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PubMed1988

T cell-mediated recognition of foreign antigen and the Ia molecule observed by stopped-flow fluorometry.

Th cell-mediated rapid recognition of foreign Ag and the Ia molecule was studied using azobenzenearsonate-L-tyrosine (ABA-L-tyrosine)-specific Th cells (I-Ak restricted), foreign Ag (ABA-L-tyrosine), and APC (H-2k). Initial transmembrane signals in Th cell hybridomas (2-45-12) and in Th cell lines (A24-17 or A33-7) were monitored by stopped-flow fluorometry with fluorescent probes. It was found that Th cells recognized foreign Ag within 1 s at 25 degrees C on the APC (B10.BR spleen cells or L cells into which I-Ak genes were transferred). Recognition of foreign Ag and the Ia molecule was shown to deliver the initial signals to Th cell hybridomas and T cell lines. First, Th cells had membrane fluidity increased and then calcium was transported from the external medium into the T cells. The initial transmembrane signals to Th cell hybridomas were inhibited by the addition of an anti-I-Ak mAb. None of the initial signals were observed in the absence of either specific foreign Ag or APC.

Animals↗

PubMed1990

Hydration feature of urinary compounds. Evidence for molecular abnormality in calcium oxalate urolithiasis.

Proton-relaxation-time measurements were performed on lyophilized urine samples collected from 11 recurrent calcium oxalate stone-formers, 9 uric acid stone-formers, 9 patients with various urological disorders, and 20 normal individuals. The T1 and T2 relaxation times were determined using a Bruker PC Multispec at 20 MHz at 37 degrees C for measurements of lyophilized sample and thereafter during gradual controlled rehydration. The prolongation of the relaxation times as a function of rehydration was found to differ significantly (P less than 0.005) between, on the one hand, the calcium oxalate stone-formers and, on the other hand the normal, uric-acid stone-formers, and patients with other urological disorders. Water compartmentalization was then calculated according to the fast proton diffusion model. At most of the experimental points during rehydration process, significantly (P less than 0.001) less water was bound to the compounds of urine from calcium oxalate stone-formers than that obtained from normal individuals. The variations in the bound hydration water may have been associated with possible changes in the structure or configuration of the compounds present in the urine of the different groups. The results reflect differences in the urinary content and/or properties of normal subjects and calcium oxalate stone-formers, and indicate that the mechanism of bound water relaxation is in some way specific to the pathophysiological state of urine.

Adult↗

PubMed2000

Induction of cytokine genes and IL-1alpha by chemical hypoxia in PC12 cells.

The role of cytokine in neuronal injury was examined in rat pheochromocytoma (PC12) cells under chemical hypoxia (i.e. KCN) and glucose deprivation. The mRNA levels of interleukin-1alpha (IL-1alpha), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were measured by reverse transcription-polymerase chain reaction (RT-PCR) in PC12 cells exposed to 0.5 mM KCN for various time intervals. Cytokine mRNA levels expressed to peak levels 30 minutes after KCN treatment and declined gradually until 240 min. The IL-1alpha activity reached the highest levels 2 hr after the same KCN treatment. Under parallel conditions, KCN increased cytosolic free calcium concentration ([Ca2+]i) in the absence of glucose. However, IL-1alpha mRNA induction by KCN was not altered under calcium-free conditions in PC12 cells, indicating its induction was Ca2+-independent. However, the phosphatidylcholine (PC)-specific phospholipase C (PLC) inhibitor D609 decreased the KCN-induced IL-1alpha mRNA and protein in PC12 cells suggests that PC-PLC might play a role in cytokine induction during hypoxia.

Animals↗

PubMed2002

A pivotal role for cADPR-mediated Ca2+ signaling: regulation of endothelin-induced contraction in peritubular smooth muscle cells.

