Incidence and risk factors for deep infection after primary shoulder arthroplasty: a meta-analysis.

OBJECTIVE: Many risk factors associated with deep infections after primary shoulder arthroplasty remain controversial and have not yet been summarized. As such, the aim of the present study was to quantitatively summarize the risk factors associated with deep infections after primary shoulder arthroplasty. MATERIALS AND METHODS: Computerized and additional manual searches on the Medline, Embase, Chinese National Knowledge Infrastructure (CNKI), and Cochrane central database for potential studies, published from inception to March 2022, were performed. All studies that assessed risk factors for deep infection after primary shoulder arthroplasty were selected without language restrictions. Eligible studies were required to fulfill quality assessment criteria from the Consort statement and to evaluate risk factors for deep infection after primary shoulder arthroplasty. Two reviewers independently extracted the relevant data, with disagreements resolved by consensus. Statistical analyses were performed using Stata version 11.0 (Statacorp LLC, College Station, TX, USA). RESULTS: Seven studies including 493,148 patients who underwent primary shoulder arthroplasty, among whom 1,314 experienced infection (0.3%), were eligible and included in this meta-analysis. Meta-analysis revealed that significantly increased risk factors for infection after primary shoulder arthroplasty included male sex (odds ratio [OR] 1.79 [95% confidence interval (CI) 1.23-2.60]), avascular necrosis (OR 2.64 [95% CI 1.61-4.34]), rotator cuff arthropathy (OR 2.14 [95% CI 1.55-2.95]), proximal humerus fracture (OR 2.68 [95% CI 1.93-3.73]), and non-union of humerus fracture (OR 5.32 [95% CI 3.52-8.02]). In contrast, advanced age was associated with a decreased likelihood for development of infection (OR 0.97 [95% CI 0.94-1]). CONCLUSIONS: Surgeons should devote close attention to the above-mentioned medical conditions to reduce deep infection after primary shoulder arthroplasty.

[Population genetic structure of wild-growing ginseng (Planax ginseng C. A. Meyer) assessed using AFLP markers].

Genetic variability in ten populations of wild-growing ginseng was assessed using AFLP markers with the application of fragment analysis on a genetic analyzer. The variation indices were high in the populations (P = 55.68%; H(S) = 0.1891) and for the species (P = 99.65%; H(S) = 0.2857). Considerable and statistically significant population differentiation was demonstrated (theta = 0.363; Bayesian approach, "full model"; F(ST) = 0.36, AMOVA). The results of AMOVA and Bayesian analysis indicate that 64.46% of variability is found within the populations. Mantel test showed no correlation between the genetic and geographic distances among the populations (r = -0.174; P = 0.817). Hierarchical AMOVA and analysis of genetic relationships based on Euclidean distances (NJ, PCoA, and MST) identified two divergent population groups of ginseng. Low gene flow between these groups (N(m) = 0.4) suggests their demographic independence. In accordance to the concept of evolutionary significant units (ESU), these population groups, in terms of the strategy and tactics for conservation and management of natural resources, should be treated as management units (MUs). The MS tree topology suggests recolonization of southern Sikhote-Alin by ginseng along two directions, from south and west.

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.

Jaks and stats as therapeutic targets.

Cytokines have critical functions in regulating immune responses. A large number of these factors bind related receptors termed the Type I and Type II families of cytokine receptors. These receptors activate Janus kinases (Jaks) and Stat family of transcription factors. The essential and specific function of Jaks and Stats is particularly well illustrated by human and mouse mutations. The possibility that these molecules could be targeted to produce novel immunosuppressive compounds is considered in this review.

Hierarchy of protein tyrosine kinases in interleukin-2 (IL-2) signaling: activation of syk depends on Jak3; however, neither Syk nor Lck is required for IL-2-mediated STAT activation.

