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PubMed2005

State-dependent compound inhibition of Nav1.2 sodium channels using the FLIPR Vm dye: on-target and off-target effects of diverse pharmacological agents.

Voltage-gated sodium channels (NaChs) are relevant targets for pain, epilepsy, and a variety of neurological and cardiac disorders. Traditionally, it has been difficult to develop structure-activity relationships for NaCh inhibitors due to rapid channel kinetics and state-dependent compound interactions. Membrane potential (Vm) dyes in conjunction with a high-throughput fluorescence imaging plate reader (FLIPR) offer a satisfactory 1st-tier solution. Thus, the authors have developed a FLIPR Vm assay of rat Nav1.2 NaCh. Channels were opened by addition of veratridine, and Vm dye responses were measured. The IC50 values from various structural classes of compounds were compared to the resting state binding constant (Kr)and inactivated state binding constant (Ki)obtained using patch-clamp electrophysiology (EP). The FLIPR values correlated with Ki but not Kr. FLIPRIC50 values fell within 0.1-to 1.5-fold of EP Ki values, indicating that the assay generally reports use-dependent inhibition rather than resting state block. The Library of Pharmacologically Active Compounds (LOPAC, Sigma) was screened. Confirmed hits arose from diverse classes such as dopamine receptor antagonists, serotonin transport inhibitors, and kinase inhibitors. These data suggest that NaCh inhibition is inherent in a diverse set of biologically active molecules and may warrant counterscreening NaChs to avoid unwanted secondary pharmacology.

Biological Assay↗
PubMed1987

The human leucocyte sodium pump in adrenocortical insufficiency.

In view of the known effects of cortisol and related compounds on sodium balance, we examined leucocyte 22Na+ efflux in patients with adrenocortical insufficiency. The ouabain-sensitive 22Na+ efflux rate constant, which reflects sodium pump activity, was lowered in secondary hypoadrenalism (mean 1.95 +/- SD 0.25 vs 2.33 +/- 0.45 h-1, P less than 0.002, n = 6). Patients with hypoadrenalism also had higher intracellular sodium content (mean 34.9 +/- SD 3.5 vs 27.8 +/- 9.5 mmol/kg dry weight, P less than 0.008) and lower intracellular potassium to sodium ratios (mean 9.7 +/- SD 0.7 vs 12.9 +/- 3.6, P less than 0.001). These defects in cellular sodium balance were corrected following replacement therapy with cortisol, prednisolone or dexamethasone. In-vitro incubation of normal leucocytes with 5-20 nmol/l dexamethasone led to an increase in the Na+/K+-ATPase activity. The effect of cortisol and related compounds on the distribution of sodium between intra- and extra-cellular fluid may be mediated by their action on the sodium pump.

Adrenal Insufficiency↗
PubMed1980

New cyclopenta[a]naphthalene derivatives. Synthesis of 2-(carbamylmethyl)-8-hydroxy-3H-cyclopental[a]naphthalene as a possible deoxyribonucleic acid binding agent.

8-Methoxy-1-oxo-2,3-dihydro-1H-cyclopenta]a]naphthalene (4) was converted to the oxalyl derivative (7) by treatment with diethyl oxalate in the presence of sodium ethoxide. Compound 7 in the form of the sodium salt was alkylated with ethyl bromoacetate in DMF to 2-(carbethoxymethyl)-8-methoxy-1-oxo-2,3-dihydro-1H-cyclopenta[a]naphthalene (8). Treatment of 8 with methanolic ammonia yielded the corresponding amide (9). Dealkylation of 8 with 48% HBr and subsequent esterification gave compound 10. Ammonolysis of 10 led to the amide 11, which after reduction and subsequent dehydration of the reduced product afforded the desired compound, 2-(carbamylmethyl)-8-hydroxy-3H-cyclopenta[a]naphthalene (2). Compound 2 was found to be mildly growth inhibitory to L1210 and CCRF--CEM leukemic cells in culture. From thermal transition temperature studies, compound 2 was found to bind to calf thymus DNA and the poly(deoxyribonucleotides), e.g., poly(dG).poly(dC), poly(dG-dC), poly(dA).poly(dT), and poly(dA-dT).

Animals↗
PubMed1996

Arsenic and chromium enhance transformation of bovine papillomavirus DNA-transfected C3H/10T1/2 cells.

