunther, and M.W. Decker, *Is epibatidine
really analgesic? Dissociation of the activity, temperature, and
analgesic effects of (+/-)-epibatidine.* Pharmacol Biochem Behav, 1995.
51(4): p. 693-698. The experiments in the present study were designed to
determine if

the activity, temperature, and analgesic effects of (+/-)-epibatidine

treatment could be dissociated. Initially (i.e., 15 min)
(+/-)-epibatidine

treatment (0.1 mumol/kg = 28 micrograms/kg, IP) impaired rotorod

performance, decreased activity, decreased temperature, and increased
jump

latency (e.g., analgesic effect). For the remaining time points measured

(i.e., 30, 60, and 120 min), activity and temperature remained

significantly reduced. In contrast, by 120 min (+/-)-epibatidine\'s
effects

on rotorod performance and analgesia (jump latency) were not observed.
When

administered after (+/-)-epibatidine (0.05 mumol/kg, IP), mecamylamine

treatment (5 mumol/kg = 1 mg/kg, IP) produced a potentiation of
analgesia.

This potentiation effect was not observed on activity and temperature

measures. The effect of (+/-)-epibatidine treatment (0.1 mumol/kg, IP)
was

also determined in mice with central nicotinic receptor blockade induced
by

treatment with chlorisondamine (23 mumol/kg = 10 mg/kg, IP). An

(+/-)-epibatidine-induced reduction in activity was not attenuated in

chlorisondamine-treated mice and only a minimal effect was observed on

(+/-)-epibatidine-induced hypothermia in chlorisondamine-treated mice.
In

contrast, in chlorisondamine-treated mice (+/-)-epibatidine\'s analgesic

effect was attenuated. Taken together, these data suggest that various

centrally mediated effects of (+/-)-epibatidine can be dissociated.

149\. Avlonitou, E. and R. Elizondo, *Effects of atropine and
pyridostigmine in heat-stressed patas monkeys.* Aviat Space Environ Med,
1988. 59(6): p. 544-8. The effects of a single intramuscular atropine
injection (0.03 mg.kg-1) and a chronic oral pyridostigmine treatment
(0.4 mg.kg-1, 3 times/day over a period of 7 d) on the thermoregulatory
effector responses of unanesthetized patas monkeys were investigated
using indirect calorimetry. The effects of atropine treatment on the
thermoregulatory effector responses of patas monkeys exposed to 25
degrees and 35 degrees C were qualitatively similar but quantitatively
greater at 35 degrees C. At 35 degrees C atropine decreased sweating
(Esw) 52%, increased rectal temperature (Tre), mean skin temperature
(Tsk), metabolic rate (MR), and whole body conductance (K), and elicited
a consistent 11% increase in heart rate (HR). Daily oral pyridostigmine
treatment to patas monkeys produced a significant 25-30% drop in serum
cholinesterase activity with no chronic effects on thermoregulatory or
cardiovascular functions. The acute effects of oral pyridostigmine
treatment in this species included transient 12% and 15% decreases in MR
and HR, respectively, and a transient 25% increase in Esw. The latter
was associated with significant acute reductions in Tre and Tsk which
lasted at least 120 min following pyridostigmine administration. It is
concluded that the patas monkey is an excellent animal model for studies
to evaluate the effects of neuroactive agents on thermoregulatory and
other physiological functions which are difficult, if not impossible, to
perform on humans.

150\. Barar, F.S. and B.R. Madan, *Tremorine-oxotremorine-induced
tremor, hypothermia and analgesia, and physostigmine toxicity, in mice
after pretreatment with beta-adrenoceptor antagonists.* J Pharm
Pharmacol, 1976. 28(4): p. 286-9. The beta-adrenoceptor blockers
propranolol, PhQA33 and LB-46 exhibited appreciable activity against
tremorine-(TMN) and oxotremorine-(OTMN) induced tremor, whereas
pronethalol, (+)-H56/28, (-)-H56/28, Ko-592 and L(+)-INPEA possessed
weak action. The two beta-blockers, namely D,L(+/-)-INPEA and D(-)-INPEA
acted as weak tremorgens. None of the above compounds suppressed the
induced peripheral cholinergic phenomena; or possessed any central
anticholinergic activity, as they were unable to afford protection
against physostigmine-induced death. Propranolol, PhQA33 and LB-46
antagonized TMN-induced hypothermia and analgesia, but were inactive
against OTMN-induced changes. A correlation of the beta-blocking and
anti-tremor activity of these agents is unlikely.

151\. Clement, J.G., *Variability of sarin-induced hypothermia in mice:
investigation into incidence and mechanism.* Biochem Pharmacol, 1991.
42(6): p. 1316-8.

152\. Francesconi, R., *et al.*, *Oral pyridostigmine administration in
rats: effects on thermoregulation, clinical chemistry, and performance
in the heat.* Pharmacol Biochem Behav, 1986. 25(5): p. 1071-5. We have
recently reported that acute intraperitoneal administration of
pyridostigmine bromide to rats resulted in significant decrements in
physical performance in the heat, adverse thermoregulatory effects, and
exacerbated elevations in several indices of heat/exercise injury. Since
it will be consumed orally as a prophylaxis for organophosphate
poisoning, pyridostigmine was dissolved in the drinking water of rats.
Consumption of pyridostigmine for 7 days (n = 34, 6.6 mg/day) resulted
in a 23% (p less than 0.001) reduction of circulating cholinesterase
when compared with a control group (n = 31) while ingestion for 14 days
(n = 35, 8.9 mg/day) elicited a 39% (p less than 0.001) inhibition of
circulating cholinesterase when compared to a second control group (n =
33). Water and food consumption, rate of weight gain, and overt behavior
were unaffected by pyridostigmine consumption. When approximately half
the animals in each group were exercised (9.14 m/min) in the heat (35
degrees C) to hyperthermic exhaustion (Tre = 42.5-43 degrees C, rats
unable to right themselves), pyridostigmine consumption for 14 days
effected a significantly (p less than 0.05) increased rate of weight
loss, but no further effects on thermoregulation or performance were
noted. Several minor increments were observed in clinical indices of
heat/exercise injury in rats consuming pyridostigmine for 14 days. These
data indicate that oral dosages of pyridostigmine can probably be
titrated to levels of cholinesterase inhibition which are efficacious in
prophylaxis against organophosphate toxicity without significant effects
on selected physiologic and metabolic processes.

153\. Gordon, C.J., *24-hour control of body temperature in the rat: II.
Diisopropyl fluorophosphate-induced hypothermia and hyperthermia.*
Pharmacol Biochem Behav, 1994. 49: p. 747-754.

154\. Epstein, Y., *et al.*, *Heat-exercise performance of
pyridostigmine-treated subjects wearing chemical protective clothing.*
Aviat Space Environ Med, 1990. 61(4): p. 310-3. Pyridostigmine bromide
is currently the pretreatment of choice for operation in a chemical
warfare (CW) environment. Under CW conditions, subjects are exposed to
thermal stress caused by CW protective clothing. This investigation was
conducted to determine if pyridostigmine affects various physiological
and biophysical parameters of human temperature regulation in subjects
wearing CW protective clothing. Pyridostigmine was administered orally
in a randomized double-blind cross-over study in four doses of 30 mg
every 8 h. An average of 33% whole blood cholinesterase inhibition was
induced in the pyridostigmine treated group 4 h after ingestion of last
tablet. The subjects were exposed to 170 min exercise-heat stress (Tdb =
33 degrees C; rh = 60%) consisting of 60 min in a sitting position and
two 50-min walks (1.39 m.s-1, 5% grade) separated by 10 min of rest.
Non-evaporative heat exchange was significantly higher, -14.0 and -10.6
W.m-2 (p less than 0.03), for the pyridostigmine-treated subjects. No
additional differences were found between treatments in the
physiological responses and heat balance parameters at the end of
exposure: heart rate (HR) was (mean +/- S.D.) 154 +/- 16 and 151 +/- 24
bpm, rectal temperature (Tre) was 39.0 +/- 0.4 and 38.9 +/- 0.2 degrees
C, heat storage over the 2 h of exercise was 62 +/- 15 and 70 +/- 15
W.m-2, and sweat rate was 832 +/- 185 and 748 +/- 52 g.h-1, in the
pyridostigmine and placebo treatments, respectively.(ABSTRACT TRUNCATED
AT 250 WORDS).

155\. Epstein, Y., *et al.*, *Effect of pyridostigmine on the
exercise-heat response of man.* Eur J Appl Physiol Occup Physiol, 1990.
61(1-2): p. 128-32. The effect of pyridostigmine on thermoregulatory
responses was evaluated during exercise and heat stress. Eight heat
acclimated, young adult male subjects received four doses of
pyridostigmine (30 mg) or identical placebo tablets every 8 h, in a
double blind, randomized, cross-over trial. A 30.3%, SD 4.6% inhibition
of the circulating cholinesterase (ChE) activity was induced in the
pyridostigmine-treated group. The subjects were exposed to 170-min
exercise and heat-stress (dry bulb temperature, 33 degrees C; relative
humidity 60%) consisting of 60 min in a sitting position and two bouts
of 50-min walking (1.39 m.s-1, 5% gradient) which were separated by
10-min rest periods. No differences were found between treatments in the
physiological responses and heat balance parameters at the end of
exposure: heart rate (fc) was 141 beats.min-1, SD 16 and 150
beats.min-1, SD 12, rectal temperature (Tre) was 38.5 degrees C, SD 0.4
degrees and 38.6 degrees C, SD 0.3 degrees, heat storage was 60 W.m-2,
SD 16 and 59 W.m-2, SD 15 and sweat rate was 678 g.h-1, SD 184 and 661
g.h-1, SD 133, in the pyridostigmine and placebo treatments,
respectively. The changes in Tre and fc over the heat-exercise period
were parallel in both study and control groups. Pyridostigmine caused a
slight slowing of fc (5 beats.min-1) which was consistent throughout the
entire exposure (P less than 0.001) but was of no clinical significance.
The overall change in fc was similar for both groups.(ABSTRACT TRUNCATED
AT 250 WORDS).

156\. Kassa, J., *et al.*, *The long-term influence of low-level sarin
exposure on behavioral and neurophysiological functions in rats.* Acta
Medica (Hradec Kralove), 2001. 44(1): p. 21-7. 1. Long term effects of
low doses of highly toxic organophosphorus agent sarin on behavioral and
neurophysiological functions were studied in rats exposed to sarin by
inhalation. The toxic effects of sarin were monitored using a functional
observational battery (FOB), an automatic measurement of motor activity
and a test of excitability of central nervous system at 3, 6 and 12
months following sarin exposure. 2. The results indicate that sarin at
symptomatic as well as asymptomatic doses (level 2 and 3) is able to
induce some neurotoxic effects (a decrease in activity and mobility, an
alteration of gait, an increase in stereotyped behavior) including an
increase in the excitability of central nervous system (an increase in
convulsive activity following the administration of
pentamethylenetetrazole) in rats at 3 months following inhalation
exposure. Some sings of increased excitability were also observed in
sarin-exposed rats following 6 or 12 months (an increase in exploratory
activity, body temperature and a hindlimb grip strength at 6 months
following exposure to sarin at asymptomatic doses, an increase in
tail-pinch response at 12 months following exposure to sarin at
symptomatic doses). 3. Therefore, nerve agents such as sarin seem to be
harmful not only at high, clinically symptomatic doses but also at low,
clinically asymptomatic doses because of long term manifestation of
alteration of neurophysiological functions in sarin-exposed rats without
disruption of cholinergic nervous system.

157\. Kolka, M.A. and L.A. Stephenson, *Human temperature regulation
during exercise after oral pyridostigmine administration.* Aviat Space
Environ Med, 1990. 61(3): p. 220-4. Four healthy males exercised in two
experiments at ambient temperatures of 22, 29, and 36 degrees C with the
relative humidity at 30% in all environments (Tdp = 3.9, 9.9, and 15.8
degrees C). One experiment in each environment was done 150 min after 30
mg oral pyridostigmine bromide (PYR) administration, and the second
experiment was done on a separate day with no medication (CON). Red
blood cell cholinesterase was 39 +/- 7% lower after PYR (11.8 vs 7.2
micromol.ml-1.min-1). Esophageal (Tes) and mean skin temperature (Tsk),
forearm blood flow (FBF), forearm sweating, and skin blood flow (SkBF)
were measured twice each minute during a 15-min rest period and during
30-min of seated cycle exercise at approximately 58% Vo2peak. Whole body
sweating was determined from weight changes before and after exercise.
PYR decreased heart rate at rest and during exercise at 29 degrees C and
36 degrees C (8bpm, p less than 0.05). Resting SkBF was 40% lower at 29
degrees C and 30% lower at 36 degrees C after PYR compared to CON (p
less than 0.05). There was no effect of PYR on heat production at rest
or during exercise. Tsk was different in the three conditions by design,
but was unchanged by PYR. Tes was not different at rest in any
condition, but was elevated during exercise at 36 degrees C (0.1 degree
C, p less than 0.05) in PYR compared to CON. These data suggest that
pyridostigmine ingestion decreased skin blood flow, which may limit
exercise thermoregulation in more severe environments.

158\. Lin, M.T., H.C. Wang, and A. Chandra, *The effects on
thermoregulation of intracerebroventricular injections of acetylcholine,
pilocarpine, physostigmine, atropine and hemicholinium in the rat.*
Neuropharmacology, 1980. 19(6): p. 561-5.

159\. Matthew, C.B., R.W. Hubbard, and R.P. Francesconi, *Atropine,
diazepam, and physostigmine: thermoregulatory effects in the
heat-stressed rat.* Life Sci, 1989. 44(25): p. 1921-7. We have
previously reported that administration of atropine (A) to unrestrained,
sedentary, heat-stressed rats resulted in a dose-dependent increase in
heating rate (rate of rise of core temperature, degree C/min).
Additionally, we have demonstrated that the decrements in treadmill
endurance and increments in heating rate of physostigmine (PH)-treated
running rats can both be restored to control levels by pretreating the
animals with A and diazepam (D). Our objective in the present work was
to determine if the administration of D + PH to A-treated unrestrained,
sedentary, heat-stressed rats (N = 16/group, 510-530 g) could improve
their thermal tolerance. The following drugs were administered singly
(at 10 min intervals) via lateral tail vein: vehicle-control (C), A (200
micrograms/kg), D (500 micrograms/kg), and PH (200 micrograms/kg). After
drug administration, the rats were heat-stressed (Tamb = 41.5 degrees C)
until a core temperature of 42.6 degrees C was attained when they were
removed to a 26 degrees C chamber. The heating rates (degrees C/min) and
tolerance times (min) of the respective groups were: C- 0.02, 235; A-
0.08, 58; A D- 0.06, 94; and A + D + PH- 0.04, 143. Administration of D
with A significantly decreased heating rate, and D + PH more than
doubled the thermal tolerance of A-treated rats. Thus, the combination
of A + D + PH not only restores PH-induced performance and
thermoregulatory decrements of rats exercised in a moderate environment,
but also reduces A-induced heat intolerance.

160\. Matthew, C.B., R.P. Francesconi, and R.W. Hubbard, *Physostigmine:
dose-response effects on endurance and thermoregulation during
exercise.* Life Sci, 1992. 50(1): p. 39-44. We previously reported that
the administration of 200 micrograms/kg of physostigmine (PH) to rats
exercising on a treadmill resulted in decrements in both endurance
(decreased running time to exhaustion) and thermoregulation. However, it
was necessary to determine the dose-response effects of PH
administration before PH-treated exercising rats could be used as a
model with which to examine the relative anticholinergic potency of
drugs. In the present work saline, 50, 100, or 200 micrograms/kg of
physostigmine salicylate (0%, 40%, 50%, and 60% whole blood
cholinesterase inhibition) was administered to rats (N = 12/group) prior
to treadmill exercise (26 degrees C, 50% rh, 11 m/min, 6 degrees
incline). The saline control group ran for 67 +/- 6 min (mean +/- SE)
with a rate of rise of core temperature of 0.051 +/- 0.007 degrees
C/min. The run times declined (80%, 64% and 48% of control) as rate of
rise of core temperature increased (116%, 180%, and 214% of control) in
a dose-dependent manner (50, 100, 200 micrograms/kg PH). Cholinergic
symptoms such as salivation, tremors, and defecation were also affected
in a dose-dependent manner by PH administration. Since cholinergic
symptoms, thermoregulatory effects, and endurance decrements all vary in
a dose-dependent manner with physostigmine administration, the
exercising rat represents a useful model for examining the relative
potency of cholinergic therapies.

161\. Maickel, R.P., *et al.*, *Antagonism of physostigmine induced
hypothermia and neuroendocrine changes following exposure to different
environmental temperatures.* Prog Neuropsychopharmacol Biol Psychiatry,
1991. 15(6): p. 873-84. 1. The magnitude of physostigmine-induced
hypothermia increased with decreasing environmental temperature. 2. The
hypothermic response was accompanied by significant changes in plasma
levels of corticosterone, glucose and fatty acids. 3. Central
cholinergic mediation appears to be a significant component of
physostigmine-induced hypothermia and neuroendocrine changes at moderate
temperature. 4. At lower ambient temperatures cholinergic blockers
produced less antagonism of physostigmine-induced effects. 5. The
decreased effectiveness of cholinergic blockers at low environmental
temperatures and the increased plasma fatty acid levels under almost all
conditions studied may be of importance in considering long term therapy
with cholinergic agonists.

162\. Monda, M., A. Viggiano, and V. De Luca, *Administration of
muscimol into the posterior hypothalamus reduces hyperthermia induced by
hippocampal neostigmine injection.* Brain Res, 2000. 887(2): p. 344-9.
The firing rate of the sympathetic nerves innervating interscapular
brown adipose tissue (IBAT), IBAT and colonic temperatures (T(IBAT) and
T(C)) and oxygen (O(2)) consumption were monitored in
urethane-anesthetized male Sprague-Dawley rats. These variables were
measured for 40 min before (baseline values) and 40 min after an
injection of neostigmine (5 x 10(-7) mol in 1 microl of saline) into the
hippocampus and a bilateral administration of a GABA(a)-agonist,
muscimol (28 ng in 0.5 microl of saline, per side) into the posterior
hypothalamus. The same variables were recorded in other rats, but the
muscimol was replaced by saline. Control animals were used with muscimol
or saline alone. The results show an increase of sympathetic firing
rate, T(IBAT), T(C) and O(2) consumption after neostigmine injection.
Muscimol significantly reduces this enhancement. The findings suggest
that hippocampus controls the sympathetic and thermogenic activation
induced by neostigmine through an influence on GABAergic tone of the
posterior hypothalamus.

163\. Prusaczyk, W.K. and M.N. Sawka, *Effects of pyridostigmine bromide
on human thermoregulation during cold water immersion.* J Appl Physiol,
1991. 71(2): p. 432-7. This study examined the effects of an oral 30-mg
dose of pyridostigmine bromide (PYR) on thermoregulatory and
physiological responses of men undergoing cold stress. Six men were
immersed in cold water (20 degrees C) for up to 180 min on two
occasions, once each 2 h after ingestion of PYR and 2 h after ingestion
of a placebo. With PRY, erythrocyte cholinesterase inhibition was 33 +/-
12% (SD) 110 min postingestion (10 min preimmersion) and 30 +/- 7% at
termination of exposure (mean 117 min). Percent cholinesterase
inhibition was significantly related to lean body mass (r = -0.91, P
less than 0.01). Abdominal discomfort caused termination in three of six
PYR experiments but in none of the control experiments (mean exposure
time 142 min). During immersion, metabolic rate, ventilatory volume, and
respiratory rate increased significantly (P less than 0.05) over
preimmersion levels and metabolic rate increased with duration of
immersion (P less than 0.01) in both treatment but did not differ
between conditions. PYR had no significant effect on rectal temperature,
mean body temperature, thermal sensations, heart rate, plasma cortisol,
or change in plasma volume. It was concluded that a 30-mg dose of PYR
does not increase an individual\'s susceptibility to hypothermia during
cold water immersion; however, in combination with cold stress, PYR may
result in marked abdominal cramping and limit cold tolerance.

164\. Roberts, D.E., *et al.*, *Pyridostigmine bromide does not alter
thermoregulation during exercise in cold air.* Can J Physiol Pharmacol,
1994. 72(7): p. 788-93. This study examined the effects of acute and
chronic pyridostigmine bromide (PB) administration on thermoregulatory
and metabolic responses to exercise in cold air (5 degrees C). Seven
healthy men completed two 7-day trials in a double-blind, crossover
experimental design: during one trial they received PB (30 mg three
times daily) and during the other trial they received placebo. For each
trial, subjects attempted four (3 h) exercise tests: low-intensity
exercise (approximately 25% VO2max) and moderate-intensity exercise
(approximately 50% VO2max), on days 2 and 3 and again on days 6 and 7.
Metabolic rate, body temperatures, and venous blood samples were
obtained before and during exercise. Red blood cell acetylcholinesterase
inhibition induced by PB increased (p \< 0.05) from 34% on day 1 to 43%
on days 3-7. Metabolic rate, body temperatures, and regional heat
conductance responses were not different between trials. Plasma glucose,
glycerol, free fatty acid, lactate, sodium, and potassium concentrations
were not different between trials. In addition, differences were not
found between acute and chronic experiments for any thermoregulatory or
metabolic responses. These findings demonstrate that the PB dosage used
by military personnel, as a pharmacological defense against nerve-agent
poisoning, should not cause any adverse thermoregulatory or metabolic
effects during moderate activity in cold climates.

165\. Sharkawi, M., *Morphine hyperthermia in the rat: its attenuation
by physostigmine.* Br J Pharmacol, 1972. 44(3): p. 544-8.

166\. Stephenson, L.A. and M.A. Kolka, *Acetylcholinesterase inhibitor,
pyridostigmine bromide, reduces skin blood flow in humans.* Am J
Physiol, 1990. 258(4 Pt 2): p. R951-7. Five subjects exercised on a
cycle ergometer for 30 min at 55% peak oxygen consumption on two
occasions in an environmental test chamber (ambient temperature = 29
degrees C; dew point temperature = 10 degrees C). Pyridostigmine bromide
(PYR), an acetylcholinesterase (AChE) inhibitor, was ingested (30 mg)
approximately 150 min before one experiment, and no drug was
administered during the other experiment (control). Red blood cell AChE
inhibition averaged 40 (+/- 7)% during PYR treatment. Esophageal
temperature (Tes), an eight site-derived mean skin temperature, forearm
blood flow (FBF; venous occlusion plethysmography), skin blood flow
(SkBF; laser-Doppler velocimetry), and metabolic rate (indirect
calorimetry) were measured. SkBF decreased 37% after PYR treatment
compared with control (P less than or equal to 0.05). The Tes threshold
for initiation of cutaneous vasodilation was 36.8 (+/- 0.3) degrees C
for the control treatment and 37.0 (+/- 0.3) degrees C for the PYR
treatment (P less than or equal to 0.01). FBF was not significantly
different between treatments, whereas heart rate was reduced by 7 and 9
beats/min during rest and exercise, respectively (P less than or equal
to 0.01). The increased threshold for initiation of cutaneous
vasodilation with AChE inhibition by PYR is compatible with nonthermal
modulation of the control of thermoregulation through increased
acetylcholine (ACh) accumulation. This could potentiate preganglionic
transmission to enhance adrenergic vasoconstrictor tone. One suggested
mechanism possible at the neuroeffector junction of the sweat gland may
be that accumulated ACh diffusion across the adventitia of adjacent
arterioles to muscarinic receptors initiates contraction of the smooth
muscle.(ABSTRACT TRUNCATED AT 250 WORDS).

167\. Wenger, C.B. and W.A. Latzka, *Effects of pyridostigmine bromide
on physiological responses to heat, exercise, and hypohydration.* Aviat
Space Environ Med, 1992. 63(1): p. 37-45. Five men underwent eight heat
stress tests (HSTs) at 35 degrees C, each consisting of four 25-min
treadmill walks (35% Vo2max), separated by 5-min rests, in four
conditions: 1) 20% relative humidity (rh), subjects euhydrated and
drinking ad libitum; 2) 20% rh, euhydrated; 3) 75% rh, euhydrated; and
4) 20% rh, hypohydrated 3% of body weight. In Conditions 2-4 subjects
drank during the walks to maintain their pre-HST weight. In each
condition we tested subjects once after 30 mg pyridostigmine bromide
(PB) by mouth and once after placebo. PB lowered heart rate a mean of 3
beats/min overall, most with hypohydration. PB did not significantly
affect rectal temperature (Tre), but reduced the rise in Tre during
hypohydrated exercise. In Condition 2, chest skin temperature decreased
more during exercise with PB. PB had no significant effect on other skin
temperatures, sweating, hematocrit, hemoglobin, total plasma protein,
osmolality, ad libitum drinking, rate of O2 uptake, or subject ratings
of temperature, discomfort, or exertion. PB thus had little effect on
physiological responses to moderate exercise-heat stress.

168\. Wenger, B., M.D. Quigley, and M.A. Kolka, *Seven-day
pyridostigmine administration and thermoregulation during rest and
exercise in dry heat.* Aviat Space Environ Med, 1993. 64(10): p. 905-11.
Seven men participated in a double-blind study of effects of
multiple-dose oral pyridostigmine bromide (PB) on physiological
responses to 4-h heat stress tests (HST\'s) in a hot dry environment, 42
degrees C, 20% relative humidity. Subjects underwent 2 7-d series of
tests, separated by 72 h, taking 30 mg PB every 8 h in one series, and
placebo in the other. Each HST began right after the 0800 dose of PB or
placebo. Subjects drank ad libitum during each HST, and performed two
55-min treadmill walks at about 40% VO2max during the last 2 h.
Inhibition of red cell cholinesterase at the start of exercise averaged
30.0% in subjects taking PB, and did not differ significantly among
HST\'s with PB. PB increased sweating and evaporative water loss by
about 4%, and lowered chest skin temperature during exercise by 0.7
degrees C; but it had no significant effect on rectal temperature, other
skin temperatures, O2 uptake, or fluid balance. PB alone had no
significant effect on heart rate (HR), but had a significant interaction
with day: although PB had essentially no effect on HR in the 1st HST,
its effect increased progressively so that HR during exercise in the 4th
HST was 8 beats.min-1 lower with PB. Multiple-dose PB had only slight
effects on responses to moderate exercise-heat stress beyond those
described after single-dose PB, and we found no adverse effects of
multiple-dose PB administration.

169\. Billet, S., N.B. Cant, and W.C. Hall, *Cholinergic projections to
the visual thalamus and superior colliculus.* Brain Res, 1999. 847(1):
p. 121-3. The parabrachial region of the brainstem reticular formation
projects to the dorsal lateral geniculate nucleus of the thalamus and to
the intermediate gray layer of the superior colliculus. We used the
retrograde axonal transport of two fluorescent labels to demonstrate
that individual parabrachial cells project to both structures. The
results suggest that cholinergic cells of the parabrachial region may
coordinate the relay of visuosensory information to the cortex with the
onset of orienting movements.

170\. Lee, P.H., M. Schmidt, and W.C. Hall, *Excitatory and inhibitory
circuitry in the superficial gray layer of the superior colliculus.* J
Neurosci, 2001. 21(20): p. 8145-53. Stratum griseum superficiale (SGS)
of the superior colliculus receives a dense cholinergic input from the
parabigeminal nucleus. In this study, we examined in vitro the
modulatory influence of acetylcholine (ACh) on the responses of SGS
neurons that project to the visual thalamus in the rat. We used
whole-cell patch-clamp recording to measure the responses of these
projection neurons to electrical stimulation of their afferents in the
stratum opticum (SO) before and during local pressure injections of ACh.
These colliculothalamic projection neurons (CTNs) were identified during
the in vitro experiments by prelabeling them from the thalamus with the
retrograde axonal tracer wheat germ agglutinin-apo-HRP-gold. In a group
of cells that included the prelabeled neurons, EPSCs evoked by SO
stimulation were significantly reduced by the application of ACh,
whereas IPSC amplitudes were significantly enhanced. Similar effects
were observed when the nicotinic ACh receptor agonist lobeline was used.
Application of the selective GABA(B) receptor antagonist
3-\[\[(3,4-dichlorophenyl)-methyl\]amino\]propyl\](diethoxymethyl)phosphinic
acid blocked ACh-induced reduction in the evoked response. In contrast,
the ACh-induced reduction was insensitive to application of the GABA(A)
receptor antagonist bicuculline. The ACh-induced reduction was also
diminished by bath application of muscimol at the low concentrations
that selectively activate GABA(C) receptors. Because GABA(C) receptors
may be specifically expressed by GABAergic SGS interneurons (Schmidt et
al., 2001), our results support the hypothesis that ACh reduces CTN
activity by nicotinic receptor-mediated excitation of local GABAergic
interneurons. These interneurons in turn use GABA(B) receptors to
inhibit the CTNs.

171\. Aizawa, H., *et al.*, *Injection of nicotine into the superior
colliculus facilitates occurrence of express saccades in monkeys.* J
Neurophysiol, 1999. 82(3): p. 1642-6. To clarify the role of cholinergic
inputs to the intermediate layer of the superior colliculus (SC), we
examined the effect of microinjection of nicotine into the SC on
visually guided saccades in macaque monkeys. After injection of 0.4-2
microl of 1-100 mM nicotine into the SC, frequency of extremely short
latency saccades (express saccades; reaction time = 70-120 ms)
dramatically increased, for the saccades the direction and amplitude of
which were represented at the location of the injection site on the
collicular map. However, no marked change was observed for the
relationship between the peak velocities and the amplitudes of saccades.
These results suggested that activation of nicotinic acetylcholine
receptors in the SC can facilitate initiation but causes no major change
in dynamics of visually guided saccades.

172\. Binns, K.E., *The synaptic pharmacology underlying sensory
processing in the superior colliculus.* Prog Neurobiol, 1999. 59(2): p.
129-59. The superior colliculus (SC) is one of the most ancient regions
of the vertebrate central sensory system. In this hub afferents from
several sensory pathways converge, and an extensive range of neural
circuits enable primary sensory processing, multi-sensory integration
and the generation of motor commands for orientation behaviours. The SC
has a laminar structure and is usually considered in two parts; the
superficial visual layers and the deep multi-modal/motor layers.
Neurones in the superficial layers integrate visual information from the
retina, cortex and other sources, while the deep layers draw together
data from many cortical and sub-cortical sensory areas, including the
superficial layers, to generate motor commands. Functional studies in
anaesthetized subjects and in slice preparations have used
pharmacological tools to probe some of the SC\'s interacting circuits.
The studies reviewed here reveal important roles for ionotropic
glutamate receptors in the mediation of sensory inputs to the SC and in
transmission between the superficial and deep layers.
N-methyl-D-aspartate receptors appear to have special responsibility for
the temporal matching of retinal and cortical activity in the
superficial layers and for the integration of multiple sensory
data-streams in the deep layers. Sensory responses are shaped by
intrinsic inhibitory mechanisms mediated by GABA(A) and GABA(B)
receptors and influenced by nicotinic acetylcholine receptors. These
sensory and motor-command activities of SC neurones are modulated by
levels of arousal through extrinsic connections containing GABA,
serotonin and other transmitters. It is possible to naturally stimulate
many of the SC\'s sensory and non-sensory inputs either independently or
simultaneously and this brain area is an ideal location in which to
study: (a) interactions between inputs from the same sensory system; (b)
the integration of inputs from several sensory systems; and (c) the
influence of non-sensory systems on sensory processing.