cADPR, a potent calcium-mobilizing intracellular messenger synthesized by ADP-ribosyl cyclases regulates openings of ryanodine receptors (RyR). Here we report that in the rat testis, a functional cADPR Ca2+ release system is essential for the contractile response of peritubular smooth muscle cells (PSMC) to endothelin (ET). We previously showed that this potent smooth muscle agonist elicits intracellular Ca2+ release in PSMC and seminiferous tubule contraction via activation of ETA and ETB receptors. ETB-R induces the mobilization of a thapsigargin-sensitive but IP3-independent intracellular Ca2+ pool. Stimulation of permeabilized PSMC with cADPR was found to elicit large Ca2+ releases blocked by either a selective antagonist of cADPR or a RyR blocker, but not by heparin. Western blotting and confocal fluorescence microscopy indicated the specific expression of type 2 RyR in perinuclear localization. ET was found to stimulate the activity of ADP-ribosyl cyclase. Microinjection of the selective cADPR antagonist 8NH2-cADPR completely abolished subsequent stimulation of Ca2+ signaling via ETA and ETB receptors. cADPR therefore appears to have an obligatory role for ETA-R and ETB-R-mediated calcium signaling in PSMC. However, ETB-R seem to be coupled exclusively to cADPR whereas ETA-R activation may be linked to IP3 and cADPR signaling pathways.

ADP-ribosyl Cyclase↗

PubMed1992

Effect of thrombin inhibitors on platelet functions: comparative analysis of DuP 714 and hirudin.

Since thrombin plays an important role in platelet-mediated arterial thrombosis, we have examined the antiplatelet activity of a synthetic thrombin inhibitor, DuP 714 (Ac-(D)Phe-Pro-boroArg), in comparison with that of the naturally occurring inhibitor hirudin. Hirudin was slightly more potent than DuP 714 in inhibiting thrombin-induced aggregation in washed human platelets (IC50s of 72 nM and 150 nM, respectively) and in inhibiting the secretion of plasminogen activator inhibitor-I from human platelets (IC50s of 300 nM and 900 nM, respectively). In contrast, DuP 714 was more potent than hirudin in inhibiting thrombin-induced [125I]fibrinogen binding to gel purified platelets, and in inhibiting thrombin-induced intracellular calcium mobilization in washed platelets. These results indicate that the tripeptide DuP 714 has comparable antiplatelet activity to the 65 amino acid hirudin. We conclude that DuP 714 may have clinical utility in the prevention of platelet-dependent, arterial thrombotic processes.

Blood Platelets↗

PubMed1975

Influence of cocaine and sodium on bretylium uptake by reserpine-treated guinea-pig left atrium.

1 The effects of cocaine and sodium on bretylium uptake into sympathetic nerve terminals were investigated in the reserpine-treated guinea-pig left atrium. The ability of bretylium pretreatment to increase the retention of noradrenaline was used as an index of bretylium uptake. Such increased retention has been assessed both by direct measurement and by the ability of tyramine to produce an inotropic response. 2 The restoration of the response to tyramine after incubation with noradrenaline was abolished when the atrium was pretreated with bretylium in the presence of cocaine. When bretylium was added before cocaine, or when alpha-methyl-noradrenaline (not a substrate for monoamine oxidase) was used for incubation, the responses to tyramine were restored in the normal way. 3 Bretylium greatly enhanced the retention of [3-H]-noradrenaline; when bretylium was added in the presence of cocaine, [3-H]-noradrenaline retention was severely impaired. 4 Pretreatment with bretylium in a low-sodium (25 mM) or sodium-free medium significantly decreased the retention of [3-H]-noradrenaline, as compared with the control. 5 Potassium deprivation did not modify the enhanced retention of [3-H]-noradrenaline induced by bretylium pretreatment. 6 Bretylium was released from the nerve terminals by exposure of the preparation to a sodium-free medium or to a solution containing calcium 50 mM, leading to a considerable decrease in [3-H]-noradrenaline retention. 7 The results are consistent with the view that both cocaine and sodium deprivation block the uptake of bretylium by the adrenergic nerve terminals, and that bretylium is probably taken up by a mechanism similar to or identical with the uptake system for noradrenaline and other amines.

Animals↗

PubMed1981

Calcium-based evaluation of dose-response curves of papaverine-like relaxants in guinea-pig taenia coli.