Tumor promoters such as phorbol esters, teleocidin and okadaic acid increase the numbers of multilayered, transformed foci produced by BPV DNA-transfected C3H/10T1/2 cells. We questioned whether arsenic and chromium, which are known human carcinogens also enhance transformation of BPV DNA-transfected C3H/10T1/2 cells. Cr(III) potassium sulfate at 100 microM enhanced transformation by 1.4-fold, but Cr(VI) as potassium chromate did not enhance transformation, although toxicity of potassium chromate may have prevented enhancement of transformation. Sodium arsenite (As(III) at 5 microM and sodium arsenate (As(V)) at 25 microM both enhanced neoplastic transformation by 6-fold. By comparison, in previous studies, sodium orthovanadate (V(IV)) or vanadyl sulfate (V(IV)) at 4 microM enhanced numbers of transformed foci by 25-50-fold. The comparatively strong enhancement of transformation by vanadium and phorbol esters suggests that neoplastic transformation may occur by mechanisms that are common to these compounds including alteration of tyrosine phosphorylation.

Animals↗
PubMed2006

Salt effects on an ion-molecule reaction--hydroxide-catalyzed hydrolysis of benzocaine.

PURPOSE: This work investigates the effect of various salts on the rate of a reaction involving a neutral species (benzocaine alkaline hydrolysis). METHODS: Benzocaine hydrolysis kinetics in NaOH solutions in the presence of different salts were studied at 25 degrees C. Benzocaine solubility in salt solutions was also determined. Solubility data were used to estimate salt effects on benzocaine activity coefficients, and pH was used to estimate salt effects on hydroxide activity coefficients. RESULTS: Salts either increased or decreased benzocaine solubility. For example, solubility increased with 1.0 M tetraethylammonium chloride (TEAC) approximately 3-fold, whereas solubility decreased approximately 35% with 0.33 M Na2SO4. Salt effects on hydrolysis rates were more complex and depended on the relative magnitudes of the salt effects on the activity coefficients of benzocaine, hydroxide ion, and the transition state. As a result, some salts increased the hydrolysis rate constant, whereas others decreased it. For example, the pseudo-first-order rate constant decreased approximately 45% (to 0.0584 h(-1)) with 1 M TEAC, whereas it increased approximately 8% (to 0.116 h(-1)) with 0.33 M Na2SO4. CONCLUSIONS: Different salt effects on degradation kinetics can be demonstrated for a neutral compound reacting with an ion. These salt effects depend on varying effects on activity coefficients of reacting and intermediate species.

Anesthetics, Local↗
PubMed1995

[Comparison of the effectiveness of sodium azide and merthiolate as bacteriocidal agents--a NMR study].

The pseudohalogenide sodium azide (NaN3) and the organometallic mercury compound merthiolate (sodium salt of ethyl mercurithiosalicylate) are commonly used as preserving solutions for biomaterials. In the present paper, these preserving solutions were compared for their efficacy in preserving porcine articular cartilage. For this purpose, porcine articular cartilage was incubated with phosphate buffer containing the corresponding amounts of NaN3 or merthiolate. Supernatants were assayed using NMR spectroscopy, and the content of soluble saccharides was determined using the dye Alcian blue. The results obtained clearly indicate that merthiolate is more effective than sodium azide. This makes the former a more suitable preserving agent for the storage of cartilage. A significant decrease in the lactate concentration, and enhancement of the acetate and the N-acetyl resonances indicate degenerative changes in the cartilage. However, when a large excess of sodium azide is used, its efficacy approaches that of merthiolate.

Animals↗
PubMed2002

Effective treatment of seborrheic dermatitis using a low dose, oral homeopathic medication consisting of potassium bromide, sodium bromide, nickel sulfate, and sodium chloride in a double-blind, placebo-controlled study.

BACKGROUND: Topical over-the-counter remedies exist to aid in the control of seborrheic dermatitis and chronic dandruff on a superficial level. Low-dose systemic oral nickel and bromide therapy has shown promise in providing improvement and eventual clearing of the disease. OBJECTIVE: The purpose of this study was to further evaluate the effect of an orally administered low-dose, homeopathic mineral therapy (Potassium bromide 1X, Sodium bromide 2X, Nickel sulfate 3X, Sodium chloride 6X) on seborrheic dermatitis and chronic dandruff. METHODS: Forty-one patients with seborrheic dermatitis and/or chronic dandruff were assigned to one of two treatment groups: Active (containing the medication) or placebo (vehicle). Study medication was administered in a placebo-controlled, randomly-selected, double-blind study for 10 weeks. At the end of 10 weeks all patients crossed over to the active medication, under a different label for an additional 10 weeks in an open study format. RESULTS: Twenty-nine patients completed the 10-week blinded portion of the study. After 10 weeks of treatment, the disease state of the active patients improved significantly over that of the placebo patients (p<0.04). The placebo patients' condition before and after crossover to active treatment was also evaluated, showing significant improvement (p<0.01) 10 weeks after crossing over to active medication. CONCLUSION: Oral therapy using a low-dose homeopathic preparation combining Potassium bromide 1X, Sodium bromide 2X, Nickel sulfate 3X, and Sodium chloride 6X, provides significant improvement in seborrheic dermatitis and dandruff after 10 weeks of dosing.

