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Effects of chronic bretylium treatment on the sympathetic neuron and the smooth musculature of the rat.

The effects of chronic i.p. injection of high doses of bretylium on sympathetic nerves on the smooth musculature of the vas deferens of adult and newborn rats were examined using fluorescence histochemistry, light and electron microscopy and organ bath physiological techniques. Bretylium treatment caused mitochondrial swelling, loss of cristae and the formation of electron-dense inclusions in the mitochondria of sympathetic neurons. However, neuron degeneration was not observed and fluorescent histochemical appearance of adrenergic neurons was normal. A small transient supersensitivity of the isolated vas deferens of bretylium-treated rats to noradrenaline, but not to acetylcholine, occurred. There was, however, considerable increase in the maximal contractile response to both noradrenaline and acetylcholine. In high calcium concentrations acetylcholine-induced contractions of vasa deferentia from bretylium-treated rats were significantly greater than control; there was no difference in magnitude of noradrenaline-induced contractions.

Acetylcholine↗

[Change from ACE inhibitor, Ca-antagonist or beta-blocker to candesartan cilexetil: better efficacy and tolerance. SWITCH study (German study segment)].

BACKGROUND AND OBJECTIVE: Lack of efficacy in the treatment of hypertension with only one drug presents a problem in general practice and often requires switching to another type of drug, because higher dosage of currently used antihypertensives increases the frequency of side effects. Angiotensin II antagonists are well tolerated and there is no evidence of dose-related increase in side effects. This study in 574 hypertensives under therapy with ACE inhibitors, beta-blockers or calcium channel blockers was undertaken to determine whether direct switching to the Angiotensin II-antagonist candesartan cilexetil at its maximal dose of 16 mg is as effective and tolerable as starting therapy with candesartan cilexetil 8 mg followed by up-titration to 16 mg after 4 weeks. PATIENTS AND METHODS: 258 men (mean age 57 +/- 11 years) and 316 women (58 +/- 12) with essential hypertension (blood pressure < 180/95 mm Hg) under ambulatory therapy with ACE-inhibitors, beta-blockers or calcium channel blockers with inadequate efficacy or tolerability were switched to monotherapy with candesartan cilexetil. Half of the patients were treated with 8 mg for 4 weeks (n = 284), the other half received 16 mg (n = 290). Both groups then were treated with candesartan cilexetil, 16 mg, for further 4 weeks. Choice of treatment was doubly blinded and randomised. RESULTS: After 4 weeks significant blood pressure reduction was observed in both treatment groups (p < 0.0001 for each pretreatment group). A tendency for more adequate blood pressure reduction under initial therapy with candesartan cilexetil 16 mg was observed. There was a small further blood pressure reduction in both treatment groups after 8 weeks. In comparison with the previous medications the proportion of patients with blood pressure reduction < 90 mm Hg diastolic was doubled in both treatment arms after 4 weeks: after initial dose of candesartan cilexetil 8 mg from 36.7% to 78.8%, after initial dose of candesartan cilexetil 16 mg from 43.9% to 81.1%. Clinically relevant side effects were not observed. CONCLUSION: Switching of antihypertensive monotherapy with ACE inhibitors, beta-blockers or calcium channel blockers to candesartan cilexetil 8 mg or 16 mg under ambulatory conditions is safe and equally well tolerated and effectively reduces blood pressure.

Adolescent↗

Effect of diltiazem on in vitro rabbit bladder function.

