Methanol elevates cytosolic calcium ions in cultured canine cerebral vascular smooth muscle cells: possible relation to CNS toxicity.

Acute exposure of cultured canine cerebral vascular smooth muscle cells to methanol (10-400 mM) results in concentration-dependent elevation of the concentration of intracellular free calcium ion ([Ca2+]i) as measured with the fluorescent indicator, fura-2, and digital imaging microscopy. The resting level of [Ca2+]i in the cerebral vascular smooth muscle cells was 89.3+/-5.3 nM. Exposure of these cells to 10 mM methanol for only 5 min resulted in significant elevation in [Ca2+]i (i.e., to 105.7+/-4.6) (p < 0.05). Methanol (10 mM) is a concentration found in the blood of victims demonstrating early CNS toxicity. Other, higher concentrations of methanol rapidly raised [Ca2+]i upwards of 60% over basal resting levels. These result suggest that methanol-induced cerebral vasospasm is a consequence of large rises in intracellular Ca2+. These events could play a crucial role in methanol-induced cerebral edema, brain hemorrhage, and cerebral and retinal infarcts, eventuating in severe deficits in brain blood flow and the known, subsequent CNS disturbances.     

Ethanol promotes rapid depletion of intracellular free Mg in cerebral vascular smooth muscle cells: Possible relation to alcohol-induced behavioral and stroke-like effects

The acute effects of ethanol on intracellular free magnesium ions ([Mg²⁺]_i) in cultured canine cerebral vascular smooth muscle cells (VSMCs) were studied by digital imaging microscopy using the Mg²⁺ fluorescent probe mag-fura-2. In 0 mM ethanol, the basal level of [Mg²⁺]_i was between 500–700 μM with a heterogeneous distribution within the cells; [Mg²⁺]_i was greater in the perinuclear than in the peripheral region. Treatment of the cells with 10, 25, and 100 mM ethanol resulted in rapid (within 30 s) concentration-dependent reduction in [Mg²⁺]_i; the greater the concentration and the greater the duration of acute exposure, the greater the fall in [Mg²⁺]_i. Exposure of cerebral VSMCs to 100 mM ethanol resulted in a 57% reduction in [Mg²⁺]_i (i.e., from 510 ± 40 to 220 ± 30 μM). These observations are consistent with the tenet that “binge drinking” of ethanol could result in cerebrovasospasm, ischemia, and rupture of cerebral blood vessels as a consequence of depletion of cerebral VSMC [Mg²⁺]_i. Deficits in [Mg²⁺]_i, O₂, and nutrient delivery could account in part for some of the behavioral actions of alcohol.     

Association of alcohol in brain injury, headaches, and stroke with brain-tissue and serum levels of ionized magnesium: a review of recent findings and mechanisms of action.