173\. Wichmann, T., R.B. Illing, and K. Starke, *Evidence for a
neurotransmitter function of acetylcholine in rabbit superior
colliculus.* Neuroscience, 1987. 23(3): p. 991-1000.
Acetylcholinesterase staining and studies on the uptake of \[3H\]choline
into the subsequent efflux of tritium from collicular slices were
carried out in order to provide evidence for a neurotransmitter function
of acetylcholine in rabbit superior colliculus. Acetylcholinesterase
staining was dense and homogeneous in superficial layers whereas the
staining was arranged in patches with slightly higher density caudally
than rostrally in the intermediate layers. The accumulation of tritium
in slices incubated with \[3H\]choline depended on time, temperature and
concentration, and was inhibited by hemicholinium-3. Accumulation was
slightly higher in caudal than in rostral slices. Electrical stimulation
enhanced tritium outflow from slices preincubated with \[3H\]choline.
Tetrodotoxin and a low calcium medium inhibited the evoked overflow
whereas hemicholinium-3 caused an enhancement. Oxotremorine decreased
the evoked overflow; atropine prevented this effect. The opioids
\[D-Ala2, MePhe4, Glycol5\]enkephalin, \[D-Ala2, D-Leu5\]enkephalin and
ethylketocyclazocine caused an inhibition. The effects of the latter two
agonists were antagonized by naloxone. The GABAB-receptor-agonist
(-)-baclofen decreased the evoked overflow at lower concentrations than
GABA, whereas the GABAA-receptor-agonist muscimol was ineffective.
Serotonin produced an inhibition which was prevented by metitepin,
alpha- and beta-adrenoceptor as well as dopamine-receptor ligands caused
no change. It is concluded that in the rabbit superior colliculus the
pattern of acetylcholinesterase staining is comparable, but not
identical to the distribution in other species. The accumulation of
\[3H\]choline, as well as the tetrodotoxin-sensitive and
calcium-dependent overflow of tritium upon electrical stimulation
(reflecting presumably release of \[3H\]acetylcholine) indicate that
acetylcholine has a neurotransmitter function in this tissue. The
release of \[3H\]acetylcholine was modulated by various transmitter
substances and related compounds. The pattern of modulation of release
differed from the pattern in other cholinergically innervated tissues.

174\. Beninato, M. and R.F. Spencer, *A cholinergic projection to the
rat superior colliculus demonstrated by retrograde transport of
horseradish peroxidase and choline acetyltransferase
immunohistochemistry.* J Comp Neurol, 1986. 253(4): p. 525-38.
Acetylcholinesterase (AChE) has been localized by histochemistry in the
superior colliculus and in the tegmentum of the caudal midbrain and
rostral pons of the rat. The pattern of AChE localization in the
superior colliculus was characterized by homogeneous staining in the
superficial layers and patchlike staining in the intermediate gray
layer. In the tegmentum, AChE was localized in the pedunculopontine
nucleus (PPN), beginning rostrally at the caudal pole of the substantia
nigra and extending caudally to the level of the parabrachial nuclei,
and in the lateral dorsal tegmental nucleus (LDTN) of the central gray.
The localization of AChE in these nuclei overlapped the distribution of
neurons stained by immunohistochemistry using an antibody to choline
acetyltransferase (ChAT), the synthesizing enzyme of the
neurotransmitter acetylcholine. Other neighboring areas that were
stained with AChE, but that did not contain ChAT-immunoreactive neurons,
included the microcellular tegmental nucleus and the ventral tegmental
nucleus. Neurons in the PPN and LDTN were determined to be potential
sources of the cholinergic projection to the intermediate gray layer of
the rat superior colliculus by double labelling with retrograde
transport of horseradish peroxidase (HRP) combined with the
immunohistochemical localization of ChAT. Three populations of neurons
were identified. A predominantly ipsilateral ChAT-immunoreactive
population was located in the pars compacta subdivision of PPN (PPNpc).
Retrograde HRP-labelled neurons in the pars dissipata subdivision of the
PPN (PPNpd), located ventral to the superior cerebellar peduncle (SCP)
at the level of the inferior colliculus, composed a second population
that was predominantly contralateral but was not ChAT immunoreactive. A
third population of retrogradely labelled neurons was predominantly
ipsilateral and ChAT immunoreactive and was located in the LDTN. These
findings compared favorably with the full extent of the projection from
this tegmental region revealed by retrograde transport of HRP from the
superior colliculus when more compatible fixation and chromogen
procedures were used. The results suggest that the PPN and the LDTN are
two sources of the cholinergic input to the superior colliculus. Since
the PPN also has extensive efferent, and afferent, connections with
basal-ganglia-related structures, this cholinergic excitatory input to
the superior colliculus, like the GABA-ergic inhibitory input from the
substantia nigra pars reticulata, may provide the basis for an
additional influence of the basal ganglia on visuomotor behavior.

175\. Weldon, D.A., L.C. Calabrese, and K.J. Nicklaus, *Rotational
behavior following cholinergic stimulation of the superior colliculus in
rats.* Pharmacol Biochem Behav, 1983. 19(5): p. 813-20. Rate which
received microinjections of carbachol into the superior colliculus
exhibited pronounced dose-dependent rotational behavior contralateral to
the site of injection (Experiment 1). Wet dog shakes were also observed
in some animals. Similar injections in the midbrain reticular formation
produced immobility with slight contralateral flexion of the neck.
Convulsions were observed in some rats after injections into either
anatomical location. In Experiment 2, circling induced by carbachol in
the superior colliculus was blocked by prior injection of either the
muscarinic receptor antagonist scopolamine or the nicotinic receptor
antagonist mecamylamine, suggesting that both nicotinic and muscarinic
receptors are involved in the effect. In Experiment 3 contralateral
rotational behavior was induced by intracollicular microinjections of
the combination of acetylcholine chloride and physostigmine. The results
suggest that collicular mediation of contralateral rotational behavior,
and perhaps orientation, might involve cholinergic receptors.

176\. Pazdernik, T.L., *et al.*, *Superior colliculus activation by
retinal nicotinic ganglion cells: a 2-deoxyglucose study.* Brain Res,
1982. 243(1): p. 197-200. Systemic injection of the acetylcholinesterase
inhibitor, di-isopropylfluorophosphate, in rats causes a marked increase
in glucose use in the superficial layers of the superior colliculus.
This activation of the superior colliculus is largely a retinal effect.
Furthermore, since this response can be blocked by intraocular as well
as systemic injections of mecamylamine, it is postulated that retinal
nicotinic receptors are involved.

177\. Kasser, R.J. and P.D. Cheney, *DFP action on rat superior
colliculus: localization and role of cholinergic receptors.*
Neurotoxicology, 1987. 8(4): p. 607-22. DFP, an irreversible
acetylcholinesterase inhibitor, markedly increases spontaneous unit
activity and reduces light-evoked responses in the superficial layers of
the rat superior colliculus (Cheney et al., 1987). The purpose of this
study was to investigate: (1) the sites of DFP action within the
retino-tectal pathway (retinal, central or both), and (2) the types of
cholinergic receptors (muscarinic, nicotinic, or both) involved. DFP
increased SGS unit activity when injected intraocularly, confining its
action to the retina, or when given systemically in bilateral enucleate
rats. Thus, DFP acts at both retinal and central sites to increase unit
activity in the SGS. Pretreating with muscarinic receptor antagonists
such as atropine or scopolamine blocked DFP\'s effects at both sites
whereas the nicotinic receptor antagonist mecamylamine was ineffective.
Moreover, DFP\'s actions were mimicked by injections of the muscarinic
receptor agonist, oxotremorine. The oxotremorine effects were also
blocked or reversed by treatment with atropine or scopolamine. We
conclude that DFP acts at both retinal and central sites to influence
SGS unit activity and, at both sites, muscarinic receptors mediate
DFP\'s effects.

178\. Kang, H.K. and T.A. Bullman, *Mortality among US veterans of the
Persian Gulf War: 7-year follow-up.* Am J Epidemiol, 2001. 154(5): p.
399-405. To assess the long-term health consequences of the 1991 Persian
Gulf War, the authors compared cause-specific mortality rates of 621,902
Gulf War veterans with those of 746,248 non-Gulf veterans, by gender,
with adjustment for age, race, marital status, branch of service, and
type of unit. Vital status follow-up began with the date of exit from
the Persian Gulf theater (Gulf veterans) or May 1, 1991 (control
veterans). Follow-up for both groups ended on the date of death or
December 31, 1997, whichever came first. Cox proportional hazards models
were used for the multivariate analysis. For Gulf veterans, mortality
risk was also assessed relative to the likelihood of exposure to nerve
gas at Khamisiyah, Iraq. Among Gulf veterans, the significant excess of
deaths due to motor vehicle accidents that was observed during the
earlier postwar years had decreased steadily to levels found in non-Gulf
veterans. The risk of death from natural causes remained lower among
Gulf veterans compared with non-Gulf veterans. This was mainly accounted
for by the relatively higher number of deaths related to human
immunodeficiency virus infection among non-Gulf veterans. There was no
statistically significant difference in cause-specific mortality among
Gulf veterans relative to potential nerve gas exposure. The risk of
death for both Gulf veterans and non-Gulf veterans stayed less than half
of that expected in their civilian counterparts. The authors conclude
that the excess risk of mortality from motor vehicle accidents that was
associated with Gulf War service has dissipated after 7 years of
follow-up.

179\. Kang, H.K. and T.A. Bullman, *Mortality among U.S. veterans of the
Persian Gulf War \[see comments\].* N Engl J Med, 1996. 335(20): p.
1498-504. BACKGROUND: Since the 1990-1991 Persian Gulf War, there has
been persistent concern that U.S. war veterans may have had adverse
health consequences, including higher-than-normal mortality. METHODS: We
conducted a retrospective cohort study of postwar mortality according to
cause among 695,516 Gulf War veterans and 746,291 other veterans. The
follow-up continued through September 1993. A stratified, multivariate
analysis (with Cox proportional-hazards models) controlled for branch of
service, type of unit, age, sex, and race in comparing the two groups.
We used standardized mortality ratios to compare the groups of veterans
with the general population of the United States. RESULTS: Among the
Gulf War veterans, there was a small but significant excess of deaths as
compared with the veterans who did not serve in the Persian Gulf
(adjusted rate ratio, 1.09; 95 percent confidence interval, 1.01 to
1.16). The excess deaths were mainly caused by accidents (1.25; 1.13 to
1.39) rather than disease (0.88; 0.77 to 1.02). The corresponding rate
ratios among 49,919 female veterans of the Gulf War were 1.32 (0.95 to
1.83) for death from all causes, 1.83 (1.02 to 3.28) for accidental
death, and 0.89 (0.45 to 1.78) for death from disease. In both groups of
veterans the mortality rates were significantly lower overall than those
in the general population. The adjusted standardized mortality ratios
were 0.44 (95 percent confidence interval, 0.42 to 0.47) for Gulf War
veterans and 0.38 (0.36 to 0.40) for other veterans. CONCLUSIONS: Among
veterans of the Persian Gulf War, there was a significantly higher
mortality rate than among veterans deployed elsewhere, but most of the
increase was due to accidents rather than disease, a finding consistent
with patterns of postwar mortality among veterans of previous wars.

180\. Bell, N.S., *et al.*, *Proposed explanations for excess injury
among veterans of the Persian Gulf War and a call for greater attention
from policymakers and researchers.* Inj Prev, 2001. 7(1): p. 4-9.
INTRODUCTION: Death rates among US veterans of the Persian Gulf War were
lower than rates among non-deployed veterans and the US population at
large, with the exception of injury deaths; returning veterans were at
significantly greater risk of injury mortality. Similar patterns of
excess injury mortality were documented among US and Australian veterans
returning from Vietnam. In spite of these consistent findings little has
been done to explain these associations and in particular to determine
whether or not, and how, war related exposures influence injury risk
among veterans returning home after deployments. HYPOTHESIZED PATHWAYS:
Several potential pathways are proposed through which injury might be
related to deployment. First, increases in injury mortality may be a
consequence of depression, post-traumatic stress disorder, and symptoms
of other psychiatric conditions developed after the war. Second,
physical and psychological traumas experienced during the war may result
in the postwar adoption of \"coping\" behaviors that also increase
injury risk (for example, heavy drinking). Third, greater injury risk
may be the indirect consequence of increased experiences of ill defined
diseases and symptoms reported by many returning veterans. Fourth,
veterans may experience poorer survivability for a given injury event
resulting in greater mortality but not morbidity. Finally, the process
that selects certain individuals for deployment may lead to a spurious
association between deployment status and injury mortality by
preferentially selecting individuals who are risk takers and/or exposed
to greater hazards. CONCLUSIONS: More research and attention from
policymakers is needed to clarify the link between deployment and
postwar increased risk of injury.

181\. 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.

182\. Mayberg, H.S., *Clinical correlates of PET- and SPECT-identified
defects in dementia.* Journal of Clinical Psychiatry, 1994. 55(11
SUPPL): p. 12-21. Functional imaging studies in patients with dementia
have focused primarily on the reliability of scan patterns to correctly
diagnose specific diseases, particularly Alzheimer\'s disease. Results
from these studies have important implications as disease-specific
treatments become available. A complementary approach is to examine the
relationships between patterns of altered brain function and specific
behaviors. This tactic has potential impact on understanding normal
brain organization as well as targeting symptom-specific treatments.
Regional abnormalities have been identified in dementia patients that
correlate with specific behavioral deficits: disturbances of language
and visuospatial function, impaired verbal fluency and selective
attention, and the presence of delusions and depression. These patterns
are seen in patients with Alzheimer\'s disease as well as with dementias
of other etiologies. The specificity of these patterns for
disease-specific and disease-independent symptoms is unknown.

183\. Eustache, F., *et al.*, *Entorhinal cortex disruption causes
memory deficit in early Alzheimer\'s disease as shown by PET.*
Neuroreport, 2001. 12(4): p. 683-685. Voxel-based mapping of the
correlations between cognitive scores and resting-state brain glucose
utilization measured by PET has recently emerged as a novel way to
reveal in living patients with Alzheimer\'s disease (AD) the neural
systems whose disruption underlies particular neuropsychological,
especially mnemonic, deficits. We have now applied this approach using a
novel cognitive paradigm designed to selectively assess verbal episodic
memory, and show that in early AD disruption of the left entorhinal
cortex underlies this memory deficit, consistent with post mortern data
showing that this brain area is affected earliest and most severely by
tau pathology in AD.

184\. Tani, Y., *et al.*, *6R-L-erythro-5,6,7,8-tetrahydrobiopterin
(R-THBP, SUN 0588) acts on the brain muscarinic and nicotinic
cholinergic receptors as evaluated by positron emission tomography (PET)
in rhesus monkey.* Adv Exp Med Biol, 1993. 338: p. 331-4.

185\. Nordberg, A., *et al.*, *Long-term tacrine treatment in three mild
Alzheimer patients: effects on nicotinic receptors, cerebral blood flow,
glucose metabolism, EEG, and cognitive abilities.* Alzheimer Dis Assoc
Disord, 1998. 12(3): p. 228-37. The effect of long-term treatment with
tacrine (tetrahydroaminoacridine) was studied in three Alzheimer
patients (aged 57, 64, and 68 years) with mild dementia. All three
patients had a Mini-Mental State Examination score of 24/30 and carried
at least one apolipoprotein E (ApoE) epsilon4 allele. Tacrine was given
in doses between 80 and to 160 mg daily for 13-31 months. A lower
tacrine concentration was observed generally in cerebrospinal fluid
(CSF) compared with plasma. The acetylcholinesterase activity in CSF
tended to be increased following longer periods of tacrine treatment,
whereas the butyrylcholinesterase activity was decreased. The three
patients repeatedly underwent positron emission tomography investigation
of cerebral blood flow, nicotinic receptors, cerebral glucose
metabolism, and electroencephalogram (EEG) and cognitive tests. Positive
influences on these parameters were observed following both short-term
and long-term treatment with tacrine. Improvement of nicotinic receptors
(measured as 11C-nicotine binding), cerebral blood flow, EEG, and some
cognitive tests (trail making test, block design test) occurred earlier
after initiation of tacrine treatment compared with the glucose
metabolism, which was increased after several months of tacrine
treatment. An improvement in attention (trail making test) was observed
following tacrine as sign for frontal lobe activation (EEG). The
functional effects of tacrine in Alzheimer patients appeared to be
related to both dose and length of cholinesterase inhibitor treatment.

186\. Ding, Y.S., *et al.*, *Mapping nicotinic acetylcholine receptors
with PET.* Synapse, 1996. 24(4): p. 403-7.

187\. Chefer, S.I., *et al.*, *2-\[18F\]F-A-85380: a PET radioligand for
alpha4beta2 nicotinic acetylcholine receptors.* Neuroreport, 1999.
10(13): p. 2715-21. 2-\[18F\]fluoro-3-(2(S)-azetidinylmethoxy)pyridine
(2-\[18F\]F-A-85380), a ligand for nicotinic acetylcholine receptors
(nAChRs) was evaluated in an in vitro binding assay with membranes of
rat brain and in vivo by PET in Rhesus monkey brain. The ligand has high
affinity for alpha4beta2 nAChRs (K(D)=50 pM), crosses the blood-brain
barrier, and distributes in the monkey brain in a pattern consistent
with that of alpha4beta2 nAChRs. The specific/non-specific binding ratio
increased steadily, reaching a value of 3.3 in the thalamus at 4 h. The
specific binding of 2-\[18F\]F-A-85380 was reversed by cytisine. These
results, in combination with the data demonstrating low toxicity of
2-\[18F\]F-A-85380, indicate that this ligand shows promise for use with
PET in human subjects.

188\. Dolci, L., *et al.*, *Synthesis of a fluorine-18 labeled
derivative of epibatidine for in vivo nicotinic acetylcholine receptor
PET imaging.* Bioorg Med Chem, 1999. 7(3): p. 467-79. Epibatidine
(exo-2-(2\'-chloro-5\'-pyridyl)-7-azabicyclo\[2.2.1\]heptane), a natural
compound isolated from the skin of the Ecuadorian poison frog
Epipedobates tricolor, is the most potent nicotinic acetylcholine
receptor (nAChR) agonist reported to date. In order to visualize and
quantify in vivo these receptors in human brain using Positron Emission
Tomography (PET), \[18F\]norchlorofluoroepibatidine
(exo-2-(2\'-\[18F\]fluoro-5\'-pyridyl)-7-azabicyclo\[2.2.1\]heptane), a
fluorine-18 (t(1/2): 110 min) radiolabeled derivative of epibatidine has
been designed. The corresponding 2\'-bromo-, 2\'-iodo- and 2\'-nitro
exo-2-(5\'-pyridyl)-7-azabicyclo\[2.2.1\]heptane analogues as labeling
precursors, as well as norchlorofluoroepibatidine as a reference
compound have been synthesized by reductive, stereoselective,
palladium-catalyzed Heck-type coupling between an N-Boc protected
azanorbornene and the corresponding halopyridine.
\[18F\]Norchlorofluoroepibatidine has been radiolabeled with fluorine-18
by nucleophilic aromatic substitution from the corresponding
Boc-protected halo- and nitro precursors using \[18F\]FK-K222 complex in
DMSO by conventional heating (at 150-180 degrees C for 10 min) or
microwave activations (at 100 Watt, for 1 to 2.5 min), followed by
TFA-removal of the protective group. Typically, using the microwave
activation procedure, 60-80 mCi (2.22-2.96 GBq) of pure
\[18F\]norchlorofluoroepibatidine could be obtained in less than 2 h
(110-115 min) from the bromo labeling precursor, with specific
radioactivities of 1.5-2.5 Ci/micromol (55.5-92.5 GBq/micromol)
calculated for End of Bombardment. The preliminary PET experiments in
baboon (Papio papio) with \[18F\]norchlorofluoroepibatidine show a high
uptake and a rapid accumulation of the radiotracer into the brain within
30 min. In the thalamus, a nAChR rich area, uptake of radioactivity
reached a maximum at 40 min (10% I.D./100 mL tissue). The ratio of
radioactivity thalamus/cerebellum (the latter being a nAChR poor area)
was 2 at 40 min and increased with time, up to 4.3 at 160 min. Its
specific regiodistribution and its high ratio of specific-to-nonspecific
binding confirm the ideal profile of \[18F\]norchlorofluoroepibatidine
as a suitable radioligand for PET imaging of nAChRs in the brain.

189\. Horti, A., *et al.*, *Fluorine-18-FPH for PET imaging of nicotinic
acetylcholine receptors.* J Nucl Med, 1997. 38(8): p. 1260-5.
Visualization of central nicotinic acetylcholine receptors (nAChRs) with
modern PET or SPECT imaging techniques has been hampered by the lack of
a radioligand with suitable in vivo binding characteristics (i.e., high
target-to-nontarget ratios and kinetics appropriate for the half-life of
the tracer and imaging modality used). This paper describes in vivo
binding, kinetics and pharmacology of a highly potent 18F-labeled analog
of epibatidine,
(+/-)-exo-2-(2-\[18F\]fluoro-5-pyridyl)-7-azabicyclo\[2.2.1\]heptane
(\[18F\]FPH), in the mouse brain with the view towards application of
this tracer for PET imaging of nAChR in human brain. METHODS:
Fluorine-18-FPH was administered intravenously to mice, and
time-activity curves were determined for several regions in the brain
and other organs. Saturation and pharmacology of \[18F\]FPH binding was
demonstrated in vivo by preinjecting unlabeled FPH or other drugs with
known pharmacological action before \[18F\]FPH was injected. The effect
of the drugs on \[18F\]FPH accumulation was evaluated. RESULTS:
\[18F\]FPH was rapidly incorporated into the mouse brain; peak activity
(2.4% of the injected dose) was measured at 5 min after intravenous
administration, followed by washout to 1.1% injected dose (ID) at 60
min. Highest concentrations of 18F occurred at 15 min in areas known to
contain high densities of nAChR inverted question marke.g., thalamus
\[9.7% of injected dose per gram tissue (ID/g inverted question mark\]
and superior colliculus (8.3% ID/g)\]. Accumulation of the 18F tracer in
hippocampus, striatum, hypothalamus and cortical areas was intermediate
(5.0, 5.6, 4.2 and 5.6% ID/g, respectively) and low in the cerebellum
(2.8% ID/g). The distribution of \[18F\]FPH in the mouse brain matched
that of other in vivo nAChR probes such as 3H-labeled epibatidine or
norchloroepibatidine, \[3H\](-)-nicotine and \[3H\]cytisine and that of
nAChR densities determined in postmortem autoradiographic studies in
rodents. Preinjection of blocking doses of unlabeled epibatidine,
(-)-nicotine, lobeline and cytisine significantly inhibited \[18F\]FPH
binding in thalamus and superior colliculus, but not in cerebellum,
whereas drugs that interact with binding sites other than acetylcholine
recognition sites of nAChR (e.g., mecamylamine, scopolamine,
N-methylspiperone and ketanserin) had no effect on \[18F\]FPH
accumulation in any of the brain regions examined. CONCLUSION:
Fluorine-18-FPH labels nAChR in vivo in the mouse brain. Because of its
high uptake into the brain and high ratios of specific-to-nonspecific
binding, this radioligand appears to be ideally suited for PET imaging
of nAChR in the mammalian brain.

190\. Ding, Y., *et al.*, *Synthesis and evaluation of
6-\[(18)F\]fluoro-3-(2(S)-azetidinylmethoxy)pyridine as a PET tracer for
nicotinic acetylcholine receptors.* Nucl Med Biol, 2000. 27(4): p.
381-9. Both ABT-594 ((R)-2-chloro-5-(2-azetidinylmethoxy)pyridine) and
A-85380 (3-\[2(S)-2-azetidinylmethoxy\]pyridine), novel nicotinic
agonists that possess potent non-opioid analgesic properties, have high
affinity for neuronal nicotinic acetylcholine receptors (nAChR) but do
not elicit the pronounced toxicity of epibatidine.
6-\[(18)F\]Fluoro-3-(2(S)-azetidinylmethoxy)pyridine
(6-\[(18)F\]fluoro-A-85380), a F-18 labeled analogue of these two
compounds, is therefore a promising radioligand for positron emission
tomography (PET) studies in humans. The use of trimethylammonium as a
leaving group in nucleophilic aromatic substitution reactions has proven
to be a versatile and efficient strategy, and offers several advantages
over other leaving groups. Here, we report the synthetic strategy for
the preparation of a precursor, as a trimethylammonium iodide salt, and
its use in the radiosynthesis to 6-\[(18)F\]fluoro-A-85380. Preliminary
compartative PET studies of 6-\[(18)F\]fluoro-A-85380 and
2-\[(18)F\]fluoro-A-85380 were carried out in baboon to examine their
suitability as tracers for studying nAChR system.

191\. Asahina, M., *et al.*, *Brain muscarinic receptors in progressive
supranuclear palsy and Parkinson\'s disease: a positron emission
tomographic study.* J Neurol Neurosurg Psychiatry, 1998. 65(2): p.
155-63. OBJECTIVES: To assess muscarinic acetylcholine receptors
(mAChRs) in the brains of patients with progressive supranuclear palsy
and Parkinson\'s disease, and to correlate the cholinergic system with
cognitive function in progressive supranuclear palsy and Parkinson\'s
disease. METHODS: Positron emission tomography (PET) and
\[11C\]N-methyl-4-piperidyl benzilate (\[11C\]NMPB) was used to measure
mAChRs in the brain of seven patients with progressive supranuclear
palsy, 12 patients with Parkinson\'s disease, and eight healthy
controls. All of the patients with progressive supranuclear palsy were
demented. The Parkinson\'s disease group consisted of 11 non-demented
patients and one demented patient. The mini mental state examination
(MMSE) was used to assess the severity of cognitive dysfunction in all
of the subjects. The modified Wisconsin card sorting test (WCST) was
used to evaluate frontal cognitive function in the non-demented patients
with Parkinson\'s disease and controls. RESULTS: The mean K3 value, an
index of mAChR binding, was significantly higher for the frontal cortex
in the patients with Parkinson\'s disease than in the controls
(p\<0.01). By contrast, the patients with progressive supranuclear palsy
had no significant changes in the K3 values of any cerebral cortical
regions. The mean score of the MMSE in the progressive supranuclear
palsy group was significantly lower than that in the control group.
Although there was no difference between the Parkinson\'s disease and
control groups in the MMSE, the non-demented patients with Parkinson\'s
disease showed significant frontal lobe dysfunction in the WCST.
CONCLUSIONS: The increased mAChR binding in the frontal cortex of the
patients with Parkinson\'s disease may reflect denervation
hypersensitivity caused by loss of the ascending cholinergic input to
that region from the basal forebrain and may be related to frontal lobe
dysfunction in Parkinson\'s disease. The cerebral cortical cholinergic
system may not have a major role in cognitive dysfunction in progressive
supranuclear palsy.

192\. Mentis, M.J., *et al.*, *Muscarinic versus nicotinic modulation of
a visual task. a pet study using drug probes.* Neuropsychopharmacology,
2001. 25(4): p. 555-64. Little is known about acetylcholine (ACh)
modulation of central visual processing in humans. Receptor densities in
visual brain regions are differentially distributed suggesting that
receptor subtypes have different functions. Using PET, we have
previously described the brain regions activated by a simple
pattern-flash stimulus in healthy elderly subjects. To evaluate
muscarinic and nicotinic contributions to ACh modulation of visual
processing, we scanned elderly subjects watching the pattern-flash
stimulus during no drug, during physostigmine augmentation, and during
scopolamine antagonism of physostigmine\'s action. These manipulations
of ACh significantly altered regional cerebral blood flow (rCBF) in
brain regions activated by the task. The pattern of rCBF values across
drug conditions suggested that muscarinic and nicotinic effects were
dissociated. Muscarinic action predominated in striate cortex (Brodmann
Area, BA 17) and lateral visual association areas (BA 18, 19), while
nicotinic action predominated in the thalamus and inferior parietal
regions (BA 39/40). Both muscarinic and nicotinic actions increased rCBF
in some regions while decreasing it in others. A parsimonious
reconciliation of these results with functional anatomy suggests that
muscarinic action modulates visual attribute processing, while nicotinic
action modulates arousal and selective attention to the visual task.

193\. Furey, M.L., *et al.*, *Cholinergic stimulation alters performance
and task-specific regional cerebral blood flow during working memory.*
Proc Natl Acad Sci U S A, 1997. 94(12): p. 6512-6. Modulation of the
cholinergic neurotransmitter system results in changes in memory
performance, including working memory (WM), in animals and in patients
with Alzheimer disease. To identify associated changes in the functional
brain response, we studied performance measures and regional cerebral
blood flow (rCBF) using positron emission tomography (PET) in healthy
subjects during performance of a WM task. Eight control subjects
received an infusion of saline throughout the study and 13 experimental
subjects received a saline infusion for the first 2 scans followed by a
continuous infusion of physostigmine, an acetylcholinesterase inhibitor,
for the subsequent 8 scans. rCBF was measured using H215O and PET in a
sequence of 10 PET scans that alternated between rest and task scans.
During task scans, subjects performed the WM task for faces.
Physostigmine both improved WM efficiency, as indicated by faster
reaction times, and reduced WM task-related activity in anterior and
posterior regions of right midfrontal gyrus, a region shown previously
to be associated with WM. Furthermore, the magnitudes of
physostigmine-induced change in reaction time and right midfrontal rCBF
correlated. These results suggest that enhancement of cholinergic
function can improve processing efficiency and thus reduce the effort
required to perform a WM task, and that activation of right prefrontal
cortex is associated with task effort.

