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PubMed1986

Effects of aminoglycoside antibiotics on the neuromuscular junction: Part I.

The effects of aminoglycoside antibiotics (AGA) including streptomycin (SM), kanamycin (KM), gentamicin (GM), dibekacin (DKB), amikacin (AMK) and sisomycin (SISO), on the neuromuscular junction were studied by in vivo and in vitro experiments. In in vitro experiments, no effect of AGA on rat phrenic nerve diaphragm preparations was observed, but the use of the antibiotics at a high concentration exerted a slight blocking effect on the neuromuscular junction. The blocking effect of SISO and DKB on the neuromuscular junction was marked. These antibiotics were definitely found to compete with eserine in terms of the blocking effect on the neuromuscular junction, but did not compete with calcium chloride. In in vitro experiments with frog sciatic nerve and musculus sartorius preparations, DKB and SISO exerted a blocking effect on the NMJ, inducing the disappearance of action potentials and the appearance of endplate potentials (EPPs). In in vitro experiments with the preparations from Rana catesbiana frogs, SM, GM, DKB and SISO exhibited an inhibiting effect on the release of acetylcholine (ACh), a chemical neurotransmitter in neuromuscular junction, resulting in a decrease in the frequency of miniature endplate potentials (mEPPS). In in vivo experiments with rabbit sciatic tibialis anterior muscle preparations, SM, GM, DKB and SISO exerted a blocking effect on the neuromuscular junction. From the facts that the effect was augmented by the use of magnesium chloride combined with these antibiotics and that the antibiotics competed with calcium chloride and potassium chloride in terms of the blocking effect on the neuromuscular junction, the effects seemed to be due to the inhibition of ACh release.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed2001

Mechanism of the inhibitory effect of histamine on amygdaloid-kindled seizures in rats.

PURPOSE: The mechanism of the inhibitory effect of histamine on amygdaloid-kindled seizures was investigated in rats. METHODS: Under pentobarbital anesthesia, rats were fixed to a stereotaxic apparatus, and bipolar electrodes were implanted into the right amygdala. A guide cannula made of stainless steel tubing was implanted into the right lateral ventricle. Electrodes were connected to a miniature receptacle, which was embedded in the skull with dental cement. EEG was recorded with an electroencephalograph; stimulation of the amygdala was applied bipolarly every day by a constant-current stimulator and continued until a generalized convulsion was obtained. RESULTS: Intracerebroventricular (i.c.v.) injection of histamine at doses of 2-10 microg resulted in a dose-related inhibition of amygdaloid-kindled seizures. I.c.v. injection of calcium chloride at doses of 10-50 microg and A23187 at doses of 2-10 microg also caused dose-dependent inhibition of amygdaloid-kindled seizures. Calcium chloride at a dose of 10 microg, which showed no significant effect on amygdaloid-kindled seizures when used alone, significantly potentiated the effect of histamine. Similar findings were observed with A23187 at a dose of 2 microg. In addition, EGTA and EGTA/AM antagonized the inhibition of kindled seizures induced by histamine. Moreover, the inhibition of kindled seizures induced by histamine was antagonized by KN62. However, calphostin C did not antagonize the inhibitory effect of histamine. CONCLUSIONS: These results indicated that histamine-induced inhibition of amygdaloid-kindled seizures may be closely associated with a calcium calmodulin-dependent protein kinase II activation pathway.

Amygdala↗
PubMed1983

Hemodynamic interactions of verapamil and isoflurane.

The hemodynamic interactions of verapamil and isoflurane were studied in eight dogs. Left ventricular function was analyzed using a right heart bypass preparation to permit rigid hemodynamic control. Hemodynamic studies were performed at 0.7, 1.05, and 1.40% isoflurane before and during the maintenance of two stable levels of verapamil, administered intravenously by combining a bolus dose (0.2 mg X kg-1) with an infusion (3.0 and 6.0 micrograms X kg-1 X min-1). Isoflurane produced a concentration-dependent depression of left ventricular function as indicated by dP/dt max, per cent systolic shortening, and left ventricular function curves. This depression was enhanced in a dose-plasma concentration-dependent manner by verapamil and was reversed by calcium chloride. Isoflurane alone and the combination of verapamil and isoflurane decreased systemic vascular resistance in a dose-dependent fashion that was antagonized partially by calcium chloride. Therefore, verapamil can enhance the hemodynamic effects of isoflurane in a dose-related manner that needs to be considered when both drugs are administered together.