Interleukin-2 (IL-2) activates several different families of tyrosine kinases, but precisely how these kinases interact is not completely understood. We therefore investigated the functional relationships among Jak3, Lck, and Syk in IL-2 signaling. We first observed that in the absence of Jak3, both Lck and Syk had the capacity to phosphorylate Stat3 and Stat5a. However, neither supported IL-2-induced STAT activation, nor did dominant negative alleles of these kinases inhibit. Moreover, pharmacological abrogation of Lck activity did not inhibit IL-2-mediated phosphorylation of Jak3 and Stat5a. Importantly, ligand-dependent Syk activation was dependent on the presence of catalytically active Jak3, whereas Lck activation was not. Interestingly, Syk functioned as a direct substrate of Jak1 but not Jak3. Additionally, Jak3 phosphorylated Jak1, whereas the reverse was not the case. Taken together, our data support a model in which Lck functions in parallel with Jak3, while Syk functions as a downstream element of Jaks in IL-2 signaling. Jak3 may regulate Syk catalytic activity indirectly via Jak1. However, IL-2-mediated Jak3/Stat activation is not dependent on Lck or Syk. While the essential roles of Jak1 and Jak3 in signaling by gammac-utilizing cytokines are clear, it will be important to dissect the exact contributions of Lck and Syk in mediating the effects of IL-2 and related cytokines.

Influence of morning or evening administration on absorption of theophylline.

BACKGROUND: Bronchoconstriction during the night causing nocturnal and early morning wheezing is recognized as a major problem for asthmatics. Oral sustained-release theophyllines (SRTs) were developed to reduce the symptoms. A circadian variation in theophylline kinetics has been demonstrated with many SRTs. The purpose of this study was to evaluate the differences in serum theophylline concentration (STC) caused by morning or evening dosing of Euphyllin Retard, a brand of SRT, for a period of 36 hours following oral administration. METHODS: A total of nine non-smoking healthy male volunteers were involved in the study, with a two-period crossover comparison. They were randomly divided into two groups. The first group took a single oral dose of 350 mg Euphyllin Retard at 8:00 A.M. and the second group took it at 8:00 P.M. Blood samples were collected during the 36 hours following administration. Two weeks later, the first group took the drug at night and the second group took it in the morning. The difference in the absorption of theophylline with daytime administration versus night-time administration was assessed using pharmacokinetic parameters derived from the plasma drug concentration vs time curve. RESULTS: The means of unextrapolated area under the concentration vs time curve (AUC) from time 0 to 24 hours (AUCUN) and of the extrapolated AUC from time 0 to infinity (AUCEX) in the night phase were higher than those in the day phase (62.403 micrograms/ml/hr vs 53.081 micrograms/ml/hr, p = 0.9186; 107.21 micrograms/ml/hr vs 98.879 micrograms/ml/hr, p = 0.8807, respectively). The mean of maximum concentration (Cmax) was higher in the night phase than that in the day phase (4.166 micrograms/dl vs 3.451 micrograms/dl, p = 0.9234). Daytime administration showed a delayed time to maximum concentration (Tmax) when compared to that of night-time administration (6.5 hr vs 5.75 hr, p = 0.6244). The terminal elimination rate constant (Kel) was lower in the day phase than in the night phase (0.053 l/hr vs 0.06 l/hr, p = 0.7601). The day phase and night phase data are combined data from the two night and two day groups. The statistical analysis of the results show that the time of administration does not influence the STC. CONCLUSIONS: No diurnal variation in theophylline kinetics was found with Euphyllin Retard. This study was performed in a limited number of normal healthy subjects, and the same result is yet to be proved in asthmatic patients and a larger population of normal subjects.

Low concentrations of ethanol deplete type-2 astrocytes of intracellular free magnesium.

The acute effects of low concentrations of ethanol on intracellular free magnesium ions ([Mg2+]i) in cultured type-2 astrocytes were studied by digital imaging microscopy using the Mg2+ fluorescent probe, mag-fura-2. In 0-mM ethanol, the basal level of [Mg+]i was 124.7+/-2.56 microM with a heterogeneous distribution within the cells. Treatment of the cells with 10 and 25 mM ethanol (10 min) resulted in rapid concentration-dependent reduction in [Mg2+]i; the greater the concentration of alcohol, the greater the depletion of [Mg2+]i. Exposure of cells to 10 and 25 mM resulted in approximately 27 and 50% reductions in [Mg2+]i, respectively. Reincubation in normal Mg2+-physiological buffer solution restored [Mg2+]i levels. These observations may suggest that acute "binge drinking" of ethanol, which often results in cerebral ischemia and stroke, may do so as a result of depletion of astrocytic [Mg2+]i, possibly producing disruption of the blood-brain barrier.