194\. Kuhl, D.E., *et al.*, *In vivo mapping of cerebral
acetylcholinesterase activity in aging and Alzheimer\'s disease.*
Neurology, 1999. 52(4): p. 691-9. OBJECTIVE: To validate an in vivo
method for mapping acetylcholinesterase (AChE) activity in human brain,
preparatory to monitoring inhibitor therapy in AD. BACKGROUND: AChE
activity is decreased in postmortem AD brain. Lacking a reliable in vivo
measure, little is known about central activity in early AD, when the
disease is commonly targeted by AChE inhibitor drug therapy. METHODS:
Intravenous N-\[11C\]methylpiperidin-4-yl propionate (\[11C\]PMP) served
as an in vivo AChE substrate. AChE activity was defined using cerebral
PET for tracer kinetic estimates of the local rate of \[11C\]PMP
hydrolysis in 26 normal controls and 14 patients with AD. Eleven AD
patients also had concomitant in vivo cerebral measures of vesicular
acetylcholine transporter (cholinergic terminal) density and glucose
metabolism. RESULTS: Cerebral AChE activity measures 1) were independent
of changes in tracer delivery to cerebral cortex; 2) agreed with
reported postmortem data concerning normal relative cerebral
distributions, absence of large age-effect in normal aging, and deficits
in AD; 3) correlated in AD cerebral cortex with concomitant in vivo
measures of cholinergic terminal deficits, but not with metabolic
deficits; and 4) agreed quantitatively with predicted level of cerebral
AChE inhibition induced by physostimine. CONCLUSIONS: This in vivo PET
method provided valid measures of central AChE activity in normal
subjects and AD patients. Applied in early AD, it should facilitate
inhibitor treatment by confirming central inhibition, optimizing drug
dosage, identifying likely responders, and testing surrogate markers of
therapeutic response.

195\. Shinotoh, H., *et al.*, *Progressive loss of cortical
acetylcholinesterase activity in association with cognitive decline in
Alzheimer\'s disease: a positron emission tomography study.* Ann Neurol,
2000. 48(2): p. 194-200. We measured brain acetylcholinesterase activity
in 30 patients with Alzheimer\'s disease (AD) and 14 age-matched
controls by positron emission tomography (PET) and using a carbon
11-labeled acetylcholine analogue. Seven AD patients had repeat PET
scans. The k3 values were calculated as an index of acetylcholinesterase
activity in a three-compartment analysis using the metabolite corrected
arterial input function. Twenty-eight of the 30 AD patients (14 each in
the early and late onset subgroups) were retained in the study so as to
equalize the range and average severity of cognitive impairment within
the early and late onset subgroups. The k3 values were significantly
reduced in the neocortex, hippocampus, and amygdala in the early onset
AD patients, although the k3 values were significantly reduced only in
the temporoparietal cortex and amygdala in the late onset AD patients.
In the longitudinal study, all 7 repeat AD patients showed further
reduction of cortical k3 values in the second PET scans, with a mean
interval of 2 years, suggesting a progressive loss of the ascending
cholinergic system from the nucleus basalis of Meynert in AD. In 37 AD
patients, there was a highly significant correlation between the
cortical k3 values and Mini-Mental State Examination scores, supporting
the cholinergic hypothesis in AD.

196\. Meltzer, C.C., *et al.*, *Serotonin in aging, late-life
depression, and Alzheimer\'s disease: The emerging role of functional
imaging.* Neuropsychopharmacology, 1998. 18(6): p. 407-430. Serotonin
(5-HT) neuron and neurotransmitter loss in normal aging and
neuropsychiatric diseases of late life may contribute to behavioral
changes commonly observed in the elderly population. Extensive evidence
implicates a deficit in serotonergic neurotransmission in the
development of major depression. It has been further suggested that the
age-related changes in 5-HT neurons may predispose the elderly to
develop depression. There is also increasing evidence that a combination
of disturbances in cholinergic and serotonergic function may play a role
in cognitive impairment in Alzheimer\'s disease (AD), with serotonergic
dysfunction potentially responsible for a significant portion of the
behavioral aspects of the disease. This implication of the 5-HT system
in aging and age-related cognitive and mood disorders rests in large
part on post mortem studies and animal models, which are limited in
their capacity to predict dynamic human biochemical-behavior
relationships or to accurately model the living human brain. Initial
applications of functional brian imaging with positron emission
tomography (PET) in the in vivo study of the brain in aging, depression,
and dementia focused on characterizing alterations in physiological
measurements of cerebral metabolism and perfusion. However, recent
advances in PET radiochemistry, instrumentation, and image processing
have paved the way for noninvasive means to test specific hypotheses
regarding the direct involvement of 5-HT neurons in the behavioral
features of aging and to define and monitor therapeutic regimens for
neuropsychiatric conditions of late life. Coupling of clinical trials in
well-characterized subject populations with PET imaging using ligands
specific for 5-HT receptor subtypes and transporter proteins promises to
increase our understanding of the role of the 5-HT system in affective
and cognitive aspects of treatment response. Longitudinal studies in
aging, late-life depression, and AD are also needed to evaluate the
complex interplay between neurodegenerative processes and serotonergic
neurotransmission.

197\. Price, J.C., *et al.*, *PET imaging of serotonin-1A receptors in
type 2 diabetes: Evidence of increased binding.* Journal of Nuclear
Medicine, 2001. 42(5 Supplement): p. 18P.

198\. Backman, L., *et al.*, *Cognitive deficits in Huntington\'s
disease are predicted by dopaminergic PET markers and brain volumes.*
Brain, 1997. 120(12): p. 2207-2217. The main aim of this study was to
investigate the relationship between dopaminergic markers and brain
volumes for striatal and cortical structures, and cognitive performance
in patients with Huntington\'s disease and control subjects. We used PET
and MRI data as predictors of performance in tasks assessing executive
function, visuospatial ability, episodic memory, verbal fluency,
perceptual speed and reasoning. The dopamine neurotransmission
parameters (D1 and D2 receptor density and dopamine transporter density)
and the volumetric measurements for caudate and putamen accounted for
substantial portions of the variance across the majority of cognitive
tasks. In addition, frontal volume showed a strong relationship with all
cognitive tasks. D1 binding and volume, measurements for the temporal
cortex and thalamic volume showed associations with a select number of
cognitive tasks. The overall data pattern is consistent with the view
that Huntington\'s disease may be characterized as a frontostriatal
dementia, in which cognitive deficits may result from pathological
changes at multiple sites in the frontostriatal circuitry.

199\. Schroeder, U., *et al.*, *Subthalamic nucleus stimulation affects
striato-anterior cingulate cortex circuit in a response conflict task: A
PET study.* Brain, 2002. 125(9): p. 1995-2004. The subthalamic nucleus
(STN) has generally been considered as a relay station within
frontal-subcortical motor control circuitry. Little is known about the
influence of the STN on cognitive networks. Clinical observations and
studies in animals suggest that the STN participates in non-motor
functions which can now be probed in Parkinson\'s disease patients with
deep brain stimulation of the STN, allowing selective and reversible
modulation of this nucleus. Using PET, we studied changes in regional
cerebral blood flow (rCBF) associated with a response conflict task
(Stroop task) in Parkinson\'s disease patients ON and OFF bilateral STN
stimulation. The Stroop task requires subjects to name the font colour
of colour words (e.g. \'blue\') printed in an incongruent colour ink
(e.g. yellow). During STN stimulation, impaired task performance
(prolonged reaction times) was associated with decreased activation in
both right anterior cingulate cortex (ACC) and right ventral striatum.
Concomitant increased activation in left angular gyrus indicative of
ongoing word processing during stimulation is consistent with an
impairment to inhibit habitual responses. ACC and ventral striatum are
part of the ACC circuit associated with response conflict tasks. The
decreased activation during STN stimulation in the ACC circuit, while
response conflict processing worsened, provides direct evidence of STN
modulating non-motor basal ganglia-thalamocortical circuitry. Impairment
in ACC circuit function could account for the subtle negative effects on
cognition induced by STN stimulation.

200\. de Leon, M.J., *et al.*, *Prediction of cognitive decline in
normal elderly subjects with
2-(18F)fluoro-2-deoxy-D-glucose/positron-emission tomography (FDG/PET).*
Proceedings of the National Academy of Sciences of the United States of
America, 2001. 98(19): p. 10966-10971. Neuropathology studies show that
patients with mild cognitive impairment (MCI) and Alzheimer\'s disease
typically have lesions of the entorhinal cortex (EC), hippocampus (Hip),
and temporal neocortex. Related observations with in vivo imaging have
enabled the prediction of dementia from MCI. Although individuals with
normal cognition may have focal EC lesions, this anatomy has not been
studied as a predictor of cognitive decline and brain change. The
objective of this MRI-guided
2-(18F)fluoro-2-deoxy-D-glucose/positron-emission tomography (FDG/PET)
study was to examine the hypothesis that among normal elderly subjects,
EC METglu reductions predict decline and the involvement of the Hip and
neocortex. In a 3-year longitudinal study of 48 healthy normal elderly,
12 individuals (mean age 72) demonstrated cognitive decline (11 to MCI
and 1 to Alzheimer\'s disease). Nondeclining controls were matched on
apolipoprotein E genotype, age, education, and gender. At baseline,
metabolic reductions in the EC accurately predicted the conversion from
normal to MCI. Among those who declined, the baseline EC predicted
longitudinal memory and temporal neocortex metabolic reductions. At
follow-up, those who declined showed memory impairment and
hypometabolism in temporal lobe neocortex and Hip. Among those subjects
who declined, apolipoprotein E E4 carriers showed marked longitudinal
temporal neocortex reductions. In summary, these data suggest that an EC
stage of brain involvement can be detected in normal elderly that
predicts future cognitive and brain metabolism reductions. Progressive
E4-related hypometabolism may underlie the known increased
susceptibility for dementia. Further study is required to estimate
individual risks and to determine the physiologic basis for METglu
changes detected while cognition is normal.

201\. Cools, R., *et al.*, *Dopaminergic modulation of high-level
cognition in Parkinson\'s disease: The role of the prefrontal cortex
revealed by PET.* Brain, 2002. 125(3): p. 584-594. This study examined
the effects of L-dopa medication in patients with Parkinson\'s disease
on cortical and subcortical blood flow changes during two tasks known to
involve frontostriatal circuitry. Eleven patients with Parkinson\'s
disease were scanned on two occasions, one ON L-dopa medication and one
OFF L-dopa medication, during performance of the Tower of London
planning task and a related test that emphasized aspects of spatial
working memory. L-dopa-induced decreases were observed in the right
dorsolateral prefrontal cortex during performance of both the planning
and the spatial working memory tasks compared with the visuomotor
control task. Conversely, L-dopa-induced blood flow increases were
observed in the right occipital lobe during the memory task relative to
the control task. Data from age-matched healthy volunteers demonstrated
that L-dopa effectively normalized blood flow in these regions in the
patient group. Moreover, a significant correlation was found between
L-dopa-induced, planning related blood flow decreases in the right
dorsolateral prefrontal cortex and L-dopa-induced changes in performance
on the planning task. These data suggest that L-dopa ameliorates
high-level cognitive deficits in Parkinson\'s disease by inducing
relative blood flow changes in the right dorsolateral prefrontal cortex.

202\. Arakawa, K., *et al.*, *Component-specific effects of
physostigmine on the cat visual evoked potential.* Exp Brain Res, 1993.
95(2): p. 271-6. Pattern visual evoked potentials (VEPs) were recorded
from the pial surface of the cat primary visual cortex prior to and
following the intravenous administration of physostigmine, an agent
which blocks the enzyme responsible for the breakdown of synaptically
released acetylcholine. The control VEP was composed of a small initial
positive deflection (P1), a subsequent large negative wave (N1) and a
second large positive wave (P2). Following physostigmine, the amplitude
of P1-N1 was diminished whereas that of N1-P2 increased. These effects
were long lasting and were blocked by prior treatment with scopolamine,
a result consistent with mediation by a muscarinic cholinergic pathway.
Waveform subtraction revealed that the physostigmine-sensitive component
had a slow, negative polarity waveform while the
physostigmine-insensitive component was also slow, but positive in
polarity. The fundamental nature of these components remains to be
assessed. Nevertheless, the results indicate that waveforms of different
polarity combine algebraically to yield the conventional VEP.

203\. Thierry, G., B. Doyon, and J.-F. Demonet, *ERP mapping in
phonological and lexical semantic monitoring tasks: A study
complementing previous PET results.* NeuroImage, 1998. 8(4): p. 391-408.
Previous PET results identified distinct neural systems involved with
phonology (vicinity of the left sylvian fissure) and lexical semantics
(left inferior temporal, left superior frontal, bilateral inferior
parietal regions). In the time domain, the phonological task was thought
to involve serial parsing of pseudo-words, whereas the lexical semantic
task would correspond to probabilistic automated access to meaning.
Auditory event-related potentials (ERPs) elicited by the same tasks were
explored on 32 channels in 12 male volunteers. Subjects had to
categorize as target a second element of a pair of pseudo-words or words
if a preceding target was detected in the first element. Depending on
the absence/presence of target in the first element, a RELEASE condition
and a HOLD condition were distinguished. RELEASE and HOLD ERPs split
earlier in the semantic task than in the phonological task (300 versus
412 ms after SOT, respectively), although words lasted longer than
pseudo-words. Corresponding paired t test maps showed a predominance of
differences over left perisylvian regions for the phonological task and
posterior bilateral regions for the semantic task. Underlying generators
were investigated using BESA (Scherg, 1990) with global task ERPs. Six
dipoles-constrained according to PET clusters-brought residual variance
down to 0.36%, from 364 to 565 ms after SOT, in both tasks. Relative
dipole amplitudes suggested a left-sided functional asymmetry for
phonology. These results support the hypothesis of left perisylvian
serial processing for phonology contrasting with bihemispheric parallel
access for semantics and substantiates BESA for temporally tackling
cognitive processes.

204\. Abdullaev, Y.G. and M.I. Posner, *Event-related brain potential
imaging of semantic encoding during processing single words.*
NeuroImage, 1998. 7(1): p. 1-13. Functional brain imaging studies with
positron emission tomography (PET) have identified blood flow changes in
widely separated areas of brain during the performance of word
processing tasks. In the present study we have utilized event-related
brain potentials (ERPs) to investigate the temporal relationships among
cortical areas previously identified by PET to be differentially
activated when performing semantic tasks with visual words. ERPs
revealed task-related differences over the central and left inferior
frontal regions around 170 and 220 ms, respectively, over a left
occipital region around 200 ms, over a large left parietotemporal region
around 600 ms, and finally over the right temporal lobe around 800 ms
after the word presentation. Analysis of topographic maps and dipole
sources as well as PET data allowed relating frontal midline positivity
around 170 ms to the anterior cingulate activation, and left inferior
frontal positivity around 220 ms to the PET activation of the left
inferior prefrontal cortex. The left parieto-temporal positivity around
600 ms seems to reflect the activity of Wernicke\'s area. The right
anterior temporal negativity beginning around 800 ms and peaking around
1100 ms may reflect the activity of the right insula. The left occipital
negativity around 200 ms is likely to reflect activation of a visual
word-form area in the left occipital lobe. These results provide the
time course for parts of the circuitry involved in semantic processing
of words and also demonstrate how combining the spatial localization of
PET with the temporal resolution of ERPs helps to understand the brain
mechanisms involved in human cognition.

205\. Gazzard, M.F. and D.P. Thomas, *A comparative study of central
visual field changes induced by sarin vapour and physostigmine eye
drops.* Exp Eye Res, 1975. 20(1): p. 15-21.

206\. Alderdice, M.T., *Physostigmine, but not neostigmine, inhibits
acetylcholine release.* Brain Res, 1979. 178(2-3): p. 596-9.

207\. Alderdice, M.T., *Further comparison of the effects of
physostigmine and neostigmine on frog neuromuscular transmission.* Clin
Exp Pharmacol Physiol, 1982. 9(1): p. 35-43. 1. The effects of
physostigmine and neostigmine were compared at frog neuromuscular
junctions using intracellular microelectrodes. 2. Both drugs increased
the amplitudes of miniature endplate potentials (MEPP) in a
dose-dependent manner because of their ability to competitively inhibit
cholinesterase activity. 3. In addition, physostigmine (1.0-20.0
mumol/l) decreased by approximately 20% the quantal content (determined
by two methods), indicating a decrease in the amount of transmitter
released upon nerve stimulation, whereas neostigmine did not alter
quantal content. 4. Because the endplate potential (EPP) amplitude is a
composite of pre- and post-junctional effects, the dose-response curve
for neostigmine on EPP amplitude approximately paralleled that obtained
on MEPP amplitude, whereas the effect of physostigmine on EPP amplitude
was depressed because of its action to decrease transmitter release. 5.
Upon washout of neostigmine, the increase in EPP amplitude reversed more
rapidly than effects on MEPP amplitude, a difference dependent on the
magnitude of quantal content and explained by an action on the
postjunctional membrane. Similar results were obtained with
physostigmine. 6. Neither drug significantly affected MEPP frequency
although small decreases were sometimes observed. 7. These results
suggest that at the frog neuromuscular junction the effects of
neostigmine are explained primarily by its inhibition of cholinesterase,
whereas physostigmine are explained primarily by its inhibition of
cholinesterase, whereas physostigmine has an additional action on nerve
terminals of decreasing nerve-stimulated release of acetylcholine.

208\. Abdel-Rahman, A., A. Shetty, and M. Abou-Donia, *Disruption of the
blood-brain barrier and neuronal cell death in cingulate cortex, dentate
gyrus, thalamus, and hypothalamus in a rat model of gulf-war syndrome.*
Neurobiol Dis, 2002. 10(3): p. 306. We investigated the effects of a
combined exposure to restraint stress and low doses of chemicals
pyridostigmine bromide (PB), N, N-diethyl-m-toluamide (DEET), and
permethrin in adult male rats, a model of Gulf-War syndrome. Animals
were exposed daily to one of the following for 28 days: (i) a
combination of stress and chemicals (PB, 1.3 mg/kg/day; DEET, 40
mg/kg/day; and permethrin, 0.13 mg/kg/day); (ii) stress and vehicle;
(iii) chemicals alone; and (iv) vehicle alone. All animals were
evaluated for: (i) the disruption of the blood-brain barrier (BBB) using
intravenous horseradish peroxidase (HRP) injections and endothelial
barrier antigen (EBA) immunostaining; (ii) neuronal cell death using H&E
staining, silver staining, and glial fibrillary acidic protein (GFAP)
immunostaining; and (iii) acetylcholinesterase (AChE) activity and
m2-muscarinic acetylcholine receptors (m2-AChR). Animals subjected to
stress and chemicals exhibited both disruption of the BBB and neuronal
cell death in the cingulate cortex, the dentate gyrus, the thalamus, and
the hypothalamus. Other regions of the brain, although they demonstrated
some neuronal cell death, did not exhibit disruption of the BBB. The
neuropathological changes in the above four brain regions were highly
conspicuous and revealed by a large number of HRP-positive neurons
(21-40% of total neurons), a decreased EBA immunostaining (42-51%
reduction), a decreased number of surviving neurons (27-40% reduction),
the presence of dying neurons (4-10% of total neurons), and an increased
GFAP immunostaining (45-51% increase). These changes were also
associated with decreased forebrain AChE activity and m2-AchR (19-25%
reduction). In contrast, in animals exposed to stress and vehicle or
chemicals alone, the above indices were mostly comparable to that of
animals exposed to vehicle alone. Thus, a combined exposure to stress
and low doses of PB, DEET, and permethrin leads to significant brain
injury. The various neurological symptoms reported by Gulf-War veterans
could be linked to this kind of brain injury incurred during the war.

209\. Abdel-Rahman, A., A.K. Shetty, and M.B. Abou-Donia, *Acute
exposure to sarin increases blood brain barrier permeability and induces
neuropathological changes in the rat brain: dose-response
relationships.* Neuroscience, 2002. 113(3): p. 721-41. We hypothesize
that a single exposure to an LD(50) dose of sarin induces widespread
early neuropathological changes in the adult brain. In this study, we
evaluated the early changes in the adult brain after a single exposure
to different doses of sarin. Adult male rats were exposed to sarin by a
single intramuscular injection at doses of 1, 0.5, 0.1 and 0.01xLD(50).
Twenty-four hours after the treatment, both sarin-treated and
vehicle-treated (controls) animals were analyzed for: (i) plasma
butyrylcholinesterase (BChE) activity; (ii) brain acetylcholinesterase
(AChE) activity, (iii) m2 muscarinic acetylcholine receptor (m2 mAChR)
ligand binding; (iv) blood brain barrier (BBB) permeability using
\[H(3)\]hexamethonium iodide uptake assay and immunostaining for
endothelial barrier antigen (EBA); and (v) histopathological changes in
the brain using H&E staining, and microtubule-associated protein (MAP-2)
and glial fibrillary acidic protein immunostaining. In animals treated
with 1xLD(50) sarin, the significant changes include a decreased plasma
BChE, a decreased AChE in the cerebrum, brainstem, midbrain and the
cerebellum, a decreased m2 mAChR ligand binding in the cerebrum, an
increased BBB permeability in the cerebrum, brainstem, midbrain and the
cerebellum associated with a decreased EBA expression, a diffuse
neuronal cell death and a decreased MAP-2 expression in the cerebral
cortex and the hippocampus, and degeneration of Purkinje neurons in the
cerebellum. Animals treated with 0.5xLD(50) sarin however exhibited only
a few alterations, which include decreased plasma BChE, an increased BBB
permeability in the midbrain and the brain stem but without a decrease
in EBA expression, and degeneration of Purkinje neurons in the
cerebellum. In contrast, animals treated with 0.1 and 0.01xLD(50) did
not exhibit any of the above changes. However, m2 mAChR ligand binding
in the brainstem was increased after exposure to all doses of the sarin.
Collectively, the above results indicate that, the early brain damage
after acute exposure to sarin is clearly dose-dependent, and that
exposure to 1xLD(50) sarin induces detrimental changes in many regions
of the adult rat brain as early as 24 hours after the exposure. The
early neuropathological changes observed after a single dose of 1xLD(50)
sarin could lead to a profound long-term neurodegenerative changes in
many regions of the brain, and resulting behavioral abnormalities.

210\. 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.

211\. Alberts, P. and V.R. Ogren, *Sarin and soman depress
\[3H\]acetylcholine secretion in guinea-pig ileum myenteric plexus.*
Acta Physiol Scand, 1985. 125(2): p. 347-8.

212\. Arroyo, G., *et al.*, *\[Nicotinic Receptor, galantamine and
Alzheimer disease\].* Rev Neurol, 2002. 34(11): p. 1057-65. Population
aging has increased and will drastically increase the prevalence of
Alzheimer disease. The disease develops inexorably towards a syndrome of
marked cognitive impairment, accompanied of emotional alterations and
profound changes of personality. The patient loses its autonomy, and
requires special attention of caregivers; this leads to a decrease of
the quality of life, not only of the patient but also of its caregivers
and family. The reduction of the number of functional nicotinic
receptors in brain keeps pace with neurological symptoms and the
severity of the disease (cholinergic theory of Alzheimer disease). There
is a pleyade of data and observations reinforcing the idea that
improving cholinergic neurotransmission is an investment in memory. Up
to now, although with limited success, this improvement has been
achieved only with the reversible inhibitors of acetylcholinesterase
tacrine, rivastigmine and donepezil, available in the clinic since a few
years. The last approved has been galantamine that in spite of being a
modest inhibitor of acetylcholinesterase, improves memory (ADAS cog
test) and slows down cognitive impairment of Alzheimer patients. To
explain this therapeutic effect, a second mechanism of action for
galantamine has been suggested, the positive allosteric modulation of
presynaptic nicotinic receptors, that will favour the release of
acetylcholine and other neurotransmitters involved in memory formation.
Furthermore, galantamine possesses neuroprotectant antiapoptotic
effects, according to recent data from our laboratory. These effects
provide new ideas and therapeutic targets that might help to find novel
and efficacious treatments for patients suffering Alzheimer disease.

213\. Akaike, A., *et al.*, *The nature of the interactions of
pyridostigmine with the nicotinic acetylcholine receptor-ionic channel
complex. II. Patch clamp studies.* Mol Pharmacol, 1984. 25(1): p.
102-12. Patch clamping of myoballs to record single channels was
performed to examine the interaction of the anticholinesterase agent
pyridostigmine (Pyr) with the acetylcholine (ACh) receptor-ion channel
complex. Single ACh channel currents were recorded from tissue-cultured
muscle cells of neonatal rats (myoballs). Pyr (50-100 microM) decreased
the frequency of channel-opening events activated by ACh, and induced a
modified form of the ACh channel currents. Channel conductance was lower
in the presence of Pyr, and channel lifetime remained unaltered or only
slightly prolonged. In addition, channel openings were frequently
interrupted by fast flickers in the presence of Pyr. Higher
concentrations (200 microM-1 mM) of the drug induced irregular waves of
bursting activity during the initial phase of the application, and,
subsequently, significantly reduced the frequency of channel openings.
Infrequent channel openings with low conductance were observed in the
patch when the micropipette was filled with Pyr alone. These results
suggest that, in addition to its anticholinesterase activity, Pyr reacts
with the ACh receptor, and both alone or in combination with ACh induces
an altered, desensitized species of the nicotinic receptor-ion channel
complex.

214\. Arendt, T., *et al.*, *Changes in acetylcholinesterase and
butyrylcholinesterase in Alzheimer\'s disease resemble embryonic
development\--a study of molecular forms.* Neurochem Int, 1992. 21(3):
p. 381-96. The pattern of molecular forms of acetylcholinesterase (AChE,
EC 3.1.1.7) and butyrylcholinesterase (BChE, EC 3.1.1.8) separated by
density gradient centrifugation was investigated in the brain and
cerebrospinal fluid in Alzheimer\'s disease (AD), in human embryonic
brain and in rat brain after experimental cholinergic deafferentation of
the cerebral cortex. While a selective loss of the AChE G4 form was a
rather constant finding in AD, a small but significant increase of G1
for both AChE and BChE was found in the most severely affected cases.
Both in normal human brain and in AD a significant relationship could be
established between the AChE G4/G1 ratio in different brain regions and
the activity of choline acetyltransferase (ChAT). A similar decrease of
the AChE G4 form as observed in AD can be induced in rat by experimental
cholinergic deafferentation of the cerebral cortex. The increase in G1
of both AChE and BChE in different brain regions in AD is quantitatively
related to the local density of neuritic plaques which are
histochemically reactive for both enzymes. In human embryonic brain, a
high abundance of G1 and a low G4/G1 ratio for both AChE and BChE was
found resembling the pattern observed in AD. Furthermore, both in
embryonic brain and in AD AChE shows no substrate inhibition which is a
constant feature of the enzyme in the adult human brain. It is,
therefore, concluded that the degeneration of the cholinergic cortical
afferentation in AD as reflected by a decrease of AChE G4 is accompanied
by the process of a neuritic sprouting response involved in plaque
formation which is probably associated with the expression of a
developmental form of the enzyme.

215\. Antonelli, T., *et al.*, *Changes in synaptosomal high affinity
choline uptake following electrical stimulation of guinea-pig cortical
slices: effect of atropine and physostigmine.* Br J Pharmacol, 1981.
74(3): p. 525-31. 1 Superfused guinea-pig cortical slices were
electrically stimulated at different frequencies and the changes in
acetylcholine (ACh) content measured. Synaptosomes were prepared at the
end of the stimulation period and high affinity choline uptake (HACU)
rate was measured. 2 The effect of increasing KC1 concentrations was
compared on ACh content of the slices and on synaptosomal HACU. 3
Electrical stimulation (2, 5, 10, 20 Hz) elicited a frequency-dependent
linear increase in synaptosomal HACU rate and a decrease in ACh content
of the slices. 4 The addition of atropine (1.5 x 10(-8) M) to the slices
enhanced and that of physostigmine (3 x 10(-5) M) reduced the
frequency-dependent increase in HACU rate. Atropine (1.5 x 10(-6) M) not
only antagonized the effect of physostigmine, but the HACU rate measured
after treatment with both drugs was larger than that found after
atropine alone. 5 These results indicate that in the cortical
cholinergic nerve endings, depolarization caused by electrical
stimulation is coupled with an increase in choline transport which can
be modulated by the addition of atropine or physostigmine. Furthermore,
within given experimental conditions a linear relationship exists
between the reciprocal of ACh content in the slices and synaptosomal
HACU.

216\. Abdel-Rahman, A., A.K. Shetty, and M.B. Abou-Donia, *Subchronic
dermal application of N,N-diethyl m-toluamide (DEET) and permethrin to
adult rats, alone or in combination, causes diffuse neuronal cell death
and cytoskeletal abnormalities in the cerebral cortex and the
hippocampus, and Purkinje neuron loss in the cerebellum.* Exp Neurol,
2001. 172(1): p. 153-71. N,N-Diethyl m-toluamide (DEET) and permethrin
have been implicated as potential neurotoxic agents that may have played
an important role in the development of illnesses in some veterans of
the Persian Gulf War. To determine the effect of subchronic dermal
application of these chemicals on the adult brain, we evaluated
histopathological alterations in the brain of adult male rats following
a daily dermal dose of DEET (40 mg/kg in 70% ethanol) or permethrin
(0.13 mg/kg in 70% ethanol) or a combination of the two for 60 days.
Control rats received a daily dermal dose of 70% ethanol for 60 days.
Animals were perfused and brains were processed for morphological and
histopathological analyses following the above regimen. Quantification
of the density of healthy (or surviving) neurons in the motor cerebral
cortex, the dentate gyrus, the CA1 and CA3 subfields of the hippocampus,
and the cerebellum revealed significant reductions in all three treated
groups compared with the control group. Further, animals receiving
either DEET or permethrin exhibited a significant number of degenerating
(eosinophilic) neurons in the above brain regions. However, degenerating
neurons were infrequent in animals receiving both DEET and permethrin,
suggesting that neuronal cell death occurs earlier in animals receiving
combined DEET and permethrin than in animals receiving either DEET or
permethrin alone. The extent of neuron loss in different brain regions
was similar among the three treatment groups except the dentate gyrus,
where neurodegeneration was significantly greater with exposure to DEET
alone. The neuron loss in the motor cerebral cortex and the CA1 subfield
of all treated groups was also corroborated by a significant decrease in
microtubule associated protein 2-immunoreactive elements (15-52%
reduction), with maximal reductions occurring in rats receiving DEET
alone; further, the surviving neurons in animals receiving both DEET and
permethrin exhibited wavy and beaded dendrites. Analysis of glial
fibrillary acidic protein immunoreactivity revealed significant
hypertrophy of astrocytes in the hippocampus and the cerebellum of all
treated groups (24-106% increase). Thus, subchronic dermal application
of DEET and permethrin to adult rats, alone or in combination, leads to
a diffuse neuronal cell death in the cerebral cortex, the hippocampal
formation, and the cerebellum. Collectively, the above alterations can
lead to many physiological, pharmacological, and behavioral
abnormalities, particularly motor deficits and learning and memory
dysfunction.