Using CaCl2 dose-contraction curves in 165.6 mM K(+)- depolarized guinea-pig taenia coli, the maximal contraction level induced by 30 mM CaCl2, and the maximal relaxation level induced by Ca-free solution were treated and 0% and 100% relaxation, respectively. Then four kinds of contraction level were compared: (1) the normal level, (2) 5.5 microns acetylcholine (ACh)-induced contraction, and (3) 1.65 mM ACh- or 3 mM BaCl2-induced contraction in normal solution; and (4) 2.2 and 30 mM CaCl2-induced contraction in K(+) depolarizing solution. The relaxing effects of papaverine, MnCl2, and isoproterenol at these different contraction levels were compared. Papaverine- and MnCl2-induced relaxation of the higher contraction levels [a part of (2), (3) and (4)] seem to reflect a mechanism of the intracellular calcium mobilization, being quite different from relaxation induced by papaverine-like relaxants and isoproterenol at the lower contraction levels [(1) and part of (2)]. The latter phenomenon seems to reflect mechanisms related to cyclic AMP. Papaverine inhibited competitively BaCl2-induced contraction only in K(+) depolarized tissues, suggesting a mechanism of direct reaction with the intracellular Ca store.

Acetylcholine↗

PubMed2006

Effect of Ca2+, Ba2+, and Sr2+ on alginate microbeads.

Microcapsules of alginate cross-linked with divalent ions are the most common system for cell immobilization. In this study, we wanted to characterize the effect of different alginates and cross-linking ions on important microcapsule properties. The dimensional stability and gel strength increased for high-G alginate gels when exchanging the traditional Ca2+ ions with Ba2+. The use of Ba2+ decreased the size of alginate beads and reduced the permeability to immunoglobulin G. Strontium gave gels with characteristics lying between calcium and barium. Interestingly, high-M alginate showed an opposite behavior in combination with barium and strontium as these beads were larger than beads of calcium-alginate and tended to swell more, also resulting in increased permeability. Binding studies revealed that different block structures in the alginate bind the ions to a different extent. More specifically, Ca2+ was found to bind to G- and MG-blocks, Ba2+ to G- and M-blocks, and Sr2+ to G-blocks solely.

Alginates↗

PubMed2004

Suppressing effects of bisphenol A on the secretory function of ovine anterior pituitary cells.

We investigated the action of bisphenol A (BPA) on cellular GH release and content, cell number, GHmRNA expression, and concentrations of cellular cyclic AMP ([cAMP]c) and calcium ion ([Ca2+]c) in primary cultured ovine anterior pituitary cells. The following results were found: (1) BPA as well as nonylphenol (NP) at 10(-6) to 10(-3) M significantly and concentration-dependently suppressed basal and GHRH-stimulated GH release, and the cellular GH content, (2) BPA suppressed the cell number in a time- and concentration-dependent manner, (3) 10(-4)M BPA suppressed GHmRNA expression to 68% of control (BPA-free), and abolished GHRH (10(-8) M)-induced increases in [cAMP]c and [Ca2+]c. From these findings we conclude that BPA possesses a suppressing action on GH synthesis and release, and this suppressing action is probably related to impairment of cellular signal transduction systems in ovine anterior pituitary cells.

Animals↗

PubMed1991

Laser-Raman Spectroscopic study of the adhesive interface between 4-MET/MMA-TBB resin and hydroxyapatite or bovine enamel.

The possible chemical interaction between synthetic hydroxyapatite or bovine enamel and a functional monomer of 4-methacryloxyethyl trimellitic acid (4-MET) diluted in methyl methacrylate (MMA) was examined by measuring the Raman spectra. It was concluded that the carboxyl group of 4-MET reacted with the calcium in the substrate to form a salt that was detected by the Raman band at around 1,380 cm-1. However, formation of the salt on the surface of the hydroxyapatite (HAP) with the carboxyl group, and polymerization of the 4-MET in the methacryl group near the surface were mutually exclusive reactions for the same 4-MET molecule.

Adhesives↗

PubMed2001

Epileptogenesis induces long-term alterations in intracellular calcium release and sequestration mechanisms in the hippocampal neuronal culture model of epilepsy.