Although there is general agreement that chronic ingestion of alcohol poses great risks for normal cardiovascular functions and peripheral-vascular homeostasis, a direct cause and effect between the real phenomena of alcohol-induced headache and risk of brain injury and stroke is not appreciated. "Binge drinking" of alcohol is associated with an ever-growing number of strokes and sudden death. It is becoming clear that alcohol ingestion can result in profoundly different actions on the cerebral circulation (e.g., vasodilation, vasoconstriction-spasm, vessel rupture), depending upon dose and physiologic state of host. Using rats, it has been demonstrated that acute, high doses of ethanol can result in stroke-like events concomitant with alterations in brain bioenergetics. We review recent in vivo findings obtained with 31P-NMR spectroscopy, optical reflectance spectroscopy, and direct in vivo microcirculatory studies on the intact brain. Alcohol-induced hemorrhagic stroke is preceded by a rapid fall in brain intracellular free magnesium ions ([Mg2+]i) followed by cerebrovasospasm and reductions in phosphocreatine (PCr)/ATP ratio, intracellular pH, and the cytosolic phosphorylation potential (CPP) with concomitant rises in deoxyhemoglobin (DH), mitochondrial reduced cytochrome oxidase aa3 (rCOaa3), blood volume, and intracellular inorganic phosphate (Pi). Using osmotic mini-pumps implanted in the third cerebral ventricle, containing 30% ethanol, it was found that brain [Mg2+]i is reduced 30% after 14 days; brain PCr fell 15%, whereas the CPP fell 40%. Such animals became susceptible to stroke from nonlethal doses of ethanol. Human subjects with mild head injury have been found to exhibit early deficits in serum ionized Mg (IMg2+); the greater the degree of early head injury (30 min-8 h), the greater and more profound the deficit in serum IMg2+ and the greater the ionized Ca (ICa2+) to IMg2+ ratio. Patients with histories of alcohol abuse or ingestion of alcohol prior to head injury exhibited greater deficits in IMg2+ (and higher ICa2+/IMg2+ ratios) and, unlike the subjects without alcohol, did not leave the hospital for at least several days. Women, for some unknown reason, exhibit a much higher incidence of morbidity and mortality from subarachnoid hemorrhage (SAH) than men. Data on 105 men and women with different types of stroke indicate that, on the average, a 20% deficit in serum IMg2+ is seen; total Mg (TMg) or blood pH is usually near normal. Women with SAH, however, exhibit much lower IMg2+ and higher ICa2+/IMg2+ ratios; the presence of ethanol in the blood is associated with even more depression in IMg2+ in SAH in women. It is possible that prior alcohol ingestion is, in large measure, responsible for a great deal of this unexplained higher incidence of SAH in women. It has recently been reported that the cyclical changes in estrogenic hormones appear to control the serum IMg2+ level in young women. A surge in estrogenic levels prior to SAH could thus precipitate, in part, the SAH. In other human studies, it has been shown that migraines and headache, dizziness, and hangover, which accompany ethanol ingestion, are associated with rapid deficits in serum IMg2+ but not in TMg. The former, and the alcohol-associated headache, can be ameliorated with IV administration of MgSO4. Premenstrual tension-headache (PTH) and its exacerbation by alcohol in women is also accompanied by deficits in IMg2+, and elevation in serum ICa2+/IMg2+; IV MgSO4 corrects the PTH and the serum deficit in IMg2+. Animal experiments show that IV Mg2+ can prevent alcohol-induced hemorrhagic stroke and the subsequent fall in brain [Mg2+]i, [PCr], pHi, and CPP. Other recent data indicate that alcohol-induced cellular loss of [Mg2+]i is associated with cellular Ca2+ overload and generation of oxygen-derived free radicals; chronic pretreatment with vitamin E prevents alcohol-induced vascular injury and pathology in the brain. (ABSTRACT TRUNCATED)     

Occupational exposure and defects of the central nervous system in offspring: review.

A study of published work was carried out in a search for evidence of a causal role for parental occupational exposure in the origin of structural and functional defects of the central nervous system (CNS) in children. Studies that consider this topic are scarce and mostly refer to broad categories of exposures and effects. Non-occupational studies referring to environmental exposure of humans and studies on experimental animals were also reviewed. The studies on animals provided straightforward evidence about morphological and behavioural abnormalities resulting from some agents used occupationally. The studies on humans yielded a scala of defects that could be ascribed to exposure to high doses of various agents in the environment. Evidence for a causal role of occupational exposure has not been found, but a highly probable influence on the developing CNS is hypothesised for lead, methyl mercury, and ionising radiation. Parental occupational exposure to cadmium, organic solvents, anaesthetics, and pesticides may also play a part in causing defects of the CNS. Well designed future research is needed to test the above hypotheses.     

Coexposure to toluene and p-xylene in man: central nervous functions.

Sixteen men were studied in an exposure chamber to assess the effect of four hours' exposure to toluene (3.25 mmol/m3), xylene (2.84 mmol/m3), a mixture of toluene and xylene (2.20 + 0.94 mmol/m3), and a control condition. With the aid of microcomputers, subjects performed tests of simple reaction time, short term memory, and choice reaction time immediately after entering the chamber, after two, and after four hours' exposure. The results indicate that the performance on the tests was unaffected by exposure. In the light of this result the risk of an acute effect on central nervous functions after exposure for four hours at concentrations that do not exceed the Swedish threshold limit values was considered to be minimal.     

