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PubMed1989

Circular dichroism of bovine serum albumin in divalent salt solutions.

The circular dichroism of bovine serum albumin in divalent salt solutions was investigated. The salts selected were magnesium chloride and calcium chloride. Their effects on the secondary and tertiary structures of the protein were compared with that of lithium chloride. It is well known that the elements Ca, Mg, and Li have many properties in common. The results show the similarity of their ions in the capacity of deforming protein, in spite of their characteristic pharmacological functions.

Calcium Chloride↗
PubMed1998

Plant derived estrogens relax rat mesenteric artery in vitro.

The aim of this study was to investigate whether plant derived estrogens have the same relaxing effects in vitro as estradiol-17beta on arterial smooth muscle. The mesenteric arterial rings of female and male Wistar rats were studied. The relaxing effects of estradiol-17beta, genistein, daidzein and beta-sitosterol were determined, with particular focus on the role of endothelium. B-sitosterol had no relaxing effect on the arteries. Estradiol-17beta, genistein and daidzein relaxed noradrenaline, potassium chloride and calcium chloride precontracted arterial rings endothelium-independently. The relaxation responses were also independent of gender. Neither the removal of endothelium, nor the inhibition prostacyclin or nitric oxide synthesis, had any effect on the relaxation responses. The exact mechanism of these findings is still unclear.

Animals↗
PubMed1977

Mechanism of crystallization of purified human midcycle cervical mucus.

The crystallization phenomenon of human midcycle cervical mucus was studied with purified mucin and sodium chloride. The drying of the purified mucin in saline was subjected to stress by the use of an air curtain, and crystallization (fern pattern) was found to follow the stress. Lithium chloride and calcium chloride were not compatible with the mucin for the formation of fernlike crystals. When a dilute solution of mucin dissolved in saline was dried, a radial crystallization pattern was observed to emanate from a single crystal in the center. Upon closer examination, the pattern was observed to emanate from the corners of the crystal.

Calcium Chloride↗
PubMed1985

Pharmacological actions of ifenprodil in the rat isolated anococcygeus muscle.

The effect of ifenprodil on the contractile responses of the rat anococcygeus muscle to noradrenaline, methoxamine, carbachol, potassium chloride and calcium chloride has been examined. Ifenprodil (10(-8)-10(-7) M) non-competitively antagonized noradrenaline and methoxamine-induced contractions while carbachol-induced responses were unaffected. Similarly, contractile responses to KCl (in tissues from reserpinized animals, or tissues bathed in Tyrode containing 10(-6) M phentolamine) and CaCl2 in tissues bathed with Ca2+-free depolarizing solution were blocked non-competitively by ifenprodil (10(-7)-10(-6) M). These findings suggest that ifenprodil possesses alpha-adrenoceptor antagonist activity, with intrinsic Ca2+ blocking properties.

Adrenergic alpha-Antagonists↗
PubMed1985

Zinc and Chlamydia trachomatis.

Zinc was noted to have significant effects upon the infection of McCoy cells by each of two strains of Chlamydia trachomatis. With a high or low Chlamydia inoculant, the number of infected cells increased up to 200% utilizing supplemental zinc (up to 1 X 10(-4) M) in the inoculation media compared with standard Chlamydia cultivation media (8 X 10(-6) M zinc). Ferric chloride and calcium chloride did not effect any such changes. Higher concentrations of zinc, after 2 hr of incubation with Chlamydia, significantly decreased the number of inclusions. This direct effect of zinc on the Chlamydia remained constant after further repassage of the Chlamydia without supplemental zinc, suggesting a lethal effect of the zinc. Supplemental zinc (up to 10(-4)M) may prove to be a useful addition to inoculation media to increase the yield of culturing for Chlamydia trachomatis. Similarly, topical or oral zinc preparations used by people may alter their susceptivity to Chlamydia trachomatis infections.

Calcium Chloride↗
PubMed1995

High and low affinity transport of L-arginine in rat brain synaptosomes.