The effect of thalidomide and 2 analogs on collagen induced arthritis.

OBJECTIVE: Thalidomide has been described as an inhibitor of both angiogenesis (which may account for its teratogenic effects on limb bud formation) and tumor necrosis factor-alpha (TNF-alpha) production. We evaluated its therapeutic potential in collagen induced arthritis (CIA), a rat model of rheumatoid arthritis (RA). METHODS: Rats were administered orally 200 mg/kg/day thalidomide (n = 10) or either of 2 analogs, EM-12 (n = 9) or supidimide (n = 9). An additional group was given thalidomide (n = 10) at 200 mg/kg twice daily, and a control group (n = 13) was given vehicle only. At completion of the protocols, serum levels of TNF-alpha and vascular endothelial growth factor (VEGF) were measured. RESULTS: Suppression of inflammatory synovitis by clinical and radiographic criteria was significantly lower in all experimental protocols except the lower dose thalidomide group. The EM-12 analog was the most efficacious, and twice daily thalidomide was better than once daily. The incidence of arthritis onset was comparable among all groups. Strong cell mediated and humoral responses to type II collagen, measured by a radiometric delayed type hypersensitivity assay and anti-type II collagen IgG ELISA, respectively, were similar in the experimental and control groups. TNF-alpha and VEGF levels were increased in all rats immunized with collagen compared to naive controls. CONCLUSION: Thalidomide and its analogs can suppress the clinical severity of rat CIA, but the mechanism of action is not a result of TNF-alpha or VEGF downregulation.

Chronic treatment of cultured cerebral vascular smooth cells with low concentration of ethanol elevates intracellular calcium and potentiates prostanoid-induced rises in [Ca2+]i: relation to etiology of alcohol-induced stroke.

The influence of chronic treatment of cultured canine cerebral vascular smooth muscle cells, with low concentrations of ethanol, on the intracellular concentrations of free calcium ([Ca2+]i) was studied by use of the fluorescent indicator, fura-2, and digital imaging microscopy. The resting level of [Ca2+]i in the cerebral vascular smooth muscle cells was 89 +/- 3.2 nM. Exposure of these cells to 10 and 25 mM ethanol for 5 days resulted in significant elevation of [Ca2+]i (mean rises to 208 +/- 11.4 and 307 +/- 14.0 nM, respectively), and potentiated the transient rise in [Ca2+]i induced by 10(-7) M PGF2 alpha. However, exposure of these cerebral cells to a high-concentration ethanol (100 mM) resulted in only a slight increase of [Ca2+]i (106 +/- 6.9 nM) and lack of effects on the [Ca2+]i response to PGF2 alpha. Irrespective of the different ethanol treatments, the subcellular distribution of [Ca2+]i was heterogeneous in all the cells tested. Our data suggest that chronic exposure of cerebral vascular smooth muscle cells to ethanol, particularly at low concentrations, results in dramatic increases in [Ca2+]i and the responses of these vascular smooth muscle cells to prostanoids. These results support an hypothesis whereby ethanol induces stroke by causing spasm and rupture of cerebral blood vessels as a consequence of large rises in intracellular Ca2+.

Low levels of serum ionized magnesium are found in patients early after stroke which result in rapid elevation in cytosolic free calcium and spasm in cerebral vascular muscle cells.