The relationship between extracellular calcium and urinary bladder function was investigated by studying the effect of the specific calcium antagonist diltiazem on the functional ability of the in vitro whole rabbit urinary bladder to empty in response to pharmacological stimulation. The bladder was found to require an extracellular calcium concentration of 4.5 X 10(-4) M to elicit near complete cholinergic-mediated emptying. Diltiazem (1 X 10(-6) - 1 X 10(-4) M) inhibition of bladder function was competitively antagonized by increasing the extracellular calcium concentration (0.45 X 10(-4) - 3.6 X 10(-4) M). In the absence of diltiazem, alterations in the extracellular calcium concentration between 0.45 X 10(-4) and 3.6 X 10(-4) M had no significant effects on bladder response to bethanechol. KCl-mediated bladder emptying was significantly more sensitive to diltiazem inhibition than was bethanechol-stimulated emptying. Even at the intermediate diltiazem concentration of 1 X 10(-5) M, increasing the extracellular calcium concentration did not completely reverse the inhibitory effect of diltiazem on bladder response to KC1. These findings are consistent with the hypothesis that diltiazem at low concentrations may inhibit extracellular calcium influx while at higher concentrations translocation of intracellular stores of calcium may be inhibited as well. The study also suggests that diltiazem is capable of inhibiting urinary bladder function and deserves further consideration as a possible therapeutic agent in certain forms of urinary bladder dysfunction.

Animals↗

In-vitro calcium phosphate growth over functionalized cotton fibers.

Biomimetic growth of calcium phosphate compound on cotton sheets treated with tetraethoxy silane and soaked in simulated body fluid solution was studied using scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX), micro-Fourier transform infrared spectroscopy (FTIR) and X-ray diffractometry (XRD). Micro-FTIR and EDAX results show that silicon was coupled to the cotton fiber when cotton was treated with tetra-ethoxy silane (TEOS) at 125 degrees C for 1 h. Calcium phosphate nucleation started to occur on the surface of TEOS-treated cotton fibers upon immersion in 1.5xSBF (simulated body fluid solution) within 3 days and after 20 days, all the fiber surfaces were found covered with a thick and porous coating of calcium phosphate. The Ca and P determined by inductively coupled plasma spectroscopy (ICP) analysis revealed that the Ca/P ratio as well as the amount of calcium phosphate coating depends on the soaking time in SBF solution.

Journal Article↗

Effects of the water content of soda lime on compound A concentration in the anesthesia circuit in sevoflurane anesthesia.

BACKGROUND: Sevoflurane anesthesia is usually performed with fresh gas flow rates greater than 2 l/min due to the toxicity of compound A in rats and limited clinical experience with sevoflurane in low-flow systems. However, to reduce costs, it would be useful to identify ways to reduce compound A concentrations in low-flow sevoflurane anesthesia. This goal of this study was to determine if compound A concentrations can be reduced by using soda lime with water added. METHODS: Low-flow sevoflurane anesthesia (fresh gas flow of 1 l/min) was performed in 37 patients using soda lime with water added (perhydrated soda lime) or standard soda lime as the carbon dioxide (CO2) absorbent. The soda lime was not changed between patients, but rather was used until CO2 rebreathing occurred. The perhydrated soda lime was prepared by spraying 100 ml distilled water onto 1 kg fresh soda lime, and water was added only when a new bag of soda lime was placed into the canister. Compound A concentrations in the circle system, soda lime temperatures, inspired and end-tidal CO2 and end-tidal sevoflurane concentrations, and CO2 elimination by the patient were measured during anesthesia. RESULTS: Compound A concentrations were significantly lower for the perhydrated soda lime (1.9 +/- 1.8 ppm; means +/- SD) than for the standard soda lime (13.9 +/- 8.2 ppm). No differences were seen between the two types of soda lime with regard to the temperature of the soda lime, end-tidal sevoflurane concentrations, or CO2 elimination. Compound A concentration decreased with the total time of soda lime use for both types of soda lime. The CO2 absorption capacity was significantly less for perhydrated soda lime than for standard soda lime. CONCLUSIONS: Compound A concentrations in the circuit can be reduced by using soda lime with water added. The CO2 absorption capacity of the soda lime is reduced by adding water to it, but this should not be clinically significant.

Anesthesia, Closed-Circuit↗

Visualization of intracellular calcium during necrotaxis.

Granulocytes are known to exhibit positive chemotaxis towards erythrocytes destroyed by laser irradiation (necrotaxis). A computerized method for visualizing intracellular calcium concentration in granulocytes during necrotaxis has been developed. Successive bursts of fluorescence were observed: 1 degree) about 10 s after the creation of the necrotactic source; 2 degrees) when the granulocyte touched the target.