The uptake of L-arginine into purified rat brain synaptosomes was investigated with respect to time and various concentrations of L-[3H]arginine. Specific uptake was found to be linear with time for up to 5 min of incubation at 37 degrees C. Electrolytes, including sodium chloride, potassium chloride, magnesium chloride and calcium chloride, inhibited uptake of 3 microM L-arginine, and the inhibitory effect increased with increased electrolyte concentration under constant osmolarity. It was found that L-arginine was transported into synaptosomes by two uptake components--a high affinity component (3.5 microM) and a low affinity component (100 microM). These two components were similar to the Ly+ system because of their extreme sensitivity to inhibition by L-lysine and L-ornithine but were distinguishable from each other by kinetic analysis of the uptake data and by their relative sensitivity to inhibition by several amino acids.

Amino Acids↗
PubMed1998

Pharmacologic actions of temiverine (p-INN) and its active metabolite, RCC-36, on isolated human urinary bladder muscle.

BACKGROUND: Temiverine (p-INN) is a newly synthesized drug that is expected to have anticholinergic action. We investigated the pharmacologic actions of temiverine and its active metabolite, RCC-36, on isolated human bladder. METHODS: Effects of temiverine and RCC-36 on the detrusor contractions induced by acetylcholine, potassium chloride (KCl), calcium chloride (CaCl2), and electric field stimulation were evaluated using the muscle-bath technique, and compared with the effects of atropine and oxybutynin. RESULTS: Atropine (10(-9) to 10(-6) mol/L), oxybutynin (10(-8) to 10(-5) mol/L), temiverine (10(-8) to 10(-5) mol/L), and RCC-36 (10(-8) to 3 x 10(-6) mol/L) caused a parallel shift to the right of the concentration-response curves to acetylcholine stimulation. The rank order of pA2 value was atropine > oxybutynin = RCC-36 > temiverine. Atropine did not suppress the maximum contractile response to acetylcholine, but the other drugs significantly suppressed this at the higher concentrations. Each drug caused a concentration-dependent inhibition of KCl (80 mmol/L)-, and CaCl2 (5 mmol/L)-induced contractile responses. Rank order of maximum inhibition was RCC-36 = temiverine > oxybutynin > atropine. Each drug caused a concentration-dependent inhibition of electric field-induced contraction with or without 10(-6) mol/L atropine pretreatment. Maximum inhibitions of temiverine and RCC-36 were significantly greater than that of oxybutynin. CONCLUSION: Atropine, oxybutynin, temiverine, and RCC-36 have different efficacies and potencies of anticholinergic and calcium antagonistic activity on isolated human detrusor muscles. Furthermore, temiverine and RCC-36 have significant inhibitory actions toward the atropine-resistant part of contractions, which may be related to the calcium antagonistic actions of these compounds.

Acetylcholine↗
PubMed1999

In vitro effects of three metallic salts and soot on the growth of five species of the dermatophytes.

The antifungal effects of zinc chloride (znCl), Calcium chloride (CaCl), magnesium chloride (MgCl) as well as soot collected from a local kitchens were tested on five isolated species of the dermatophytes from cases of human dermatophytoses. Of these three metallic salts, ZnCl showed the highest inhibitory effect on the dermatophytes. At a concentration of 0.01 M it showed 100% inhibition on these fungi. Although the other two metallic salts showed some inhibitory effect CaCl at concentration between 0.01 M and 1 M showed only 50% inhibitory effect while MgCl at concentration between 0.1 M and 1 M showed less than 50% inhibitory effect on the fungi. Also soot inhibited the growth of all the dermatophytes tested at concentration between 0.1 and 2%. Trichophyton rubrum and Microspum audouinii appeared to be most sensitive to these compounds compared to the other three species which include T. mentagrophytes, T. tonsurans and M. gypseum.

Arthrodermataceae↗
PubMed1975

Insulin and proinsulin release during calcium infusion in a patient with islet-cell tumor.