217\. Aizenman, E., G.G. Bierkamper, and E.F. Stanley, *Botulinum toxin
prevents stimulus-induced backfiring produced by neostigmine in the
mouse phrenic nerve-diaphragm.* J Physiol, 1986. 372: p. 395-404. The
origin of motor nerve antidromic activity (backfiring) induced by
anticholinesterase treatment was examined in the mouse phrenic
nerve-hemidiaphragm preparation. Botulinum toxin was used to determine
whether backfiring is due to (a) a direct effect of the cholinesterase
inhibitor on the nerve terminal, or (b) an indirect effect via the
prolongation of the action of acetylcholine. In previously untreated
control preparations, neostigmine produced spontaneous and
stimulus-induced antidromic activity in the phrenic nerve when rapidly
introduced into the diaphragm via its vasculature. This activity could
be reversibly blocked by d-tubocurarine and decamethonium, but not by
atropine. Neostigmine-induced backfiring did not occur in preparations
in which transmitter release was blocked with botulinum toxin. Infusion
of a small bolus of a high concentration of acetylcholine following
neostigmine treatment resulted in a short-term increase in the incidence
of antidromic activity, followed by block, in both controls and
botulinum toxin-treated preparations. It is concluded that transmitter
release is necessary for the production of backfiring following
cholinesterase inhibition since neostigmine alone does not elicit
antidromic activity in botulinum toxin-treated preparations at
concentrations which are effective in controls. Our results support the
hypothesis that the effects of neostigmine on the motoneurone terminal
are mediated by the prolonged action of acetylcholine that occurs with
inhibition of acetylcholinesterase.

218\. Amaki, Y., K. Kobayashi, and M. Kaneko, *\[The effect of
neostigmine and edrophonium on the MEPP frequency\].* Masui, 1989.
38(6): p. 760-6. Edrophonium may have stronger effect on presynaptic
site than neostigmine. Comparative study was done by measuring the
miniature endplate potential (MEPP) frequency which reflect the
presynaptic activity using rat phrenic nerve hemidiaphragm preparation.
Preparations were placed in an organ bath, filled with Krebs solution
and bubbled with 95% O2 and 5% CO2 at room temperature. A glass
electrode filled with 3M KCL was inserted as close to the endplate as
possible and 20 spots of MEPP were recorded for each drug. For recorded
MEEP with amplification, frequency was analyzed with the signal
processor. The highest effective level was different, being 10(-6)M in
neostigmine and 10(-5)M in edrophonium. At these levels, MEPP
frequencies of both drugs showed increases 2-4 times as high as the
level in the control group and the degrees of increase were about the
same with the two drugs. Our study revealed that both drugs have a
strong effect on presynaptic sites and that these stimulating effects of
edrophonium and neostigmine were about the same degree if 10 times as
high a dosage as that of the former is used.

219\. Anderson, R.J., *et al.*, *Decreased tetanic contracture of rat
skeletal muscle induced by pyridostigmine.* J Toxicol Environ Health,
1986. 18(2): p. 221-30. The purpose of this study was to compare the
effects of various regimens of pyridostigmine administration on the
contractile strength of skeletal muscle. Rats were exposed to
pyridostigmine according to 3 dosing schedules: 2 mg/kg ip daily, 5
mg/kg X d by sc infusion, and 25 mg/kg X d by sc infusion. After 1, 4,
10, and 20 d of exposure, measurements were made of muscle tension
during tetanic stimulation, and of muscle mass, erythrocyte
acetylcholinesterase activity, and body weight. Pyridostigmine produced
a dose-dependent decrement in the contracture generated during tetanic
stimulation. Peak effect was observed after 4 d of exposure but remained
depressed after 20 d. The magnitude of the decrement correlated with the
frequency of the tetanic stimulation (from 20 to 100 Hz). Muscle tension
at the end of the tetanic stimulus was affected to a greater extent than
the initial tension. The 25-mg/kg infusion of pyridostigmine
significantly depressed erythrocyte acetylcholinesterase activity
throughout the study and also decreased body weight on d 1-4. No change
in muscle mass was observed in any treatment group. These results show
that pyridostigmine exposure results in decrements in skeletal muscle
contracture that are dose-dependent, frequency-dependent, and
time-dependent. The effect is probably not the result of muscle wasting
and does not correlate well with erythrocyte acetylcholinesterase
inhibition. The results are consistent with effects of pyridostigmine,
both presynaptically and postsynaptically, at the neuromuscular junction
that affect neurotransmitter release and receptor responsiveness.

220\. Anderson, R.J., *Effect of pyridostigmine on tetanic contracture
of innervated and denervated rat skeletal muscle.* Neurotoxicology,
1987. 8(4): p. 553-60. Pyridostigmine has been reported to cause
histological damage and decrements in tetanic contracture of skeletal
muscle. Cholinesterase inhibition, decreased transmitter release and
desensitization of postsynaptic receptors have all been proposed as
mechanisms for these effects. The purpose of the present study was to
determine whether the effect of pyridostigmine to decrease tetanic
contracture was due to pre- or postsynaptic action. Rats with innervated
or chronically denervated hindlimbs were given pyridostigmine (25 mg/kg)
s.c. by infusion for four days. Tetanic contracture (20, 50, 100 Hz) was
evoked by sciatic nerve stimulation in the innervated muscles and direct
muscle stimulation in the denervated muscles. Pyridostigmine
significantly decreased the contracture of innervated triceps surae
muscle in a frequency dependent manner. Post-tetanic potentiation was
reduced from control muscle responses. In denervated muscle
pyridostigmine had no effect on contracture or post-tetanic
potentiation. These results show that innervation is required for the
detrimental effect of pyridostigmine and that the drug has no effect on
skeletal muscle.

221\. Anderson, R.J. and W.L. Chamberlain, *Pyridostigmine-induced
decrement in skeletal muscle contracture is not augmented by soman.*
Neurotoxicology, 1988. 9(1): p. 89-96. Previous studies have reported
that pyridostigmine induces a decrement in contractile force generated
during tetanic stimulation of skeletal muscle. Although our studies
suggested that pyridostigmine affected release of transmitter from the
motor nerve terminal, we could not exclude the possibility that the
drug\'s action was due to depolarization blockade of the muscle brought
on by excessive transmitter in the synaptic cleft. The purpose of this
study was to determine whether the effect of combined treatment with
pyridostigmine and an irreversible cholinesterase inhibitor (soman)
would potentiate the decrement in muscle contracture observed with
pyridostigmine alone. As reported previously, pyridostigmine (25 mg/kg)
significantly reduced muscle contracture during tetanic stimulation
(20-100 Hz), and soman (0.075 mg/kg) increased muscle contracture.
Combined treatment with pyridostigmine and soman produced a decrease in
muscle contracture equivalent to the effect of pyridostigmine alone.
Since there was no evidence of depolarization blockade of the muscle
despite aggressive treatment with two cholinesterase inhibitors, these
results support the view that pyridostigmine has a significant
presynaptic action to decrease neurotransmitter release. This action
opposes the drug\'s inhibition of cholinesterase, and the net effect of
combined treatment with pyridostigmine and soman is a muscle response
which is largely unchanged from the effect of pyridostigmine alone.

222\. Damodaran, T.V., *et al.*, *Sarin causes early differential
alteration and persistent overexpression in mRNAs coding for glial
fibrillary acidic protein (GFAP) and vimentin genes in the central
nervous system of rats.* Neurochem Res, 2002. 27(5): p. 407-15.
Neurotoxic effects of single dose of 0.5 x LD50 sarin
(O-isopropylmethylphosphonoflouridate) on central nervous system (CNS)
of male Sprague-Dawley rats were studied. We investigated the mRNA
expression of the astroglial marker genes glial fibrillary acidic
protein (GFAP) and vimentin to evaluate the fate of astroglial and
neuronal cells, because reactive gliosis is very often used to assess
the extent of CNS damage. Rats were treated with 50 microg/kg/ml of
sarin and terminated at the time-points 1 and 2 hours and 1, 3, and 7
days post-treatment. Control rats were treated with normal saline. Total
RNA was extracted and Northern blots were hybridized with cDNA probes
for GFAP and vimentin, as well as 28S RNA (control). The data obtained
indicate that a single dose of sarin (0.5 x LD50) showed induction in
the transcript levels of GFAP and vimentin in the cortex, cerebellum,
brainstem and midbrain, and spinal cord. The induction showed distinct
spatial-temporal differences for each tissue studied. Both GFAP and
vimentin were induced at 1 hour in all the tissues studied except
brainstem, where moderate and high levels of GFAP induction were noted
at 1 and 3 days. Overexpressed transcript levels of GFAP and vimentin
remained high in more responsive tissues such as the brainstem and
midbrain. Other tissues, such as the cortex, spinal cord, and cerebellum
showed a more downward trend for either GFAP or vimentin, or both,
transcript levels at 7 days. It is noteworthy that both cortex (318 +/-
12%) and spinal cord (368 +/- 12%) showed relatively higher induction of
GFAP, whereas cortex alone showed the highest level of overexpressed
vimentin transcript levels (284 +/- 11%). Overall it is also clear that
both GFAP and vimentin are needed for the effective recovery involving
co-ordinated alternating up- and downregulation of these two key
astrocyte genes, depending on tissue specificity. The changes seen in
the transcript levels of GFAP and vimentin may be the result of
astrocyte dysfunction and loss, accompanied by compensatory
proliferation and dedifferentiation of the astroglia. These changes
could affect the neuronal cell types, thus altering the neuron-glia
homeostasis.

223\. 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.

224\. Somani, S.M., R.K. Kutty, and G. Krishna, *Eseroline, a metabolite
of physostigmine, induces neuronal cell death.* Toxicol Appl Pharmacol,
1990. 106(1): p. 28-37. The toxic effects of physostigmine, an
anticholinesterase drug, and its metabolite eseroline were investigated
in three neuronal cell culture systems, mouse neuroblastoma N1E-115, rat
glioma C6, and neuroblastoma-glioma hybrid NG 108-15. Physostigmine and
eseroline (0.5 nM) elicited a time-dependent leakage of lactic acid
dehydrogenase (LDH) from all three cell types. An increased release of
\[14C\]adenine nucleotides was also detected from cells when they were
prelabeled with \[14C\]adenine. Eseroline was comparatively more toxic
than the parent compound, physostigmine. Eseroline elicited a dose- and
time-dependent leakage of LDH and release of adenine nucleotides from
the neuronal cells. A nonneuronal cell line, rat liver ARL-15, was
comparatively the most resistant cell type to eseroline toxicity. The
concentrations of eseroline needed for 50% release of adenine
nucleotides or 50% leakage of LDH from NG-108-15 and N1E-115 cells in 24
hr ranged from 40 to 75 microM. The concentrations of eseroline needed
to obtain similar responses in C6 and ARL-15 cells were much higher and
ranged from 80 to 120 microM. Phase contrast microscopy showed extensive
damage to three neuronal cell lines at concentrations of eseroline as
low as 75 microM. The loss of ATP from N1E-115 cells exceeded 50% when
they were treated with 0.3 mM eseroline for 1 hr\--at which time the
leakage of LDH was not detectable. It seems that eseroline causes
neuronal cell death by a mechanism involving loss of cell ATP. Thus, the
formation of eseroline may contribute to the toxic effect of
physostigmine.

225\. Spencer, P.S., *et al.*, *Self-reported exposures and their
association with unexplained illness in a population-based case-control
study of Gulf War veterans.* J Occup Environ Med, 2001. 43(12): p.
1041-56. Many factors have been considered as possible causes of the
unexplained illness reported by veterans of the Gulf War (GW). In this
study, we report an analysis of risk factors and unexplained illness in
a population-based sample of GW veterans who underwent clinical
evaluation. Multiple risk factors were compared in 241 veterans who met
criteria for unexplained illness and 113 healthy controls. Results
suggest that GW unexplained illness is most highly associated with
combat conditions, heat stress, and having sought medical attention
during the GW. When controlling for multiple simultaneous exposures
during the GW, interactions around pyridostigmine bromide, insecticides
and repellents, and stress were not significant. These results indicate
that most unexplained illness in GW veterans cannot be explained by
neurotoxic effects of exposures to chemicals that inhibit cholinesterase
activity.

226\. Abu-Qare, A.W. and M.B. Abou-Donia, *Combined exposure to sarin
and pyridostigmine bromide increased levels of rat urinary
3-nitrotyrosine and 8-hydroxy-2\'-deoxyguanosine, biomarkers of
oxidative stress.* Toxicol Lett, 2001. 123(1): p. 51-8. In this study
concentrations of markers of oxidative stress 3-nitrotyrosine and
8-hydroxy-2\'-deoxyguanosine (8-OhdG) were determined in rat urine
following a single oral dose of pyridostigmine bromide (PB) 13 mg/kg and
a single intramuscular dose of sarin 80 microg/kg alone or in
combination. Urine samples were collected 16, 24, 48, 72, and 96 h
following dosing. Control urine samples of five rats treated with normal
saline were also collected at the same time intervals. A combined dose
of PB and sarin significantly increased levels of 3-nitrotyrosine and
(8-OhdG) starting 48 h after dosing. An increase in the concentration of
these markers was not detected following a single dose of PB or sarin
alone. Maximal increase in 3-nitrotyrosine and 8-OhdG was detected 48 h
after administration of a combination PB and sarin. The results indicate
that concurrent exposure to PB and sarin could generate free radical
species that may cause oxidative stress in rats. The results may have
significant impact if veterans were expose to sarin following an oral
dose of PB.

227\. Abu-Qare, A.W., H.B. Suliman, and M.B. Abou-Donia, *Induction of
urinary excretion of 3-nitrotyrosine, a marker of oxidative stress,
following administration of pyridostigmine bromide, DEET
(N,N-diethyl-m-toluamide) and permethrin, alone and in combination in
rats.* Toxicol Lett, 2001. 121(2): p. 127-34. In this study, we
determined levels of 3-nitrotyrosine in rat urine following
administration of a single oral dose of 13 mg/kg pyridostigmine bromide
(PB) (3-dimethylaminocarbonyloxy-N-methylpyridinum bromide), a single
dermal dose of 400 mg/kg N,N-diethyl-m-toluamide (DEET) and a single
dermal dose of 1.3 mg/kg permethrin, alone and in combination. Urine
samples were collected from five treated and five control rats at 4, 8,
16, 24, 48, and 72 h following dosing. Solid-phase extraction coupled
with high-performance liquid chromatography with ultraviolet detection
at 274 nm was used for the determination of tyrosine and
3-nitrotyrosine. A single oral dose of PB and a single dermal dose of
DEET or their combination significantly (P\<0.05) increased levels of
3-nitrotyrosine starting 24 h after dosing compared with control urine
samples. The maximum increase of 3-nitroytyrosine was detected 48 h
after combined administration of PB and DEET. The ratio of
3-nitrotyrosine to tyrosine in urine excreted 48 h after dosing was
0.19+/-0.04, 0.20+/-0.05, 0.28+/-0.03, 0.32+/-0.04, 0.19+/-0.05,
0.42+/-0.04, 0.27+/-0.03, 0.36+/-0.04, and 0.48+/-0.04 following
administration of water, ethanol, PB, DEET, permethrin, PB+DEET,
PB+permethrin, DEET+permethrin, and PB+DEET+permethrin, respectively.
The results indicate that an oral dose of PB and a dermal administration
of DEET, alone and in combination, could generate free radical species,
and thus increase levels of 3-nitrotyrosine in rat urine. Induction of
3-nitrotyrosine, a marker of oxidative stress, following exposure to
these compounds could be significant in understanding the proposed
enhanced toxicity following combined exposure to these compounds.

228\. Artru, A.A. and J.D. Michenfelder, *A re-examination of
physostigmine-induced cerebral protection in the hypoxic mouse. A
critical assessment of the model.* Stroke, 1980. 11(2): p. 197-9. Using
the hypoxic (FiO2 = 0.05) mouse model as originally described, the
survival time following pretreatment with physostigmine was examined.
The maximum increase in survival time was 87% following a physostigmine
dose of 0.4 mg/kg. This increase was considerably less than that
previously reported for this drug in a hypoxic mouse study wherein the
standard method for exposing mice to hypoxia was altered. We speculate
that this alteration in methodology resulted in small variations in FiO2
sufficient to account for the differences between these studies.

229\. Andjelkovic, D.S. and M.R. Terzic, *The effects of physostigmine
on the oxygen uptake in rat brain tissue.* Experientia, 1979. 35(3): p.
372-3. Physostigmine in a dose of 0.1 mg/kg i.v. expressly stimulated
the oxygen uptake in the rat cerebral cortex. This effect was blocked by
propranolol and seems be mediated by catecholamines. Since atropine also
antagonized the stimulant effect of physostigmine, it appears that the
action of physostigmine is primarily cholinergic and that the adrenergic
effect is a secondary phenomenon. The higher dose of physostigmine (0.4
mg/kg i.v.) caused a depression of rat brain oxygen uptake.

230\. Rupreht, J., H.J. Schneck, and B. Dworacek,
*\[Physostigmine\--recent pharmacologic data and their significance for
practical use\].* Anaesthesiol Reanim, 1989. 14(4): p. 235-41.
Physostigmine is widely used for treatment of the central
anticholinergic syndrome during recovery from anaesthesia. The drug is
also very useful in treatment of intoxicated patients, in
differential-diagnostic procedures of coma of unknown origin, and in
restoration of vigilance after prolonged sedation for mechanical
ventilation. Besides the specific central cholinergic action of
physostigmine, several new pharmacological actions have now been
established. Analgesic action is dependent on the interaction with the
5-HT (serotoninergic) system and is independent of narcotic or
cholinergic agonists. The antianalgetic stress hormone, ACTH, also does
not interfere with this action. Physostigmine does not interfere with
the anaesthetic state when given during general anaesthesia. It
attenuates several withdrawal states, especially alcohol delirium,
opiate and nitrous oxide withdrawal syndromes. The drug may offer a
protective mechanism against hypoxic damage of the brain and may also be
beneficial in amnestic syndromes and sleep disorders. Physostigmine
produces central and peripheral cardiovascular stimulation. It has been
shown that physostigmine can be useful in prevention and treatment of
postanaesthetic behavioural disturbances following anaesthesia with
propofol. Number of indications for use of physostigmine has increased
considerably.

231\. Bradley, R.J., R. Sterz, and K. Peper, *Agonist and inhibitory
effects of pyridostigmine at the neuromuscular junction.* Brain Res,
1986. 376(1): p. 199-203. When all of the AChE at the endplate is
irreversibly inhibited by phospholine iodide the ionophoretically
induced ACh endplate currents are increased more than 10-fold in
amplitude. The reversible AChE inhibitor pyridostigmine only increases
the current to about half this value because its effects are obscured by
receptor blocking. It was found that pyridostigmine can activate the
receptor ion channels when released by ionophoresis at the endplate,
thus suggesting that agonist-like desensitization could contribute to
the blocking effects.

232\. Zheng, J.Q., *et al.*, *Neostigmine competitively inhibited
nicotinic acetylcholine receptors in sympathetic neurons.* Life Sci,
1998. 62(13): p. 1171-8. The present experiment investigates the effect
of neostigmine on nicotinic acetylcholine receptors (nAChRs) in the
cultured neurons from neonatal rat superior cervical ganglia (SCG).
Using whole-cell patch clamp techniques, we found that the amplitudes of
the currents induced by 50 microM dimethylphenylpiperazinium (DMPP) were
21.5+/-10.7%, 52.9+/-9.2% and 86.9+/-4.9% depressed at the increased
concentrations of neostigmine 100, 200 and 400 microM, respectively. The
inhibition of neostigmine decreased gradually with the increased
concentration of nicotine from 10 to 160 microM. Lineweaver-Burk\'s
double-reversible plot illustrated that neostigmine blocked neuronal
nAChRs in a competitive manner. Hyperpolarization of membrane potential
from -40 mV to -100 mV did not significantly influence the blockade of
neostigmine. Neostigmine could not accelerate the decay of the
DMPP-induced currents, neither evoke any detectable currents in SCG
neurons. The results indicate that neostigmine depress neuronal nAChRs
in a competitive, concentration-dependent and voltage-independent
manner, and can not facilitate desensitization of the receptors. The
present data suggest that neostigmine blocks neuronal nAChRs by
interacting with the ACh binding sites of the receptors.

233\. Shaw, K.P., *et al.*, *The reversible cholinesterase inhibitor
physostigmine has channel-blocking and agonist effects on the
acetylcholine receptor-ion channel complex.* Mol Pharmacol, 1985. 28(6):
p. 527-38. The actions of the carbamate cholinesterase inhibitors,
physostigmine (Phy) and physostigmine methiodide (MetPhy), were studied
on the acetylcholine receptor-ion channel complex (AChR) of skeletal
muscles. Low concentrations of these agents produced cholinesterase
inhibition which resulted in potentiation of nerve-elicited muscle
twitches and an increased peak amplitude and prolongation of the decay
time constant (tau EPC) of endplate currents (EPCs) elicited in frog
(Rana pipiens) sartorius muscles. However, increasing concentrations of
Phy depressed the peak amplitude and shortened the decay phase of the
EPC with an apparent loss in the voltage dependence of tau EPC. At
higher concentrations and depolarized potentials, EPC decays were double
exponential. The effects of both Phy and MetPhy on the postsynaptic AChR
complex were also evident in preparations pretreated with
diisopropylfluorophosphate. Under these conditions, a linear
relationship between the reciprocal of tau EPC and the concentration of
these agents was observed. Single channel studies revealed that Phy
(20-600 microM) shortened channel lifetime and decreased channel
conductance at very high concentrations. In addition, Phy (0.5 microM)
induced the appearance of channel openings with conductance similar to
that of acetylcholine. High concentrations (greater than 50 microM) of
this agent activated channel openings with decreased conductance.
Similar results were obtained with MetPhy. Thus, the reversible
cholinesterase inhibitors Phy and MetPhy altered the properties of the
AChR by interacting as agonists capable of inducing desensitization and
blockade.

234\. Zheng, J.Q., *et al.*, *Physostigmine blocked nicotinic
acetylcholine receptors in rat sympathetic ganglion neurons.* Zhongguo
Yao Li Xue Bao, 1997. 18(6): p. 508-11. AIM: To study the blocking
mechanism of physostigmine (Phy) on nicotinic acetylcholine receptors
(NAChR) in sympathetic neurons. METHODS: The whole-cell patch-clamp
technique was used to observe the effects of Phy on NAChR in the
cultured sympathetic neurons from neonatal rat superior cervical ganglia
(SCG). RESULTS: Phy 5 -20 mumol.L-1 inhibited neuronal NAChR in a
concentration-dependent manner and accelerated the desensitization of
NAChR. Changing the membrane potential from -50 to -90 mV did not affect
the blocking effect of Phy. Phy 200 mumol.L-1 did not induce any
noticeable response in SCG neurons. CONCLUSION: Phy blocked NAChR in the
sympathetic ganglion neurons by interacting with the allosteric sites
out of the binding sites and the open ionic channels of the receptors.
Phy did not possess excitative effect on NAChR in SCG neurons.

235\. Navratil, J., *et al.*, *The labilizing and stabilizing effect of
physostigmine on CNS.* Acta Univ Palacki Olomuc Fac Med, 1990. 126: p.
39-44. The authors observed biphasic changes of dopamine, norepinephrine
and serotonin in the rat brain stem after a single application of
physostigmine (1 mg/kg of body weight). The results confirm the
hypothesis of \"labilization and stabilization\" of membranes.

236\. Navratil, J., *et al.*, *Increase of nonesterified fatty acids in
CNS after physostigmine application.* Homeost Health Dis, 1991. 33(4):
p. 172-3.

237\. Tapia, R., *Release and uptake of glutamate as related to
excitotoxicity.* Rev Bras Biol, 1996. 56 Su 1 Pt 1: p. 165-74. It has
been established that neurons exposed to high concentrations of
glutamate or other excitatory amino acids degenerate and die. Neuronal
damage appears to be due to the activation of different types of
glutamate receptors, among which the ionotropic N-methyl-D-aspartate
(NMDA) type seems particularly involved, since its channel is permeable
to Ca2+ and an increase in the cytoplasmic concentration of this cation
promotes a chain of events leading to cell death. The mechanism of such
glutamate receptor-mediated neurodegeneration has been defined as
excitotoxicity, and several pieces of evidence suggest that this
mechanism might contribute to the neuronal death associated with certain
neurological disorders, such as ischemia, cerebral trauma and some
chronic neurodegenerative diseases. A relevant question is whether the
origin of endogenous extracellular glutamate is important for the
induction of excitotoxicity. An excess of glutamate release, or a
deficiency in its clearance from the synaptic cleft, which depends
mainly on its transport by high affinity carriers, are potential sources
for the accumulation of extracellular glutamate. In the present article
some experimental results from our laboratory, aimed at obtaining
information on this question, are reviewed. These experiments include
the use of 4-aminopyridine, a convulsant drug that enhances the release
of glutamate, and of some inhibitors of glutamate transport, in vivo and
in neuronal cell cultures. The results obtained indicate that an
increase of endogenous extracellular glutamate due to these procedures
is not sufficient to induce neuronal death, at least under the
experimental conditions used.

238\. Pitt, D., P. Werner, and C.S. Raine, *Glutamate excitotoxicity in
a model of multiple sclerosis.* Nat Med, 2000. 6(1): p. 67-70. Glutamate
excitotoxicity mediated by the AMPA/kainate type of glutamate receptors
damages not only neurons but also the myelin-producing cell of the
central nervous system, the oligodendrocyte. In multiple sclerosis,
myelin, oligodendrocytes and some axons are lost as a result of an
inflammatory attack on the central nervous system. Because glutamate is
released in large quantities by activated immune cells, we expected that
during inflammation in MS, glutamate excitotoxicity might contribute to
the lesion. We addressed this by using the AMPA/kainate antagonist NBQX
to treat mice sensitized for experimental autoimmune encephalomyelitis,
a demyelinating model that mimics many of the clinical and pathologic
features of multiple sclerosis. Treatment resulted in substantial
amelioration of disease, increased oligodendrocyte survival and reduced
dephosphorylation of neurofilament H, an indicator of axonal damage.
Despite the clinical differences, treatment with NBQX had no effect on
lesion size and did not reduce the degree of central nervous system
inflammation. In addition, NBQX did not alter the proliferative activity
of antigen-primed T cells in vitro, further indicating a lack of effect
on the immune system. Thus, glutamate excitotoxicity seems to be an
important mechanism in autoimmune demyelination, and its prevention with
AMPA/kainate antagonists may prove to be an effective therapy for
multiple sclerosis.

239\. Chen, Q., C.L. Veenman, and A. Reiner, *Cellular expression of
ionotropic glutamate receptor subunits on specific striatal neuron types
and its implication for striatal vulnerability in glutamate
receptor-mediated excitotoxicity.* Neuroscience, 1996. 73(3): p. 715-31.
Glutamate receptors are composed of subtype-specific subunits. Variation
in the precise subunit composition of a receptor may result in
significant functional differences. Thus, a precise knowledge of subunit
composition on striatal neurons is a prerequisite for understanding the
selective vulnerability of striatal neurons to excitatory amino acids.
In the present study, we used an immunohistochemical double-labelling
approach to localize ionotropic glutamate receptor subunits (NMDAR1,
GluR1, GluR2/3, GluR4 and GluR5/6/7) on specific striatal neuron
populations. Our results showed that striatal cholinergic and
somatostatin interneurons were not labelled for the
alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate, receptor subunits
GluR1, GluR2/3 and GluR4. Most cholinergic and somatostatin interneurons
(83.3% to 100%), however, were double-labelled for the
N-methyl-D-aspartate receptor subunit NR1 and kainic acid receptor
subunits GluR5/6/7. All parvalbumin interneurons were labelled for GluR1
and GluR4, and 96% GluR1 positive and 95% GluR4 positive neurons were
also double-labelled as parvalbumin interneurons. About half of all
parvalbumin interneurons co-localized with GluR2/3, and over 97% were
labelled for NR1 and GluR5/6/7. Among striatal projection neurons,
enkephalin-positive (mainly striatopallidal) neurons, striatonigral
neurons (mainly substance P-positive) and calbindin-positive matrix
neurons were not immunostained for GluR1 or GluR4. In contrast, 95% to
100% of each of these types of projection neurons were double-labelled
for NR1, GluR2/3 and GluR5/6/7. Our results demonstrate that striatal
neuron types differ in their expression of ionotropic glutamate receptor
subunits and subtypes. The clear difference between striatal
interneurons and projection neurons in ionotropic glutamate receptor
subtypes/subunits supports the idea that differential glutamate receptor
expression mechanism may account for the selective vulnerability of
striatal projection neurons to excitotoxicity, and that glutamate
receptor-mediated excitotoxicity may be involved in the striatal
neurodegenerative diseases.

240\. Paschen, W., *Glutamate excitotoxicity in transient global
cerebral ischemia.* Acta Neurobiol Exp (Warsz), 1996. 56(1): p. 313-22.
The glutamate excitotoxicity hypothesis of ischemic cell damage holds
that cell damage caused by transient cerebral ischemia is triggered by
glutamate, released during ischemia from the intracellular compartment
into the synaptic cleft: high extracellular glutamate levels activate
ionotropic glutamate receptors, thus inducing an overflow of calcium
ions into the neurones and a calcium-induced activation of catabolic
processes. However, several arguments (and much of the evidence) in
favour of this hypothesis do not bear closer examination. On the other
hand, evidence is accumulating that, after transient ischemia, calcium
fluxes through ionotropic glutamate receptors of the non-NMDA type may
play a major role in the manifestation of ischemic cell damage. Calcium
fluxes through non-NMDA receptors are determined by mRNA editing of
non-NMDA receptor subunits: calcium fluxes are blocked in the presence
of an edited subunit. A possible role of mRNA editing in the development
of ischemic cell damage is discussed.

241\. Zhang, J., *et al.*, *Secondary excitotoxicity contributes to
dopamine-induced apoptosis of dopaminergic neuronal cultures.* Biochem
Biophys Res Commun, 1998. 248(3): p. 812-6. Dopamine (DA) and related
catechols may contribute to selective degeneration of dopaminergic
neurons in the substantia nigra in Parkinson\'s disease. To investigate
whether DA induces apoptosis of dopaminergic neurons, we characterized
the effects of various concentrations of exogenous DA on a substantia
nigra/neuroblastoma hybrid cell line (MES 23.5 or MES). The hybrid MES
cells were maintained in the presence of 50 microM glutamate in
logarithmic growth on poly-D-lysine-precoated T-75 flasks and plated
either onto petri dishes with glass coverslips for morphological studies
or onto 6-well plates for quantification of apoptosis by flow cytometry.
The results showed that DA exposure (0.5-20 microM) induced time- and
dose-dependent apoptotic cell death of MES cells. To further analyze the
mechanism responsible for DA-mediated apoptosis, we repeated the
experiments at 20 microM DA in the presence or absence of 40 microM
nomifensine, a DA re-uptake inhibitor, and 25 microM
2-amino-5-phosphonopentanoic acid (AP5), an N-methyl-D-aspartate (NMDA)
receptor antagonist. The data indicate that both compounds significantly
prevented DA-induced apoptosis of MES cells and that combination of AP5
and nomifensine provided greater protection against DA toxicity than AP5
alone. These results suggest for the first time that DA-induced
apoptosis in dopaminergic neurons is partially attributable to increased
vulnerability of these cells to non-toxic levels of excitatory amino
acids, i.e., secondary excitotoxicity.

