Serum ionized magnesium and calcium levels in adult patients with seizures.

OBJECTIVE: Prior studies have been equivocal about whether or not serum levels of the divalent ions calcium and magnesium are altered during different types of seizures. Magnesium is a potential modulator of seizure activity because of its ability to antagonize the excitatory calcium influx through the N-methyl-D-aspartate (NMDA) receptor. We hypothesize that serum ionized levels of calcium (Ca(2+)) and magnesium (Mg(2+)) would be altered significantly during certain types of seizures. MATERIAL AND METHODS: A convenience sample of seizure patients presenting to an emergency department (ED) were enrolled in this prospective study. Novel ion-selective electrodes were used to measure Ca(2+) and Mg(2+). Data were reported as mean values+/-standard deviations. Group comparisons were analyzed by ANOVA with post-hoc testing using the Bonferroni, or the Fisher exact test, where appropriate, alpha = 0.05 (two-tailed). RESULTS: Forty-nine patients with seizure and 32 healthy racially matched controls were included in the study. Seizure patients had a significantly (p<0.001) lower mean Mg(2+), but not total serum Mg and a significantly (p<0.001) higher Ca(2+)/Mg(2+) ratio than that in controls. CONCLUSIONS: We were able to show significantly lower Mg(2+) and higher ionized Ca(2+)/Mg(2+) ratios in seizure patients compared with a racially matched control group.

Ionized magnesium levels and the ratio of ionized calcium to magnesium in asthma patients before and after treatment with magnesium.

OBJECTIVE: Prior studies have been equivocal about the efficacy of magnesium therapy in acute asthma exacerbations. We hypothesize that pretreatment ionized magnesium (Mg(2+)) levels and/or the ratio of ionized calcium to ionized magnesium (Ca(2+)/Mg(2+)) may have been confounding variables in these previous studies. Here, we report on the incidence of abnormal divalent ion levels in our asthma population. MATERIAL AND METHODS: The study was designed as a randomized, double-blind, placebo-controlled trial of intravenous magnesium. Inclusion criteria were: age >18 years, percentage predicted forced expiratory volume (FEV(1)) <75 % after an initial beta-agonist. African-American patients (AA) at an urban university hospital were randomized to 2 g IV Mg or placebo. Mg(2+) and Ca(2+)/Mg(2+) levels were measured pre- and post-infusion. Data were reported as means+/-SD. Student's t-test and Fisher's exact test were used where appropriate (alpha = 0.05, two tailed). RESULTS: Fifty-five AA patients (mean age of 42.7 years+/-15.6 years, range 18-75 years) were studied. A significantly (p<0.05) lower level of Mg(2+) was found in asthma (AS) patients compared with that in the AA group, by 0.03 mmol/L (95 % CI, 0.007-0.053 mmol/L). The AS group had a mean increase in Ca(2+)/Mg(2+) ratios over the AA group, of 0.27 (95 % CI, 0.16-0.38); 100 % of patients with abnormal divalent ion levels were corrected with IV magnesium. CONCLUSIONS: We identified a subgroup of asthmatic patients with significant abnormalities in their divalent ion concentrations, which was corrected with IV magnesium.

Skeletal muscle magnesium content in identical twins, discordant for type 2 diabetes.

BACKGROUND: Low magnesium (Mg) status has been implied as a factor in the development of type 2 diabetes mellitus (DM) and its complications. We therefore studied Mg-status in identical twins, discordant for type 2 DM and in matched controls. Through correlation analysis, possible associations between Mg-status and glucose uptake were evaluated. METHODS: Plasma Mg concentration was measured in 12 monozygote twin pairs, discordant for type 2 DM and in 12 matched controls. Muscle Mg content was measured in 10 persons from each group. An oral glucose tolerance test and a euglycaemic, hyperinsulinaemic clamp were utilized. RESULTS: Neither muscle Mg content nor plasma Mg concentration differed among groups. Plasma Mg concentration decreased during the euglycaemic, hyperinsulinaemic clamp. In the control group, muscle Mg content correlated positively with insulin stimulated glucose disposal rate (r=0.77, p<0.01) and negatively with two hour plasma glucose concentration during an oral glucose tolerance test (OGTT) (r=- 0.64, p<0.05). In the control group, the two hour plasma glucose concentration during an oral glucose tolerance test correlated with the decrease in plasma Mg concentration (r=- 0.80, p<0.002) and with the change in muscle Mg content (r=0.90, p<0.0005) induced by the clamp. None of these associations were found in the two twin groups. CONCLUSIONS: Normal plasma Mg concentration and muscle Mg content were found in persons with type 2 DM and in persons, who were heavily predisposed to the development of type 2 DM, indicating a normal whole-body Mg content. However, the missing associations between measures of glucose disposal and changes in both plasma Mg concentration and muscle Mg content in the two twin groups indicates, that physiological mechanisms, which partly regulates insulin sensitivity and Mg status in healthy individuals are either exhausted or fully utilized in both type 2 DM and in genetically identical twins without DM.