Ninety-eight patients admitted to the emergency rooms of three urban hospitals with a diagnosis of either ischemic stroke or hemorrhagic stroke exhibited early and significant deficits in serum ionized Mg2+ (IMg2+), but not total Mg, as measured with a unique Mg2+-sensitive ion-selective electrode. Twenty-five percent of these stroke patients exhibited >65% reductions in the mean serum IMg2+ found in normal healthy human volunteers or patients admitted for minor bruises, cuts or deep lacerations. The stroke patients also demonstrated significant elevation in the serum ionized Ca2+ (ICa2+)/IMg2+ ratio, a sign of increased vascular tone and cerebrovasospasm. Exposure of primary cultured canine cerebral vascular smooth muscle cells to the low concentrations of IMg2+ found in the stroke patients, e.g. 0.30-0.48 mM, resulted in rapid and marked elevations in cytosolic free calcium ions ([Ca2+]i) as measured with the fluorescent probe, fura-2, and digital image analysis. Coincident with the rise in [Ca2+]i, many of the cerebral vascular cells went into spasm. Reintroduction of normal extracellular Mg2+ ion concentrations failed to either lower the [Ca2+]i overload or reverse the rounding-up of the cerebral vascular cells. These results suggest that changes in Mg2+ metabolism play important roles in stroke syndromes and in the etiology of cerebrovasospasm associated with cerebral hemorrhage.

Extracellular Mg2+ regulates intracellular Mg2+ and its subcellular compartmentation in fission yeast, Schizosaccharomyces pombe.

Effects of extracellular magnesium ions ([Mg2+]o) on intracellular free Mg2+ ([Mg2+]i) and its subcellular distribution in single fission yeast cells, Schizosaccharomyces pombe, were studied with digital-imaging microscopy and an Mg2+ fluorescent probe (mag-fura-2). Using 0.44 mM [Mg2+]o, [Mg2+]i in yeast cells was 0.91 +/- 0.08 mM. Elevation of [Mg2+]o to 1.97 mM induced rapid (within 5 min) increments in [Mg2+]i (2.18 +/- 0.11 mM). Lowering [Mg2+]o to 0.06 mM, however, exerted no significant effects on [Mg2+]i (0.93 +/- 0.14 mM), at least for periods of up to 30 min. Irrespective of the [Mg2+]o used, the subcellular distribution of [Mg2+]i remained heterogeneous, i.e. where the sub-plasma membrane region > cytoplasm > nucleus. [Mg2+] in all three subcellular compartments increased significantly, two- to threefold, concomitant with [Mg2+] when placed in 1.97 mM [Mg2+]o. We conclude that [Mg2+]i in fission yeast is maintained at a physiologic level when [Mg2+]o is low, but intracellular free Mg2+ rapidly rises when [Mg2+]o is elevated. Like most eukaryotic cells, yeast may have a Mg2+ transport system(s) which functions to maintain gradients of Mg2+ from the outside to inside the cell and among its subcellular compartments.

Exposure of piglet coronary arterial muscle cells to low alcohol results in elevation of intracellular free Ca2+: relevance to fetal alcohol syndrome.

Chronic exposure of cultured piglet neonatal coronary arterial smooth muscle cells to low concentrations of ethanol (46-115 mg/dl) for 7 days resulted in concentration-dependent elevation in intracellular free Ca2+ ions ([Ca2+i); these rises (22-56%) in [Ca2+]i were not reversible upon short-term exposure to normal, Ca2(+)-containing physiological salt solution. These findings help to provide a rational basis for why ethanol can result in the well-known fetal alcohol syndrome, including cardiac defects and in-utero death.

Acute cocaine results in rapid rises in intracellular free calcium concentration in canine cerebral vascular smooth muscle cells: possible relation to etiology of stroke.

Effects of cocaine on intracellular free calcium concentration ([Ca2+]i) in cultured canine cerebral vascular smooth muscle cells were studied using digital imaging microscopy and the calcium molecular fluorescent indicator, fura-2. Acute treatment of cerebral vascular smooth muscle cells with cocaine HCl, from a low concentration of 10(-9) M up to 10(-5) M, induced significant increases of [Ca2+]i. Irrespective of the changes in [Ca2+]i, the subcellular distribution of [Ca2+]i appeared heterogeneous in both normal and cocaine-treated cells. These results suggest that cocaine induces cerebral vasospasm by a rapid elevation of [Ca2+]i in vascular smooth muscle cells; these ionic events could play a crucial role in the pathogenesis of cocaine-induced cerebral ischemia and stroke.