242\. Hugon, J., J.M. Vallat, and M. Dumas, *\[Role of glutamate and
excitotoxicity in neurologic diseases\].* Rev Neurol (Paris), 1996.
152(4): p. 239-48. Glutamate is one of the major excitatory
neurotransmitter in the central nervous system. Glutamate acts on 4
different post synaptic receptors; NMDA (N-Methyl-D-aspartate) AMPA
(alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid), Kainate and
metabotropic receptors. The three former receptors are linked to
membrane ion channels whereas metabotropic receptors are coupled with a
G protein. Glutamate is involved in the physiologic processes of
learning, memory and motricity. Glutamate is also a potent neurotoxin
responsible for toxic neuronal death of post synaptic neurons. This
action has been denominated excitotoxicity and occurs as a consequence
of a prolonged or a strong activation of glutamate post-synaptic
receptors. The rise in intracellular calcium seems to play a major role
in the pathological events following excitotoxicity. The pathophysiology
of several acute or chronic neurological disorders has been linked to
excitotoxicity. This excitotoxic process could be present in acute
neuronal death observed in stroke, hypoglycemia and traumatisms of the
central nervous system and in chronic neuronal degeneration observed in
Amyotrophic Lateral Sclerosis (ALS), Alzheimer\'s disease, Parkinson\'s
disease, Huntington\'s disease and neuro AIDS. A better knowledge of the
cellular events induced by excitotoxicity will allow to consider new
therapeutic approaches in various neurological disorders.

243\. Shaw, P.J. and P.G. Ince, *Glutamate, excitotoxicity and
amyotrophic lateral sclerosis.* J Neurol, 1997. 244 Suppl 2: p. S3-14.
The \"glutamate hypothesis\" is one of three major pathophysiological
mechanisms of motor neurone injury towards which current research effort
into amyotrophic lateral sclerosis (ALS) is directed. There is great
structural and functional diversity in the glutamate receptor family
which results from combinations of 14 known gene products and their
splice variants, with or without additional RNA editing. It is possible
that motor neurones express a unique molecular profile of glutamate
receptors. Abnormal activation of glutamate receptors is one of five
main candidates as a final common pathway to neuronal death. In
classical acute excitotoxicity, there is influx of Na+ and CI-, and
destabilisation of intracellular Ca2+ homeostasis, which activates a
cascade of harmful biochemical events. The concept of secondary
excitotoxicity, where cellular injury by glutamate is triggered by
disturbances in neuronal energy status, may be particularly relevant to
a chronic neurodegenerative disease such as ALS. Data are now beginning
to emerge on the fine molecular structure of the glutamate receptors
present on human motor neurones, which have a distinct profile of AMPA
receptors. Two important molecular features of motor neurones have been
identified that may contribute to their vulnerability to
neurodegeneration. The low expression of calcium binding proteins and
the low expression of the GluR2 AMPA receptor subunit by vulnerable
motor neurone groups may render them unduly susceptible to
calcium-mediated toxic events following glutamate receptor activation.
Eight lines of evidence that indicate a disturbance of glutamatergic
neurotransmission in ALS patients are reviewed. The links between
abnormal activation of glutamate receptors and other potential
mechanisms of neuronal injury, including activation of calcium-mediated
second messenger systems and free radical mechanisms, are emphasised.
Riluzole, which modulates the glutamate neurotransmitter system, has
been shown to prolong survival in patients with ALS. Further research
may allow the development of subunit-specific therapeutic targeting of
glutamate receptors and modulation of \"downstream\" events within motor
neurones, aimed at protecting vulnerable molecular targets in specific
populations of ALS patients.

244\. *US links motor neurone disease with Gulf War service.* BMJ, 2002.
324: p. 65. ALS.

245\. Nguimfack Mbodie, P.C., *\[Do the glutamate excitotoxicity theory
and potential free radicals implication in schizophrenia
aetiopathogenesis provide a new enlightenment to links between: genome,
environment and biology in the determinism of that disorder?\].*
Encephale, 2002. 28(2): p. 147-53. The aetiopathogenesis of
schizophrenia constitutes nowadays one of the major points of interest
for researchers on this cosmopolitan disorder which involves about 1% of
the world population and which significantly alters the social
functioning of the individual. Numerous studies have focused on the role
played by genome, environmental factors and biology in the development
of symptoms. The neurodevelopmental theory is an illustration with the
perinatal period considered as the main provider of environmental
factors (hypertension, infections, bleedings during pregnancy, acute and
chronic fetal distress.). Many authors found significant associations
between such factors, the occurrence of brain lesions and finally
schizophrenic symptoms. Although no convincing genetic model had been
established to date for schizophrenia, nevertheless it appears that a
predisposition not inheritable under the mendelian mode exists and
authors showed that disease gets more and more severe over schizophrenic
descendants. The risk to be schizophrenic being a first degree relative
of the schizophrenic person is about ten time superior than in general
population. Indeed, this risk is also about ten time superior in
biological parents of schizophrenic adoptees than in biological parents
of healthy adoptees. Studies done in monozygotic comparing to dizygotic
twins are in favour of an important role played by genetic factors more
than socioeducational or psychological factors. Concerning biology, the
dopaminergic hypothesis remains shared by numerous authors although
direct links with incriminated factors are not well established. Now is
suspected the glutamate excitotoxicity with implication of free radicals
in schizophrenia. These free radicals are products of various enzymatic
activations led by overstimulation of post synaptic receptors (NMDA and
AMPA) by the excess glutamate. Therefore, according to that concept,
some amino acids as glutamate and derivatives could have through free
radicals a noxious effect on neuronal synapses. This could be due to a
failing of their recapture at the presynaptic level in addition to a
dysfunctioning of the antioxidizing system (glutathion, carnosine,
superoxide dismutase, aspartate) to which dopamine and other monoamines
might participate. The question is whether or not this theory
contributes to shed light on links between: genome, environmental
factors and biology in schizophrenia. Through the review and discussion
of genetical aspects of schizophrenia, environmental factors and the
biological aspect, we intend to revive debate on that question. The
articles and authors were selected with regard to the aptness of their
publications on that subject, their evolving ideas and finally the
interest of their works for neurosciences. This new approach perhaps is
opening the way to new therapeutic perspectives in the treatment of
schizophrenia based on the antioxidizing substances as shown for some
neurological diseases (amyotrophic lateral sclerosis, Parkinson\'s
disease and Huntington\'s chorea) for which experiments are going on.

246\. Doble, A., *The role of excitotoxicity in neurodegenerative
disease: implications for therapy.* Pharmacol Ther, 1999. 81(3): p.
163-221. Glutamic acid is the principal excitatory neurotransmitter in
the mammalian central nervous system. Glutamic acid binds to a variety
of excitatory amino acid receptors, which are ligand-gated ion channels.
It is activation of these receptors that leads to depolarisation and
neuronal excitation. In normal synaptic functioning, activation of
excitatory amino acid receptors is transitory. However, if, for any
reason, receptor activation becomes excessive or prolonged, the target
neurones become damaged and eventually die. This process of neuronal
death is called excitotoxicity and appears to involve sustained
elevations of intracellular calcium levels. Impairment of neuronal
energy metabolism may sensitise neurones to excitotoxic cell death. The
principle of excitotoxicity has been well-established experimentally,
both in in vitro systems and in vivo, following administration of
excitatory amino acids into the nervous system. A role for
excitotoxicity in the aetiology or progression of several human
neurodegenerative diseases has been proposed, which has stimulated much
research recently. This has led to the hope that compounds that
interfere with glutamatergic neurotransmission may be of clinical
benefit in treating such diseases. However, except in the case of a few
very rare conditions, direct evidence for a pathogenic role for
excitotoxicity in neurological disease is missing. Much attention has
been directed at obtaining evidence for a role for excitotoxicity in the
neurological sequelae of stroke, and there now seems to be little doubt
that such a process is indeed a determining factor in the extent of the
lesions observed. Several clinical trials have evaluated the potential
of antiglutamate drugs to improve outcome following acute ischaemic
stroke, but to date, the results of these have been disappointing. In
amyotrophic lateral sclerosis, neurolathyrism, and human
immunodeficiency virus dementia complex, several lines of circumstantial
evidence suggest that excitotoxicity may contribute to the pathogenic
process. An antiglutamate drug, riluzole, recently has been shown to
provide some therapeutic benefit in the treatment of amyotrophic lateral
sclerosis. Parkinson\'s disease and Huntington\'s disease are examples
of neurodegenerative diseases where mitochondrial dysfunction may
sensitise specific populations of neurones to excitotoxicity from
synaptic glutamic acid. The first clinical trials aimed at providing
neuroprotection with antiglutamate drugs are currently in progress for
these two diseases.

247\. Rodriguez, M.C., J.A. Obeso, and C.W. Olanow, *Subthalamic
nucleus-mediated excitotoxicity in Parkinson\'s disease: a target for
neuroprotection.* Ann Neurol, 1998. 44(3 Suppl 1): p. S175-88. Dopamine
deficiency causes disinhibition and overactivity of the subthalamic
nucleus (STN). Output neurons from the STN are excitatory and use
glutamate as a neurotransmitter. They project to the external and
internal segments of the globus pallidum (GPe and GPi), the substantia
nigra pars reticulata (SNr), and the pedunculopontine nucleus (PPN). In
addition, STN neurons provide excitatory innervation to dopaminergic
(DA) neurons in the substantia nigra pars compacta (SNc) that contain
glutamate receptors. Stimulation of the STN induces bursting activity in
SNc dopaminergic neurons. This raises the possibility that the
disinhibition of STN neurons that occurs as a result of a dopamine
lesion might induce excitotoxic damage in target structures, including
the SNc. In addition, the reduction in complex I activity found in the
nigra in Parkinson\'s disease (PD) may cause mitochondrial dysfunction
and make SNc dopaminergic neurons vulnerable to even physiologic
concentrations of glutamate. We postulate that the dopamine loss that
occurs in PD produces augmented STN activity which, in turn, causes
further damage to vulnerable dopaminergic neurons, thereby creating a
scenario for an increasing cycle of neuronal loss in the SNc. In
addition, STN overactivity could, in theory, cause damage to the GPi,
SNr, and PPN and thereby account for the development of parkinsonian
features that do not respond to levodopa in patients with advanced
disease. This hypothesis suggests that pharmacologic or surgical
therapies that reduce STN neuronal overactivity or block glutamate
receptors in the SNc and other target structures might be
neuroprotective and might slow or halt the progression of
neurodegeneration in PD.

248\. Beal, M.F., *Excitotoxicity and nitric oxide in Parkinson\'s
disease pathogenesis.* Ann Neurol, 1998. 44(3 Suppl 1): p. S110-4. A
potential role for excitotoxic processes in Parkinson\'s disease (PD)
has been strengthened by the recent observations that there appears to
be a mitochondrially encoded defect in complex I activity of the
electron transport chain. An impairment of oxidative phosphorylation
will enhance vulnerability to excitotoxicity. Substantia nigra neurons
possess N-methyl-D-aspartate receptors and there are glutamatergic
inputs into the substantia nigra from both the cerebral cortex and the
subthalamic nucleus. After activation of excitatory amino acid
receptors, there is an influx of calcium followed by activation of
neuronal nitric oxide (NO) synthase, which can then lead to the
generation of peroxynitrite. Consistent with such a mechanism, studies
of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine neurotoxicity in both
mice and primates have shown that inhibition of neuronal NO synthase
exerts neuroprotective effects. Studies utilizing excitatory amino acid
receptor antagonists have been inconsistent in mice but show significant
neuroprotective effects in primates. These results raise the prospect
that excitatory amino acid antagonists for neuronal NO synthase
inhibitors might be useful in the treatment of PD.

249\. Zhang, J., *et al.*, *Secondary excitotoxicity contributes to
dopamine-induced apoptosis of dopaminergic neuronal cultures.* Biochem
Biophys Res Commun, 1998. 248(3): p. 812-6. Dopamine (DA) and related
catechols may contribute to selective degeneration of dopaminergic
neurons in the substantia nigra in Parkinson\'s disease. To investigate
whether DA induces apoptosis of dopaminergic neurons, we characterized
the effects of various concentrations of exogenous DA on a substantia
nigra/neuroblastoma hybrid cell line (MES 23.5 or MES). The hybrid MES
cells were maintained in the presence of 50 microM glutamate in
logarithmic growth on poly-D-lysine-precoated T-75 flasks and plated
either onto petri dishes with glass coverslips for morphological studies
or onto 6-well plates for quantification of apoptosis by flow cytometry.
The results showed that DA exposure (0.5-20 microM) induced time- and
dose-dependent apoptotic cell death of MES cells. To further analyze the
mechanism responsible for DA-mediated apoptosis, we repeated the
experiments at 20 microM DA in the presence or absence of 40 microM
nomifensine, a DA re-uptake inhibitor, and 25 microM
2-amino-5-phosphonopentanoic acid (AP5), an N-methyl-D-aspartate (NMDA)
receptor antagonist. The data indicate that both compounds significantly
prevented DA-induced apoptosis of MES cells and that combination of AP5
and nomifensine provided greater protection against DA toxicity than AP5
alone. These results suggest for the first time that DA-induced
apoptosis in dopaminergic neurons is partially attributable to increased
vulnerability of these cells to non-toxic levels of excitatory amino
acids, i.e., secondary excitotoxicity.

250\. Hugon, J., J.M. Vallat, and M. Dumas, *\[Role of glutamate and
excitotoxicity in neurologic diseases\].* Rev Neurol (Paris), 1996.
152(4): p. 239-48. Glutamate is one of the major excitatory
neurotransmitter in the central nervous system. Glutamate acts on 4
different post synaptic receptors; NMDA (N-Methyl-D-aspartate) AMPA
(alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid), Kainate and
metabotropic receptors. The three former receptors are linked to
membrane ion channels whereas metabotropic receptors are coupled with a
G protein. Glutamate is involved in the physiologic processes of
learning, memory and motricity. Glutamate is also a potent neurotoxin
responsible for toxic neuronal death of post synaptic neurons. This
action has been denominated excitotoxicity and occurs as a consequence
of a prolonged or a strong activation of glutamate post-synaptic
receptors. The rise in intracellular calcium seems to play a major role
in the pathological events following excitotoxicity. The pathophysiology
of several acute or chronic neurological disorders has been linked to
excitotoxicity. This excitotoxic process could be present in acute
neuronal death observed in stroke, hypoglycemia and traumatisms of the
central nervous system and in chronic neuronal degeneration observed in
Amyotrophic Lateral Sclerosis (ALS), Alzheimer\'s disease, Parkinson\'s
disease, Huntington\'s disease and neuro AIDS. A better knowledge of the
cellular events induced by excitotoxicity will allow to consider new
therapeutic approaches in various neurological disorders.

251\. Olney, J.W., *et al.*, *Excitotoxicity of L-dopa and 6-OH-dopa:
implications for Parkinson\'s and Huntington\'s diseases.* Exp Neurol,
1990. 108(3): p. 269-72. Despite several decades of research aimed at
elucidating the mechanisms underlying neuronal degeneration in
Parkinson\'s and Huntington\'s diseases, these mysteries remain
unfathomed. The brain contains high concentrations of the putative
transmitters, glutamate and aspartate, which have neurotoxic
(excitotoxic) potential and are thought to cause neuronal degeneration
in certain acute neurological disorders. However, no mechanism has been
identified by which these diffusely distributed agents might cause the
regionally selective patterns of neuronal degeneration characterizing
Parkinson\'s and Huntington\'s diseases. Here we report that L-DOPA, the
natural precursor to dopamine, is a weak excitotoxin and its
ortho-hydroxylated derivative, 6-OH-DOPA, is a powerful excitotoxin. We
propose that an excitotoxic process mediated by L-DOPA or an acidic
derivative such as 6-OH-DOPA might be responsible for degeneration of
nigral neurons in Parkinson\'s disease or striatal neurons in
Huntington\'s disease.

252\. Greene, J.G. and J.T. Greenamyre, *Bioenergetics and glutamate
excitotoxicity.* Prog Neurobiol, 1996. 48(6): p. 613-34. Bioenergetic
defects and abnormalities in glutamate neurotransmission have both been
proposed to play important roles in neurological diseases of varying
chronology, etiology and pathology. Recent experimental evidence
suggests an intimate relationship between these two systems. Metabolic
inhibition predisposes neurons to glutamate-mediated \"excitotoxic\"
damage. The exact mechanism of this increased susceptibility is yet to
be defined, but may involve, singly or in combination, decreased
voltage-dependent Mg2+ blockade of the N-methyl-D-aspartate (NMDA)
subtype of glutamate receptor, abnormalities in cellular Ca2+
homeostasis, or elevated production of reactive oxygen species. It is
believed that enhancement of excitotoxicity by impaired metabolism may
be a ubiquitous mechanism of neuronal death in neurological disease.
Further elucidation of the exact mechanism of this enhancement may lead
to the discovery of new targets for therapeutic intervention.

253\. Tymianski, M., *et al.*, *Characterization of neuroprotection from
excitotoxicity by moderate and profound hypothermia in cultured cortical
neurons unmasks a temperature-insensitive component of glutamate
neurotoxicity.* J Cereb Blood Flow Metab, 1998. 18(8): p. 848-67.
Although profound hypothermia has been used for decades to protect the
human brain from hypoxic or ischemic insults, little is known about the
underlying mechanism. We therefore report the first characterization of
the effects of moderate (30 degrees C) and profound hypothermia (12
degrees to 20 degrees C) on excitotoxicity in cultured cortical neurons
exposed to excitatory amino acids (EAA; glutamate, N-methyl-D-aspartate
\[NMDA\], AMPA, or kainate) at different temperatures (12 degrees to 37
degrees C). Cooling neurons to 30 degrees C and 20 degrees C was
neuroprotective, but cooling to 12 degrees C was toxic. The extent of
protection depended on the temperature, the EAA receptor agonist
employed, and the duration of the EAA challenge. Neurons challenged
briefly (5 minutes) with all EAA were protected, as were neurons
challenged for 60 minutes with NMDA, AMPA, or kainate. The protective
effects of hypothermia (20 degrees and 30 degrees C) persisted after
rewarming to 37 degrees C, but rewarming from 12 degrees C was
deleterious. Surprisingly, however, prolonged (60 minutes) exposures to
glutamate unmasked a temperature-insensitive component of glutamate
neurotoxicity that was not seen with the other, synthetic EAA; this
component was still mediated via NMDA receptors, not by ionotropic or
metabotropic non-NMDA receptors. The temperature-insensitivity of
glutamate toxicity was not explained by effects of hypothermia on
EAA-evoked \[Ca2+\]i increases measured using high- and low-affinity
Ca2+ indicators, nor by effects on mitochondrial production of reactive
oxygen species. This first characterization of excitotoxicity at
profoundly hypothermic temperatures reveals a previously unnoticed
feature of glutamate neurotoxicity unseen with the other EAA, and also
suggests that hypothermia protects the brain at the level of neurons by
blocking, rather than slowing, excitotoxicity.

254\. Budd, S.L. and D.G. Nicholls, *Mitochondria, calcium regulation,
and acute glutamate excitotoxicity in cultured cerebellar granule
cells.* J Neurochem, 1996. 67(6): p. 2282-91. Exposure of cultured
cerebellar granule cells to 100 microM glutamate plus glycine in the
absence of Mg2+ causes calcium loading of the in situ mitochondria and
is excitotoxic, as demonstrated by a collapse of the cellular ATP/ADP
ratio, cytoplasmic Ca2+ deregulation (the failure of the cell to
maintain a stable cytoplasmic free Ca2+ concentration), and extensive
cell death. Glutamate-evoked Ca2+ deregulation is exacerbated by the
mitochondrial respiratory chain inhibitor rotenone. Cells maintained by
glycolytic ATP, i.e., in the presence of the mitochondrial ATP synthase
inhibitor oligomycin, remain viable for several hours but are still
susceptible to glutamate; thus, disruption of mitochondrial ATP
synthesis is not a necessary step in glutamate excitotoxicity. In
contrast, the combination of rotenone (or antimycin A) plus oligomycin,
which collapses the mitochondrial membrane potential, therefore
preventing mitochondrial Ca2+ transport, allows glutamate-exposed cells
to maintain a high ATP/ADP ratio while accumulating little 45Ca2+ and
maintaining a low bulk cytoplasmic free Ca2+ concentration determined by
fura-2. It is concluded that mitochondrial Ca2+ accumulation is a
necessary intermediate in glutamate excitotoxicity, whereas the
decreased Ca2+ flux into cells with depolarized mitochondria may reflect
a feedback inhibition of the NMDA receptor mediated by localized Ca2+
accumulation in a microdomain accessible to the mitochondria.

255\. Nicholls, D.G. and S.L. Budd, *Mitochondria and neuronal glutamate
excitotoxicity.* Biochim Biophys Acta, 1998. 1366(1-2): p. 97-112. The
role of mitochondria in the control of glutamate excitotoxicity is
investigated. The response of cultured cerebellar granule cells to
continuous glutamate exposure is characterised by a transient elevation
in cytoplasmic free calcium concentration followed by decay to a plateau
as NMDA receptors partially inactivate. After a variable latent period,
a secondary, irreversible increase in calcium occurs (delayed calcium
deregulation, DCD) which precedes and predicts subsequent cell death.
DCD is not controlled by mitochondrial ATP synthesis since it is
unchanged in the presence of the ATP synthase inhibitor oligomycin in
cells with active glycolysis. However, mitochondrial depolarisation (and
hence inhibition of mitochondrial calcium accumulation) without parallel
ATP depletion (oligomycin plus either rotenone or antimycin A) strongly
protects the cells against DCD. Glutamate exposure is associated with an
increase in the generation of superoxide anion by the cells, but
superoxide generation in the absence of mitochondrial calcium
accumulation is not neurotoxic. While it is concluded that mitochondrial
calcium accumulation plays a critical role in the induction of DCD we
can find no evidence for the involvement of the mitochondrial
permeability transition.

256\. Castilho, R.F., M.W. Ward, and D.G. Nicholls, *Oxidative stress,
mitochondrial function, and acute glutamate excitotoxicity in cultured
cerebellar granule cells.* J Neurochem, 1999. 72(4): p. 1394-401. On
exposure to glutamate, cultured rat cerebellar granule cells undergo a
delayed Ca2+ deregulation (DCD), which precedes and predicts cell death.
We have previously shown that mitochondria control the sensitivity of
the neurons to DCD. Mitochondrial depolarization by rotenone/oligomycin
before glutamate addition is strongly neuroprotective, and the
indication is therefore that mitochondrial Ca2+ loading leads to a
delayed loss of bioenergetic function culminating in DCD and cell death.
In this report it is shown that superoxide (O2.-) generation in intact
cells, monitored by oxidation of hydroethidine to ethidium, was enhanced
by glutamate only when mitochondria were polarized. Production of
superoxide was higher in the subset of cells undergoing DCD. In the
presence of rotenone and oligomycin, addition of glutamate did not
result in increased superoxide generation. Menadione-generated
superoxide enhances the DCD of cells exposed to glutamate; in contrast,
glutamate-induced DCD was potently inhibited by the presence of the
cell-permeant antioxidant manganese(III) tetrakis(4-benzoic acid)
porphyrin. An inverse correlation is observed between the cytoplasmic
free Ca2+ maintained in individual cells in the presence of glutamate
and the ability of these cells to restore basal Ca2+ when NMDA receptors
are inhibited and mitochondrial Ca2+ is released. It is concluded that
mitochondrial Ca2+ accumulation and reactive oxygen species each
contribute to DCD, probably related to damage to a process controlling
Ca2+ efflux from the cell.

257\. Schubert, D. and D. Piasecki, *Oxidative glutamate toxicity can be
a component of the excitotoxicity cascade.* J Neurosci, 2001. 21(19): p.
7455-62. Along with ionotropic and metabotropic glutamate receptors, the
cystine/glutamate antiporter x(c)(-) may play a critical role in CNS
pathology. High levels of extracellular glutamate inhibit the import of
cystine, resulting in the depletion of glutathione and a form of cell
injury called oxidative glutamate toxicity. Here we show that a portion
of the cell death associated with NMDA receptor-initiated excitotoxicity
can be caused by oxidative glutamate toxicity. In primary mouse cortical
neurons the cell death resulting from the short-term application of 10
&mgr;m glutamate can be divided into NMDA and NMDA receptor-independent
phases. The NMDA receptor-independent component is associated with high
extracellular glutamate and is inhibited by a variety of reagents that
block oxidative glutamate toxicity. These results suggest that oxidative
glutamate toxicity toward neurons lacking functional NMDA receptors can
be a component of the excitotoxicity-initiated cell death pathway.

258\. Moghaddam, B., *et al.*, *Glucocorticoids mediate the
stress-induced extracellular accumulation of glutamate.* Brain Res,
1994. 655(1-2): p. 251-4. The hippocampal damage caused by stress has
been attributed to an increased glutamatergic tone brought about by
secretion of glucocorticoids. Although exposure to stress has been shown
to increase the outflow of glutamate, direct involvement of
glucocorticoid in this phenomenon has not been examined. The present
study demonstrates that adrenalectomy attenuates the stress-induced
outflow of glutamate in the hippocampus and prefrontal cortex and that
glucocorticoid replacement abolishes this attenuation.

259\. Tombaugh, G.C., *et al.*, *Glucocorticoids exacerbate hypoxic and
hypoglycemic hippocampal injury in vitro: biochemical correlates and a
role for astrocytes.* J Neurochem, 1992. 59(1): p. 137-46. The acute
secretion of glucocorticoids is critical for responding to physiological
stress. Under normal circumstances these hormones do not cause acute
neuronal injury, but they have been shown to enhance ischemic and
seizure-induced neuronal injury in the rat brain. Using fetal rat
hippocampal cultures, we asked whether hypoxic and hypoglycemic cell
damage in vitro could be exacerbated by direct exposure to
corticosterone (CORT). Each of these insults alone damaged neuronal
cells, whereas 4-6 h of hypoxic treatment could damage age-matched
astrocytes if glucose was reduced or omitted. Ischemic-like injury to
both cell types could be attenuated by pretreatment with high (30 mM)
glucose. Exposure to 100 nM CORT did not affect cell viability under
control conditions but enhanced both hypoxic and hypoglycemic neuronal
injury. In both cases, pretreatment with high glucose abolished this
CORT-mediated synergy. In astrocyte cultures, CORT exacerbated both
hypoxic and hypoglycemic injury and this effect was also attenuated by
high-glucose pretreatment. Identical 24-h CORT treatment caused a 13%
reduction in glucose uptake in astrocytes and a 38% reduction in
glycogen content, without affecting the level of intracellular glucose.
Thus, CORT could endanger both neurons and astrocytes in mixed
hippocampal cultures and this effect emerged only under conditions of
substrate depletion. The metabolic disruption in astrocytes by CORT
further suggests that the ability of CORT to exacerbate neuronal injury
may be due in part to impaired glial cell function.

260\. Suchecki, D., P.A. Tiba, and S. Tufik, *Paradoxical sleep
deprivation facilitates subsequent corticosterone response to a mild
stressor in rats.* Neurosci Lett, 2002. 320(1-2): p. 45-8. The
pituitary-adrenal responsiveness to a mild stressor was assessed in rats
that were deprived of paradoxical sleep (PS) and in controls that were
not deprived. Animals were either individually- or group-deprived for 96
h and hormone levels were assessed at 0, 5, 20 or 60 min after a saline
injection+novelty and compared with rats which were not deprived. Both
types of PS deprivation resulted in elevated adrenocorticotropin levels
at 0 min, which peaked at 5 min in all animals. Individually-deprived
rats exhibited the highest corticosterone (CORT) levels at 0 min. Peak
levels were higher and occurred earlier in PS-deprived than in control
rats (5 vs. 20 min, respectively). At 20 min, CORT levels had already
returned to unstressed levels in PS-deprived rats, but not in control
rats. These data indicate that PS deprivation induces facilitation of
the adrenocortical response to a mild stressor, but do not suggest that
PS deprivation changes the negative feedback sensitivity to CORT.

261\. Hairston, I.S., *et al.*, *Sleep deprivation elevates plasma
corticosterone levels in neonatal rats.* Neurosci Lett, 2001. 315(1-2):
p. 29-32. Plasma corticosterone (CORT) levels were measured after short
periods of sleep deprivation in rats at postnatal days 12, 16, 20, and
24. There was an age-dependent increase in basal CORT levels and sleep
deprivation significantly elevated CORT at all ages compared to
non-sleep deprived controls. The levels of CORT after sleep deprivation
in P16, P20 and P24 animals were similar, resulting in an age-dependent
decrease of the magnitude of the response. Sleep deprived P12 animals
had lower levels of CORT. However, the observed response to sleep
deprivation suggests that sleep loss is a significant stressor at this
age. These observations suggest that younger animals are more sensitive
to the effects of mild sleep deprivation than older ones.

262\. Maelicke, A., *et al.*, *Physostigmine and neuromuscular
transmission.* Ann N Y Acad Sci, 1993. 681: p. 140-54. Single channel
studies carried out in cultured rat myoballs and cultured hippocampal
neurons, and ion flux studies performed on Torpedo electrocyte membrane
vesicles, showed that physostigmine (Phy), a well-established
acetylcholinesterase inhibitor, interacts directly with nicotinic
acetylcholine receptors (nAChR). Low concentrations (0.1 microM) of Phy
activate the receptor integral channel, whereas higher concentrations
blocked the channel in its opened state. In contrast to channel
activation by acetylcholine (ACh) and classical cholinergic agonists,
however, Phy was capable of activating the nAChR channel even when the
ACh binding sites were blocked by competitive antagonists, such as
alpha-neurotoxins and d-tubocurarine, or when the nAChR was desensitized
by preincubation with high concentrations of ACh. The binding site at
which Phy binds and activates the nAChR was mapped. It was located
within the N-terminal extracellular region of the alpha-polypeptide, in
close proximity to the binding site of the natural transmitter. These
data identify a novel binding site at nAChRs from many species and
tissues that may be involved in receptor regulatory processes.