Magnesium reduces insulin-stimulated glucose uptake and serum lipid concentrations in type 1 diabetes.

A magnesium (Mg) deficit has been described in patients with type 1 diabetes, and it has been related to the development of cardiovascular disease. We tested the hypothesis that type 1 diabetic patients have deficits in dietary Mg intake and that proper long-term (24 weeks) oral Mg supplementation would reduce cardiovascular risk factors. Therefore, the Mg status, dietary Mg intake, and the effect of Mg supplementation were evaluated in 10 type 1 diabetic patients and 5 control subjects. Muscle Mg content was decreased by 7% in the type 1 diabetic patients, and it increased by 5% after 24 weeks of oral MgO supplementation. Acute and chronic Mg supplementation decreased serum total cholesterol, serum low-density lipoprotein (LDL)-cholesterol, and apolipoprotein B. Insulin-stimulated glucose uptake decreased by 35% after 24 weeks of oral MgO supplementation. Eight of 10 patients with type 1 diabetes had a daily intake of Mg below 90% of the recommended daily allowance. In conclusion, a Mg deficit was found in type 1 diabetic patients. The deficit might be due partly to a relatively Mg-deficient diet. Mg repletion was associated with a decrease in atherogenic lipid fractions and a reduced insulin-stimulated glucose uptake.

Circulating divalent cations in asymptomatic ovarian hyperstimulation and in vitro fertilization patients.

It is known that ovarian hyperstimulation and in vitro fertilization are accompanied by a steady increase in circulating estrogen and progesterone far beyond what is normal for young women. We have recently demonstrated that the biologically active fractions of calcium and magnesium in blood are altered depending on when in the menstrual phase a blood sample is drawn in normal cycling women. The serum ionized Ca/Mg ratio is also altered in accordance with the menstrual cycles. This suggests that the sex steroid hormones may modulate serum levels of ionized Mg and the ionized Ca/Mg ratio. We therefore studied the relationships between sex steroid hormones and the concentrations of ionized magnesium and calcium in the blood of hyperstimulated patients. We were able to demonstrate that with each increment in estrogen, a decrease in ionized Mg occurred, and as the progesterone rose in the blood, the ionized Ca/Mg ratio increased. Our results support the idea that serum estrogen and progesterone levels in women modulate the blood levels of circulating ionized Mg and the serum ionized Ca/Mg ratio.

Importance of magnesium ions in development of tolerance to ethanol: studies on cultured cerebral vascular smooth muscle cells, type-2 astrocytes and intact rat brain.