Calcium and calcium-dependent processes have been hypothesized to be involved in the induction of epilepsy. It has been shown that epileptic neurons have altered calcium homeostatic mechanisms following epileptogenesis in the hippocampal neuronal culture (HNC) and pilocarpine models of epilepsy. To investigate the mechanisms causing these alterations in [Ca2+]i homeostatic processes following epileptogenesis, we utilized the HNC model of in vitro 'epilepsy' which produces spontaneous recurrent epileptiform discharges (SREDs). Using [Ca2+]i imaging, studies were initiated to evaluate the mechanisms mediating these changes in [Ca2+]i homeostasis. 'Epileptic' neurons required much longer to restore a glutamate induced [Ca2+]i load to baseline levels than control neurons. Inhibition of Ca2+ entry through voltage and receptor gated Ca2+ channels and stretch activated Ca2+ channels had no effect on the prolonged glutamate induced increase in [Ca2+]i in epileptic neurons. Employing thapsigargin, an inhibitor of the sarco/endoplasmic reticulum calcium ATPase (SERCA), it was shown that thapsigargin inhibited sequestration of [Ca2+]i by SERCA was significantly decreased in 'epileptic' neurons. Using Ca2+ induced Ca2+ release (CICR) cell permeable inhibitors for the ryanodine receptor (dantrolene) and the IP3 receptor (2-amino-ethoxydiphenylborate, 2APB) mediated CICR, we demonstrated that CICR was significantly augmented in the 'epileptic' neurons, and determined that the IP3 receptor mediated CICR was the major release mechanism altered in epileptogenesis. These data indicate that both inhibition of SERCA and augmentation of CICR activity contribute to the alterations accounting for the impaired calcium homeostatic processes observed in 'epileptic' neurons. The results suggest that persistent changes in [Ca2+]i levels following epileptogenesis may contribute to the long-term plasticity changes manifested in epilepsy and that understanding the basic mechanisms mediating these changes may provide an insight into the development of novel therapeutic approaches to treat epilepsy and prevent or reverse epileptogenesis.

Animals↗

PubMed2006

Store-operated calcium channels and pro-inflammatory signals.

In non-excitable cells such as T lymphocytes, hepatocytes, mast cells, endothelia and epithelia, the major pathway for calcium [Ca2+] entry is through store-operated Ca2+ channels in the plasma membrane. These channels are activated by the emptying of intracellular Ca2+ stores, however, neither the gating mechanism nor the downstream targets of these channels has been clear established. Here, I review some of the proposed gating mechanisms of store-operated Ca2+ channels and the functional implications in regulating pro-inflammatory signals.

Animals↗

PubMed1985

Mediation of muscarinic stimulation of pepsinogen secretion in the frog.

The in vitro release of pepsinogen secretion by the isolated esophagus of the American bullfrog was studied with an improved model system. The tissue was mounted in a double chamber that preserves mucosal polarity and provides both control and test segments, each 1 cm2 from the same tissue. Pepsinogen secretion was severalfold higher than previously found with mucosal strips and could be sustained for several hours. Bethanechol (BCh) caused concentration-dependent (0.1-50 microM) pepsinogen secretion with a Vmax of 74 +/- 12 micrograms X mg prot-1 X h-1 or 50-60% of total pepsinogen; Km was 3 microM and 500 microM BCh stimulated at less than the Vmax value. Atropine specifically blocked BCh and pA2 = 9.3. In the presence of 100 microM isobutylmethyxanthine, BCh produced a dose-dependent increase in tissue cAMP but not cGMP. BCh remained effective in Ca2+-free medium. In calcium-free medium EGTA concentration dependently (0.2-5 mM) suppressed the pepsinogen response to BCh. The evidence thus far suggests that cholinergic stimulation of pepsinogen secretion in the tissue acts via both cAMP and Ca2+. More specific studies would be required for absolute confirmation of either or both apparent mechanisms and to resolve how they interact.

1-Methyl-3-isobutylxanthine↗

PubMed1997

[Heat production from reaction of inhalation anesthetics with dry soda lime].

UNLABELLED: There are some case reports about excessive heat production in the absorbent canister when sevoflurane or enflurane are washed into a circle containing dried soda lime. This observation was often made in the DRAGER ISO 8 circle system with the gas inlet upstream of the soda lime canister with the gas-flow from bottom to top. METHODS: The temperature in the center of an absorbent canister was measured 3.0 cm and 7.5 cm above the bottom. Soda lime (DRAGERSORB 800) was dried in an O2 stream for 2-3 days until there was no further loss in weight. 5 Vol% of desflurane, enflurane, isoflurane and sevoflurane in 2 1/min O2 or 4 Vol% of halothane in 2.5 I/min O2 were continuously fed into the canister. The concentration of the respective inhalational agents were measured after the soda lime canister using a DATEX Capnomac. Experiments were performed at ambient temperatures of 20-22 degrees C. RESULTS: A considerable temperature increase was achieved with all anaesthetics. The highest temperatures were measured at the upper sensor with 56-58 degrees C for desflurane, 76-80 degrees C for enflurane and isoflurane, 84-88 degrees C for halothane and 126-130 degrees C for sevoflurane. IR-detection for some agents was considerably delayed or the time course indicated that other compounds might have formed which absorb at the wavelength monitored. DISCUSSION: The high temperatures indicate the degradation rather than absorption of the volatile anaesthetics. CO is known to be degradation product of all currently used volatile anaesthetics except sevoflurane. Sevoflurane, however, produced the highest temperatures passing through dried soda lime. There are no reports about new specific breakdown products for sevoflurane on dried soda lime.