263\. Aiello-Malmberg, P., *et al.*, *Effects of morphine, physostigmine
and raphe nuclei stimulation on 5-hydroxytryptamine release from the
cerebral cortex of the cat.* Br J Pharmacol, 1979. 65(4): p. 547-55. 1.
The release of 5-hydroxytryptamine (5-HT) from the cerebral cortex and
caudate nucleus of brainstem-transected cats and from the cerebral
cortex of rats anaesthetized with urethane was determined by
radioenzymatic and biological assay. 2. The stimulation of nucleus
linearis intermedius of raphe doubles the basal 5-HT release in the
caudate nucleus and increases it 3 fold in the cerebral cortex. The
effects of the electrical stimulation of the raphe are potentiated by
chlorimipramine. 3. Brain 5-HT release is greatly increased by morphine
hydrochloride (6 mg/kg i.v.) and by physostigmine (100 microgram/kg
i.v.), but not by DL-DOPA (50 mg/kg i.v.). 4. It is suggested that the
5-HT releasing action of physostigmine can contribute to some of its
pharmacological effects such as the analgesic effect so far attributed
exclusively to its indirect cholinomimetic activity. 5. The 5-HT
releasing action of physostigmine seems unrelated to its
anticholinesterase activity.

264\. Anden, N.E., *Effects of oxotremorine and physostigmine on the
turnover of dopamine in the corpus striatum and the limbic system.* J
Pharm Pharmacol, 1974. 26(9): p. 738-40.

265\. Choi, J.Y., J.C. Morris, and C.Y. Hsu, *Aging and cerebrovascular
disease.* Neurol Clin, 1998. 16(3): p. 687-711. Aging influences
cerebrovascular disease expression by a variety of mechanisms.
Age-related changes in cerebral autoregulation, cellular metabolism, the
blood-brain barrier, and autonomic function may leave the
cerebrovascular system vulnerable to injury. Certain cerebrovascular
disease, such as atrial fibrillation, watershed infarctions, carotid
artery atherosclerosis, cerebral hemorrhages, subdural hematomas, and
transient global amnesia manifest in the elderly. Vascular dementia and
white matter disease are better understood with newer neuroimaging
studies, careful neuropsychological and histopathologic examinations.
Atherosclerosis and cerebral amyloid angiopathy may have larger roles
than previously understood in Alzheimer\'s disease.

266\. Banks, W. and A. Kastin, *Aluminum increases permeability of the
blood brain barrier to labeled DSIP and ß-endorphin: possible
implications for senile and dialysis dementia.* Lancet, 1983: p.
1227-1229.

267\. Sinton, C.M., *et al.*, *Stressful manipulations that elevate
corticosterone reduce blood brain barrier permeability to pyridostigmine
in the rat.* Toxicology and Applied Pharmacology, 2000. 165: p. 99-105.

268\. Telang, F.W., *et al.*, *Pyridostigmine, a carbamate
acetylcholinesterase AChE inhibitor and reactivator, is used
prophylactically against chemical warfare agents.* Nucl Med Biol, 1999.
26(2): p. 249-50.

269\. Jones, K.H., *et al.*, *Subchronic effects following a single
sarin exposure on blood-brain and blood-testes barrier permeability,
acetylcholinesterase, and acetylcholine receptors in the central nervous
system of rat: a dose-response study.* J Toxicol Environ Health A, 2000.
61(8): p. 695-707. Subchronic neurotoxic effects of sarin (O-isopropyl
methylphosphonofluoridate) treatment at various doses in male Sprague
Dawley rats were studied. The animals were treated with a single
intramuscular (im) injection of 0.01, 0.1, 0.5, or 1 x LD50 (100
microg/kg). The animals were maintained for 90 d thereafter.
\[3H\]Hexamethonium iodide was used to monitor the changes in
blood-brain barrier (BBB) permeability in cortex, brainstem, midbrain,
and cerebellum. Brainstem exhibited a significant decrease
(approximately 58% of control) in uptake of \[3H\]hexamethonium iodide
at 1 x LD50 dose. No significant changes were observed in BBB
permeability in cortex, midbrain, and cerebellum at any dose. Plasma
butyrylcholinesterase (BChE) activity remained unchanged, reflecting
recovery of the enzyme activity from the initial inhibition following
single exposure of 1 x LD50 sarin. Acetylcholinesterase (AChE) activity
in the cortex remained inhibited (approximately 29%), whereas in the
brainstem there was an increase (approximately 20%) at 1 x LD50 dose of
sarin. The m2-selective muscarinic acetylcholine receptor (m2-mAChR)
ligand binding was inhibited significantly at 1 x LD50 in the cortex,
whereas brainstem showed significantly increased (approximately 45%)
ligand binding at 1 x LD50 dose. Nicotinic acetylcholine receptor
(nAChR), on the other hand, showed a biphasic response in ligand binding
in the cortex with a decrease (approximately 30%) at 0.01 x LD50 but an
increase (approximately 40%) at 1 x LD5O. Brainstem did not show any
significant change in nAChR ligand binding. These results suggest that
single exposure of sarin could lead to changes that may play an
important role in neuropathological abnormalities in the central nervous
system.

270\. Lallement, G., *et al.*, *Heat stress, even extreme, does not
induce penetration of pyridostigmine into the brain of guinea pigs.*
Neurotoxicology, 1998. 19(6): p. 759-66. Stress due to forced swimming
was recently shown to allow penetration of pyridostigmine (PYR) into the
brain of mice. Accordingly, it was suggested that in troops exposed to
emotional stress under conditions of war, as during the Gulf War, the
BBB may have unexpectedly become permeable to PYR thus leading to an
increased frequency of CNS symptoms. In this study, the entry of PYR
into the brain was investigated in guinea pigs subjected to different
heat stress levels. In a first group, guinea pigs were maintained at
room temperature for 2 hours, their core temperature remaining stable at
about 39.8 degrees C. In a second group, animals were placed in a
climatic chamber in order to keep their core temperature at 41.5 degrees
C for 2 hours. In a third group, animals were subjected to a high
ambient temperature (42.6 degrees C) during about 2 hours and developed
heatstroke symptoms, their core temperature progressively increasing and
reaching around 44.3 degrees C. In each group, the stress of the animals
was assessed by measuring the increase of plasma cortisol level. PYR
(0.2 mg/kg, s.c.) was injected 90 minutes after beginning the
experiment. Penetration of the drug into the brain was examined by
measurement of acetylcholinesterase (AChE) activity in the cortex, the
striatum and the hippocampus of the animals 30 minutes after PYR
administration. A passage of this drug into the brain was also evaluated
autoradiographically after i.v. injection of tritiated PYR 90 minutes
after the beginning of the experiment (100 microCi/animal). Whatever the
group examined, no entry of PYR into the CNS could be detected. Exposure
to an ambient temperature at 42.6 degrees C for 2 hours resulted by
itself in a partial inhibition of cerebral AChE activity. Our results,
which agree with previous data obtained in humans exposed to heat
stress, are opposite to the recent research showing a central passage of
PYR in mice following a forced swim stress test. This demonstrated that
the penetration of PYR into the brain of rodents under stress depends on
the experimental conditions used (animal species, nature of the
stressor, etc.). Extrapolations to humans of results primarily obtained
in rodents about central passage of a drug under stress must thus be
done very carefully.

271\. Friedman, A., *et al.*, *Pyridostigmine brain penetration under
stress enhances neuronal excitability and induces early immediate
transcriptional response.* Nat Med, 1996. 2(12): p. 1382-5.
Pyridostigmine, a carbamate acetylcholinesterase (AChE) inhibitor, is
routinely employed in the treatment of the autoimmune disease myasthenia
gravis. Pyridostigmine is also recommended by most Western armies for
use as pretreatment under threat of chemical warfare, because of its
protective effect against organophosphate poisoning. Because of this
drug\'s quaternary ammonium group, which prevents its penetration
through the blood-brain barrier, the symptoms associated with its
routine use primarily reflect perturbations in peripheral nervous system
functions. Unexpectedly, under a similar regimen, pyridostigmine
administration during the Persian Gulf War resulted in a greater than
threefold increase in the frequency of reported central nervous system
symptoms. This increase was not due to enhanced absorption (or decreased
elimination) of the drug, because the inhibition efficacy of serum
butyryl-cholinesterase was not modified. Because previous animal studies
have shown stress-induced disruption of the blood-brain barrier, an
alternative possibility was that the stress situation associated with
war allowed pyridostigmine penetration into the brain. Here we report
that after mice were subjected to a forced swim protocol (shown
previously to simulate stress), an increase in blood-brain barrier
permeability reduced the pyridostigmine dose required to inhibit mouse
brain AChE activity by 50% to less than 1/100th of the usual dose. Under
these conditions, peripherally administered pyridostigmine increased the
brain levels of c-fos oncogene and AChE mRNAs. Moreover, in vitro
exposure to pyridostigmine increased both electrical excitability and
c-fos mRNA levels in brain slices, demonstrating that the observed
changes could be directly induced by pyridostigmine. These findings
suggest that peripherally acting drugs administered under stress may
reach the brain and affect centrally controlled functions.

272\. Tian, H., *et al.*, *Neither forced running nor forced swimming
affect acute pyridostigmine toxicity or brain-regional cholinesterase
inhibition in rats.* Toxicology, 2002. 176(1-2): p. 39-50.
Stress-induced change in the distribution of the drug pyridostigmine
(PYR) has been proposed as a contributing factor to unexplained
illnesses in Persian Gulf War veterans. We evaluated the effects of two
stress models, forced running and forced swimming, on acute PYR (30
mg/kg, p.o.) toxicity and cholinesterase (ChE) inhibition in the blood
and selected brain regions of young adult male Sprague-Dawley rats (6
weeks of age). Plasma corticosterone levels were measured at 0, 1 and 3
h after termination of forced swimming or forced running to confirm the
induction of stress. PYR was given either immediately before stress (15
min swimming; 20 min running) or immediately after stress (15 min
swimming; 90 min running) and cholinergic toxicity and ChE inhibition
were evaluated at 1, 2 or 4 h after PYR exposure. Additionally, rats
were subjected to either swimming (15 min) or running (90 min) stress,
anesthetized, injected with horseradish peroxidase (HRP, 100 mg/kg,
transcardial) and brain-regional HRP activity measured as an indicator
of altered blood-brain barrier integrity. Both forced swimming and
forced running resulted in significant elevations of plasma
corticosterone levels. PYR caused cholinergic toxicity at all
time-points evaluated. Swimming and running stress had little influence
on expression of PYR-induced toxicity, however. Blood ChE activity was
generally inhibited 77-91% at 1-4 h after PYR, but rats pretreated with
PYR prior to forced swimming showed lesser inhibition (64%) 1 h after
dosing, possibly because of swimming-induced hypothermia and delayed
absorption of the drug. Minimal changes in ChE activity were noted in
frontal cortex, cerebellum and hippocampus following PYR exposure
(maximal inhibition 28%), and neither swimming nor running stress
affected the degree of inhibition. Neither stress model increased HRP
accumulation in any brain region. The results suggest that stress
associated with forced running or forced swimming has little effect on
acute PYR toxicity, entry of PYR into the brain or PYR-induced
brain-regional ChE inhibition.

273\. Cook, W.O., *et al.*, *Comparison of effects of anatoxin-a(s) and
paraoxon, physostigmine and pyridostigmine on mouse brain cholinesterase
activity.* Toxicon, 1988. 26(8): p. 750-3. Anatoxin-a(s), an alkaloid
neurotoxin from the freshwater cyanobacterium, Anabaena flos-aquae
NRC-525-17, was compared to paraoxon, physostigmine and pyridostigmine
for effects on brain cholinesterase after i.p. injection into Balb/c
mice. The duration of clinical signs in mice injected with anatoxin-a(s)
persisted longer than in mice given the carbamates and was comparable
with that of paraoxon. Anatoxin-a(s) did not inhibit brain
cholinesterase activity suggesting that this toxin is unable to cross
the blood-brain barrier.

274\. Lin, J., J.J. Freeman, and J.W. Kosh, *Antagonism of acute
physostigmine and neostigmine toxicity in mice by hemicholinium-3.* Res
Commun Chem Pathol Pharmacol, 1987. 56(1): p. 137-40. Hemicholinium-3
(HC-3) was administered intraperitoneally to mice concurrently with the
intraperitoneal administration of physostigmine or neostigmine. HC-3
increased the LD50 values for both physostigmine and neostigmine but did
not alter the effect on brain ACh levels produced by these agents. Since
HC-3 does not cross the blood brain barrier after intraperitoneal
administration, the antidoting action of HC-3 is peripherally mediated
and does not solely involve an inhibition of ACh synthesis. The increase
in brain acetylcholine caused by neostigmine was related to a reduction
in acetylcholinesterase activity, providing evidence that
intraperitoneally administered neostigmine crosses the blood-brain
barrier.

275\. Kamikawa, T., *et al.*, *Effects of coenzyme Q10 on exercise
tolerance in chronic stable angina pectoris.* Am J Cardiol, 1985. 56(4):
p. 247-51. The effects of coenzyme Q10(CoQ10) on exercise performance
were studied in 12 patients, average age 56 years, with stable angina
pectoris. The study involved a double-blind, placebo-controlled,
randomized, crossover protocol, using multistage treadmill exercise
tests. CoQ10(150 mg/day in 3 daily doses) was administered orally for 4
weeks, tended to reduce anginal frequency from 5.3 +/- 4.9 to 2.5 +/-
3.3 attacks for 2 weeks and nitroglycerin consumption from 2.6 +/- 2.8
to 1.3 +/- 1.7 tablets for 2 weeks compared with patients receiving the
placebo, but the reduction was not statistically significant. Exercise
time increased from 345 +/- 102 seconds with placebo to 406 +/- 114
seconds during CoQ10 treatment (p less than 0.05). The time until 1 mm
of ST-segment depression occurred increased from 196 +/- 76 seconds with
placebo to 284 +/- 104 seconds during CoQ10 treatment (p less than
0.01). During the exercise test, ST-segment depression, heart rate and
pressure-rate product at the same and at the maximal workload showed no
significant difference between patients after placebo and CoQ10
administration. The average CoQ10 plasma concentration increased from
0.95 +/- 0.48 microgram/ml to 2.20 +/- 0.98 microgram/ml after CoQ10
treatment. This increase was significantly related to the increase in
exercise duration (r = 0.68, p less than 0.001). Only 1 patient had a
loss of appetite, but continued therapy. This study suggests that CoQ10
is a safe and promising treatment for angina pectoris.

276\. Overvad, K., *et al.*, *Coenzyme Q10 in health and disease.* Eur J
Clin Nutr, 1999. 53(10): p. 764-70. The literature concerning the
importance of coenzyme Q10 in health and disease has been reviewed.
Usual dietary intake together with normal in vivo synthesis seems to
fulfil the demands for Q10 in healthy individuals. The importance of Q10
supplementation for general health has not been investigated in
controlled experiments. The literature allows no firm conclusions about
the significance of Q10 in physical activity. In different
cardiovascular diseases, including cardiomyopathy, relatively low levels
of Q10 in myocardial tissue have been reported. Positive clinical and
haemodynamic effects of oral Q10 supplementation have been observed in
double-blind trials, especially in chronic heart failure. These effects
should be further examined. No important adverse effects have been
reported from experiments using daily supplements of up to 200 mg Q10
for 6-12 months and 100 mg daily for up to 6 y.

277\. Thorsteinsdottir, B., *et al.*, *No difference in ubiquinone
concentration of muscles and blood in fibromyalgia patients and healthy
controls.* Clin Exp Rheumatol, 1998. 16(4): p. 513-4.

278\. Lister, R.E., *An open, pilot study to evaluate the potential
benefits of coenzyme Q10 combined with Ginkgo biloba extract in
fibromyalgia syndrome.* J Int Med Res, 2002. 30(2): p. 195-9. An open,
uncontrolled study was undertaken to measure the subjective effects of
coenzyme Q10 combined with a Ginkgo biloba extract in volunteer subjects
with clinically diagnosed fibromyalgia syndrome. Anecdotal reports from
patients with fibromyalgia syndrome have claimed benefits from the use
of these supplements. The aim of this study was to determine if these
reports could be substantiated in a pilot clinical trial. Patient
questioning had determined that poor quality of life was a major factor
in the condition and a quality-of-life questionnaire was used to measure
potential benefit. Subjects were given oral doses of 200 mg coenzyme Q10
and 200 mg Ginkgo biloba extract daily for 84 days. Quality of life was
measured, using the well-validated Dartmouth Primary Care Cooperative
Information Project/World Organization of Family Doctors (COOP/WONCA)
questionnaire that measures seven different subjective responses, at 0-,
4-, 8-, and 12-week intervals. The subjects were asked for an overall
self-rating at the end of the study. A progressive improvement in the
quality-of-life scores was observed over the study period and at the
end, the scores showed a significant difference from those at the start.
This was matched by an improvement in self-rating with 64% claiming to
be better and only 9% claiming to feel worse. Adverse effects were
minor. A controlled study is now planned.

279\. Sneed, P.A., *Treatment of fibromyalgia with ubiquinone 10 and
succinic acid.* Official Gazette of the United States Patent and
Trademark Office Patents, 2002. 1255(3): p. No Pagination. A method is
described for using a combination of ubiquinone 10 and succinic acid in
the treatment of human patients afflicted with fibromyalgia to alleviate
one or more symptoms associated with that disease state. Fibromyalgia
positive patients treated buccally, sublingually or by oral ingestion
administration of ubiquinone 10 and succinic acid enjoy a reduction in
clinical symptoms of the disease.

280\. Krieger, A.C. and N.S. Redeker, *Obstructive sleep apnea syndrome:
its relationship with hypertension.* J Cardiovasc Nurs, 2002. 17(1): p.
1-11. Respiratory disturbance during sleep, in its most common form,
obstructive sleep apnea, is a prevalent condition in the general
population. During the past several years, researchers have investigated
a possible link between obstructive sleep apnea and hypertension. This
article discusses the available epidemiologic and clinical evidence
supporting this link. The possible mechanisms leading to the development
of hypertension in patients with obstructive sleep apnea will also be
discussed, as well as the effects of therapeutic interventions on
arterial blood pressure. Last, the clinical importance of such
interaction, which may lead to excessive cardiovascular morbidity and
mortality, will be addressed.

281\. Blankfield, R.P. and S.J. Zyzanski, *Bilateral leg edema,
pulmonary hypertension, and obstructive sleep apnea: a cross-sectional
study.* J Fam Pract, 2002. 51(6): p. 561-4. This study was undertaken to
clarify whether pulmonary hypertension is a useful marker for underlying
obstructive sleep apnea in patients with edema. Twenty-eight ambulatory
adults with bilateral leg edema and a normal echocardiogram were
enrolled. Sixteen subjects had pulmonary hypertension, and 12 subjects
had normal pulmonary artery pressures. Spirometry, pulse oximetry on
room air, and polysomnography were obtained for each subject. Ten of 16
(63%) pulmonary hypertension subjects and 9 of 12 (75%) nonpulmonary
hypertension subjects had obstructive sleep apnea (P =.48). Eleven of 16
(69%) pulmonary hypertension subjects and 11 of 12 (92%) nonpulmonary
hypertension subjects were obese (P =.20). If these results are
generalizable, obstructive sleep apnea is frequently associated with
bilateral leg edema and obesity, regardless of the presence of pulmonary
hypertension. Thus, especially in obese patients, bilateral leg edema
may be a useful clinical marker for underlying obstructive sleep apnea.

282\. Yamakawa, H., *et al.*, *Pulmonary hypertension in patients with
severe obstructive sleep apnea.* Psychiatry Clin Neurosci, 2002. 56(3):
p. 311-2. Thirty-seven patients (35 men and two women) with obstructive
sleep apnea-hypopnea syndrome (OSAHS) without any known cardiovascular
and lung diseases were examined by Doppler echocardiography. Eight of
the 37 (21.6%) patients experienced daytime pulmonary hypertension (PH),
and all of them had severe OSAHS with an apnea-hypopnea index of \> 30.
The study suggested that one-third of patients with severe OSAHS had
daytime PH.

283\. Lee, A.G., *et al.*, *Sleep apnea and intracranial hypertension in
men.* Ophthalmology, 2002. 109(3): p. 482-5. PURPOSE: To investigate
sleep apnea as an associated finding in idiopathic intracranial
hypertension (IIH) in men. DESIGN: Multicenter, retrospective,
noncomparative interventional case series. METHODS: Retrospective review
of all men with the diagnosis of IIH seen within the last 5 years at
three tertiary care academic ophthalmologic institutions. Cases with
sleep apnea (SA) and IIH were identified and reviewed. RESULTS:
Thirty-two cases of IIH in men were reviewed. Six cases with SA met the
modified Dandy criteria for the diagnosis of IIH. Of these six patients,
one received acetazolamide alone, four received acetazolamide and
continuous positive airway pressure (CPAP), and one was treated with
CPAP alone. All patients had preserved central acuity (20/20 or better
in both eyes), enlarged blind spots, and optic disc edema in both eyes.
Five patients had normal visual fields after treatment, and one patient
had residual visual field loss. Three patients had normal optic nerve
examinations, with resolution of the optic disc edema at last follow-up.
After resolution of the optic disc edema, these three patients were
maintained on CPAP but discontinued acetazolamide. Two patients had
persistent but improved papilledema and are under continued treatment
with acetazolamide and CPAP. One patient had optic disc pallor in both
eyes and is stable. CONCLUSIONS: SA was a common finding in men meeting
the modified Dandy criteria for IIH in adults. Treatment of sleep apnea
with nocturnal oxygenation may improve the signs and symptoms of IIH in
affected men.

284\. Blankfield, R.P., A.A. Tapolyai, and S.J. Zyzanski, *Left
ventricular dysfunction, pulmonary hypertension, obesity, and sleep
apnea.* Sleep Breath, 2001. 5(2): p. 57-62. The purpose of this study
was to determine the frequency of central and obstructive sleep apnea in
adult patients who have echocardiographic evidence of left ventricular
dysfunction and pulmonary hypertension. Subjects with left ventricular
dysfunction, pulmonary hypertension (pulmonary artery systolic pressure
\>30 mm Hg) and no lung disease were evaluated for risk factors
associated with pulmonary hypertension. Of eight eligible adults, six
completed the study. Subjects were from suburban and inner city family
practices. Spirometric assessment, pulse oximetry on room air,
rheumatologic evaluation, polysomnography, and additional history were
taken. All six subjects had sleep apnea (apnea-plus-hypopnea index, or
AHI, \> or = 20): obstructive, central, or mixed. All were obese, and
almost all the subjects had a restrictive pattern on spirometry, which
is consistent with obesity. All had a pulmonary artery systolic blood
pressure of 35 mm Hg or greater. None had daytime hypoxemia or collagen
vascular disease, and none had ever used appetite suppressants. This
study found a strong association between pulmonary hypertension and
obstructive or central sleep apnea in obese patients with congestive
heart failure (CHF). We propose that a pulmonary artery systolic
pressure of 35 mm Hg or greater in ambulatory patients with CHF may
signify an increased risk of sleep apnea.

285\. Silverberg, D.S., A. Iaina, and A. Oksenberg, *Treating
obstructive sleep apnea improves essential hypertension and quality of
life.* Am Fam Physician, 2002. 65(2): p. 229-36. About one half of
patients who have essential hypertension have obstructive sleep apnea,
and about one half of patients who have obstructive sleep apnea have
essential hypertension. A growing body of evidence suggests that
obstructive sleep apnea is a major contributing factor in the
development of essential hypertension. Despite many patients with
obstructive sleep apnea having clear symptoms of the disorder, an
estimated 80 to 90 percent of cases are undiagnosed. When physicians
routinely seek the diagnosis of obstructive sleep apnea by asking
patients (especially those with hypertension) three basic sleep-related
questions about snoring, excessive daytime sleepiness and reports of
witnessed apneic events, the number of cases diagnosed and treated
increases by about eightfold. Eliminating snoring and occurrences of
apneic-hypopneic episodes will dramatically improve patients\' quality
of sleep and eliminate excessive daytime sleepiness, which has a
detrimental effect on general functioning. Increased alertness will
reduce the likelihood that patients will be involved in motor vehicle
crashes. In most studies in which blood pressure was measured following
treatment for obstructive sleep apnea, daytime and nighttime blood
pressure levels were found to decrease significantly. This decrease in
blood pressure may also reduce the likelihood of cardiovascular
complications. The key to the diagnosis of obstructive sleep apnea is
physician knowledge about the disorder. The dramatic improvement in
quality of life that occurs when patients are successfully treated for
obstructive sleep apnea makes detecting and treating this disorder
imperative.

286\. Chaska, B.W., *Obstructive sleep apnea and essential
hypertension\--is there a link?* Am Fam Physician, 2002. 65(2): p. 182,
185-6.

287\. Alchanatis, M., *et al.*, *Daytime pulmonary hypertension in
patients with obstructive sleep apnea: the effect of continuous positive
airway pressure on pulmonary hemodynamics.* Respiration, 2001. 68(6): p.
566-72. BACKGROUND: Limited information exists regarding the development
of pulmonary hypertension in patients with obstructive sleep apnea (OSA)
in the absence of lung and heart comorbidity. OBJECTIVES: The aims of
this study were to investigate whether OSA patients without any other
cardiac or lung disease develop pulmonary hypertension, and to assess
the effect of continuous positive airway pressure (CPAP) treatment on
pulmonary artery pressure (P(PA)). METHODS: Twenty-nine patients aged 51
+/- 10 years with OSA and 12 control subjects were studied with
pulsed-wave Doppler echocardiography for estimation of P(PA) before and
after 6-month effective treatment with CPAP. RESULTS: A significantly
higher mean P(PA) was found in OSA patients as compared to control
subjects (17.2 +/- 5.2 vs. 12.1 +/- 1.9 mm Hg, p \< 0.001). Six out of
the 29 OSA patients had mild pulmonary hypertension (P(PA) \> or = 20 mm
Hg). Significant differences were observed between pulmonary
hypertensive and normotensive OSA patients with respect to age (62 +/- 4
vs. 48 +/- 15 years, respectively, p \< 0.05), body mass index (41 +/- 7
vs. 32 +/- 4 kg/m(2), p \< 0.02) and daytime P(a)O(2) (81 +/- 9 vs. 92
+/- 9 mm Hg, p \< 0.05). CPAP treatment was effective in reducing mean
P(PA) in both groups of pulmonary hypertensive and normotensive OSA
patients (decreases in P(PA) from 25.6 +/- 4.0 to 19.5 +/- 1.5 mm Hg, p
\< 0.001; from 14.9 +/- 2.2 to 11.5 +/- 2.0 mm Hg, respectively, p \<
0.001). CONCLUSIONS: A proportion (20.7%) of OSA patients without any
other lung or heart disease and characterized by older age, greater
obesity and lower daytime oxygenation develop mild pulmonary
hypertension which has been partially or completely reversed after
6-month CPAP treatment. In conclusion, OSA alone constitutes an
independent risk factor for the development of pulmonary hypertension.

288\. Richert, A., K. Ansarin, and A.S. Baran, *Sleep apnea and
hypertension: pathophysiologic mechanisms.* Semin Nephrol, 2002. 22(1):
p. 71-7. This article reviews the pathophysiology of hypertension (HTN)
in obstructive sleep apnea (OSA). The article is divided into 3
sections. The first section describes epidemiologic studies of the
relationship of sleep-related breathing disorders, including OSA, to HTN
and argues that OSA contributes to the genesis of HTN. The second
section describes the known immediate physiologic consequences of 3
components of OSA that may contribute to the genesis of persistent
systemic HTN. The 3 components are (1) the large negative intrathoracic
pressure changes associated with OSA, (2) intermittent hypoxemia, and
(3) arousal from sleep. The last section reviews current physiologic
models of essential HTN genesis and attempts to integrate them with the
suspected HTN-generating aspects of OSA. In its summary, the authors
conclude that OSA contributes to the genesis of HTN and advise
physicians not to ignore the contribution of frequently comorbid non-OSA
factors, such as obesity, to the genesis of OSA-related HTN.

289\. Lavie, P., *et al.*, *Obstructive sleep apnea and hypertension:
from correlative to causative relationship.* J Clin Hypertens
(Greenwich), 2001. 3(5): p. 296-301. Sleep-disordered breathing,
manifested by repetitive episodes of partial or complete cessation of
breathing during sleep associated with brief arousal and autonomic
activation, is estimated to affect as many as 4% of adult men and 2% of
adult women. Studies conducted during the 1980s revealed a strong
association between sleep-disordered breathing and hypertension. The
results of these early studies, which relied on relatively small samples
of patients, have been confirmed in recent years by large-scale
epidemiologic studies that are controlled for all possible confounding
factors. This paper reviews the evidence suggesting a causative
relationship between hypertension and disordered breathing in sleep. The
authors discuss the possible underlying mechanisms of the two entities
and address the clinical implications of this relationship. They
conclude by recommending a proactive approach to the diagnosis of
breathing disorders in sleep, in order to prevent the cardiovascular
sequelae of this syndrome.

290\. von Kanel, R., *et al.*, *The hypercoagulable state in sleep apnea
is related to comorbid hypertension.* J Hypertens, 2001. 19(8): p.
1445-51. OBJECTIVE: Obstructive sleep apnea (OSA) is associated with
increased prevalence of atherosclerotic disease. A hypercoagulable state
thought to underly atherosclerosis has been described in both OSA and
systemic hypertension. We wondered about the respective contribution of
apnea and hypertension to a hypercoagulable state. DESIGN: Eighty-seven
subjects with symptoms suggestive of OSA, mean age 47 years (range 32-64
years), underwent polysomnography and blood pressure (BP) screening. OSA
was diagnosed when respiratory disturbance index (RDI) \> or = 15.
Subjects having systolic BP (SBP) \> 140 mmHg and/or diastolic BP (DBP)
\> 90 mmHg were classified as having hypertension. Three
hypercoagulability markers were measured: thrombin/antithrombin III
complex (TAT), fibrin D-dimer (DD), and von Willebrand factor antigen
(vWF:ag). RESULTS: Analysis of variance and multiple linear regression
were performed on the following four subject groups: (1) normotensive
non-apneics (n = 19), (2) normotensive apneics (n = 38), (3)
hypertensive non-apneics (n = 11), and (4) hypertensive apneics (n =
19). OSA (groups 2 and 4) had no significant main effect on hemostasis.
Hypertensives (groups 3 and 4) had higher plasma levels of TAT
(median/inter-quartile range, 148/59-188 versus 77/53-108 pmol/l; P =
0.009) and of DD (376/265-721 versus 303/190-490 ng/ml; P = 0.040) than
normotensives (groups 1 and 2). Across all subjects, SBP was the only
significant predictor of TAT (P = 0.001) and of DD (P = 0.004), whereas
DBP was the only significant predictor of vWF:ag (P = 0.029). These
findings persisted even after controlling for gender, age, body mass
index, RDI, mean SaO2, and hematocrit. CONCLUSION: Hypercoagulability in
OSA is mediated by comorbid hypertension and might account for high
cardiovascular morbidity in OSA in general.