The infusion of calcium results in the release of gastrin, calcitonin, and serotonin from certain nonbeta islet cell tumors of the pancreas, medullary carcinomas of the thyroid, and carcinoid tumors, respectively. In this study, intravenous infusion of either calcium chloride or calcium aluconate in a patient with an islet-cell carcinoma resulted in a simultaneous rise in plasma immunoreactive insulin and proinsulin, and concurrent hypoglycemia. After resection of the tumor, calcium infusion caused no change in these parameters. Similarly, calcium infusion caused no change in plasma insulin or glucose in normal volunteers. The response of this tumor suggests that calcium infusion may be a useful provocative test to detect insulin-secreting neoplasia. A derangement of the stimulus-secretion coupling mechanism for insulin in the tumor cells may be responsible for their abnormal sensitivity to calcium ion.

Adenoma, Islet Cell↗
PubMed1985

Interaction of warfarin with human serum albumin. A stoichiometric description.

Reversible binding of warfarin to defatted serum albumin was studied by equilibrium dialysis at pH 7.4, in a 66 mM sodium phosphate buffer at 37 degrees. The binding isotherm could be described by two stoichiometric binding constants, K1 in the range 141,000 to 192,000 M-1 and K2 at 39,000 to 57,000 M-1. At least two additional molecules could be bound but gave indeterminate binding constants. The product K3 X K4 was about 4.7 X 10(7) M-2. Different site models were possible, either one high affinity and several low affinity sites, or two high affinity sites, cooperative, independent, or anticooperative, together with two low affinity sites. Binding affinity for the first warfarin molecule did not vary with pH in the interval from 6 to 9. The affinity decreased with increasing concentrations of sodium sulfate, sodium chloride, and calcium chloride, depending upon ionic strength. Specific effects of chloride and calcium ions were not observed. Light absorption spectra indicated that the warfarin anion was bound to albumin. All observations were consistent with a binding process involving albumin and the warfarin anion, without participation of hydrogen ions and not influenced by the N-B conformational transition of albumin.

Electrolytes↗
PubMed1995

Calcium-activated chloride conductance is not increased in pancreatic duct cells of CF mice.

Calcium-activated anion secretion is elevated in the pancreatic ductal epithelium of transgenic cf/cf mice which lack the cystic fibrosis transmembrane conductance regulator (CFTR). To elucidate whether this effect is due to increased activity of calcium-activated chloride channels, we have studied the relationship between CFTR and calcium-activated chloride currents in pancreatic duct cells isolated from Cambridge cf/cf mice. CFTR chloride currents activated by cAMP were detected in 59% (29/49) of wild-type cells and in 50% (20/40) of heterozygous cells. However, we could not detect any CFTR currents in the homozygous cf/cf cells (0/25). The maximum CFTR current density measured at a membrane potential of 60 mV was 23.5 +/- 2.8 pA/pF (n = 29) in wild-type cells, and about half that value, i.e. 12.4 +/- 1.6 pA/pF (n = 20) in heterozygotes (P = 0.004). Calcium-activated chloride currents were detected in 73% (24/33) of wild-type, 75% (21/28) of heterozygous and in 58% (7/12) of homozygous cf/cf cells. There was no significant difference between the steady-state calcium-activated current densities in the three genotypic groups; the current measured at 60 mV being 527 +/- 162 pA/pF (n = 24) from wild-type, 316 +/- 35 pA/pF (n = 21) from heterozygote and 419 +/- 83 pA/pF (n = 7) from homozygous cells. Our data suggest that lack of CFTR does not enhance the calcium-activated chloride conductance in murine pancreatic duct cells.

Animals↗
PubMed2001

Antioxidant status affects color stability and tenderness of calcium chloride-injected beef.