Animals↗
PubMed2002

Suppressive role of endogenous regucalcin in the regulation of nitric oxide synthase activity in heart muscle cytosol of normal and regucalcin transgenic rats.

The role of regucalcin, a regulatory protein of Ca2+ signaling, in the regulation of nitric oxide (NO) synthase activity in the cytosol of rat heart muscle was investigated. The addition of calcium chloride (5-20 microM) into the enzyme reaction mixture containing the heart cytosolic protein caused a significant increase in NO synthase activity. The Ca2+ effect was significantly inhibited by trifluoperazine (TFP; 20 or 50 microM), an antagonist of calmodulin, indicating the existence of Ca2+/calmodulin-dependent NO synthase activity in rat heart muscle cytosol. NO synthase activity was significantly decreased by the addition of regucalcin (10(-9) or 10(-8) M). This effect was also seen in the presence of calcium chloride (10 microM), TFP (50 microM) or EGTA (1 mM), a chelator of Ca2+. Meanwhile, the effect of regucalcin (10(-8) M) in decreasing NO synthase activity was not seen in the presence of Nw-nitro-L-arginine methylester (NAME; 10(-6) or 10(-5) M), an inhibitor of the enzyme. The presence of anti-regucalcin monoclonal antibody (25 or 50 ng/ml) in the enzyme reaction mixture caused a significant increase in NO synthase activity. This effect was completely abolished by the addition of regucalcin (10(-7) M). NO synthase activity was not significantly changed in the heart muscle cytosol of transgenic rats overexpressing endogenous regucalcin as compared with that of wild-type rats. However, the effect of calcium (10 micro M) addition in increasing NO synthase activity was significantly weakened in the heart muscle cytosol of regucalcin transgenic rats. The present study demonstrates that endogenous regucalcin has a suppressive effect on NO synthase activity in the heart muscle cytosol of rats.

Animals↗
PubMed1987

Bupivacaine accentuates the cardiovascular depressant effects of verapamil in conscious dogs.

To investigate the potential risk of regional anaesthesia in a verapamil-treated patient, the cardiovascular effects of a combined intravenous infusion of bupivacaine and verapamil were studied in seven conscious and chronically instrumented dogs and were compared to those obtained when each drug was infused separately in the same animals. During verapamil infusion, the decrease in arterial pressure and myocardial contractile force, and the increase in heart rate were constant during the infusion. The lengthening of PR interval correlated with the plasma level of verapamil. During infusion of bupivacaine alone, there was an increase in heart rate and arterial pressure at the end of infusion, whereas the initial depression of myocardial contractile force was compensated. PR interval remained unchanged throughout the infusion of bupivacaine. During combined infusion of bupivacaine and verapamil, there was a time-dependent decrease in heart rate, arterial pressure and myocardial contractile force. A further increase in PR interval correlated with verapamil plasma concentrations which were higher than when verapamil was infused alone; bupivacaine plasma levels were in the same range as during bupivacaine infusion alone. Short periods of second-degree atrioventricular block developed in three out of the seven dogs but no relation was found between QT interval and heart rate in the whole group. Calcium chloride during bupivacaine-verapamil returned heart rate, arterial pressure and myocardial contractile force to their control values within 5 min. Atrioventricular block disappeared and PR interval was shortened following administration of calcium. Bupivacaine and verapamil have additive cardiovascular effects which lead to atrioventricular conduction dysfunction. The effects, at these doses, are reversed by calcium chloride.

Animals↗
PubMedDate not supplied

Bovine serum albumin-loaded pectinate beads as colonic peptide delivery system: preparation and in vitro characterization.