Analog-Digital Conversion↗

A simple spectrophotometric assay for micromolar amounts of lanthanum in the presence of calcium and phosphate.

A sensitive spectrophotometric assay for micromolar amounts of lanthanum in the presence of calcium and phosphate (as hydroxyapatite) was developed utilizing the change in absorption (at 652 nm) when the dye arsenazo III was complexed with lanthanum. Arsenazo III was used at a level of 25 microM and the solution pH was maintained at 3.1 with 0.2 M sodium acetate. Lanthanum concentrations down to 0.5 microM could be reliably assayed. Calcium ion did not complex well with arsenazo III at pH 3.1. With calcium present at 100 microM and lanthanum at 10 microM, the assay was 115 times more sensitive for lanthanum. The assay is simple, rapid, reproducible and, unlike the assay using radioactive lanthanum, can be performed at any time.

Arsenazo III↗

Local calcium transients regulate the spontaneous motility of dendritic filopodia.

During development, dendrites, and in particular dendritic filopodia, undergo extensive structural remodeling, presumably to help establish synaptic contacts. Here, we investigated the role of calcium signaling in dendritic plasticity by simultaneously recording calcium dynamics and filopodial growth in rat hippocampal slice cultures. Local calcium transients occurred in dendritic filopodia and shafts, often at putative synaptic sites. These events were highly correlated with filopodial motility: comparatively rare when individual filopodia emerged from the dendrite, they became more frequent after filopodia started growing, finally causing them to halt. Accordingly, an experimental reduction of the frequency of local calcium transients elicited filopodial growth and, conversely, calcium uncaging reduced filopodial motility. Our observations suggest that low levels of local calcium transients facilitate filopodial outgrowth, whereas high levels inhibit the formation of filopodia and stabilize newly formed ones. This process may facilitate synapse formation and may serve as a homeostatic mechanism distributing synapses evenly along developing dendrites.

2-Amino-5-phosphonovalerate↗

The interaction of the NK1 receptor antagonist CP-96,345 with L-type calcium channels and its functional consequences.

1. We investigated the effects of the non-peptide NK1 receptor antagonist, CP-96,345, its inactive enantiomer CP-96,344, and the racemic mixture (+/-)-CP-96,345, on the binding of [3H]-nimodipine and [3H]-diltiazem to L-type calcium channels in rat cerebral cortex membranes. In isolated peripheral tissues containing tachykinin receptors, the effects of (+/-)-CP-96,345 have been compared with those of diltiazem. 2. In guinea-pig trachea, (+/-)-CP-96,345 produced antagonism of responses to the selective NK1 agonists [Sar9, Met(O2)11]SP and substance P-methyl ester that was apparently competitive in nature (pKB 7.0-7.5), while in guinea-pig ileum the antagonism was not surmountable. 3. The reduction of maximum responses by (+/-)-CP-96,345 in the guinea-pig ileum was not selective; it was obtained with muscarinic agonists and other agents, and was also observed in the portal vein of the rat where NK1 receptors are not present. 4. The tissue-specific reduction of maximum responses by (+/-)-CP-96,345 in ileum was reproduced by diltiazem. 5. (+/-)-CP-96,345 produced a concentration-dependent enhancement of [3H]-nimodipine binding to rat cerebral cortex membranes with a maximal stimulation of 186 +/- 29% above control (EC50 83.2 nM). Scatchard analysis revealed that (+/-)-CP-96,345 increased the affinity of [3H]-nimodipine for its binding sites without affecting Bmax (control: KD = 0.32 nM; with 100 nM (+/-)-CP-96,345: KD = 0.074 nM). 6. CP-96,345, CP-96,344, and the racemate all inhibited [3H]-diltiazem binding in rat cerebral cortex membranes with Ki values of 22.5 nM, 34.5 nM and 29.9 nM respectively; a similar value was obtained for diltiazem itself (33.6 nM). In comparison, CP-96,345 and ( +/- )-CP-96,345 inhibited the binding of[125I]-Bolton-Hunter-conjugated substance P in this tissue with Ki values of 59.6 nM and 82.0 nM respectively, while CP-96,344 had no measurable affinity (IC50> 10 microM).7. Substance P and a range of ligands selective for NK1, NK2, or NK3 receptors had no significant effect at 10 microM on either [3H]-diltiazem or [3H]-nimodipine binding.8. The results indicate that in addition to possessing affinity for the NK1 receptor, the non-peptide antagonist, CP-96,345, displays high affinity for [3H]-diltiazem binding sites on L-type calcium channels.The functional effect that may be observed in integrated models will be a consequence of either property, or be a composite effect of NK1 receptor antagonism and L-channel blockade.