291\. Sanders, M.H., *Article reviewed: Association of sleep-disordered
breathing, sleep apnea, and hypertension in a large community-based
study.* 2000. 1(4): p. 327-328.

292\. Otsuka, K., *\[Nasal CPAP treatment and hypertension and altered
cardiovascular variability associated with obstructive sleep apnea
(OSA)\].* Nippon Rinsho, 2001. 59(5): p. 983-91. In the past 5 years
several epidemiologic studies have demonstrated that sleep-related
breathing disorders are an independent risk factor for hypertension,
probably resulting from a combination of repetitive episodes of hypoxia,
hypercapnea, arousals, and a striking surge in sympathetic excitation,
and altered baroreflex control during sleep. Obstructive sleep apnea
(OSA) may lead to the cardiac arrhythmias and myocardial ischemia and it
is a possible risk factor for stroke. We confirmed that nasal CPAP has
been shown to lower blood pressure in some hypertensive OSA patients.
Early recognition and treatment of sleep-apnea may improve
cardiovascular function.

293\. Narkiewicz, K. and V.K. Somers, *Obstructive sleep apnea as a
cause of neurogenic hypertension.* Curr Hypertens Rep, 1999. 1(3): p.
268-73. Abnormalities in neural circulatory control may contribute
importantly to the hypertensive state. The sympathetic nervous system in
particular is a key mechanism for increasing blood pressure. Patients
with obstructive sleep apnea have increased sympathetic activity.
Obesity or other coexisting disease states do not explain the heightened
sympathetic drive. This review examines the evidence linking sleep apnea
with hypertension and the possible role of excessive sympathetic drive
as a mediator of higher blood pressure in sleep apnea. Abnormalities in
reflex circulatory control that could act to increase sympathetic
activity in sleep apnea are also discussed.

294\. Shiomi, T., *et al.*, *Primary pulmonary hypertension with central
sleep apnea: sudden death after bilevel positive airway pressure
therapy.* Jpn Circ J, 2000. 64(9): p. 723-6. An obese 23-year-old man
with sleep-disordered breathing and primary pulmonary hypertension (PPH)
had been administered oral beraprost sodium, anticoagulant warfarin, and
home oxygen therapy, at another hospital as treatment for the PPH, but
he had not experienced any symptomatic improvement. The patient had a
body mass index of 32.4kg/m2, and complained of fatigue, shortness of
breath on exertion, excessive daytime sleepiness, and snoring. Arterial
blood gas analysis showed a PaO2 and a PaCO2 of 70.9 and 31.2mmHg,
respectively. A polysomnographic study revealed central sleep apnea with
an apnea-hypopnea index (AHI) of 29.7episodes/h. The patient showed
improvement of daytime sleepiness after starting nocturnal nasal bilevel
positive airway pressure (BiPAP) therapy for the central sleep apnea,
but his pulmonary hypertension, measured in the daytime, worsened. The
patient died suddenly while walking to the bathroom in the morning 1
month after initiation of BiPAP therapy. It is necessary to consider the
possibility of sudden death when nasal BiPAP therapy is given to a PPH
patient with central sleep apnea.

295\. Hudgel, D.W., *Beyond systemic hypertension: understanding cardiac
dysfunction in obstructive sleep apnea.* Respiration, 2000. 67(4): p.
360-1.

296\. Salo, T.M., *et al.*, *Comparison of autonomic withdrawal in men
with obstructive sleep apnea syndrome, systemic hypertension, and
neither condition.* Am J Cardiol, 2000. 85(2): p. 232-8. Obstructive
sleep apnea syndrome is characterized by obesity, nocturnal breathing
abnormalities, arterial hypertension, and an increased number of
cardiovascular events. Sympathetic activity is increased during
nocturnal apneic episodes, which may mediate the cardiovascular
complications of sleep apnea. We studied 15 male subjects with
obstructive sleep apnea syndrome and associated hypertension, 54
subjects with mild to moderate essential hypertension, and 25 healthy
normotensive men. Cardiovascular autonomic control was assessed using
frequency domain measures of heart rate variability (HRV) during a
controlled breathing test and during orthostatic maneuver. Compared with
normotensive and hypertensive groups, total power and low- and
high-frequency components of HRV during controlled breathing were
significantly (analysis of variance, p\<0.0001) lower in the obstructive
sleep apnea syndrome. During the orthostatic maneuver, the change in
total power of HRV was different between the 3 groups (analysis of
variance, p = 0.004). The total power of HRV tended to increase in the
normotensive (4.11+/-12.29 ms2) and in hypertensive (2.31+/-12.65 ms2)
groups, but decreased (1.13+/-1.23 ms2) in the hypertensive group with
obstructive sleep apnea syndrome. According to multivariate regression
analysis, age and sleep apnea were the major independent determinants of
HRV. This study found that an abnormal response to autonomic nervous
tests characterizes hypertension in overweight subjects with obstructive
sleep apnea syndrome. This could be due to autonomic withdrawal or
supersaturation of the end-organ receptors by excessive and prolonged
sympathetic stimulation. Our results also show the reduced response of
orthostatic maneuver and controlled breathing in the hypertensive group
compared with the normotensive group.

297\. Willis, R.A., *et al.*, *The effect of ethanol and/or food
restriction on coenzyme Q in liver in rats.* Mol Aspects Med, 1997. 18
Suppl: p. S205-11. The individual and combined effects of ethanol and
lovastatin on rats and their prevention by supplemental coenzyme Q10
(CoQ10) was studied. The ethanol and lovastatin findings are reported
elsewhere. This paper focuses on the food restriction which occurred in
rats fed 35% of energy as ethanol and those control rats pair-fed to the
35% of energy as ethanol group. Six groups of rats received 35% of
energy as ethanol (with or without lovastatin and/or CoQ10 treatment).
One group served as a 0% ethanol ad libitum control and one 0% ethanol
control group was pair-fed to the 35% ethanol group. Rats receiving 35%
of energy as ethanol and their pair-fed controls consumed 83% of the
energy/day consumed by the ad libitum controls. This was consistent
regardless of lovastatin or CoQ10 treatment. Weight gains were 84% of
control. The energy reduction was consistently associated with a
substantial (48%+) increase in liver CoQ9 concentrations. Reports by
others of associations between food restriction and increased longevity
in rodents has focused on a decrease in oxidant damage in tissues of
food restricted animals. The increase in CoQ levels in the food
restricted animals would result in an increase in antioxidant protection
and might explain the observed increases in longevity.

298\. Silvestre-Aillaud, P., *et al.*, *Could coenzyme Q10 and
L-carnitine be a treatment for diabetes secondary to 3243 mutation of
mtDNA?* Diabetologia, 1995. 38(12): p. 1485-6.

299\. Kucharska, J., *et al.*, *Deficit of coenzyme Q in heart and liver
mitochondria of rats with streptozotocin-induced diabetes.* Physiol Res,
2000. 49(4): p. 411-8. Mitochondrial dysfunction and oxidative stress
participate in the development of diabetic complications, however, the
mechanisms of their origin are not entirely clear. Coenzyme Q has an
important function in mitochondrial bioenergetics and is also a powerful
antioxidant. Coenzyme Q (CoQ) regenerates alpha-tocopherol to its active
form and prevents atherogenesis by protecting low-density lipoproteins
against oxidation. The aim of this study was to ascertain whether the
experimentally induced diabetes mellitus is associated with changes in
the content of endogenous antioxidants (alpha-tocopherol, coenzymes Q9
and Q10) and in the intensity of lipoperoxidation. These biochemical
parameters were investigated in the blood and in the isolated heart and
liver mitochondria. Diabetes was induced in male Wistar rats by a single
intravenous injection of streptozotocin (45 mg x kg(-1)), insulin was
administered once a day for 8 weeks (6 U x kg(-1)). The concentrations
of glucose, cholesterol, alpha-tocopherol and CoQ homologues in the
blood of the diabetic rats were increased. The CoQ9/cholesterol ratio
was reduced. In heart and liver mitochondria of the diabetic rats we
found an increased concentration of alpha-tocopherol, however, the
concentrations of CoQ9 and CoQ10 were decreased. The formation of
malondialdehyde was enhanced in the plasma and heart mitochondria. The
results have demonstrated that experimental diabetes is associated with
increased lipoperoxidation, in spite of the increased blood
concentrations of antioxidants alpha-tocopherol and CoQ. These changes
may be associated with disturbances of lipid metabolism in diabetic
rats. An important finding is that heart and liver mitochondria from the
diabetic rats contain less CoQ9 and CoQ10 in comparison with the
controls. We suppose that the deficit of coenzyme Q can participate in
disturbances of mitochondrial energy metabolism of diabetic animals.

300\. Liou, C.W., *et al.*, *Correction of pancreatic beta-cell
dysfunction with coenzyme Q(10) in a patient with mitochondrial
encephalomyopathy, lactic acidosis and stroke-like episodes syndrome and
diabetes mellitus.* Eur Neurol, 2000. 43(1): p. 54-5.

301\. Kishi, T., *et al.*, *Bioenergetics in clinical medicine. XI.
Studies on coenzyme Q and diabetes mellitus.* J Med, 1976. 7(3-4): p.
307-21. The activity of the succinate dehydrogenase-coenzyme Q10
reductase from 120 diabetic patients was significantly lower (P less
than 0.001) and the per cent deficiency was significantly higher (P less
than 0.001) than that of the controls. The diabetes of 37 patients was
controlled by diet; the enzyme activity was lower (P less than 0.001)
and the deficiency was higher (P less than 0.02) than for controls. In
decreasing effectiveness, Dymelor, Glyburide, Phenformin and Tolazamide
inhibited the COQ10-enzyme, NADH-oxidase. Tolbutamide, Glypizide, and
Chlorpropamide were noninhibitory to succinoxidase and NADH-oxidase.
Patients receiving Tolazamide and Phenformin showed a higher incidence
(P less than 0.001 to P less than 0.05) of COQ10-deficiency than
patients controlled by diet or normal controls. Certain diabetic
patients controlled by diet may have a deficiency of COQ10 which may be
enhanced by the inhibition by certain commonly used antidiabetic drugs
of COQ10-enzymes. A deficiency of COQ10 in the pancreas could impair
bioenergetics, the generation of ATP, and the biosynthesis of insulin.

302\. Iwasaki, Y., N. Wakata, and M. Kinoshita, *Parkinsonism induced by
pyridostigmine.* Acta Neurol Scand, 1988. 78(3): p. 236.

303\. Iwasaki, Y., M. Kinoshita, and N. Wakata, *\[A case report of
pyridostigmine induced parkinsonism\].* Rinsho Shinkeigaku, 1988. 28(6):
p. 662-4.

304\. Gothoni, P., M. Lehtinen, and M. Fincke, *Drugs for Parkinson\'s
disease reduce tremor induced by physostigmine.* Naunyn Schmiedebergs
Arch Pharmacol, 1983. 323(3): p. 205-10. The effects of anticholinergic
and dopaminergic drugs used for Parkinson\'s disease were studied on the
tremor induced by physostigmine (0.3-3.0 mg/kg) in rats. For the
measurement of tremor a new electronic device was employed. Atropine
(0.3-1.2 mg/kg) and biperiden (0.01-1.0 mg/kg) reduced the
physostigmine-induced tremor in a dose-related manner and could abolish
it. Biperiden was less potent than atropine. Methylatropine in a dose of
1.2 mg/kg slightly inhibited the tremor. Amantadine (0.3-3.0 mg/kg)
reduced the tremor but only to a certain degree. Bromocriptine (0.1-10.0
mg/kg) reduced it in a manner that was not dose-related. Pimozide
potentiated the tremor in the dose of 0.2 mg/kg but not in larger doses.
At the onset of the tremor, a small decrease in rectal temperature
occurred. The hypothermia lasted significantly longer than the tremor.
Neither the anticholinergic nor the dopaminergic anti-Parkinson drugs
altered the hypothermic effect of physostigmine. The results show that
those anti-Parkinson drugs, which act by increasing the dopaminergic
activity can counteract the tremor induced by physostigmine. However,
these drugs are clearly less active than th anticholinergic
anti-Parkinson drugs.

305\. Friedrich, M.J., *Pesticide study aids Parkinson research.* Jama,
1999. 282(23): p. 2200.

306\. Semchuk, K.M., E.J. Love, and R.G. Lee, *Parkinson\'s disease and
exposure to agricultural work and pesticide chemicals.* Neurology, 1992.
42(7): p. 1328-35. This population-based case-control study of 130
Calgary residents with neurologist-confirmed idiopathic Parkinson\'s
disease (PD) and 260 randomly selected age- and sex-matched community
controls attempted to determine whether agricultural work or the
occupational use of pesticide chemicals is associated with an increased
risk for PD. We obtained by personal interviews lifetime occupational
histories, including chemical exposure data, and analyzed the data using
conditional logistic regression for matched sets. In the univariate
analysis, a history of field crop farming, grain farming, herbicide use,
or insecticide use resulted in a significantly increased crude estimate
of the PD risk, and the data suggested a dose-response relation between
the PD risk and the cumulative lifetime exposure to field crop farming
and to grain farming. However, in the multivariate analysis, which
controlled for potential confounding or interaction between the exposure
variables, previous occupational herbicide use was consistently the only
significant predictor of PD risk. These results support the hypothesis
that the occupational use of herbicides is associated with an increased
risk for PD.

307\. Ritz, B. and F. Yu, *Parkinson\'s disease mortality and pesticide
exposure in California 1984-1994.* Int J Epidemiol, 2000. 29(2): p.
323-9. BACKGROUND: In the last two decades reports from different
countries emerged associating pesticide and herbicide use with
Parkinson\'s disease (PD). California growers use approximately 250
million pounds of pesticides annually, about a quarter of all pesticides
used in the US. METHODS: We employed a proportional odds mortality
design to compare all cases of PD recorded as underlying (1984-1994) or
associated causes (1984-1993) of death occurring in California with all
deaths from ischaemic heart disease (ICD-9 410-414) during the same
period. Based on pesticide use report data we classified California
counties into several pesticide use categories. Agricultural census data
allowed us to create measures of percentage of land per county treated
with pesticides. Employing logistic regression models we estimated the
effect of pesticide use controlling for age, gender, race, birthplace,
year of deaths, and education. RESULTS: Mortality from PD as the
underlying cause of death was higher in agricultural pesticide-use
counties than in non-use counties. A dose response was observed for
insecticide use per county land treated when using 1982 agricultural
census data, but not for amounts of restricted pesticides used or length
of residency in a country prior to death. CONCLUSIONS: Our data show an
increased PD mortality in California counties using agricultural
pesticides. Unless all of our measures of county pesticide use are
surrogates for other risk factors more prevalent in pesticide use
counties, it seems important to target this prevalent exposure in rural
California in future studies that use improved case finding mechanisms
and collect pesticide exposure data for individuals.

308\. Taylor, M.C., *et al.*, *Paraoxonase polymorphisms, pesticide
exposure and Parkinson\'s disease in a Caucasian population.* J Neural
Transm, 2000. 107(8-9): p. 979-83. Parkinson\'s disease (PD) has been
associated with exposure to pesticides and oxidative injury. The
involvement of paraoxonase in both pesticide metabolism and lipid
peroxidation suggests that it may play a role in the pathogenesis of PD.
We examined the frequency of polymorphic alleles of the PON1 and PON2
genes in a sample of caucasian subjects with PD. The frequency
distribution of these genotypes did not differ significantly between
patients and controls, including those who had reported exposure to
pesticides.

309\. Berger, A., *Parkinson\'s disease linked with pesticide.* Bmj,
2000. 321(7270): p. 1175A.

310\. Adam, D., *Pesticide use linked to Parkinson\'s disease.* Nature,
2000. 408(6809): p. 125.

311\. Betarbet, R., *et al.*, *Chronic systemic pesticide exposure
reproduces features of Parkinson\'s disease.* Nat Neurosci, 2000. 3(12):
p. 1301-6. The cause of Parkinson\'s disease (PD) is unknown, but
epidemiological studies suggest an association with pesticides and other
environmental toxins, and biochemical studies implicate a systemic
defect in mitochondrial complex I. We report that chronic, systemic
inhibition of complex I by the lipophilic pesticide, rotenone, causes
highly selective nigrostriatal dopaminergic degeneration that is
associated behaviorally with hypokinesia and rigidity. Nigral neurons in
rotenone-treated rats accumulate fibrillar cytoplasmic inclusions that
contain ubiquitin and alpha-synuclein. These results indicate that
chronic exposure to a common pesticide can reproduce the anatomical,
neurochemical, behavioral and neuropathological features of PD.

312\. Helmuth, L., *Neuroscience. Pesticide causes Parkinson\'s in
rats.* Science, 2000. 290(5494): p. 1068.

313\. Karen, D.J., *et al.*, *Striatal dopaminergic pathways as a target
for the insecticides permethrin and chlorpyrifos.* Neurotoxicology,
2001. 22(6): p. 811-7. Because insecticide exposure has been linked to
both Parkinsons disease and Gulf War illness, the neurotoxic actions of
pyrethroid and organophosphate insecticides on behavior and striatal
dopaminergic pathways were investigated in C57BL/6 mice treated with
permethrin (three i.p. doses at 0.2-200 mg/kg) or chlorpyrifos (three
s.c. doses at 25-100 mg/kg) over a 2-week period. Permethrin altered
maximal \[3H\]dopamine uptake in striatal synaptosomes from treated
mice, with changes in Vmax displaying a bell-shaped curve. Uptake was
increased to 134% of control at a dose of 1.5 mg/kg. At higher doses of
PM (25 mg/kg), dopamine uptake declined to a level significantly below
that of control (50% of control at 200 mg/kg, P \< 0.01). We also
observed a small, but statistically significant decrease in
\[3H\]dopamine uptake by chlorpyrifos, when given at a dose of 100
mg/kg. There was no significant effect on the Km for dopamine transport.
Evidence of cell stress was observed in measures of
mitochondrialfunction, which were reduced in mice given high-end doses
of chlorpyrifos and permethrin. Although cytotoxicity was not reflected
in decreased levels of striatal dopamine in either 200 mg/kg PM or 100
mg/kg CPF treatment groups, an increase in dopamine turnover at 100
mg/kg CPF was indicated by a significant increase in titers of the
dopamine metabolite, 3,4-dihydroxyphenylacetic acid. Both permethrin and
chlorpyrifos caused a decrease in open field behavior at the highest
doses tested. Although frank Parkinsonism was not observed, these
findings confirm that dopaminergic neurotransmission is affected by
exposure to pyrethroid and organophosphorus insecticides, and may
contribute to the overall spectrum of neurotoxicity caused by these
compounds.

314\. Ambrani, L.M. and M.H. Van Woert, *Modification of the
tremorigenic activity of physostigmine.* Br J Pharmacol, 1972. 46(2): p.
344-7.

315\. Ambani, L.M., M.H. Van Woert, and M.B. Bowers, Jr., *Physostigmine
effects on phenothiazine-induced extrapyramidal reactions.* Arch Neurol,
1973. 29(6): p. 444-6.

316\. Haley, R.W., *et al.*, *Evaluation of neurologic function in Gulf
War veterans. A blinded case-control study.* Jama, 1997. 277(3): p.
223-30. OBJECTIVE: To determine whether Gulf War-related illnesses are
associated with central or peripheral nervous system dysfunction.
DESIGN: Nested case-control study. PARTICIPANTS: Twenty-three veterans
with factor analysis-derived syndromes (the cases), 10 well veterans
deployed to the Gulf War (the deployed controls), and 10 well veterans
not deployed to the Gulf War (the nondeployed controls). METHOD: With
investigators blinded to group identities, participants underwent
objective neurophysiological, audiovestibular, neuroradiological,
neuropsychological, and blood tests. MAIN OUTCOME MEASURES: Evidence of
neurologic dysfunction. RESULTS: Compared with the 20 controls, the 23
cases had significantly more neuropsychological evidence of brain
dysfunction on the Halstead Impairment Index (P=.01), greater interside
asymmetry of the wave I to wave III interpeak latency of brain stem
auditory evoked potentials (P=.02), greater interocular asymmetry of
nystagmic velocity on rotational testing, increased asymmetry of
saccadic velocity (P=.04), more prolonged interpeak latency of the
lumbar-to-cerebral peaks on posterior tibial somatosensory evoked
potentials (on right side, P=.03, and on the left side, P=.005), and
diminished nystagmic velocity after caloric stimulation bilaterally (P
values range from .02 to .04). Cases (n=5) with syndrome 1 (\"impaired
cognition\") were the most impaired on brain stem auditory evoked
potentials (P=.005); those (n=13) with syndrome 2 (\"confusion-ataxia\")
were the most impaired on the Halstead Impairment Index (P=.006),
rotational testing (P=.01), asymmetry of saccadic velocity (P=.03), and
somatosensory evoked potentials (P\< or =.01); and those (n=5) with
syndrome 3 (\"arthro-myo-neuropathy\") were the most impaired on caloric
stimulation (P\< or =.01). CONCLUSIONS: The 3 factor-derived syndromes
identified among Gulf War veterans appear to represent variants of a
generalized injury to the nervous system.

317\. Hom, J., R.W. Haley, and T.L. Kurt, *Neuropsychological correlates
of Gulf War syndrome.* Arch Clin Neuropsychol, 1997. 12: p. 531-544.

318\. McDowell, I., *Alzheimer\'s disease: insights from epidemiology.*
Aging (Milano), 2001. 13(3): p. 143-62. While a complete understanding
of the pathogenesis of Alzheimer\'s disease (AD) remains elusive, many
conclusions can be drawn from the numerous epidemiological studies
undertaken to date. Prevalence and incidence estimates show consistency,
following a roughly exponential pattern with a doubling of both
parameters roughly every five years after age 65. Roughly 7% of the
population aged 65 and over has AD. The clinical course of the disease
is reasonably well established and mortality rates rise with increasing
levels of cognitive deficit. Four risk factors for AD are firmly
established: increasing age, the presence of the
apolipoproteinE-epsilon4 allele, familial aggregation of cases, and
Down\'s syndrome. Numerous other associations have been shown in some
studies, but not in others. For example, women generally appear at
higher risk than men, as do people with lower levels of education;
depression is probably prodromal; head injury is an established risk
factor, and may interact with the apoE gene; several occupational
exposures appear hazardous, and exposure to aluminum in the water supply
confers excess risk. Hypertension and other vascular symptoms appear to
predispose to AD, which is now seen as nosologically closer to vascular
dementia than was previously believed. Several apparently protective
factors have been identified, although preventive trials based on these
have so far shown minimal effectiveness. The use of non-steroidal
anti-inflammatory drugs to treat arthritis is associated with a reduced
risk of AD, as is estrogen use by post-menopausal women. Physical
activity appears beneficial, as does a diet with high levels of vitamins
B6, B12 and folate. while red wine in moderate quantities appears
protective. This review concludes with a discussion of the strengths and
limitations of current epidemiological methods for studying Alzheimer\'s
disease.

319\. Snowdon, D.A., *et al.*, *Linguistic ability in early life and
cognitive function and Alzheimer\'s disease in late life. Findings from
the Nun Study.* Jama, 1996. 275(7): p. 528-32. OBJECTIVE\--To determine
if linguistic ability in early life is associated with cognitive
function and Alzheimer\'s disease in late life. DESIGN\--Two measures of
linguistic ability in early life, idea density and grammatical
complexity, were derived from autobiographies written at a mean age of
22 years. Approximately 58 years later, the women who wrote these
autobiographies participated in an assessment of cognitive function, and
those who subsequently died were evaluated neuropathologically.
SETTING\--Convents in the United States participating in the Nun Study;
primarily convents in the Milwaukee, Wis, area. PARTICIPANTS\--Cognitive
function was investigated in 93 participants who were aged 75 to 95
years at the time of their assessments, and Alzheimer\'s disease was
investigated in the 14 participants who died at 79 to 96 years of age.
MAIN OUTCOME MEASURES\--Seven neuropsychological tests and
neuropathologically confirmed Alzheimer\'s disease. RESULTS\--Low idea
density and low grammatical complexity in autobiographies written in
early life were associated with low cognitive test scores in late life.
Low idea density in early life had stronger and more consistent
associations with poor cognitive function than did low grammatical
complexity. Among the 14 sisters who died, neuropathologically confirmed
Alzheimer\'s disease was present in all of those with low idea density
in early life and in none of those with high idea density.
CONCLUSIONS\--Low linguistic ability in early life was a strong
predictor of poor cognitive function and Alzheimer\'s disease in late
life.

320\. *Head injury linked to increased risk of Alzheimer\'s disease.*
FDA Consum, 2001. 35(1): p. 8.

321\. Plassman, B.L., *et al.*, *Documented head injury in early
adulthood and risk of Alzheimer\'s disease and other dementias.*
Neurology, 2000. 55(8): p. 1158-66. BACKGROUND: The association between
antecedent head injury and AD is inconsistent. OBJECTIVE: To examine the
association between early adult head injury, as documented by military
hospital records, and dementia in late life; and to evaluate the
interaction between head injury and APOE epsilon4 as risk factors for
dementia. METHODS: The study had a population-based prospective
historical cohort design. It included men who were World War II Navy and
Marine veterans, and were hospitalized during their military service
with a diagnosis of either a nonpenetrating head injury or another
unrelated condition. In 1996 to 1997, military medical records were
abstracted to document the occurrence and details of closed head injury.
The entire sample was then evaluated for dementia and AD using a
multistage procedure. There were 548 veterans with head injury and 1228
without head injury who completed all assigned stages of the study. The
authors estimated risk of dementia, specifically AD, using proportional
hazards models. RESULTS: Both moderate head injury (hazard ratio \[HR\]
= 2.32; CI = 1.04 to 5.17) and severe head injury (HR = 4.51; CI = 1.77
to 11.47) were associated with increased risk of AD. Results were
similar for dementia in general. The results for mild head injury were
inconclusive. When the authors stratified by the number of APOE epsilon4
alleles, they observed a nonsignificant trend toward a stronger
association between AD and head injury in men with more epsilon4
alleles. CONCLUSIONS: Moderate and severe head injuries in young men may
be associated with increased risk of AD and other dementias in late
life. However, the authors cannot exclude the possibility that other
unmeasured factors may be influencing this association.

322\. SoRelle, R., *Head injuries linked to Alzheimer\'s disease.*
Circulation, 2000. 102(18): p. E9036-7.

323\. Sobel, R.K., *Alzheimer\'s first blow. Head injuries raise risk.*
US News World Rep, 2000. 129(18): p. 54.

324\. Norris, F.H., *et al.*, *Trial of oral physostigmine in
amyotrophic lateral sclerosis.* Clin Pharmacol Ther, 1993. 54(6): p.
680-2. We evaluated a double-blind, placebo-controlled, and
double-crossover trial of oral physostigmine salicylate for a 9-month
period in 13 of 25 patients with sporadic amyotrophic lateral sclerosis
(ALS). A large dropout rate of 48% was secondary to eight deaths and
four exclusions attributed to the incapability to swallow the tablets
(physostigmine) and capsules (lecithin) or to attend the clinic.
Parameters used for assessment of the drug efficacy included body
weight, ALS score, Jamar grip strength, forced vital capacity, and
maximum voluntary ventilation. It revealed slight benefit in reduced
loss of grip strength compared with the pretrial and placebo periods.
However, the rates of decline for body weight, ALS score, forced vital
capacity, maximum voluntary ventilation, and megascore did not differ
significantly between the pretrial, placebo, and physostigmine periods.
We therefore concluded that overall no significant alteration in the
clinical course was gained by oral physostigmine therapy in the 13
patients with ALS who were included in this study.

325\. Aquilonius, S.M., *et al.*, *Cholinesterase inhibitors lack
therapeutic effect in amyotrophic lateral sclerosis. A controlled study
of physostigmine versus neostigmine.* Acta Neurol Scand, 1986. 73(6): p.
628-32. Seven patients with amyotrophic lateral sclerosis participated
in a double-blind cross-over trial of oral physostigmine and neostigmine
(10 and 45 mg/day, respectively, for 3 days). Six of the patients were
also given intravenous injections (1 and 1.5 mg, respectively) of the
drugs in an open trial. No significant effects on muscle strength or
neurophysiological parameters were observed.

326\. Backman, S.B., *et al.*, *Different properties of the bradycardia
produced by neostigmine and edrophonium in the cat.* Can J Anaesth,
1996. 43(7): p. 731-40. PURPOSE: The bradycardia produced by neostigmine
and edrophonium was examined according to its relation to cholinesterase
inhibition and to its sensitivity to block by muscarinic receptor
antagonists. For comparison, the ability of muscarinic antagonists to
block the bradycardia produced by electrical stimulation of the vagus
nerve was determined. METHODS: Cats were anaesthetized, vagotomized and
propranolol-treated. Heart rate was continuously recorded. Erythrocyte
cholinesterase activity of arterial blood was measured using a
radiometric technique. The right vagus nerve was isolated for electrical
stimulation. The muscarinic antagonists used were atropine,
glycopyrrolate, pancuronium, gallamine, and AFDX-116. RESULTS:
Neostigmine produced a dose-dependent decrease in cholinesterase
activity which reached a plateau at a cumulative dose of 0.16 mg.kg-1
(ED50 0.009 +/- 0.003 mg.kg-1). Neostigmine produced a dose-dependent
decrease in heart rate with the dose-response relationship (ED50 0.1 +/-
0.01 mg.kg-1; P = 0.0006) shifted to the right of that for the
inhibition of cholinesterase activity. In contrast to the
anticholinesterase effect, the bradycardic effect did not reach a
plateau and continued to increase even at doses at which the
cholinesterase inhibition was maximal. The maximal decrease in heart
rate when the heart was still in sinus rhythm was by 81 +/- 13 bpm (49
+/- 7% of baseline), which was produced by a dose of 0.32 mg.kg-1.
Edrophonium produced dose-dependent decreases in cholinesterase activity
and heart rate, which were highly correlated (correlation coefficient r
= 0.99, P \< 0.0001). The ED50 of the reduction in heart rate (0.9 +/-
0.18 mg.kg-1) and cholinesterase activity (0.89 +/- 0.12 mg.kg-1)
produced by edrophonium were similar. Moreover, the reduction in heart
rate and cholinesterase activity produced by edrophonium reached a
plateau at the same dose (6.4 mg.kg-1). At this dose, heart rate
decreased by 22 +/- 2 bpm (14.6 +/- 0.9% of baseline). Compared to the
bradycardia produced by stimulation of the vagus nerve, that produced by
neostigmine was blocked by muscarinic antagonists at significantly lower
doses while that produced by edrophonium was blocked at similar doses.
CONCLUSIONS: The neostigmine-induced bradycardia is poorly correlated
with cholinesterase inhibition compared to that produced by edrophonium,
and has a higher sensitivity to muscarinic receptor antagonists compared
to that produced by edrophonium or vagus nerve stimulation. These
results are consistent with the hypothesis that the neostigmine-induced
bradycardia is, in part, the result of neostigmine directly activating
cholinergic receptors within the cardiac parasympathetic pathway. The
bradycardia produced by edrophonium may be accounted for solely by an
anticholinesterase action.