Objective of this study was to prepare a drug delivery system for therapeutic peptides that are degraded in the upper part of the gastrointestinal tract due to degradation activity of the enzymes. Delivering peptide to the colon in which enzymatic activity is low is next hope for absorption of these agents. Pectin, a naturally occurring water soluble polysaccharide, as a matrix for peptide delivery was studied. Degradation of pectin by the colonic enzymes makes it suitable for colon-specific delivery of drugs. Bovine serum albumin (BSA) was used as a model peptide. Calcium pectinate beads were prepared by extruding BSA-loaded pectin solution to an agitating calcium chloride solution, and gelled spheres were formed instantaneously by an ionotropic gelation reaction. The effect of several factors such as concentration of pectin, concentration of calcium chloride, and total drug loading on the pattern of drug release in the dissolution medium was studied. Prepared beads showed good resistance in the release medium. The entrapment efficiency of the beads was high (between 63% and 99%). Entrapment efficiency of BSA was reversely dependent to the amount of the drug loaded in the beads. The amount of BSA loaded on the beads affects pattern of drug release. The concentration of the pectin showed the highest impact on the rate of drug release. Presence of the pectiolytic enzymes facilitated the drug release from the beads.

Calcium Chloride↗
PubMed1997

ATP-sensitive voltage- and calcium-dependent chloride channels in sarcoplasmic reticulum vesicles from rabbit skeletal muscle.

Chloride channels in the sarcoplasmic reticulum (SR) are thought to play an essential role in excitation-contraction (E-C) coupling by balancing charge movement during calcium release and uptake. In this study the nucleotide-sensitivity of Cl- channels in the SR from rabbit skeletal muscle was investigated using the lipid bilayer technique. Two distinct ATP-sensitive Cl- channels that differ in their conductance and kinetic properties and in the mechanism of ATP-induced channel inhibition were observed. The first, a nonfrequent 150 pS channel was inhibited by trans (luminal) ATP, and the second, a common 75 pS small chloride (SCl) channel was inhibited by cis (cytoplasmic) ATP. In the case of the SCl channel the ATP-induced reversible decline in the values of current (maximal current amplitude, Imax and integral current, I') and kinetic parameters (frequency of opening FO, probability of the channel being open PO, mean open TO and closed Tc times) show a nonspecific block of the voltage- and Ca2+-dependent SCl channel. ATP was a more potent blocker from the cytoplasmic side than from the luminal side of the channel. The SCl channel block was not due to Ca2+ chelation by ATP, nor to phosphorylation of the channel protein. The inhibitory action of ATP was mimicked by the nonhydrolyzable analogue adenylylimidodiphosphate (AMP-PNP) in the absence of Mg2+. The inhibitory potency of the adenine nucleotides was charge dependent in the following order ATP4- > ADP3- > > > AMP2-. The data suggest that ATP-induced effects are mediated via an open channel block mechanism. Modulation of the SCl channel by [ATP]cis and [Ca2+]cis indicates that (i) this channel senses the bioenergetic state of the muscle fiber and (ii) it is linked to the ATP-dependent cycling of the Ca2+ between the SR and the sarcoplasm.

Adenosine Triphosphate↗
PubMed1986

Calcium influence on neuronal sensitivity to ethanol in selectively bred mouse lines.

Sensitivity to ethanol, as measured by blood ethanol concentration at loss of righting reflex, was increased significantly in SS but not LS mice following intracerebroventricular (ICV) administration of calcium chloride or A23187, a calcium ionophore. Magnesium chloride or lanthanum chloride, ICV, did not alter sensitivity to ethanol in either SS or LS mice, further indicating a specificity for calcium cation. Calcium was without effect on sensitivity to halothane narcosis in LS or SS mice. Endogenous brain calcium content was similar in these mouse lines, and ethanol administration either in vivo or in vitro did not alter brain calcium concentration. These results indicate that differences in brain sensitivity to ethanol are mediated, in part, by genetic differences in calcium-related processes and support the hypothesis that ethanol-induced narcosis may be due to alterations in calcium metabolism in the CNS.