This study was designed to examine the roles of intracellular free magnesium ion concentration ([Mg(2+)](i)) in ethanol-induced intoxication and development of tolerance in cultured canine cerebral vascular smooth muscle cells and astrocytes as well as intact rat brain. The basal, resting level of [Mg(2+)](i) in cerebrovascular cells was 732.5 +/- 82.4 microM. Exposure of cultured canine cerebral vascular smooth muscle cells to ethanol (10 and 25 mM) for 24 h reduced the concentrations of [Mg(2+)](i) to 521.1 +/- 59.6 microM, and 308.2 +/- 37.8 microM, respectively. However, exposure of these cultured vascular cells to the same concentrations of ethanol, after initial pretreatment with ethanol for 24 h, failed to interfere with the levels of [Mg(2+)](i). Measurement of [Mg(2+)](i) at 48 h and 72 h indicated that the decreased levels of [Mg(2+)](i) induced by ethanol at 24 h treatment returned toward baseline. Similar experiments were performed in cultured type-2 astrocytes isolated from neonatal rat brain. The basal level of [Mg(2+)](i) in type-2 astrocytes was about 125 microM. Incubation of these cells with 10 mM ethanol for 10 min resulted in a 27% reduction in the level of [Mg(2+)](i), whereas incubation with 25 mM ethanol resulted in almost a 50% reduction in [Mg(2+)](i). The decreased levels of [Mg(2+)](i) lasted around 30 min, until the measurement finished. Continuous incubation of these cultured astrocytes, with ethanol (either 10 mM or 25 mM), for more than 24 h, indicated that the concentrations of [Mg(2+)](i) in type-2 astrocytes were equivalent to those at basal, resting levels. In vivo 31P-NMR spectroscopy, performed on intact rat brains, indicated that an initial administration of 4 mg/kg ethanol ( approximately 20-25 mM blood alcohol level) resulted (after 20-40 min of exposure) in severe deficits in whole brain [Mg(2+)](i) (550 +/- 33 microM to 358 +/- 24 microM). Repeated injections of ethanol (4 mg/kg) over the next 24-72 h resulted in progressively diminishing effects on brain [Mg(2+)](i). These experimental data indicate that chronic ethanol treatment can induce a tolerance to depletion of [Mg(2+)](i) in cerebrovascular smooth muscle cells, type-2 astrocytes as well as intact rat brain. The results suggest that [Mg(2+)](i) might play a major role in alcohol-induced tolerance in the brain.

Serum ionized magnesium in post-traumatic headaches.

The objective of this study was to determine the values of serum ionized magnesium, total magnesium, and ionized calcium/ionized magnesium ratios in children with headaches. One hundred thirty-five children with primary complaints of headaches were classified according to the criteria of the International Headache Society. Blood samples were obtained and tested for ionized magnesium (IMg(2+)), total magnesium, ionized calcium (ICa(2+)), and pH. The ICa(2+)/IMg(2+) ratio was calculated. Nine children were given a diagnosis of post-traumatic headache. Six of them had statistically significant (P <.05) lowered IMg(2+) levels and high ICa(2+)/IMg(2+) ratios. Abnormalities in serum IMg(2+) concentrations and ICa(2+)/IMg(2+) ratios were found in children with post-traumatic headaches, but total magnesium levels were normal.

Importance of extracellular Ca2+ and intracellular Ca2+ release in ethanol-induced contraction of cerebral arterial smooth muscle.

The present study was designed to investigate the roles of extracellular Ca2+ ([Ca2+]0) influx and intracellular free Ca2+ ([Ca2+]i) release in ethanol-induced contractions of isolated canine cerebral arteries and primary cultured, cerebral vascular smooth muscle cells. Ethanol (20-200 mM) produced significant contractions in isolated canine basilar arterial rings in a concentration-dependent manner. Removal of [Ca2+]0 and pretreatment of canine basilar arterial rings with verapamil (an antagonist of voltage-gated Ca2+ channels), thapsigargin (a selective antagonist of the sarcoplasmic reticulum Ca2+ pump), caffeine plus ryanodine (a specific antagonist of ryanodine-sensitive Ca2+ release), or heparin (an inositol 1,4,5,-trisphosphate [InsP3]-mediated Ca2+ release antagonist) markedly attenuated (approximately 50%-80%) ethanol-induced contractions. The absence of [Ca2+]0 and preincubation of primary single smooth muscle cells obtained from canine basilar arteries with verapamil, thapsigargin, heparin, or caffeine plus ryanodine markedly attenuated (approximately 50%-80%) the transient and sustained elevations in [Ca2+]i induced by ethanol. Results of the present study suggest to us that both Ca2+ influx through voltage-gated Ca2+ channels and Ca2+ release from intracellular stores (both InsP3 sensitive and ryanodine sensitive) are required for ethanol-induced contractions of isolated canine basilar arteries.

Antioxidants prevent depletion of [Mg2+]i induced by alcohol in cultured canine cerebral vascular smooth muscle cells: possible relationship to alcohol-induced stroke.