Dairy, magnesium, and calcium intake in relation to insulin sensitivity: approaches to modeling a dose-dependent association

Dairy intake has been inversely associated with insulin resistance, which may be partly due to the specific effects of calcium and magnesium. Data from the Insulin Resistance Atherosclerosis Study (1992-1999) for 1,036 US adults without diabetes at baseline were examined to evaluate the cross-sectional association of habitual dairy, magnesium, and calcium intake with insulin sensitivity at baseline and after 5 years of follow-up. Insulin sensitivity was directly measured with a validated, 12-sample, insulin-enhanced, intravenous glucose tolerance test with minimal model analysis. Dietary intake was assessed by a validated food frequency interview, and dietary supplement dose was confirmed by reviewing the supplement label. Several statistical approaches were used to ensure appropriate modeling of the dose-dependent association. No association was found between dairy intake and insulin sensitivity (p=0.41); however, associations were positive for magnesium and calcium intake (p=0.016) after adjusting for demographic, nondietary lifestyle and dietary factors, and food groups. Furthermore, magnesium intake was associated with insulin sensitivity in a threshold fashion, with a Bayesian method-estimated threshold (325 mg) (beta=0.0607/100 mg, p=0.0008 for <325 mg of magnesium/day; and beta=-0.001/100 mg, p=0.82 for >or=325 mg of magnesium/day). This study suggests that magnesium and calcium intake specifically, but not dairy intake, is associated with insulin sensitivity.     

Roles of tyrosine kinase-, 1-phosphatidylinositol 3-kinase-, and mitogen-activated protein kinase-signaling pathways in ethanol-induced contractions of rat aortic smooth muscle: possible relation to alcohol-induced hypertension.

Insights into the relations between and among ethanol-induced contractions in rat aorta, tyrosine kinases (including src family of cytoplasmic tyrosine kinases), 1-phosphatidylinositol 3-kinases (PI-3Ks), mitogen-activated protein kinases (MAPKs), and regulation of intracellular free Ca(2+) ([Ca(2+)](i)) were investigated in the present study. Ethanol-induced concentration-dependent contractions in isolated rat aortic rings were attenuated greatly by pretreatment of the arteries with low concentrations of an antagonist of protein tyrosine kinases (genistein), an src homology domain 2 (SH2) inhibitor peptide, a highly specific antagonist of p38 MAPK (SB-203580), a potent, selective antagonist of two specific MAPK kinases-MEK1/MEK2 (U0126)-and a selective antagonist of mitogen-activated protein kinase kinase (MAPKK) (PD-98059), as well as by treatment with wortmannin or LY-294002 (both are selective antagonists of PI-3Ks). Inhibitory concentration 50 (IC(50)) levels obtained for these seven antagonists were consistent with reported inhibition constant (Ki) values for these tyrosine kinase, MAPK, and MAPKK antagonists. Ethanol-induced transient and sustained increases in [Ca(2+)](i) in primary single smooth muscle cells from rat aorta were markedly attenuated in the presence of genistein, an SH2 domain inhibitor peptide, SB-203580, U0126, PD-98059, wortmannin, and LY-294002. A variety of specific antagonists of known endogenously formed vasoconstrictors did not inhibit or attenuate either the ethanol-induced contractions or the elevations of [Ca(2+)](i). Results of the present study support the suggestion that activation of tyrosine kinases (including the src family of cytoplasmic tyrosine kinases), PI-3Ks, and MAPK seems to play an important role in ethanol-induced contractions and the elevation of [Ca(2+)](i) in smooth muscle cells from rat aorta. These signaling pathways thus may be important in hypertension in human beings associated with chronic alcohol consumption.     

Central nervous system serotonin and personality as variables contributing to excessive alcohol consumption in non-human primates.