Animals↗
PubMed1987

Fractionation of proteins in reconstituted skim-milk.

Properties of the dried milk were improved when calcium chloride was added to the raw milk prior to the drying process. Solubility, wettability, undenatured whey protein and protein retention indices of the dried milk were increased and the reconstitutability was improved. The elution profile of the protein fractions on a Sephadex G-100 column differed according to the forward preheating of the raw milk and the addition of calcium chloride.

Animals↗
PubMed1998

Calcium ions potentiate lipoxygenase activity of cytochrome c at the physiological pH.

Cytochrome c oxidized linoleic acid, with lipoxygenase-like activity, to 9-hydroperoxy-10,12-octadecadienoic acid (9-HpODE), 13-hydroperoxy-9,11-octadecadienoic acid (13-HpODE), 13-keto-9,11-octadecadienoic acid (13-KODE), and hydroxy-epoxy-octadecenoic acids (HEPOs) in the presence of calcium chloride at the physiological pH of 7.5. In the absence of calcium, the reaction was remarkably reduced. Statistically significant increase of 13-KODE was observed 1 min after the reaction was started by adding calcium. Calcium chloride at concentrations as low as 30 microM caused statistically significant increase of 13-KODE production in the presence of 1 microM cytochrome c, and cytochrome c at concentrations as low as 10 nM caused statistically significant increase of 13-KODE production in the presence of 2 mM calcium. The addition of bleomycin enhanced the reaction, and the addition of adriamycin attenuated the reaction. The same oxidized linoleic acid products were found in the submitochondrial particles which were hydrolyzed by phospholipase-A2 in the presence of calcium. Lipoxygenase activity potentiated by calcium may be involved in the lipid peroxidation in the submitochondrial particles hydrolyzed by phospholipase-A2.

Animals↗
PubMedDate not supplied

A simple gravimetric method for estimation of plasma fibrinogen.

The study was undertaken to evaluate a new method of estimating plasma fibrinogen levels, gravimetrically using calcium chloride in albino rats. The results were compared with the standard method in which thrombin was used as clotting agent and were found to be statistically not significant. Advantages of using calcium chloride instead of thrombin have been discussed.

Animals↗
PubMed1986

Heat shock response of the rat lens.

The sequence relationship between the small heat shock proteins and the eye lens protein alpha-crystallin (Ingolia, T. D., and E. E. Craig, 1982, Proc. Natl. Acad. Sci. USA, 79: 2360-2364) prompted us to subject rat lenses in organ culture to heat shock and other forms of stress. The effects on protein synthesis were followed by labeling with [35S]methionine and analysis by one- and two-dimensional gel electrophoresis and fluorography. Heat shock gave a pronounced induction of a protein that could be characterized as the stress protein SP71. This protein probably corresponds to the major mammalian heat shock protein hsp70. Also two minor proteins of 16 and 85 kD were induced, while the synthesis of a constitutive heat shock-related protein, P73, was considerably increased. The synthesis of SP71 started between 30 and 60 min after heat shock, reached its highest level after 3 h, and had stopped again after 8 h. In rat lenses that were preconditioned by an initial mild heat shock, a subsequent shock did not cause renewed synthesis of SP71. This effect resembles the thermotolerance phenomenon observed in cultured cells. The proline analogue azetidine-2-carboxylic acid, zinc chloride, ethanol, and calcium chloride did not, under the conditions used, induce stress proteins in the rat lens. Sodium arsenite, however, had very much the same effects as heat shock. Calcium ionophore A23187 specifically and effectively induced the synthesis of the glucose-regulated protein GRP78. No special response to stress on crystallin synthesis was noticed.

Animals↗
PubMed1989

[Effect of nisoldipine on cultured myocardial cells of neonatal rats].