Low serum concentrations of Mg(2+) ions have been reported, recently, in patients with coronary disease, atherosclerosis, and stroke as well as in patients with cerebral hemorrhage. The aim of the present study was to determine whether potent antioxidants [alpha-tocopherol and pyrrolidine dithiocarbamate (PDTC)] can prevent or ameliorate intracellular Mg(2+) ([Mg(2+)](i)) depletion associated with cerebral vascular injury induced by alcohol. Exposure of cultured canine cerebral vascular smooth muscle cells to alcohol (10-100 mM) for 24 h induced marked depletion in [Mg(2+)](i) (i.e., approximately 30-65%, depending upon alcohol concentration). Treatment of the cultured cells with either PDTC (0.1 microM) or alpha-tocopherol (15 microM) for 24 h, alone, failed to interfere with basal [Mg(2+)](i) levels. However, preincubation of the cells with either alpha-tocopherol or PDTC for 24 h completely inhibited the depletion of [Mg(2+)](i) induced by exposure to 10-100 mM ethanol. These results indicate that alpha-tocopherol and PDTC prevent decreases in [Mg(2+)](i) produced by ethanol. Moreover, these new results suggest that such protective effects of alpha-tocopherol and PDTC on cerebral vascular cells might be useful therapeutic tools in prevention and amelioration of cerebral vascular injury and stroke in alcoholics.

Divalent cations in women with PCOS: implications for cardiovascular disease.

Polycystic ovary syndrome (PCOS) patients are known to have a high incidence of insulin resistance and glucose intolerance and tend to be at eventual high risk of hypertension, diabetes mellitus and cardiovascular disease. It has been repeatedly shown that a low serum ionized magnesium (Mg2+) and a high ionized calcium to magnesium (Ca2+/Mg2+) ratio is often associated with insulin resistance, cardiovascular problems, diabetes mellitus and hypertension. We were therefore interested in assessing the serum divalent cation profile of PCOS patients compared with that of normal women of similar age. We found significantly lower serum Mg2+ and total magnesium and a significantly higher serum Ca2+/Mg2+ ratio in the PCOS patients compared with the controls. No correlation was found, however, between the serum concentrations of steroid hormones (estrogen, progesterone and testosterone), or any of the cations in the PCOS patients or the controls.

Extracellular magnesium regulates nuclear and perinuclear free ionized calcium in cerebral vascular smooth muscle cells: possible relation to alcohol and central nervous system injury.

Quantitative digital imaging microscopy, confocal laser scanning microscopy (CLSM), and multiple molecular fluorescent probes were utilized to test the hypothesis that cerebral vascular muscle cell nuclear ([Ca(2+)](n)), perinuclear ([Ca(2+)](pn)), and cytoplasmic free calcium ([Ca(2+)](i)) levels are regulated by the concentration of extracellular free magnesium ions ([Mg(2+)](o)). Primary cultured canine cerebral vascular smooth muscle cells were loaded with either fura-2/AM, indo-1/AM, or fluo-3/AM, and the subcellular Ca(2+) responses to stepwise reduction in [Mg(2+)](o) (i.e., from 1.36 to 0.17 mM) were analyzed over time. With normal 1.36 mM [Mg(2+)](o)-containing incubation media, basal mean [Ca(2+)](i) was 89.6+/-15 nM. Lowering [Mg(2+)](o) to 1.07, 0.88, 0.48, and 0.17 mM resulted in rapid (<4 min) increments in [Ca(2+)](i) going to 213+/-43, 368+/-67, 471+/-77, and 642+/-98 nM, respectively; the longer the exposure time (up to 30 min) to lowered [Mg(2+)](o), the higher the [Ca(2+)](i). Restoration of [Mg(2+)](o) to normal caused decreases in [Ca(2+)](i) to 215.9+/-42.3 nM, but only complete removal of [Ca(2+)](o) returned [Ca(2+)](i) to basal levels. Results show that basal [Ca(2+)](pn) (282+/-92 nM) exceeds basal cytoplasmic Ca(2+) (61+/-27.8 nM) and [Ca(2+)](n) (20+/-7.6 nM). However, reduction of normal [Mg(2+)](o) to 0.48 mM resulted in dramatic, rapid rises in all subcellular compartments, where [Ca(2+)](pn) (1503+/-102 nM)>cytoplasmic Ca(2+) (688+/-49 nM) approximately equal to [Ca(2+)](n) (674+/-12 nM). Nuclear Ca(2+) rose dramatically (e.g., 35-40 times basal levels). Both verapamil (1 microM) and Ni(2+) (5 mM) prevented, completely, the rises in Ca(2+) in all compartments, suggesting that Mg(2+)-dependent Ca(2+) accumulation may be dependent on nuclear, endoplasmic reticulum-Golgi, and cytoplasmic L-type voltage membrane-regulated Ca(2+) channels. The normally low [Ca(2+)](n) suggests that Ca(2+) does not transport passively across the nuclear membrane in cerebral vascular smooth muscle cells. These results may help to explain much of the impact of hypomagnesemic states on cerebral-central nervous system pathobiology, and, particularly, alcohol-induced strokes.