Extracellular Mg2+ modulates intracellular Ca2+ in acutely isolated hippocampal CA1 pyramidal cells of the guinea-pig.

Using digital imaging microscopy and fluorescent probes, isolated hippocampal CA1 pyramidal neurons of the guinea-pig were used to examine the roles of [Mg2+]o in regulation of [Ca2+]i and [Mg2+]i. Low extracellular Mg ([Mg2+]o) (0.3 mM) significantly increased [Ca2+]i compared to 1.2 and 4.8 mM [Mg2+]o. In contrast, [Mg2+]i levels remained relatively constant, irrespective of alterations of [Mg2+]o. The sustained rise in [Ca2+]i induced by low [Mg2+]o was reduced 70% by 1 microM verapamil and 42% by 1 mM Ni2+, and completely abolished by 5 mM Ni2+. The data suggest that [Mg2+]o regulates [Ca2+]i in hippocampal neurons, probably by modulating Ca2+ entry via voltage-sensitive Ca2+ channels, which may play important roles in epileptogenesis, memory, learning and brain trauma. Furthermore, the results demonstrate that intracellular Mg2+ concentration does not follow passively the concentration of Mg2+ in the extracellular solution.

Suppression of collagen-induced arthritis by an angiogenesis inhibitor, AGM-1470, in combination with cyclosporin: reduction of vascular endothelial growth factor (VEGF).

Pannus formation characterized by neovascularization is a prominent pathologic finding in both rheumatoid arthritis (RA) and rat collagen-induced arthritis (CIA). CIA is a T-cell-dependent process induced by immunization of inbred LOU rats with native type II collagen in incomplete Freund's adjuvant. AGM-1470 is a highly specific inhibitor of new blood vessel formation by its effects on endothelial cell migration, endothelial cell proliferation, and capillary tube formation. Cyclosporin A (CSA) is an immunomodulating agent that inhibits IL-2 and other cytokine production involved in early antigen activation of T-cells. In this study the effects of single and combination therapy with AGM-1470 (27 mg/kg alternate days) and low-dose CSA (4 mg/kg/day continuous infusion via osmotic pump) on established CIA (total n = 62) were examined. At Day 18 post arthritis onset, clinical arthritis was significantly reduced in rats treated with single-agent AGM-1470 (1.88 +/- 0.33) or combination therapy (1.13 +/- 0.32) (P < 0.00001 and 0.000001, respectively) versus control. Single-agent CSA-treated rats, even if given CSA beginning on the day of immunization, did not attenuate arthritis severity. THe longitudinal mean arthritis score of combination-treated rats was significantly lower than that of rats receiving AGM-1470 (P < 0.0001), reflecting a more moderate early disease course in combination-treated rats. Disease severity in rats treated with single-agent CSA was not significantly different from control rats. Mean WBC counts, differentials, and delayed-type hypersensitivity responses were similar in all groups. CII antibody levels were lower in AGM-1470 protocols compared to CSA or controls. Flow cytometry of peripheral blood, spleen, and lymph nodes demonstrated decreased levels of CD4+ cells in rats given CSA. TNF-alpha levels remained elevated, even in treated rats, while vascular endothelial growth factor levels were reduced in rats receiving AGM-1470 compared to both arthritic controls and naive rats. Both single-agent and combination therapies were well tolerated. This is the first study to examine the effects of AGM-1470 together with CSA. Combination therapy was more effective than single-agent therapy. The results suggest that the use of interventions with distinct mechanisms of action may be efficacious in the treatment of RA.

Alcohols induce rapid depletion of intracellular free Mg2+ in cerebral vascular muscle cells: relation to chain length and partition coefficient.