Nisoldipine 1.5 nmol/L, 0.75, and 1.5 mumol/L depressed the beat rates of myocardial cells. The effects were negatively correlated with dosages. Nisoldipine depressed the beat rates increased by calcium chloride 0.15 mmol/L or isoprenaline 1.5 mumol/L and atropine 20 nmol/L. Conversely, the above drugs reversed the beat rates depressed by nisoldipine. Therefore, the acting nisoldipine may be antagonistic to calcium chloride, isoprenaline and atropine. In addition, nisoldipine 10 nmol/L may significantly aggravated the damage to myocardial cells induced by mitomycin C 5 micrograms/ml in which elevated lactate dehydrogenase appeared.

Animals↗
PubMedDate not supplied

Alginate-diltiazem hydrochloride beads: optimization of formulation factors, in vitro and in vivo availability.

Alginate beads containing diltiazem hydrochloride (DTZ) were prepared by the ionotropic gelation method. The effects of various factors (alginate concentration, additives type, calcium chloride concentration and curing time) on the efficiency of drug loading were investigated. The formulation containing a mixture of 0.8% methylcellulose (MC) and 4% alginate cured in 2% calcium chloride for 6 h was chosen as the best formula regarding the loading efficiency. The release rate of DTZ from various beads formulations was investigated. The release of drug from alginate beads followed two mechanisms; by diffusion and relaxation of the polymer at pH 1.2, whilst diffusion and erosion are at pH 6.8. The in vitro release of DTZ from MC-alginate beads showed an extended release pattern which was compared with that from commercially available sustained-release (Dilzem SR) and fast release tablets (Dilzem). Thermal analysis revealed that the drug was molecularly dispersed in the beads matrix. Although the release characteristics of DTZ from Dilzem SR and MC-alginate beads were completely different, the bioavailability of DTZ in dogs was comparable as measured by AUC, MRT and relative bioavailability. The absolute bioavailability of MC-alginate beads and Dilzem SR was 88 and 93%, respectively.

Alginates↗
PubMed2006

The use of a setting accelerator and its effect on pH and calcium ion release of mineral trioxide aggregate and white Portland cement.

The aim of this study was to evaluate the influence of adding 10% calcium chloride (CaCl2) indicated as setting accelerator on pH and release of calcium ions of commercially available materials: ProRoot MTA (tooth-colored formula), MTA Branco, and White Portland cement. The products were mixed for 30 seconds in previously established ratios. To estimate the values of pH and release of calcium ion, polyethylene tubes were filled with the materials and immersed in containers with 10 ml of deionized water. The analyses were performed baseline, 30 minutes, 60 minutes, and 24 hours after the mixing process. A pHmeter (MicroNal B 371, Sao Paulo, SP, Brazil) and an atomic absorption spectrophotometer (Model GBC 904; CG Corp, Melbourne, Australia) equipped with a hollow cathode lamp were used to determine the data. According to the Student's t-test, the comparison between the pure and the CaCl2-added products indicated that the presence of this substance increased immediately the pH, although the results were very similar when they were analyzed in the other intervals. In addition, products with CaCl2 released more calcium ions than pure materials in the 24-hour period (p<0.05). The results revealed that the addition of CaCl2 to MTA enhanced the physicochemical properties of this product. Thus, materials mixed with CaCl2 became easier to handle and needed lower amounts of water in the mixing process.

Aluminum Compounds↗
PubMed2004

Culture of chondrocytes in alginate beads.

A classic method for the encapsulation and culture of chondrocytes in alginate beads is described. Chondrocytes are released from cartilage matrix by collagenase/dispase digestion and mixed with a solution of 1.25% alginic acid until a homogenous suspension is obtained. The suspension is drawn into a syringe and pushed gently through a needle, so that drops fall into a solution of calcium chloride. Beads form instantaneously and further polymerize after 5 min in the calcium chloride solution. Chondrocytes from any species, including human osteoarthritic chondrocytes, can be cultured with this technique. Under these conditions, chondrocytes maintain a high degree of differentiation. Beads can be dissolved by chelation of calcium with EDTA. In this way, chondrocytes can be recovered and further separated from the matrix by centrifugation. Almost all molecular and biochemical techniques, as well as a number of biological assays, are compatible with the culture of chondrocytes in alginate.

Alginates↗