Importance of PKC and PI3Ks in ethanol-induced contraction of cerebral arterial smooth muscle.

We investigated the relationships of two potential intracellular signaling pathways, protein kinase C (PKC) and phosphatidylinositol 3-kinases (PI3Ks), to ethanol-induced contractions in cerebral arteries. Ethanol (20-200 mM) induces concentration-dependent constriction in isolated canine basilar arteries that is inhibited in a concentration-dependent manner by pretreatment of these vessels with 10(-9)-10(-3) M Gö-6976 (an antagonist selective for PKC-alpha and PKC-betaI), 10(-10)-10(-4) M bisindolylmaleimide I (a specific antagonist of PKC), and 10(-10)-10(-4) M wortmannin or 10(-8)-10(-2) M LY-294002 (selective antagonists of PI3Ks). Ethanol-induced increases in intracellular Ca(2+) concentration (from approximately 100 to approximately 500 nM) in canine basilar smooth muscle cells are also suppressed markedly (approximately 20-70%) in the presence of a similar concentration range of Gö-6976, bisindolymaleimide I, wortmannin, or LY-294002. This study suggests that activation of PKC isoforms and PI3Ks appears to be an important signaling pathway in ethanol-induced vasoconstriction of cerebral blood vessels.

Antioxidants prevent ethanol-induced contractions of canine cerebral vascular smooth muscle: relation to alcohol-induced brain injury.

The present study was designed to test the hypothesis that alpha-tocopherol (Vit. E) and pyrrolidine dithiocarbamate (PDTC) might exert direct effects on alcohol-induced contractions of canine basilar cerebral arteries. After precontraction of arterial ring segments with ethanol, PDTC (10(-8)-10(-6) M) and Vit. E (10(-6)-10(-4) M) induced concentration-dependent relaxations of cerebral arteries, compared to untreated controls. The effective concentrations producing approximately 50% of the maximal relaxation responses (EC(50) values) were about 2.48+/-0.09 x 10(-7) M for PDTC, and 1.87+/-0.10 x 10(-5) mM for Vit. E, respectively. Preincubation of these arterial rings with EC(50)'s of PDTC or Vit. E for 40 min attenuate markedly the contractions produced by alcohol, at concentrations of 1-400 mM. However, both PDTC and Vit.E do not relax equi-potent precontractions induced by either KCl or prostaglandin F(2alpha) (PGF(2alpha)) or inhibit their contractions. These data suggest that alcohol-induced contractions of cerebral arteries are mediated via excitation-contraction coupling pathways different from those used by KCl or receptor-mediated agonists such as PGF(2alpha). The present results, when viewed in light of other recently published data, suggest that antioxidants may prove useful in the amelioration and treatment of alcohol-induced brain damage and strokes.

Sex steroid hormones exert biphasic effects on cytosolic magnesium ions in cerebral vascular smooth muscle cells: possible relationships to migraine frequency in premenstrual syndromes and stroke incidence.