Adult↗
PubMed2005

Effects of selected biocides used in the disinfection of cooling towers on toxicity and bioaccumulation in Artemia larvae.

This paper reports the acute toxicity of three biocides used in the disinfection of cooling towers, tetrakis(hydroxymethyl) phosphonium chloride (THPC), trichloroisocyanuric acid (TCIC), and sodium bromide (NaBr), on Artemia larvae, their effect on the phototactic response of this organism, and the potential of bioaccumulation in this species. The 24-h median lethal concentration (LC50) values for these biocides with respect to 24-, 48-, and 72-h-old Artemia, determined by static bioassays, showed the following rank order of toxicity: THPC < TCIC < NaBr. An age-dependent increase in sensitivity was seen for each compound. All three biocides reduced the phototactic response of 24-h-old Artemia larvae in 24-h static bioassays; the median inhibitory concentration ratios obtained were 30 to 40 times lower than their respective 24-h LC50 values. The results suggest that phototaxis bioassays could provide the speed and simplicity required for screening many potential pollutants for harmful effects. The bioconcentration factors obtained for Artemia larvae exposed to 10% LC50 for 168 h in renewal assays were 93.75, 1.67, and 0.23 for THPC, TCIC, and NaBr, respectively. This shows these biocides pose no bioaccumulation risk in this organism, although the value of 93.75 obtained for THPC is close to the threshold above which such a risk is considered to exist.

Animals↗
PubMed1985

Enhanced reactivation of UV-irradiated adenovirus 2 in HeLa cells treated with non-mutagenic chemical agents.

Treatment of HeLa cells with ethanol and sodium arsenite, compounds which are known to elicit the heat-shock response, before infection with UV-irradiated adenovirus 2 has been found to result in the enhanced reactivation of the damaged virus in a manner similar to that obtained by pre-irradiation or heating of the cells. Enhanced reactivation may be the result of the inhibition of DNA synthesis caused by these agents since hydroxyurea also produced a significant enhancement.

Adenoviruses, Human↗
PubMed1992

[Chronic bromide intoxication caused by bromide-containing combination drugs].

A 49-year-old woman who had noted increasing fatigue and found it difficult to concentrate became confused and uncoordinated with rapid speech. Anxious and suffering from insomnia she had for 6 weeks taken a prescription-free bromide-containing drug mixture (daily 0.09 g potassium bromide and 1.8 g sodium bromide), to a total bromide intake of 60 g. The admission diagnosis of chronic bromism was confirmed by a markedly increased serum bromide concentration (325 mg/l). Once she had stopped taking the drug and had increased her salt intake she became symptom-free within 8 days. The case demonstrates that, while chronic bromism has become rare, it should still be included in the differential diagnosis, even after intake of supposedly harmless medication.

Bromides↗
PubMed1999

Ammonium and sodium urates precipitating from synthetic urine and fine structure of urate renal calculi.

A study of ammonium and sodium urate precipitation in vitro and the fine structure of several urate renal calculi was carried out to contribute to an understanding of the participation of ammonium and sodium urates in urolithiasis. Ammonium urate precipitated in vitro in two different morphologies: a typical spherulite morphology formed at high supersaturation and disorganized needle-like crystals formed at low supersaturation. In all cases sodium urate precipitated in vitro as bundles of curved fibrils, its crystallization being inhibited by calcium in concentrations between 20 and 60 mg/l depending on the sodium urate supersaturation. From a collection of 1300 renal calculi, only three had ammonium urate as their main component (0.2%), three were mixed calculi (0.2%) consisting of ammonium urate and calcium oxalate (two) or uric acid (one), and in one calculus ammonium urate was present as a minor component. Only in a mixed calculus of uric acid and calcium oxalate was sodium urate detected in a very low quantity. The study of the fine structure of the renal calculi constituted mainly by ammonium urate demonstrated similar patterns in which spherulites, needle-like individual crystals and an amorphous mass of ammonium urate with abundant organic matter in non-organized structures coexist. As minor components, struvite or calcium oxalate crystals were found. A general mechanism of the formation of such calculi is proposed.