Anesthetics, Inhalation↗

PubMed2002

Diamond-like carbon coatings with Ca-O-incorporation for improved biological acceptance.

Diamond-like carbon (DLC) coatings were modified by doping the thin films with Ca-O compounds. Raman spectroscopy indicates growth of sp(2)-hybridised, ordered regions in size and/or number within the amorphous carbon-hydrogen network as a result of the Ca-O-incorporation. CaCO(3) was identified by X-ray induced photoelectron spectroscopy. Proliferation and morphology of L929 mouse fibroblasts reveal improved biocompatibility of Ca-O-modified DLC.

Animals↗

PubMed1991

Counteraction by nicardipine of the ionophoretic effect of gliclazide: a mitochondrial study.

Using mitochondria, we demonstrate that gliclazide exhibits a calcium ionophoretic activity. Indeed, gliclazide induces a decrease in the respiratory control of mitochondria; this effect is increased by addition of Ca2+ and corrected by ruthenium red, all characteristics of a calcium ionophoretic compound. Our results, on a biological membrane, confirm previous findings in vitro. Moreover, we show that nicardipine counteracts the action of gliclazide. Besides the effect on ATP-stimulated K+ channels, the ionophoretic effect of gliclazide may play a role in its hypoglycaemic effect, thus this counteracting action of nicardipine might induce drug interaction during the concomitant clinical administration of gliclazide and nicardipine.

Animals↗

PubMed2000

Evaluation of argatroban and DUP 714 as anticoagulants for blood gas, electrolyte and ionized calcium analyses.

The objective of this study was to determine if the thrombin inhibitors Argatroban and DUP 714 could anticoagulate whole blood without influencing the analyses of blood gases, electrolytes, ionized calcium or CO-oximetry. The anticoagulant potency of DUP 714 (0.5-68 micromol/l) and Argatroban (1.5-390 micromol/l) was evaluated using the activated partial thromboplastin time (APTT), prothrombin time (PT) and whole blood clot time (WBCT). APTT and the PT were measured using a Behring Fibrintimer. APTT was found to be more sensitive to prolongation by both of the thrombin inhibitors than were the PT or WBCT assays. DUP 714 was found to a more potent anticoagulant than Argatroban. DUP 714 anticoagulated specimens (>2.2 micromol/l) did not clot for at least 2 days, whereas Argatroban preserved specimens (390 micromol/l) clotted within 5.5 h of collection. No statistically significant changes in the measurement of pH, PCO2, PO2, Na, K, ionized calcium, oxyhaemoglobin, deoxyhaemoglobin, methaemoglobin or carboxyhaemoglobin (measured using a Corning 288 Blood Gas/Electrolyte Analyzer and a Coming 270 CO-oximeter) were detected in DUP 714 (34 micromol/l) or Argatroban (390 micromol/l) anticoagulated whole blood specimens. In conclusion, DUP 714 and Argatroban are suitable anticoagulants for preserving blood prior to blood gas and electrolyte analyses.

Antithrombins↗

PubMed1963

Microdetermination of calcium and magnesium in biological materials.

The use of the dye calcon (1-(2 hydroxy-1-naphthylazo)-2-naphthol-4 sulphonic acid) for the estimation of calcium using E.D.T.A. and a commercial photoelectric titrimeter is described. The interfering effects of magnesium and phosphate have been overcome. The method has been extended to estimations on biological materials. Results on 55 sera show that the E.D.T.A./calcon method gave slightly lower results (-0.15 mg./100 ml. +/- 0.029) than the oxalate precipitation method. Magnesium may also be estimated by incorporating the use of Eriochrome black T.

Azo Compounds↗