327\. Baraka, A., *Safe reversal. 2. Atropine-neostigmine mixture: an
electrocardiographic study.* Br J Anaesth, 1968. 40(1): p. 30-6.

328\. Baraka, A., *Safe reversal. 1. Atropine followed by neostigmine.
An electrocardiographic study.* Br J Anaesth, 1968. 40(1): p. 27-9.

329\. Beebe, D.S., S.J. Shumway, and R. Maddock, *Sinus arrest after
intravenous neostigmine in two heart transplant recipients.* Anesth
Analg, 1994. 78(4): p. 779-82.

330\. Bramich, N.J., J.A. Brock, and G.D. Hirst, *Potentiation by
neostigmine of responses to vagal nerve stimulation in the sinus venosus
of the toad.* Auton Neurosci, 2000. 82(3): p. 109-14. The effects of the
cholinesterase inhibitor neostigmine on the responses to vagus nerve
stimulation of isolated sinus venosus/atrial preparations of the toad,
Bufo marinus, were examined. In control solutions, trains of stimuli
applied to the vagus nerve led to a decrease in heart rate that was
susceptible to muscarinic receptor blockade. Membrane potential
recordings made from sinus venosus cells showed that the responses to
trains of stimuli, delivered at frequencies of less than 10 Hz, were
little changed by the addition of neostigmine. However, the responses to
longer trains of stimuli at 10 Hz (30 versus 10 s) were potentiated and
the nature of the membrane potential changes was altered. The results
suggest that, due to the activity of cholinesterases, acetylcholine
(ACh) released from parasympathetic nerves normally has little access to
the muscarinic receptors in the pacemaker region, which are linked to
potassium channels.

331\. Cavaliere, F., *et al.*, *\[Effectiveness of low-dose neostigmine
in the treatment of sinus tachycardia during aortocoronary bypass\].*
Minerva Anestesiol, 1993. 59(7-8): p. 361-5. Thirteen patients, who
underwent surgery for myocardial revascularization, were given
neostigmine, 0.25-1 mg i.v., in order to control intraoperative sinus
tachycardia. Three minutes after the administration, the drug was
effective in all patients, by decreasing the heart rate and improving
the indices of the risk of myocardial ischemia. Seven minutes later the
heart rate decreased further. In 5 patients out of 13, short periods of
bradycardia were observed during surgical compression or dislocation of
the heart. As cardiac rate decreased, P and T waves and ST, QT, and QTc
intervals lengthened without reaching pathological levels. Three minutes
after neostigmine, the stroke volume was unchanged in spite of the
decrease of the heart rate; consequently cardiac index decreased. Seven
minutes later a slight increase of the stroke volume balanced the
further decrease of the heart rate; so cardiac index did not change any
more. Arterial pressure decreased slightly after neostigmine whereas
systemic vascular resistance was unaffected. Also central venous
pressure did not change while pulmonary capillary wedge pressure showed
a small increase 10 minutes after neostigmine. Finally airway resistance
did not change significantly.

332\. Constable, P.D., *et al.*, *Atrial fibrillation associated with
neostigmine administration in three cows.* J Am Vet Med Assoc, 1990.
196(2): p. 329-32. Three cattle with gastrointestinal disease were given
neostigmine as part of the treatment for postsurgical ileus. All 3
cattle developed atrial fibrillation either before or during neostigmine
administration, and 2 converted to normal sinus rhythm after cessation
of treatment.

333\. Dowdy, E.G., *Effect of neostigmine on cardiac rhythm in
digitalized dogs. Guest discussion.* Anesth Analg, 1971. 50(6): p.
1088-9.

334\. Fuchs-Buder, T., *et al.*, *Antagonism of vecuronium-induced
neuromuscular block in patients pretreated with magnesium sulphate:
dose-effect relationship of neostigmine.* Br J Anaesth, 1999. 82(1): p.
61-5. We have investigated the dose-effect relationship of neostigmine
in antagonizing vecuronium-induced neuromuscular block with and without
magnesium sulphate (MgSO4) pretreatment. Neuromuscular block was
assessed by electromyography with train-of-four (TOF) stimulation.
First, we determined neostigmine-induced recovery in patients pretreated
with MgSO4 (group A) or saline (group B) (n = 12 each). The height of
T1, 5 min after neostigmine, was 43 (7)% in group A and 65 (6)% in group
B (P \< 0.01). Respective values after 10 min were 59 (7)% and 83 (5)%
(P \< 0.01). TOF ratio, 5 min after neostigmine, was 29 (6)% in group A
and 29 (5)% in group B. Respective values after 10 min were 38 (11)% and
51 (7)% (P \< 0.01). To gain insight into the mechanisms leading to
delayed recovery after MgSO4, we calculated assisted recovery, defined
as neostigmine-induced recovery minus mean spontaneous recovery.
Spontaneous recovery was assessed in another 24 patients. Patients in
group C received MgSO4/vecuronium and patients in group D vecuronium
only (n = 12 each). Five minutes after neostigmine, assisted recovery
was 22 (7)% in the MgSO4 pretreated patients and 28 (6)% in controls (P
\< 0.05). Ten minutes after neostigmine, values were 24 (7)% and 22
(6)%. Maximum assisted recovery was not influenced by MgSO4 pretreatment
(27 (6)% in group A and 32 (6)% in group B) and time to maximum effect
was comparable between groups: 6 (4-10) min and 7 (5-8) min,
respectively. We conclude that neostigmine-induced recovery was
attenuated in patients treated with MgSO4. This was mainly a result of
slower spontaneous recovery and not decreased response to neostigmine.

335\. Eldor, J. and D.Z. Frankel, *Use of low-dose neostigmine
intravenously in the treatment of supraventricular tachycardia: an
immediate bradycardic effect.* Resuscitation, 1989. 18(1): p. 103-10.
Neostigmine was first used 50 years ago to treat sinus tachycardia and
paroxysmal auricular tachycardia. Then there were reports of successful
treatment by neostigmine of other forms of supraventricular
tachycardias. However, reports of sudden death using neostigmine for
reversal of neuromuscular blockade at the end of an operation, which
were not properly treated with atropine abandoned the use of neostigmine
as an antiarrhythmic drug. Low-dose neostigmine intravenously was used
in the treatment of supraventricular tachycardia in three patients
described herein. It gave an immediate bradycardic effect in all three
patients. The use of a low-dose neostigmine intravenously for an
immediate treatment of supraventricular tachycardia is a novel
suggestion. It has to be further evaluated and compared to the
conventional drugs used like digoxin, verapamil, propranolol or esmolol.

336\. Ivankovic, A.D., *et al.*, *Effect of neostigmine on cardiac
rhythm in digitalized dogs.* Anesth Analg, 1971. 50(6): p. 1079-88.

337\. Kadoya, T., *et al.*, *Development of rapid atrial fibrillation
with a wide QRS complex after neostigmine in a patient with intermittent
Wolff-Parkinson-White syndrome.* Br J Anaesth, 1999. 83(5): p. 815-8. We
report the case of a 67-yr-old man with intermittent
Wolff-Parkinson-White (WPW) syndrome in whom neostigmine produced
life-threatening tachyarrhythmias. The patient was scheduled for
microsurgery for a laryngeal tumour. When he arrived in the operating
room, the electrocardiogram showed normal sinus rhythm with a rate of 82
beat min-1 and a narrow QRS complex which remained normal throughout the
operative period. On emergence from anaesthesia, the sinus rhythm (87
beat min-1) changed to atrial fibrillation with a rate of 80-120 beat
min-1 and a normal QRS complex. We did not treat the atrial fibrillation
because the patient was haemodynamically stable. Neostigmine 1 mg
without atropine was then administered to antagonize residual
neuromuscular block produced by vecuronium. Two minutes later, the
narrow QRS complexes changed to a wide QRS complex tachycardia with a
rate of 110-180 beat min-1, which was diagnosed as rapid atrial
fibrillation. As the patient was hypotensive, two synchronized DC
cardioversions of 100 J and 200 J were given, which restored sinus
rhythm. No electrophysiological studies of anticholinesterase drugs have
been performed in patients with WPW syndrome. We discuss the use of
these drugs in this condition.

338\. Kjellberg, M. and T. Tammisto, *Heart-rate changes after atropine
and neostigmine given for the reversion of muscle paralysis.* Acta
Anaesthesiol Scand, 1970. 14(3): p. 203-10.

339\. Klingenmaier, C.H., *et al.*, *Reversal of neuromuscular blockade
with a mixture of neostigmine and glycopyrrolate.* Anesth Analg, 1972.
51(3): p. 468-72.

340\. Pratila, M.G. and V. Pratilas, *A case of tachydysrhythmia.
Refractory to propranolol and responsive to neostigmine.* Anaesthesia,
1977. 32(10): p. 1017-9. A case of atrial fibrillation, with a fast
ventricular response, which developed under enflurane anaesthesia is
described in a patient previously treated with digitalis and
propranolol. The intravenous administration of propranolol was
ineffective whereas that of neostigmine and atropine, 2 hours later, was
successful and reduced the ventricular rate to normal values.

341\. Raymond, C.W., *Letter: Neostigmine methysulfate in the treatment
of cardiac arryhthmia induced by perphenazine-amitriptyline.* Can Med
Assoc J, 1976. 114(2): p. 102-7.

342\. Rosner, V., E.R. Kepes, and F.F. Foldes, *The effects of atropine
and neostigmine on heart rate and rhythm. Recommendation for their use
to reverse residual neuromuscular block.* Br J Anaesth, 1971. 43(11): p.
1066-74.

343\. Schultheis, L.W., *et al.*, *High dose neostigmine treatment of
malignant sinus tachycardia.* Pacing Clin Electrophysiol, 1997. 20(5 Pt
1): p. 1369-72. Sinus tachycardia caused by circulating catecholamines
in the setting of congestive heart failure may impair systemic perfusion
because of decreased diastolic filling time. We report the case of a
patient with Wolff-Parkinson-White syndrome with angina and cardiogenic
shock who improved dramatically following administration of neostigmine.
Cardiac output, blood pressure, and stroke volume increased as heart
rate was reduced. A previous attempt at heart rate control, in the same
patient, using a low dose beta-antagonist, precipitated hemodynamic
collapse. The remarkable recovery of our patient suggests that
acetylcholinesterase inhibitors may warrant further investigation in
patients with severe sinus tachycardia.

344\. Tetzlaff, J.E. and M.J. Choban, *Asystole following neostigmine
administration during carotid sinus stimulation.* J Clin Anesth, 1990.
2(3): p. 212-3.

345\. Wetterslev, J., *et al.*, *Split-dose atropine versus
glycopyrrolate with neostigmine for reversal of gallamine-induced
neuromuscular blockade.* Acta Anaesthesiol Scand, 1991. 35(5): p.
398-401. The effects of a split-dose of atropine sulphate versus a
single dose of glycopyrrolate given with neostigmine for the reversal of
gallamine-induced neuromuscular blockade were studied in 55 patients
undergoing gynaecological surgery. The patients were randomized to
receive either a single dose of glycopyrrolate (7 micrograms.kg-1) or
two doses of atropine (8 micrograms.kg-1 each), given with an interval
of 1 min. There were no differences between the two methods with respect
to percentage heart rate changes, salivation or arousal time. Four
patients demonstrated cardiac arhythmias in the atropine group, whereas
none occurred in the glycopyrrolate group (P less than 0.05). It is
concluded that a split-dose of atropine has similar chronotropic effects
to a single dose of glycopyrrolate for the reversal of gallamine-induced
neuromuscular blockade. However, the finding of a higher incidence of
cardiac arrhythmias in the atropine group suggests that this reversal
regime should be reserved for patients without cardiac disease.

346\. Bigger, J.T., Jr., *et al.*, *Is physostigmine effective for
cardiac toxicity of tricyclic antidepressant drugs.* Jama, 1977.
237(13): p. 1311.

347\. Das, P.K. and S.K. Bhattacharya, *Studies on the effect of
physostigmine on experimental cardiac arrhythmias in dogs.* Br J
Pharmacol, 1972. 44(3): p. 397-403.

348\. Dysken, M.W. and D.S. Janowsky, *Dose-related
physostigmine-induced ventricular arrhythmia: case report.* J Clin
Psychiatry, 1985. 46(10): p. 446-7. Physostigmine, a centrally-acting
cholinesterase inhibitor, has been shown to have potential as a
treatment for primary degenerative dementia. In an experiment to test
its effects, i.v. physostigmine caused the onset of unifocal premature
contractions and runs of bigeminy in an 85-year-old man.

349\. Elonen, E., *Effect of beta-adrenoceptor blocking drugs,
physostigmine, and atropine on the toxicity of doxepin in mice.* Med
Biol, 1975. 53(4): p. 231-7. Large doses of doxepin given intravenously
to animals cause tachyarrhythmias, and still higher doses lead to a
progressive and, finally, lethal bradycardia. The effect of pretreatment
with five different beta-adrenoceptor blocking drugs (propranolol,
alprenolol, practolol, metoprolol or tolamolol), p physostigmine, or
atropine on these toxic actions of doxepin was investigated. Mice were
sedated with diazepam. Doxepin was injected i.v. 0.1 mg every 15 sec
until death. ECG was recorded at 10 sec after every injection. All five
beta-blockers injected i.p. 30 min before doxepin inhibited the
doxepin-induced tachyarrhythmias. None of the drugs prevented or
postponed the death of mice. Large doses of beta-blockers
dose-dependently enhanced the doxepin-induced bradycardia and
accelerated death. The cardioselective beta-blocking drugs practolol and
metoprolol proved less active in enhancing bradycardia than the third
cardio-selective drug, tolamolol, and non-selective propranolol and
alprenolol. This difference may have resulted from properties other than
beta-blockade since practolol and metoprolol lack the
\"cardiodepressant\" and local anaesthetic properties. Since
physostigmine and atropine did not modify the doxepin effects the
anticholinergic property may not be important in the severe cardiotoxic
effects of doxepin.

350\. Goldberger, A.L. and G.P. Curtis, *Immediate effects of
physostigmine on amitryptyline-induced QRS prolongation.* J Toxicol Clin
Toxicol, 1982. 19(5): p. 445-54. Prior studies have suggested that
physostigmine may be useful in reversing QRS prolongation due to
amitriptyline toxicity. To investigate this question, we devised a
pharmacologic model in rabbits utilizing an initial intravenous bolus of
amitriptyline (4-6 mg) followed by a constant amitriptyline infusion
(0.2-0.4 mg/min) empirically titered to maintain the QRS at 50% or more
of control value for at least 5 min. Intravenous physostigmine (2 mg)
sufficient to produce muscle fasciculations and significant (P less than
0.01) slowing of sinus rate was then administered to six animals. No
significant change in QRS duration was noted at 1, 3, and 5 min
intervals following physostigmine. Although no immediate antidotal
effect of physostigmine on amitriptyline-induced QRS prolongation could
be demonstrated, these results do not exclude a possible interaction
between the membrane effects of the tricyclic antidepressants (and
related agents) and the vagal branch of the autonomic nervous system.

351\. James, T.N., F. Urthaler, and R.K. Lewis, *Termination of
\"ventricular\" arrhythmias from digoxin by selective production of
complete atrioventricular block with physostigmine in the dog.* J
Pharmacol Exp Ther, 1979. 211(3): p. 561-70. In previous studies we have
come to doubt that ventricular rhythms of an automatic nature will arise
spontaneously from the peripheral Purkinje system. In 20 anesthetized
dogs, digoxin was administered i.v. (0.1-1.0 mg/kg) and in 12 dogs by
selectively perfusing the atrioventricular (AV) node artery (2 ml; 40
microgram/ml). We obtained the following results. First, selective
pharmacological production of complete AV block (acetylcholine or
physostigmine) interrupts the \"ventricular\" arrhythmias considered
characteristic of digitalis intoxication.Second, digitalis arrhythmias
are difficult to produce when this type of complete heart block had been
previously established. Third, abolition of ventricular arrhythmias by
selective pharmacological production of heart block can be reversed
(i.e., the arrhythmia restored) with atropine. Fourth, rapid pacing of
the ventricles during complete heart block in dogs poisoned with
digitalis can eventually induce ventricular arrhythmias, but not
quickly. We interpret that these digitalis arrhythmias originated within
the acetylcholine-sensitive portion of the AV node-His bundle region.

352\. Janowsky, D.S., *et al.*, *Effects of physostigmine on pulse,
blood pressure, and serum epinephrine levels.* Am J Psychiatry, 1985.
142(6): p. 738-40. In this study, the infusion of physostigmine in 14
patients with affective disorder who were pretreated with
methscopolamine caused significant and often profound increases in the
patients\' epinephrine levels, pulse rates, and blood pressure. Since
physostigmine is being used experimentally in the treatment of elderly
subjects who have Alzheimer\'s disease, these cardiovascular effects may
have clinical importance.

353\. Kulig, K. and B.H. Rumack, *Physostigmine and asystole.* Ann Emerg
Med, 1981. 10(4): p. 228-20.

354\. Maister, A.H., *Atrial fibrillation following physostigmine.* Can
Anaesth Soc J, 1983. 30(4): p. 419-21. The side effects of
physostigmine, a tertiary amine anticholinesterase agent commonly used
to treat postanaesthetic depression caused by a large number of drugs,
are thought to be fairly mild. A case is reported in which atrial
fibrillation followed a total dose of 2 mg of this drug to a patient who
was slow in recovering from a general anaesthetic. The arrhythmia was
self-limiting and required no therapy. Indications and precautions for
the use of physostigmine are discussed.

355\. Pentel, P. and C.D. Peterson, *Asystole complicating physostigmine
treatment of tricyclic antidepressant overdose.* Ann Emerg Med, 1980.
9(11): p. 588-90. Physostigmine is a commonly used therapy for the
anticholinergic manifestations of tricyclic antidepressant (TCA)
overdose. We describe two patients with TCA toxicity who developed
asystole following the administration of physostigmine to treat
seizures.

356\. Schmidt, W. and K. Lang, *Life-threatening dysrhythmias in severe
thioridazine poisoning treated with physostigmine and transient atrial
pacing.* Crit Care Med, 1997. 25(11): p. 1925-30. OBJECTIVES: To
describe clinical, electrocardiographic, and blood chemistry findings in
a case of high-dosage thioridazine self-poisoning, focusing on the
cellular mechanisms of the cardiovascular toxicity. DESIGN: Case report
and literature review. SETTING: Intensive care unit (ICU) of a district
hospital in Germany. PATIENT: A 68-yr-old male patient admitted to the
ICU for treatment of a severe thioridazine intoxication. INTERVENTIONS:
Prevention of absorption (gastric lavage), mechanical ventilation,
fluids, alkalinization, catecholamines, drugs (physostigmine,
neostigmine), direct current cardioversion/defibrillation, and transient
pacemaker (atrial stimulation). MEASUREMENTS AND MAIN RESULTS: Central
nervous, cardiovascular, and gastrointestinal systems indicated the
adverse side effects of thioridazine intoxication over a period of 9
days. During high toxic thioridazine plasma concentrations (6061 to 6480
ng/mL), a life-threatening crisis occurred due to malignant ventricular
arrhythmias followed by bradycardia (e.g., sinus node arrest). The
electrocardiogram showed delays in all parts of the conduction system of
the heart, including prolonged repolarization for several days, which
disappeared completely when thioridazine plasma concentrations were
within the therapeutic range. CONCLUSIONS: An individual therapeutic
approach is needed in cases of thioridazine overdose. The primary aim is
to stabilize the cardiac rhythm and the circulation.

357\. Schulz, M. and A. Schmoldt, *Successful physostigmine treatment of
acute dothiepin intoxication.* Pharmazie, 1994. 49(8): p. 614. We report
on a severe intoxication with the tricyclic antidepressant dothiepin. A
treatment with a continuous infusion of physostigmine was successful and
improved the ventricular arrhythmia. Dothiepin and its active
metabolites were determined in plasma and urine by HPLC.

358\. Seitz, R., M. Wehr, and N. Pallmert, *\[Arrhythmias in poisoning
with tricyclic antidepressive agents. Successful treatment with
physostigmine and sodium\].* Dtsch Med Wochenschr, 1982. 107(28): p.
1096-9. A 26-year-old female patient was admitted 30 minutes after
suicidal poisoning with 750 mg amitriptyline (Saroten), 4800 mg
dibenzepine (Noveril retard) and 2100 mg thioridazine (Melleril). Life
threatening disturbances of cardiac rhythm occurred in addition to coma,
pulmonary insufficiency requiring assisted ventilation, and hypotension.
Physostigmine only caused temporary improvement and the arrhythmias only
resolved completely after additional administration of NaCl.

359\. Snyder, B.D., L. Blonde, and W.R. McWhirter, *Reversal of
amitriptyline intoxication by physostigmine.* Jama, 1974. 230(10): p.
1433-4.

360\. Tobis, J. and B.N. Das, *Cardiac complications in amitriptyline
poisoning. Successful treatment with physostigmine.* Jama, 1976.
235(14): p. 1474-6.

361\. Wright, S.P., *Usefulness of physostigmine in imipramine
poisoning. A dramatic response in a child resistant to other therapy.*
Clin Pediatr (Phila), 1976. 15(12): p. 1123-8.

362\. Weisdorf, D., *et al.*, *Physostigmine for cardiac and neurologic
manifestations of phenothiazine poisoning.* Clin Pharmacol Ther, 1978.
24(6): p. 663-7. Psychotropic drugs such as the phenothiazine
neuroleptics and tricyclic antidepressants are known to cause
electrocardiographic abnormalities as well as a central anticholinergic
syndrome. Physostigmine is known to reverse the central muscarinic
anticholinergic manifestations by inhibition of the enzyme
cholinesterase. An unusual case of trifluoperazine overdose, in which
the patient presented with cardiac arrhythmias and a central
anticholinergic syndrome, is presented. Treatment with physostigmine
reversed the central anticholinergic syndrome as well as the
electrocardiographic abnormalities. Effects of phenothiazines on
altering cardiac status are also discussed.

363\. Dorner, G., *et al.*, *Further evidence of teratogenic effects
apparently produced by neurotransmitters during brain differentiation.*
Endokrinologie, 1977. 70(3): p. S326-330.

364\. Dorner, G., K. Hecht, and G. Hinz, *Teratopsychogenetic effects
apparently produced by nonphysiological neurotransmitter concentrations
during brain differentiation.* Endokrinologie, 1976. 68(1): p. 1-5.

365\. Dorner, G. and G. Hinz, *Apparent effects of neurotransmitters on
sexual differentiation of the brain without mediation of sex hormones.*
Endokrinologie, 1978. 71(1): p. S104-108.

366\. Landauer, W., *Cholinomimetic teratogens: Studies with chicken
embryos.* Teratology, 1975. 12: p. 125-146.

367\. Al-Shekhlee, A., N. Robin, and H.J. Kaminski,
*Pyridostigmine-induced microcephaly.* Neurology, 2001. 56(11): p.
1606-7.

368\. Cowan, D.N., *et al.*, *The risk of birth defects among children
of Persian Gulf War veterans.* N Engl J Med, 1997. 336(23): p. 1650-6.
BACKGROUND: There has been suspicion that service in the Persian Gulf
War affected the health of veterans adversely, and there have been
claims of an increased rate of birth defects among the children of those
veterans. METHODS: We evaluated the routinely collected data on all live
births at 135 military hospitals in 1991, 1992, and 1993. The data base
included up to eight diagnoses from the International Classification of
Diseases, 9th Revision, Clinical Modification (ICD-9-CM) for each birth
hospitalization, plus information on the demographic characteristics and
service history of the parents. The records of over 75,000 newborns were
evaluated for any birth defect (ICD-9-CM codes 740 to 759, plus
neoplasms and hereditary diseases) and for birth defects defined as
severe on the basis of the specific diagnoses and the criteria of the
Centers for Disease Control and Prevention. RESULTS: During the study
period, 33,998 infants were born to Gulf War veterans and 41,463 to
non-deployed veterans at military hospitals. The overall risk of any
birth defect was 7.45 percent, and the risk of severe birth defects was
1.85 percent. These rates are similar to those reported in civilian
populations. In the multivariate analysis, there was no significant
association for either men or women between service in the Gulf War and
the risk of any birth defect or of severe birth defects in their
children. CONCLUSIONS: This analysis finds no evidence of an increase in
the risk of birth defects among the children of Gulf War veterans.

369\. Penman, A.D., R.S. Tarver, and M.M. Currier, *No evidence of
increase in birth defects and health problems among children born to
Persian Gulf War Veterans in Mississippi.* Mil Med, 1996. 161(1): p.
1-6. The Department of Veterans\' Affairs, Jackson, Mississippi, and the
Mississippi State Department of Health conducted a collaborative
investigation of an apparent increase in the numbers of birth defects
and other health problems among children born to veterans of two
Mississippi National Guard units who had served in the Persian Gulf War.
The medical records of all children conceived by and born to veterans of
the two units after deployment were reviewed; observed numbers of birth
defects and other health problems were compared with expected numbers
using rates from birth defect surveillance systems and previous surveys.
The total number of all types of birth defects was not greater than
expected, but whether the number of specific birth defects was greater
than expected could not be determined. The frequency of premature birth,
low birth weight, and other health problems appeared similar to that in
the general population.

370\. Araneta, M.R., *et al.*, *Birth defects prevalence among infants
of Persian Gulf War veterans born in Hawaii, 1989-1993.* Teratology,
2000. 62(4): p. 195-204. BACKGROUND: Gulf War veterans (GWVs) have
expressed concern about possible teratogenic exposures. However,
epidemiologic studies on birth defects prevalence among their progeny
have been limited to military hospitals, anomalies diagnosed among
newborns, or self-reported data. To measure the prevalence of selected
birth defects among infants of GWVs and nondeployed veterans (NDVs) in
Hawaii, using birth defects surveillance records. METHODS: Personal
identifiers of 684,645 GWVs and 1,587,102 NDVs and their families were
matched against birth certificate records of 99,545 live births reported
to the State of Hawaii Department of Health between 1989 and 1993 to
identify births to military personnel. These births were matched with
records from the Hawaii Birth Defects Program. RESULTS: A total of
17,182 military infants (3,717 GWV infants and 13,465 NDV infants) were
identified. Of these, 367 infants (2.14/100 live births) were identified
with one or more of 48 major birth defects diagnoses. The prevalence of
the 48 birth defects were similar for GWV and NDV infants during the
prewar and postwar periods, and among GWV infants who were conceived
before and after the Gulf war. CONCLUSIONS: The results must be
interpreted with caution because of the small number of affected infants
in each birth defects category. This study demonstrated the feasibility
of measuring birth defects prevalence among military infants through
multiple data linkage. Further, it included live births to parents who
had separated from the military, births in civilian hospitals, and birth
defects diagnosed through the first year of life.

371\. Doyle, P., E. Roman, and N. Maconochie, *Birth defects among
children of Persian Gulf War veterans.* N Engl J Med, 1997. 337(16): p.
1175-6.

372\. Zhou, Y.X., *et al.*, *Detection of the organophosphorus nerve
agent sarin by a competitive inhibition enzyme immunoassay.* Arch
Toxicol, 1995. 69(9): p. 644-8. Two artificial antigens,
NalphaNepsilon-di(O,O-diisopropyl) phosphoryl L-lysine (DIP)- bovine
serum albumin (BSA) conjugate (DIP-BSA) and DIP-KLH (keyhole limpet
hemocyanin), were synthesized. Antibodies against sarin (O-isopropyl
methylphosphonofluoridate) were obtained after immunization of rabbits
with DIP-KLH conjugate. A competitive inhibition enzyme immunoassay
(CIEIA) was developed to detect the organophosphorus nerve agent sarin.
The antibody solutions could be inhibited by sarin as low as 10(-6)
mol/l, and the standard curve was linear over 3 orders of magnitude. The
coefficients of intraassay and interassay variation of this method were
5.4-6.2% (n = 11) and 8.0-9.5% (n = 6) at a sarin concentration range of
10(-3)-10(-6) mol/l, respectively. The recovery of sarin in water
samples at the concentration of 5 x 10(-5) mol/l was in the range of
96.8-102.5%. The specificity of the antiserum was assessed by comparing
the inhibition induced by sarin with soman, Vx, isopropyl alcohol and
isopropyl methyl phosphonic acid. The results showed that less than 5
mmol/l soman, 2 mmol/l Vx, 16 mmol/l isopropyl alcohol and 8 mmol/l
isopropyl methyl phosphonic acid did not influence the determination of
sarin in water samples.

372.5 Kang H, Magee C, Mahan C, et al. Pregnancy outcomes among U.S.
Gulf War veterans: a population-based survey of 30,000 veterans. *Ann
Epidemiol*. 2001;11:504-11.

PURPOSE: We evaluated an association between veterans\' Gulf War service
and reported adverse pregnancy outcomes. METHODS: We conducted a health
survey in which selected reproductive outcomes of a population-based
sample of 15,000 Gulf War veterans representing four military branches
and three unit components (active, reserve, and National Guard) were
compared to those of 15,000 non-Gulf veteran controls. RESULTS: Male
Gulf veterans, compared with their non-Gulf veteran controls, reported a
significantly higher rate of miscarriage (odds ratio \[OR\] = 1.62; 95%
confidence interval \[CI\] = 1.32-1.99). Female Gulf veterans also
reported more miscarriages than their respective controls, although
their excess was not statistically significant (OR= 1.35; CI =
0.97-1.89). Both men and women deployed to the Gulf theater reported
significant excesses of birth defects among their liveborn infants.
These excess rates also extended to the subset of \"moderate to severe\"
birth defects \[males: OR= 1.78 (CI = 1.19-2.66); females: OR = 2.80 (CI
= 1.26-6.25)\]. No statistically significant differences by deployment
status were found among men or women for stillbirths, pre-term
deliveries or infant mortality. CONCLUSION: The risk of veterans
reporting birth defects among their children was significantly
associated with veteran\'s military service in the Gulf War. This
observation needs to be confirmed by a review of medical records to rule
out possible reporting bias.