Animals↗

Inhibition of cAMP accumulation via recombinant human serotonin 5-HT1A receptors: considerations on receptor effector coupling across systems.

Inhibition of forskolin-stimulated cyclic AMP accumulation was measured in two stable HeLa cell lines HA6 and HA7 expressing different levels of recombinant human 5-HT1A receptors. These cells were studied previously to characterize another second messenger system activated by 5-HT1A receptors, i.e. calcium mobilization. The pharmacological characterization of the inhibition of cyclic AMP accumulation was made using agonists (5-HT, 8-OH-DPAT, buspirone, MDL 73005) and putative antagonists (SDZ 216-525, NAN-190, WAY-100135, pindolol, propranolol, WAY 100635). It is shown that 5-HT, 8-OH-DPAT, buspirone, MDL 73005 behaved as full (or nearly full) and potent agonists, whereas SDZ 216-525, NAN-190 and WAY-100135 displayed a limited (and similar) degree of intrinsic activity at human 5-HT1A receptors; on the other hand pindolol, propranolol and WAY 100635 behaved as "silent" antagonists. The effects were quantitatively and qualitatively very similar in both cells lines for all drugs tested, suggesting that the coupling between 5-HT1A receptors and inhibition of cyclic AMP accumulation in HeLa cells is very tight. There were, however, significant variations in both the level of agonism and the potency of a number of compounds when calcium mobilization and the inhibition of cyclic AMP accumulation were compared. Especially in HA7 cells which express lower receptor levels, a number of drugs failed to display agonism (e.g. buspirone or MDL 73005), whereas in HA6 cells they acted as partial agonists. Together, the data show that functional responses mediated by the same receptor can vary rather dramatically depending on receptor density and/or on the effector system involved. Interestingly, 5-HT1A receptor-mediated inhibition of adenylate cyclase activity measured in calf hippocampal membranes shows very similar degrees of potency and intrinsic activity for a number of compounds that have been tested on the inhibition of cyclic AMP accumulation in HeLa cells, suggesting that the very tight coupling observed in the recombinant system may apply to native 5-HT1A receptors.

Adenylyl Cyclases↗

Detection of Ca(2+)-transients elicited by flash photolysis of DM-nitrophen with a fast calcium indicator.

A confocal spot detection optical setup was used to record fluorescence signals in response to calcium pulses, elicited by flash photolysis of DM-nitrophen, with the calcium indicators CaOrange-5N and Fluo-3. Our results yield the following conclusions: [Ca2+] changes are almost perfect spikes at pCa 9 and broader transients followed by a step at pCa 7. The [Ca2+] spikes were used to measure the dissociation rate constant of the Ca2+ dyes. Experiments at pCa 7 were used to verify the kinetic rate constants of the dyes and to obtain those of DM-nitrophen. The association rate constant of this compound was found to be more than one order of magnitude faster than that suggested previously. CaOrange-5N was able to track changes in [Ca2+] more accurately than Fluo-3. This latter dye introduced severe distortions which preclude a quantitative deconvolution of the fluorescence transients into changes in the free [Ca2+].