Non-human primates will readily consume an alcohol solution for its reinforcing effects when such a solution is palatable, with some subjects consuming alcohol to excess. In this review, we discuss variables that contribute to high alcohol consumption and the behaviours that are correlated with it in a non-human primate model. Developmental and behavioural correlates of central nervous system (CNS) serotonergic activity, as measured by concentrations of the serotonin metabolite 5-hydroxyindol-3-ylacetic acid (5-HIAA) in the cerebrospinal fluid (CSF), were used to investigate neurogenetic influences on alcohol consumption, as well as personality traits that characterize excessive alcohol intake. Inter-individual differences in CSF 5-HIAA concentrations were shown to have trait-like qualities, and with stable inter-individual differences across time and settings. Research has shown numerous similarities between human and non-human primates with respect to Type I- and II-like alcohol abuse and their associated behaviours. In the present series of studies, features characteristic of Type I alcohol misuse, such as high levels of anxiety, hypothalamic-pituitary-adrenal output, and situational stress predicted high alcohol intake. Primates with low CSF 5-HIAA concentrations also exhibited behaviours characteristic of Type II alcohol abuse. Principal among the traits that these subjects exhibited were deficits in impulse control. For example, subjects with low CSF 5-HIAA concentrations engaged in spontaneous behaviours that bring reinforcement but placed them at risk, such as entering food baited traps, jumping from dangerous heights to get from one tree to another, and consuming large amounts of alcohol. They can be characterized by other Type II-like deficits, such as impaired social competence, social alienation, and unrestrained, violent aggression. Non-human primates with low CSF 5-HIAA concentrations also exhibited high intrinsic tolerance following modest intakes of alcohol, and high rates of aggression during intoxication. High preferences for sweet solutions were shown to predict excessive alcohol consumption. Maternal and paternal genetic influences played major roles in producing low CNS serotonin function and excessive alcohol consumption. These genetic influences on serotonin function were exacerbated by early rearing experiences, particularly parental deprivation.     

Auditory event-related potential (P300) in relation to peripheral nerve conduction in workers exposed to lead, zinc, and copper: effects of lead on cognitive function and central nervous system.

By measuring auditory event-related potential (P300 and N100) and peripheral nerve conduction velocities, the subclinical effects of lead, zinc, and copper on the central and peripheral nervous system were examined in 22 male gun metal foundry workers exposed to these metals. Their blood lead (BPb) concentrations ranged from 12 to 59 micrograms/dl (median 30). Control subjects were 14 healthy workers, employed at the same factory, who had never been occupationally exposed to these metals. In the gun metal foundry workers, the latencies of P300 and N100 were significantly prolonged; the latency of P300 was significantly correlated with BPb concentrations and other indicators of lead absorption. Similarly, the maximal motor and sensory conduction velocities in the radial and median nerves were significantly slowed, and were significantly correlated with indicators of lead absorption. The data suggest that lead exposure at low levels affects cognitive and central auditory nervous system function together with peripheral nerve conduction.     

Vitamin K2 (menatetrenone) induces iNOS in bovine vascular smooth muscle cells: no relationship between nitric oxide production and gamma-carboxylation

It has been recently reported that vitamin K2 (menaquinone-4: menatetrenone, VK2) has an anti-atherogenic effect as well as the ability to produce clotting factors and improve osteoporosis. However, the mechanism by which VK2 acts on atherosclerosis is still unclear. In this paper, we investigated the effects of vitamin K and its side chain on NO production as an anti-atherogenic substance in a cultured vascular system. Treatment of bovine vascular smooth muscle cells (SMC) with VK2 (30 microM) caused a time-dependent (24-72 h) increase in the nitrite (NO2) level in the conditioned medium, but not in bovine vascular endothelial cells. Classical NOS inhibitor (L-nitro arginine) and iNOS-specific inhibitors completely blocked the increased nitrite level induced by VK2 treatment, but D-nitro arginine could not it. Immunostaining and Western blotting analysis showed that VK2 induced iNOS protein in the SMC. VK2 has a naphtoquinone nucleus, which is identical in menadione (VK3), and an unsaturated side chain, which is called geranylgeraniol (GGO). To determine whether the structure of VK2 was related to an increasing nitrite level, we investigated the nitrite level in conditioned medium treated with VK3 or GGO. Neither VK3 nor GGO treatment of SMC increased the nitrite level. In addition, warfarin, an inhibitor of VK2-dependent gamma-carboxylation, did not affect the increased nitrite level induced by VK2 in SMC. In conclusion, VK2 caused NO production through iNOS induction in bovine SMC, that was not related to the structure of VK2, naphtoquinone nucleus or its side chain, independently of gamma-carboxylation.     