Clinically, it is known that: (1) magnesium (Mg) supplementation relieves premenstrual problems (e.g., migraine, bloating and edema) occurring in the late luteal phase of the menstrual cycle; and (2) migraine syndromes, particularly in women, are associated with deficits in brain and serum ionized Mg levels. We investigated whether concentrations of sex steroid hormones, found in the serum during the menstrual cycle of women, are associated with changes in the levels of cytosolic free magnesium ions ([Mg2+]i in single cultured canine cerebral vascular smooth muscle cells. The resting level of [Mg2+]i in these cells was 645 +/- 89 microM before exposure to sex steroid hormones. Exposure of these vascular cells to a low concentration of estrogen (10 pg/ml) failed to interfere with the levels of [Mg2+]i. However, exposure to estrogen, at concentrations ranging from 40 to 200 pg/ml, induced significant loss of [Mg2+]i in a concentration-dependent manner. At a concentration of 200 pg/ml estrogen, the level of [Mg2+]i decreased approximately 30% in comparison with controls. Progesterone produced biphasic effects on the levels of [Mg2+]i, depending on its concentration. Exposure of the cultured cells to a low concentration of progesterone (0.5 ng/ml) resulted in an increased level of [Mg2+]i (from 690 +/- 50 microM to 753 +/- 56 microM, p < 0.05). However, when these cells were exposed to higher concentrations of progesterone (i.e., from 5.0 to 20 ng/ml), the cellular levels of [Mg2+]i were decreased significantly. The higher the estrogen or progesterone concentration, the lower the levels of [Mg2+]i. In contrast, testosterone, a male hormone, didn't produce any significant alteration in [Mg2+]i levels in these cerebral vascular smooth muscle cells. These data indicate that low, physiological concentrations of female sex hormones, estrogen and progesterone, help cerebral vascular smooth cells sustain normal concentrations of [Mg2+]i, which are beneficial to vascular function, whereas high levels of estrogen and progesterone deplete, significantly, [Mg2+]i in cerebral vascular smooth muscle cells, possibly resulting in cerebrovasospasms and reduced cerebral blood flows related to premenstrual syndromes, migraine and stroke risk. Our findings could provide new insight into the mechanism whereby migraine occurs frequently in the late luteal phase in the premenstrual syndrome. In addition, our results demonstrate that female sex steroids but not testosterone (in physiologic concentrations) can exert direct effects on [Mg2+]i in cerebral vascular cells.

Use of protein kinase C inhibitors results in rapid [Mg(2+)](i) mobilization in primary cultured rat aortic smooth muscle cells: are certain kinase C isoforms natural homeostatic regulators of cystolic free Mg(2+).

The effects of five different protein kinase C inhibitors--calphostin C, chelerythrine, bisindolylmaleimide I, staurosporine and Gö6979--on intracellular free magnesium ([Mg(2+)](i)) content and mobilization were investigated in primary, cultured rat aortic smooth muscle cells. All these protein kinase C inhibitors significantly and rapidly increased [Mg(2+)](i) both in normal media (1.2 mM Mg(2+)) and in Mg(2+) free media. These data suggest that the increments of [Mg(2+)](i), induced by the diverse protein kinase C inhibitors, are derived from the release of bound intracellular Mg(2+) and that activation of protein kinase C isozymes are normally responsible for helping to maintain basal levels of [Mg(2+)](i) in rat aortic smooth muscle cells.

Ethanol-induced contractions in cerebral arteries: role of tyrosine and mitogen-activated protein kinases.

BACKGROUND AND PURPOSE: The relationship between alcohol consumption and stroke appears complex; moderate ingestion is associated with reduced stroke risk, while heavy intake is associated with increased stroke risk. Ethanol has been shown both experimentally and epidemiologically to induce hemorrhagic and ischemic strokes, which are associated with cerebral vasoconstriction. Ethanol is known to induce contraction in isolated cerebral arteries and intact microvessels from diverse mammalian animals. The relationships between ethanol-induced contractions in cerebral arteries, intracellular free Ca(2+) ([Ca(2+)](i)), tyrosine kinases (including the src family), and mitogen-activated protein kinases (MAPK) were investigated in the present study. METHODS: Cerebral arterial muscle tension and [Ca(2+)](i) were quantified by an isometric contraction technique and direct visualization of Ca(2+) in single cells. RESULTS: Ethanol induces concentration-dependent contractions in intact canine basilar arteries, which are attenuated significantly by pretreatment of the arteries with low concentrations of an antagonist of protein tyrosine kinases (genistein); an src homology 2 (SH2) domain inhibitor peptide; a highly specific antagonist of p38 MAPK (SB-203580); a potent, selective antagonist of MEK1/MEK2 (U0126); and a selective antagonist of mitogen-activated protein kinase kinase (MAPKK) (PD-98059). IC(50) levels obtained for these 5 antagonists are consistent with reported K:(i) values for these tyrosine kinase, MAPK, and MAPKK antagonists. Ethanol induces transient and sustained increases in [Ca(2+)](i) in primary single smooth muscle cells from canine basilar arteries, which are markedly attenuated in the presence of genistein, an SH2 domain inhibitor peptide, SB-203580, U0126, and PD-98059. Several specific antagonists of known endogenously formed vasoconstrictors do not inhibit or attenuate either the ethanol-induced contractions or the elevation of [Ca(2+)](i). CONCLUSIONS: The present study suggests that activation of protein tyrosine kinases (including the src family) and MAPK appear to play important roles in the ethanol-induced contractions and the elevation of [Ca(2+)](i) in smooth muscle cells from canine basilar arteries. The results could be used to suggest that selective antagonists of protein tyrosine kinases and MAPK may be useful both prophylactically and therapeutically in alcohol-induced strokes.