Acute effects of a series of alcohols (methanol, ethanol, n-butanol) on intracellular free magnesium concentration ([Mg2+]i) in canine cerebral vascular smooth muscle cells was studied using mag-fura-2 and digital imaging microscopy. In 1.2 mM [Mg2+]o, basal [Mg2+]i was 500 +/- 30 microM. Exposure of cells to a low concentration (25 mM) of ethanol, but not methanol, for only 30 s resulted in significant loss of [Mg2+]i. Exposure to 100 mM methanol, ethanol, and butanol for 30 s resulted in a relative order of potency for [Mg2+]i depletion, where butanol >> ethanol > methanol. The heterogeneous and relative subcellular compartmented concentrations of [Mg2+]i, where perinuclear > nuclear >> peripheral (cytosolic) region, was not significantly altered by the alcohols. The degree of cellular depletion of [Mg2+]i was directly a function of each alcohol's partition coefficient and chain length. The latter is suggestive of the probability that alcohols promote intracellular depletion of Mg2+ by partitioning in membranes and disordering lipid bilayers.

Production of no-carrier-added 64Cu from zinc metal irradiated under boron shielding.

BACKGROUND: Positron emission tomography offers advantages for radioimmunodiagnosis of cancer but requires radionuclides of appropriate half-life that have high specific activity and high radio-purity. This work was designed to develop a viable method to produce and purify 64Cu, which has high specific activity, for positron emission tomography. METHODS: 64Cu was produced at the University of Missouri Research Reactor by the nuclear reaction, 64Zn(n,p)64Cu. Highly pure zinc metal (99.9999%) was irradiated in a specially designed boron nitrite lined container, which minimized thermal neutron reactions during irradiation. A new two-step procedure was developed to chemically separate the no-carrier-added 64Cu from the zinc metal target. RESULTS: 64Cu recovery for 24 runs averaged 0.393 (+/- 0.007) mCi per milligram of zinc irradiated. The boron-lined irradiation container reduced unwanted zinc radionuclides 14.3-fold. Zinc radionuclides and non-radioactive zinc were separated successfully from the 64Cu. The new separation technique was fast (2 hours total time) and highly efficient for removing the zinc. The zinc separation factor for this technique averaged 8.5 x 10(-8), indicating less than 0.0000085% of the zinc remained after separation. Thus far, the highest 64Cu specific activity at end of irradiation was 683 Ci/mg Cu, with an average of 512 Ci/mg Cu for the last six analyzed runs. CONCLUSION: The boron-lined irradiation container has sufficient capacity for 75-fold larger-sized zinc targets (up to 45 g). The new separation technique was excellent for separating 64Cu, which appears to be a radionuclide with great potential for positron emission tomography.

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 microM 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 microM). 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.

Cocaine induces rapid loss of intracellular free Mg2+ in cerebral vascular smooth muscle cells.

Acute exposure of cultured canine cerebral vascular smooth muscle cells to low concentrations of cocaine HCl (10(-9) to (10(-7) M) resulted in significant, rapid (1 min) loss of intracellular free Mg ions ([Mg2+]i); these reductions (12-25%) in [Mg2+]i were reversible upon exposure to normal, Mg(2+)-containing physiological salt solution. These findings help to provide a rational basis for why cocaine can result in cerebrovasospasm and stroke.

Mg2+ and caffeine-induced intracellular Ca2+ release in human vascular endothelial cells.

Interaction of ionized magnesium ([Mg2+]o) and caffeine in regulation of intracellular free calcium concentration ([Ca2+]i) in human aortic endothelial cells was studied using fura-2 and digital imaging microscopy. In 1.2 mM [Mg2+]o, basal [Ca2+]i was 73.7 +/- 22.4 nM, with a heterogeneous distribution within the cells. No significant changes of basal [Ca2+]i were found either when cells were treated with 10 mM caffeine or when [Mg2+]o was lowered from 1.2 mM to 0.3 mM. However, a combined superfusion of the cells with 0.3 mM [Mg2+]o and 10 mM caffeine resulted in a significant elevation of [Ca2+]i to 382.8 +/- 57.1 nM, probably by release of Ca2+ from internal stores, which was attenuated by NiCl2 (1 mM). These results suggest that a Ca(2+)-induced Ca2+ release mechanism is involved in regulation of [Ca2+]i in endothelial cells, which may be either regulated or modulated by Mg2+.