The objectives of this study were to determine whether vitamin E supplementation influences color and tenderness of beef injected with calcium chloride. Market heifers (n = 12) were fed a standard finishing diet with minimal levels of vitamin E (NE group). Another 12 market heifers were fed the NE diet with the inclusion of 1,000 IU/d of DL-alpha-tocopherol per animal for the last 125 d on feed (E group). Animals were slaughtered after 125 d on the diets and upon reaching an ultrasound backfat thickness > 10 mm. Half of the longissimus muscles from each treatment group (NE and E) were pumped to 10% over the original weight with 250 mM CaCl2 (Ca) at 24 h postmortem. Remaining muscles (NE and E) were pumped to 10% over the original weight with water (NC) at 24 h postmortem. After equilibrating overnight, steaks (2.54 cm) were overwrapped with O2-permeable film and stored for 7 d after injection. Hunter "L," "a," and "b" values were obtained each day of storage. Trained panelists evaluated color on d 1, 4, and 7 after injection. 2-Thiobarbituric acid-reactive substances (TBARS) values were measured on d 1 and 7 after injection. Warner-Bratzler (W-B) shear force values and trained sensory panel evaluations at 1, 3, and 7 d after injection were obtained. Immunoblotting techniques were used to monitor the 30-kDa degradation product of troponin-T at 1, 3, and 7 d after injection. At 4 d after injection, E/Ca steaks were the least discolored (P < 0.05). The E/Ca steak TBARS values were not significantly different from values for NE/NC steaks at 7 d after injection, whereas NE/Ca steaks had greater (P < 0.05) TBARS values after 7 d following injection compared with all other groups. Treatment with Ca resulted in higher off-flavor scores (P < 0.05). The E/Ca samples had the most rapid tenderization and proteolysis of all treatment groups. Warner-Bratzler shear values were lower in the E/Ca samples than in the E/NC samples at 1, 3, and 7 d after injection (P < 0.05). No difference in shear force was noted between NE/Ca and NE/NC samples at any time point. No difference in sensory tenderness was noted between NE/Ca and NE/NC samples at 1 d after injection. However, Ca-injected samples (NE/Ca and E/Ca) were rated as being significantly more tender than their uninjected counterparts (NE/NC and E/NC) at 3 and 7 d after injection. Injection of CaCl2 may result in more rapid and immediate tenderization if beef from animals supplemented with vitamin E is used. Vitamin E incorporation into muscle tissue may potentiate the action of exogenously added calcium by protecting the calpains from oxidation.

Animals↗
PubMed1992

The effect of calcium on pulmonary vascular resistance and right ventricular function.

Calcium chloride is frequently administered to patients immediately after separation from cardiopulmonary bypass to improve the contractile state of the myocardium. Animal studies suggest that calcium chloride may produce increases in pulmonary vascular resistance, which can precipitate right ventricular failure. In an attempt to determine the effect of calcium chloride administration after cardiopulmonary bypass on right ventricular function, this study was designed to evaluate patients with normal and elevated pulmonary vascular resistance. Fifty patients scheduled for elective cardiac surgery were prospectively studied for changes in ionized calcium levels before and after bypass. The impact of calcium administration on right ventricular function was assessed by a pulmonary artery catheter modified for the measurement of right ventricular ejection fraction. In all patients the level of ionized calcium decreased during bypass from a mean of 4.91 to 4.29 mg.dl-1. However, the infusion of calcium chloride (10 mg.kg-1) after bypass resulted in increasing the ionized calcium levels to prebypass levels. Administration of calcium chloride after bypass to patients with normal right ventricular function resulted in a transient increase in both cardiac output and right ventricular ejection fraction without any change in pulmonary vascular resistance. Eight patients with both elevated pulmonary vascular resistance and depressed right ventricular function were evaluated to determine the effect of calcium chloride after bypass on pulmonary vascular resistance and right ventricular ejection fraction. Administration of calcium chloride (10 mg.dl-1) to these patients did not result in any significant increase in pulmonary vascular resistance or depression of right ventricular performance. More important, in these patients, right ventricular ejection fraction and cardiac output were significantly increased after calcium chloride administration. In summary, the results of this study fail to demonstrate any increase in pulmonary vascular resistance or deterioration of right ventricular function with the administration of calcium chloride (10 mg.kg-1) after bypass in patients with elevated pulmonary vascular resistance.

Aged↗
PubMed2006

Influence of indomethacin in the rat aneurysm model.