Chemical Precipitation↗
PubMed1986

[Carcinogenic hazard of small doses of nitrite in connection with the endogenous synthesis of nitroso compounds].

Carcinogenic risk of small doses of precursors of nitroso compounds, i.e. sodium nitrite (total dose in 0.2-2.0 g/mouse in drinking water) and morpholine (total dose is 0.23 g/mouse in bread) was studied in 520 CBA and 290 C57Bl mice during 96 weeks. It is shown that under these conditions the carcinogenic effect was more pronounced in CBA mice: there was a significant increase in general incidence of tumours, particularly liver tumours and hemoblastoses in CBA females and malignant liver tumours in CBA males.

Animals↗
PubMed1976

[Mechanism of excitation and contraction uncoupling in frog and guinea pig myocardial fibers during block of slow sodium-calcium channels by compound D-600].

In the experiments performed on the strips of frog's atria and ventricle it was found that rhythmic stimulation facilitated the dissociation of excitation--contraction coupling caused by D-600 compound through the block of calcium channels, i.e. D-600 prevented the increase of tension in the series of contractions or even converted positive staircase into negative one. The first contraction was affected by D-600 to a lesser degree than the subsequent ones. Guinea pig's left auriculus depressing action of D-600 was found to be strongly dependent on the frequency of stimulation. The data obtained were analysed by the model of excitation--contraction coupling. It was concluded that in amphibians as well as in mammalians tension amplitude during rhythmic activity of myocardial cells is determined mainly by Ca inflow through the excitable calcium channels. On the other hand the amplitude of the first contraction depends on the storage content determined by Ca inflow through the Ca channels open at rest. These latter channels have low sensitivity to D-600 and are better presented in atrium than in ventriculum. An increase of contraction tension observed in low sodium medium took place also under the action of D-600; in this case Ca storage was replenished by the Na-Ca exchange diffusion.

Action Potentials↗
PubMed1990

Stress protein synthesis in human keratinocytes treated with sodium arsenite, phenyldichloroarsine, and nitrogen mustard.

Cells from bacteria to man respond to sublethal thermal and certain chemical stresses by synthesis of heat shock, or stress, proteins. The human epidermal keratinocyte is a target for a variety of cytotoxic substances. One response of cells exposed to such agents may be the synthesis of stress proteins. Human epidermal keratinocytes were treated thermally (43 degrees C) or chemically with sodium arsenite and the skin irritants phenyldichloroarsine and mechlorethamine. Proteins synthesized by keratinocytes were radiolabeled with [35S]methionine, separated on polyacrylamide gels under denaturing conditions, and visualized by fluorography. Quantitation by computer-assisted densitometry of fluorograms revealed different patterns of synthesis of two heat shock proteins (hsp's) with apparent molecular weights of 70 and 90 kDa after treatment with heat, sodium arsenite, phenyl-dichloroarsine, or mechlorethamine. Sodium arsenite induced the highest levels of synthesis of these two proteins, approximately 10-fold and 3-fold increases in hsp-70 and hsp-90, respectively. Phenyldichloroarsine at 0.5 microM produced a 2-fold increase in hsp-70 but no significant increase in hsp-90. Mechlorethamine, in contrast, had an apparent inhibitory effect on hsp-70 synthesis. These results suggest that some but not all skin irritants induce the synthesis of heat shock proteins in human keratinocytes.

Arsenic↗
PubMed2004

Spectrophotometric determination of hydroxylamine and its derivatives in pharmaceuticals.

A sensitive spectrophotometric method for the determination of hydroxylamine is described. The method is based on the oxidation of hydroxylamine to nitrite using sodium arsenate under alkaline condition. The formed nitrite is determined based on the diazo coupling reaction between p-nitroaniline and N-(1-naphthyl)ethylenediamine dihydrochloride [NEDA]. The system obeys Beer's law over the concentration range 0-7 microg of hydroxylamine at 545 nm and the colour is stable for 3 h. The molar absorptivity of the colour system is found to be 6.7 x 10(4) l mol(-1) cm(-1). The relative standard deviation is 1.2% for ten determinations at 4 microg of hydroxylamine. Interferences due to various foreign ions have been studied and the method has been applied to the determination of hydroxylamine and its derivatives used in pharmaceutical formulations after hydrolysis.

Arsenates↗