Acetates↗

2-Aminoethoxydiphenyl borate (2-APB) stimulates a conformationally coupled calcium release pathway in the NG115-401L neuronal cell line.

We report in this study a 2-aminoethoxydiphenyl borate (2-APB) activated Ca2+ pathway in NG115-401L (401L) neuronal cells bearing resemblance to hormonal and ryanodine receptor activated pathways. We observed that 2-APB, in contrast to much earlier work, did not inhibit store operated Ca2+ channel (SOC) function, but rather induced potent Ca2+ discharge responses that robustly activated SOC-mediated Ca2+ influx. Further, these studies intriguingly revealed that the 2-APB-induced Ca2+ release pathway likely couples conformationally to targets in the plasma membrane, as membrane permeabilization or actin perturbation abolished the ability of the compound to stimulate Ca2+ signals. These findings suggest that conformationally sensitive complexes form between endoplasmic reticulum and plasma membrane components that not only regulate Ca2+ influx, previously proposed as the conformational coupling hypothesis, but are also required to promote Ca2+ release from intracellular stores. These observations further characterize the 401L neuronal cell line as having unique characteristics that may prove useful in gaining insight into the nature of the coupling mechanism linking Ca2+ release to Ca2+ influx.

Animals↗

Role of calcium in mediating the biphasic contraction of the rabbit urinary bladder.

1. The response of the urinary bladder to field stimulation is biphasic in nature consisting of an initial phasic contraction followed by a prolonged tonic phase which lasts for the duration of the stimulation. 2. The phasic response is mediated by the release of neurohumoral transmitters, primarily acetylcholine (via muscarinic receptor stimulation) and ATP (via purinergic receptor stimulation). The tonic component is mediated entirely via muscarinic receptor stimulation. 3. The present study investigates the dependence on extracellular calcium of the phasic and tonic contractile responses to field stimulation, bethanechol, and ATP. The results can be summarized as follows: 4. Field stimulation (2 and 32 Hz) and bethanechol evoke a biphasic contractile response whereas ATP evokes only a phasic response. 5. There were no significant effects of either calcium channel blockers or calcium fee EGTA medium on either spontaneous contraction or basal tension of muscle strips. 6. The calcium channel antagonists diltiazem and verapamil inhibited both the phasic and tonic responses induced by field stimulation (both 2 and 32 Hz) in a dose dependent manner. 7. For both 2 and 32 Hz stimulation, the ED50 s for the inhibition of the tonic phases of the responses to field stimulation were significantly lower than the ED50s for the inhibition of the phasic responses. 8. The tonic phase of the responses to field stimulation were inhibited to a significantly greater degree than the phasic responses by incubation in calcium-free medium containing EGTA. 9. Both the phasic and tonic components of the response to bethanechol stimulation were inhibited equally, and followed a similar time course as the tonic component of field stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

Outcome studies in diabetic nephropathy.

Outcome studies in diabetic nephropathy have focused on strategies to prevent progression of diabetic nephropathy, the leading cause of ESRD in the United States. Once diabetics develop overt nephropathy, prognosis is poor. Risk factors for diabetic nephropathy are discussed, and include hyperglycemia, hypertension, angiotensin II, proteinuria, dyslipidemia, smoking, and anemia. Major outcomes as well as outcome studies in diabetic nephropathy for patients with microalbuminuria and macroalbuminuria are reviewed. Furthermore, the role of therapy with angiotensin converting enzyme inhibitors, angiotensin II receptor blockers, calcium channel blockers, and mineralocorticoid receptor antagonists as well as selected combination therapy are discussed. Recommendations for therapy with ace inhibitors and angiotensin II receptor blockers are made based on this evidence.

Albuminuria↗

Role of the phospholipase C-inositol 1,4,5-trisphosphate pathway in calcium release-activated calcium current and capacitative calcium entry.