The objective of this study was to trigger the formation of rat abdominal aortic aneurysm by applying calcium chloride periarterially and then to detect the degree of prevention of aneurysm occurrence by oral introduction of indomethacin in some of the rats. Thirty-one rats were divided into three groups. The infrarenal aorta above the iliac bifurcation of rats was treated with sodium chloride in group 1 (control, n = 7), calcium chloride in group 2 (n = 12), and calcium chloride-indomethacin in group 3 (n = 12) periarterially. The rats of each group were randomly selected at the end of the first, second, and third weeks postoperatively; and vessel diameters of abdominal aortas were measured by digital photography using a micrometer. Aneurysmal development was not observed in any of the rats in the control group. None of the comparisons was statistically significant (p > 0.05). Aneurysmal development was observed in all of the rats in the calcium chloride group. Results from the first, second, and third weeks postoperatively were statistically significant (p < 0.05). A middle aneurysmal development was observed in all rats in the calcium chloride-indomethacin group. Only results from the second and third weeks postoperatively were statistically significant (p < 0.05). Measurements in groups 2 and 3 were statistically significant when compared to group 1 (p < 0.001). However, the mean increase in the indomethacin-treated group (group 3) was only 26.1%. The macroscopic appearance of the control group and an aneurysm induced by calcium-chloride application are shown.

Administration, Oral↗
PubMed1994

Electrodiffusive movements of chloride ion in sodium-free solution: a possible calcium activated chloride conductance in cultured heart cells.

OBJECTIVE: The aim was to demonstrate that removal of extracellular sodium (Na+o), a manoeuvre known to increase intracellular calcium (Cai2+), can activate a chloride ion conductance in cultured embryonic chick heart cells. METHODS: Intracellular chloride activity (aiCl) and membrane potentials (Em) were simultaneously measured using chloride selective and 3 M KCl filled microelectrodes. RESULTS: In Na+o-free and K+o-free solutions, a frusemide (0.3 mM) resistant decrease in aiCl of 10 mM was recorded within 10 min, along with a 5 mV hyperpolarisation, and the calculated chloride equilibrium potential (ECl) followed the change of Em, suggesting a possibly enhanced chloride conductance. When cells were exposed to Na+o-free solution, the decrease of aiCl by approximately 10 mM was associated with an initial depolarisation, followed by a hyperpolarisation to approximately -60 mV (more negative than ECl). Subsequent removal of K+o depolarised Em to -39 mV (approximately ECl), and no further loss of aiCl occurred. Restoration of K+o caused a hyperpolarisation of Em (more negative than ECl) and a continuing decline of aiCl. Prevention of K+o induced hyperpolarisation by addition of 1 mM Ba2+ stopped the decline of aiCl (Em approximately ECl), suggesting that following Na+o removal, alteration of the driving force for chloride led to a chloride efflux via an enhanced chloride conductance. When Em depolarised to -50 mV after 3 min exposure to 0.1 mM ouabain, removal of [Cl-]o caused a further depolarisation and readdition of [Cl-]o induced an 18 mV hyperpolarisation. This chloride induced hyperpolarisation was blocked by removal of [Ca2+]o (+1 mM EGTA). CONCLUSIONS: The increase in chloride conductance observed under conditions that are known to raise Ca2+i supports the presence of a Ca2+i activated chloride conductance in cultured chick heart cells.

Animals↗
PubMed2003

The interacting binding domains of the beta(4) integrin and calcium-activated chloride channels (CLCAs) in metastasis.