Defects of the central nervous system in Finland: I. Variations in time and space, sex distribution, and parental age.

Data from the Finnish Register of Congenital Malformations for the years 1965-73 were used in a search for associations between environmental influences and defects of the central nervous system (CNS). The material consisted of 710 cases of CNS defects and their matched-pair controls. Moreover, and 'internal' control group of 259 cases of polydactyly and their matched-pair controls were used. The first report gives information on variations in time and space, sex distribution, and parental age. A higher incidence of anencephaly was noted in the eastern part of the country, but no significant secular or seasonal variations were found. The sex ratio (M/F) was lower than expected in the groups of anencephaly and CNS defects as a whole. High parental age turned out to be a risk factor in the group of all CNS defects, mainly owing to the subgroup of hydrocephaly. The dangers of observational studies due to confounding factors are discussed.     

Autonomic nervous system dysfunction in workers exposed to lead, zinc, and copper in relation to peripheral nerve conduction: a study of R-R interval variability.

Quantitative assessment of the autonomic neurotoxicity due to lead was undertaken by measuring variability in the electrocardiographic R-R interval (CVRR) in 16 male workers exposed to lead, zinc, copper, and tin and in 16 unexposed control subjects. Two component coefficients of variation in the R-R interval, the C-CVRSA (respiratory sinus arrhythmia) and C-CVMWSA (Mayer wave related sinus arrhythmia), were examined; these indices are considered to reflect parasympathetic and sympathetic activities, respectively. Maximal motor and sensory conduction velocities (MCV and SCV) in the median nerve were also measured. In the 16 exposed workers, blood lead concentrations ranged from 16 to 60 (mean 34) micrograms/dl. The CVRR and C-CVRSA were found to be significantly reduced in the workers with elevated lead, zinc, and copper absorption as compared to unexposed control subjects; also, the MCV and SCV were significantly slowed. The C-CVMWSA was not significantly reduced, and was positively related to plasma zinc concentrations. No significant relationships were found between indicators of lead and copper absorption and these electrophysiological measurements. These data suggest that subclinical toxicity of lead occurs in the parasympathetic component of the autonomic nervous system as well as in the peripheral nerves. Zinc may antagonize the autonomic nervous dysfunction caused by lead.     

Dose dependent effects of chronic exposure to toluene on neuronal and glial cell marker proteins in the central nervous system of rats.

The dose dependent effects of chronic exposure to toluene on the neuronal marker proteins (gamma-enolase, calbindin-D28k) and glial cell marker proteins (alpha-enolase, creatine kinase-B, and beta-S100 protein) were investigated in the central nervous system (CNS) of rats. Three groups of animals were exposed to 100 ppm, 300 ppm, or 1000 ppm toluene vapour eight hours a day, six days a week for 16 weeks. The contents of the marker substances were determined with enzyme immunoassays. A significant increase in the three glial cell marker proteins was noted in the cerebellum after exposure to 100 ppm toluene; a more pronounced increase occurred at the higher toluene concentrations. beta-S100 protein also exhibited a dose dependent increase in the brainstem and spinal cord. On the other hand, the two neuronal cell markers did not show a quantitative decrease in the CNS. This means that the development of gliosis, rather than neurone death, is induced by chronic exposure to toluene. The significant biochemical changes induced around the threshold limit value and the concentration dependent alterations suggest that these nerve specific marker proteins may be used to evaluate solvent related damage to the CNS.     