We investigated the putative roles of phospholipase C, polyphosphoinositides, and inositol 1,4,5-trisphosphate (IP(3)) in capacitative calcium entry and calcium release-activated calcium current (I(crac)) in lacrimal acinar cells, rat basophilic leukemia cells, and DT40 B-lymphocytes. Inhibition of phospholipase C with blocked calcium entry and I(crac) activation whether in response to a phospholipase C-coupled agonist or to calcium store depletion with thapsigargin. Run-down of cellular polyphosphoinositides by concentrations of wortmannin that block phosphatidylinositol 4-kinase completely blocked calcium entry and I(crac). The membrane-permeant IP(3) receptor inhibitor, 2-aminoethoxydiphenyl borane, blocked both capacitative calcium entry and I(crac). However, it is likely that 2-aminoethoxydiphenyl borane does not inhibit through an action on the IP(3) receptor because the drug was equally effective in wild-type DT40 B-cells and in DT40 B-cells whose genes for all three IP(3) receptors had been disrupted. Intracellular application of another potent IP(3) receptor antagonist, heparin, failed to inhibit activation of I(crac). Finally, the inhibition of I(crac) activation by or wortmannin was not reversed or prevented by direct intracellular application of IP(3). These findings indicate a requirement for phospholipase C and for polyphosphoinositides for activation of capacitative calcium entry. However, the results call into question the previously suggested roles of IP(3) and IP(3) receptor in this mechanism, at least in these particular cell types.

Androstadienes↗

Microglial calcium responses to platelet-activating factor are inhibited by analogue CAS 99103-16-9 and dihydropyridine PCA 4248 but not by ginkgolide A.

Calcium signals evoked in N9 microglial cells were monitored using the calcium indicator dye Fluo-4 in a fluorescence imaging plate reader. Platelet activating factor in the range 100 nM to 20 microM elicited graded calcium responses. The analogue CAS 99103-16-9 inhibited the evoked calcium rise with an apparent KB of 1.3 +/- 0.4 microM. The dihydropyridine PCA 4248 inhibited the evoked calcium rise with an apparent KB of 1.2 +/- 0.2 microM. Ginkgolide A at concentrations up to 18 microM had no effect on the evoked calcium rise. While CAS 99103-16-9 and PCA 4248 appear to be simple competitive inhibitors of platelet-activating factor responses, the efficacy of ginkgolide in more complex pharmacological situations may result from an action at a site other than the platelet-activating factor receptor.

Aniline Compounds↗

Where plants make oxygen: a structural model for the photosynthetic oxygen-evolving manganese cluster.

In the photosynthetic evolution of oxygen, water oxidation occurs at a catalytic site that includes four manganese atoms together with the essential cofactors, the calcium and chlorine ions. A structural model and a determination of the manganese oxidation states based on x-ray absorption spectroscopy are presented. The salient features, in both higher plants and cyanobacteria, are a pair of di-mu-oxo bridged manganese binuclear clusters linked by a mono-mu-oxo bridge, one proximal calcium atom, and one halide. In dark-adapted samples, manganese occurs in oxidation states (III) and (IV). Data from oriented membranes display distinct dichroism, precluding highly symmetrical structures for the manganese complex.

Bridged-Ring Compounds↗

In vivo effect of fluoride-substituted apatite on rat bone.

Different types of calcium phosphate compounds are commercially available for medical and dental applications as bone substitute materials. Biological apatites contain several kinds of minor elements such as carbonate (CO3), magnesium (Mg), and fluoride (F) in enamel, dentin, and bone. It has been shown that F ion and F-substituted apatite promoted osteoblast proliferation and inhibited osteoclast cell activity. The purpose of this study was to investigate the in vivo rat tibia activity on F-substituted apatite (FAp). Apatites of unsintered calcium deficient apatite (CDA), and FAps, with low, medium, and high F concentrations, were implanted in rat tibia for 1 and 2 weeks. Implanted tissues were embedded in paraffin blocks, stained with hematoxylin-eosin and histomorphometrically observed. Results showed that low F concentration induced better and faster new bone formation in vivo compared to CDA. Therefore the results suggested that F as a minor element in bone rendered a suitable effect on bone formation in vivo.

Animals↗