CLCA (chloride channel, calcium-activated) proteins are novel pulmonary vascular addresses for blood-borne, lung-metastatic cancer cells. They facilitate vascular arrest of cancer cells via adhesion to beta4 integrin and promote early, intravascular, metastatic growth. Here we identify the interacting binding domains of endothelial CLCA proteins (e.g. hCLCA2, mCLCA5, mCLCA1, and bCLCA2) and beta4 integrin. Endothelial CLCAs share a common beta4-binding motif (beta4BM) in their 90- and 35-kDa subunits of the sequence F(S/N)R(I/L/V)(S/T)S, which is located in the second extracellular domain of the 90-kDa CLCA and near the N terminus of the 35-kDa CLCA, respectively. Using enzyme-linked immunosorbent, pull-down, and adhesion assays, we showed that glutathione S-transferase fusion proteins of beta4BMs from the 90- and 35-kDa CLCA subunits bind to the beta4 integrin in a metal ion-dependent manner. Fusion proteins from fibronectin and the integrins beta1 and beta3 served as negative controls. beta4BM fusion proteins competitively blocked the beta4/CLCA adhesion and prevented lung colonization of MDA-MB-231 breast cancer cells. A disrupted beta4BM in hCLCA1, which is not expressed in endothelia, failed to interact with beta4 integrin. The corresponding CLCA-binding domain of the beta4 integrin is localized to the specific determining loop (SDL). Again enzyme-linked immunosorbent, pull-down, and adhesion assays were used to confirm the interaction with CLCA proteins using a glutathione S-transferase fusion protein representing the C-terminal two-thirds of beta4 SDL (amino acids 184-203). A chimeric beta4 integrin in which the indicated SDL sequence had been replaced with the corresponding sequence from the beta1 integrin failed to bind hCLCA2. The dominance of the CLCA ligand in beta4 activation and outside-in signaling is discussed in reference to our previous report that beta4/CLCA ligation elicits selective signaling via focal adhesion kinase to promote metastatic growth.

Amino Acid Motifs↗
PubMed1994

Cyclopiazonic acid stimulates Ca2+ influx through non-specific cation channels in endothelial cells.

A non-specific cation channel in cultured human umbilical vein endothelial cells was obtained by cell-attached patch-clamp study. This channel showed a conductance of 28 pS when both pipette and bath contained 140 mM potassium chloride. when pipette solution was changed into 140 sodium chloride with 5 mM calcium chloride, the conductance was 26 pS. when 120 mM calcium chloride was used as the only cation in the pipette, a conductance of 6 pS was obtained. Bath application of cyclopiazonic acid, an inhibitor of the sarcoplasmic reticulum Ca2+ pump in smooth muscle and other tissues, dose dependently activates this non-specific cation channel. It is assumed that cyclopiazonic acid by blockade of the refilling of Ca2+ stores depletes the rapidly exchanging intracellular Ca2+ stores and this action stimulates Ca2+ influx through the non-specific cation channels in human umbilical vein endothelial cells.

Calcium↗
PubMed1999

Hyperpolarization-activated inward potassium and calcium-sensitive chloride currents in beating pacemaker insect neurosecretory cells (dorsal unpaired median neurons).

Hyperpolarization-activated inward currents were studied in single adult cockroach Periplaneta americana pacemaker neurosecretory cells, identified as dorsal unpaired median neurons using the whole-cell patch-clamp technique. Under current clamp, injection of negative current produced a hyperpolarization of the cell membrane with a sag in the membrane potential toward the resting value. Under voltage clamp, the whole-cell current-voltage relationship exhibited an unexpected biphasic aspect. The global hyperpolarization-activated inward current could be dissociated by means of 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid and tetraethylammonium chloride sensitivity, ionic selectivity, voltage dependence and activation threshold as inward potassium and calcium-sensitive chloride currents. The inward potassium current was activated around -80 mV. The reversal potential followed the potassium equilibrium potential when the extracellular potassium concentration was raised. This current was not dependent on the external sodium concentration and was sensitive to 10 mM tetraethylammonium chloride or 5 mM barium chloride. The hyperpolarization-activated inward calcium-sensitive chloride current was activated in a range of potential 20 mV more positive than the potassium current. The estimated reversal potential (-71 mV) was very close to the equilibrium potential for chloride ions ( 73 mV). Intracellularly applied 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and external application of 1 mM zinc chloride, calcium-free saline or high concentrations of intracellular 1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetate blocked the inward chloride current. Current-clamp experiments indicated that the inward potassium current accounted for inward rectification of dorsal unpaired median neurons. Our findings report, for the first time in pacemaker neurosecretory cells, the co-existence of two distinct hyperpolarization-activated inward currents which have specialized function in pacemaker activity.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