Tension headaches and muscle tension: is there a role for magnesium?

Although many theories and hypotheses have been offered for the etiology of tension-type headache (TH), no one previous hypothesis seems to adequately explain TH. This may, in large measure, account for why it is often difficult to effectively treat TH. Herein, we review current and old hypotheses of TH and offer a new hypothesis which is consistent with what is known about TH. We show that magnesium (Mg) metabolism may be pivotal in both the etiology and treatment of TH. Measurement of serum ionized Mg2+ (IMg2+) levels and brain intracellular free Mg2+ ([Mg2+]i) appear to offer excellent methods for establishing the validity of our hypothesis. Since approximately 70% of patients who have a TH exhibit muscular tightness and tenderness, it is distinctly possible that problems in Mg metabolism and dietary intake are the links to concomitant muscle tension and TH. The significance of release of pain mediators, muscle cramps, muscle strains (and damage) and muscle tension to TH, and its relationship to Mg metabolism, are reviewed. These are all associated with a Mg-deficient state. It seems clear from the available data that TH's are more associated with muscle tension or scalp tension than any other headache type. From the data available, Mg supplementation appears to be of great benefit in many of these situations. We believe there is a great need for clinicians to examine Mg2+ metabolism, bioavailable Mg2+ in muscle tissues and blood, and the effectiveness of Mg salts (in a double-blinded, placebo-controlled manner) in subjects with TH and muscle tension.

Low [Mg(2+)](o) induces contraction and [Ca(2+)](i) rises in cerebral arteries: roles of ca(2+), PKC, and PI3.

Removal of extracellular Ca(2+) concentration ([Ca(2+)](o)) and pretreatment of canine basilar arterial rings with either an antagonist of voltage-gated Ca(2+) channels (verapamil), a selective antagonist of the sarcoplasmic reticulum Ca(2+) pump [thapsigargin (TSG)], caffeine plus a specific antagonist of ryanodine-sensitive Ca(2+) release (ryanodine), or a D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)]- mediated Ca(2+) release antagonist (heparin) markedly attenuates low extracellular Mg(2+) concentration ([Mg(2+)](o))-induced contractions. Low [Mg(2+)](o)-induced contractions are significantly inhibited by pretreatment of the vessels with Gö-6976 [a protein kinase C-alpha (PKC-alpha)- and PKC-betaI-selective antagonist], bisindolylmaleimide I (Bis, a specific antagonist of PKC), and wortmannin or LY-294002 [selective antagonists of phosphatidylinositol-3 kinases (PI3Ks)]. These antagonists were also found to relax arterial contractions induced by low [Mg(2+)](o) in a concentration-dependent manner. The absence of [Ca(2+)](o) and preincubation of the cells with verapamil, TSG, heparin, or caffeine plus ryanodine markedly attenuates the transient and sustained elevations in the intracellular Ca(2+) concentration ([Ca(2+)](i)) induced by low-[Mg(2+)](o) medium. Low [Mg(2+)](o)-produced increases in [Ca(2+)](i) are also suppressed markedly in the presence of Gö-6976, Bis, wortmannin, or LY-294002. The present study suggests that both Ca(2+) influx through voltage-gated Ca(2+) channels and Ca(2+) release from intracellular stores [both Ins(1,4,5)P(3) sensitive and ryanodine sensitive] play important roles in low-[Mg(2+)](o) medium-induced contractions of isolated canine basilar arteries. Such contractions are clearly associated with activation of PKC isoforms and PI3Ks.