Dose dependent effects of chronic exposure to toluene on neuronal and glial cell marker proteins in the central nervous system of rats.

The dose dependent effects of chronic exposure to toluene on the neuronal marker proteins (gamma-enolase, calbindin-D28k) and glial cell marker proteins (alpha-enolase, creatine kinase-B, and beta-S100 protein) were investigated in the central nervous system (CNS) of rats. Three groups of animals were exposed to 100 ppm, 300 ppm, or 1000 ppm toluene vapour eight hours a day, six days a week for 16 weeks. The contents of the marker substances were determined with enzyme immunoassays. A significant increase in the three glial cell marker proteins was noted in the cerebellum after exposure to 100 ppm toluene; a more pronounced increase occurred at the higher toluene concentrations. beta-S100 protein also exhibited a dose dependent increase in the brainstem and spinal cord. On the other hand, the two neuronal cell markers did not show a quantitative decrease in the CNS. This means that the development of gliosis, rather than neurone death, is induced by chronic exposure to toluene. The significant biochemical changes induced around the threshold limit value and the concentration dependent alterations suggest that these nerve specific marker proteins may be used to evaluate solvent related damage to the CNS.     

Diet, alcohol, body mass, and social factors in relation to blood pressure: the Caerphilly Heart Study.

Data for 387 men who had completed seven-day weighed dietary records as part of the Caerphilly Heart Study were examined for relations of alcohol, diet, body mass index (BMI), and other variables to blood pressure. These included age, smoking, exercise, and social class. For men not on antihypertensive treatment (n = 356) regression analysis showed that age (p less than 0.001), BMI (p less than 0.05), and alcohol intake (p less than 0.01) were significantly related to systolic blood pressure, and BMI (p less than 0.001) and alcohol intake (p less than 0.01) to diastolic blood pressure. In addition, protein intake (p less than 0.05) was significantly and inversely related to the risk of being hypertensive, but other dietary variables were not related to blood pressure. For men on antihypertensive treatment (n = 31) significant inverse correlations were observed between diastolic blood pressure and the intakes of potassium (p less than 0.01), fibre (p less than 0.001), polyunsaturated fat (p less than 0.01), and a number of other dietary variables. Reasons for these differences are discussed.     

The assessment of OPV vaccines by the monkey neurovirulence test: why and how to qualify the experts in histology of central nervous system

Prior to batch release of oral poliovirus vaccines (OPV) for marketing purpose, the World Health Organisation (WHO) and European pharmacopoeia require a monkey neurovirulence test to be performed both by the manufacturer and the relevant National Control Laboratory (NCL) to assess vaccine safety as regards neurovirulence.Due to the subjectivity of histological examination and of neural lesions scoring, the French NCL has set up a proficiency testing procedure to qualify a new expert.     

Prevalence of gallstone disease in relation to smoking, alcohol use, obesity, and glucose tolerance: a study of self-defense officials in Japan.

Risk factors of gallstone disease were investigated in male self-defense officials who received, between October 1986 and December 1990, a retirement health examination at the Self-Defense Forces Fukuoka Hospital, Fukuoka, Japan. Gallbladder ultrasonography, successfully performed with 2,739 of 2,756 men, found 61 men with gallstones and 38 men with previous removal of the gallbladder; the overall prevalence of gallstone disease was 3.6%. Multiple logistic regression analysis assessed the risk of gallstone disease in relation to smoking, alcohol use, body mass index, glucose tolerance, and rank. Alcohol use was associated with a decreased risk, and body mass index was positively related to gallstone disease. Men with impaired glucose tolerance had a slightly elevated risk, whereas diabetes mellitus was not associated with gallstone disease. Analysis for prevalent gallstones and the postcholecystectomy state showed an inverse association of alcohol use with the latter; a positive association with impaired glucose tolerance was also confined primarily to the latter condition. These findings provide little support for a protective effect of alcohol use in the formation of gallstones. It was inconclusive whether impaired glucose tolerance was associated selectively with postcholecystectomy.