Low [Mg(2+)](o) induces contraction of cerebral arteries: roles of tyrosine and mitogen-activated protein kinases.

The present study was designed to investigate the mechanism of action of low extracellular magnesium ion concentration ([Mg(2+)](o)) on isolated canine basilar arteries and single cerebral vascular smooth muscle cells from these arteries. Low-[Mg(2+)](o) medium (0-0.6 mM) produces endothelium-independent contractions in isolated canine basilar arteries in a concentration-dependent manner; the lower the concentration of [Mg(2+)](o), the stronger the contractions. The low-[Mg(2+)](o) medium-induced contractions are significantly attenuated by pretreatment of the arteries with low concentrations of either SB-203580, U-0126, PD-98059, genistein, or an Src homology 2 (SH2) domain inhibitor peptide. IC(50) levels obtained for these five antagonists are consistent with reported inhibitor constant (K(i)) values for these tyrosine kinase and mitogen-activated protein kinase (MAPK) antagonists. Low-[Mg(2+)](o) medium (0-0.6 mM) produces transient intracellular calcium ion concentration ([Ca(2+)](i)) peaks followed by a slow, sustained, and elevated plateau of [Ca(2+)](i) in primary single smooth muscle cells from canine basilar arteries. Low-[Mg(2+)](o) medium induces rapid and stable increases in [Ca(2+)](i); these increases are inhibited markedly in the presence of either SB-203580, U-0126, PD-98059, genistein or a SH2 domain inhibitor peptide. Several specific antagonists of known endogenously formed vasoconstrictors do not inhibit or attenuate either the low-[Mg(2+)](o)-induced contractions or the elevation of [Ca(2+)](i). The present study suggests that activation of several cellular signaling pathways, such as protein tyrosine kinases (including the Src family) and MAPK, appears to play important roles in low-[Mg(2+)](o)-induced contractions and the elevation of [Ca(2+)](i) in smooth muscle cells from canine basilar arteries.

Antioxidants prevent elevation in [Ca(2+)](i) induced by low extracellular magnesium in cultured canine cerebral vascular smooth muscle cells: possible relationship to Mg(2+) deficiency-induced vasospasm and stroke.

Low serum concentrations of Mg(2+) ions have been reported, recently, in patients with coronary disease, atherosclerosis and stroke as well as in patients with cerebral hemorrhage. The aim of the present study was to determine whether potent antioxidants [alpha-tocopherol and pyrrolidine dithiocarbamate (PDTC)] can prevent or ameliorate intracellular Ca(2+) ([Ca(2+)](i)) overload associated with cerebral vascular injury induced by low extracellular free Mg(2+) ([Mg(2+)](o)). Exposure of cultured canine cerebral vascular smooth muscle cells to low [Mg(2+)](o) (0.15-0.6 mM) vs. normal [Mg(2+)](o) (1.2 mM) for either 10 min or 2 h induced concentration-dependent rises in [Ca(2+)](i). Treatment of the cultured cells with either PDTC (0.1 microM) or alpha-tocopherol (15 microM) for 24 h, alone, failed to interfere with basal [Ca(2+)](i) levels. However, preincubation of the cells with either alpha-tocopherol or PDTC for 24 h completely inhibited the elevation of [Ca(2+)](i) induced by exposure to low [Mg(2+)](o), not only for 10 min, but also for 2 h. These results indicate that alpha-tocopherol and PDTC prevent rises in [Ca(2+)](i) produced by low [Mg(2+)](o), which probably result from low [Mg(2+)](o)-induced lipid peroxidation of cerebral vascular smooth muscle cell membranes. Moreover, these new results suggest that such protective effects of alpha-tocopherol and PDTC on cerebral vascular cells might be useful therapeutic tools in cerebral vascular injury associated with low [Mg(2+)](o) and accumulation of [Ca(2+)](i).