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Effects of mercury compounds on the spontaneous and potassium-evoked release of [3H]dopamine from mouse striatal slices.

The effects of mercury compounds on the spontaneous and potassium-evoked release of [3H]dopamine from mouse striatal slices have been examined. All mercury compounds examined produced concentration-dependent increases in the spontaneous release of [3H]dopamine, with an order of potency of methylmercury greater than mercuric (Hg2+) mercury greater than p-choloromercuribenzene sulfonic acid. Methylmercury had no effect on the 25 mM potassium evoked release of [3H]dopamine in the presence of 1.3 mM calcium. However, in calcium-free conditions, methylmercury significantly increased the potassium-evoked release of [3H]dopamine. Mercuric mercury significantly reduced the 25 mM potassium evoked release of [3H]dopamine in the presence of 1.3 mM calcium, and this response was not reversible with brief washing of the tissue. In calcium-free conditions, mercuric mercury significantly elevated the evoked release of [3H]dopamine, similar to the result obtained with methylmercury. It is suggested that mercury compounds alter dopaminergic synaptic function, possibly by disrupting calcium homeostasis or calcium-dependent processes, and that methylmercury and mercuric mercury can have differential effects to alter dopaminergic neurotransmission.

4-Chloromercuribenzenesulfonate↗

Experimental bases for ascorbic acid therapy of poisoning by hexavalent chromium compounds.

The most frequent outcome of the usually transdermal absorption of hexavalent chromium compounds is uraemia due to tubular necrosis. We have confirmed earlier observations that this can be prevented by the immediate application of ascorbic acid (AA) with the aim of reducing Cr(VI) to Cr(III). The spontaneous reducing capacity of samples of serum and plasma for Cr(VI) compounds was polarographically determined to be about 2 ppm. Addition of AA in doses of 50 to 1000 ppm led to a rapid and dose-dependent reduction of chromium(VI), which was studied on the concentration level of 5 ppm. For example in the presence of 1000 ppm AA, five ppm chromium(VI) fade to 0.7 ppm within 20 min and to undetectable concentrations after 40 min. These experiments demonstrate the effectiveness of AA for the treatment of Cr(VI) poisoning. Reduction is increased and accelerated by AA and the resulting Cr(III)-protein complexes are non-toxic and can be excreted with the urine. Early and repeated high i.v. doses of AA are recommended as the therapy of choice for Cr(VI) poisoning. In cases of delayed medical treatment, AA should be immediately applied orally.

Ascorbic Acid↗

Gallocyanin chromalum as a nuclear stain in cytology. I. A cytophotometric comparison of the Husain-Watts Gallocyanin chromalum staining protocol with the Feulgen procedure.

In the present study, the staining characteristics of the Gallocyanin chromalum technique devised by Husain and Watts are compared with the Feulgen reaction. Liver imprints, blood smears, and cervical smears were fixed in ethanol and stained with either the Husain and Watts Gallocyanin chromalum reagent or the Feulgen-Schiff reagent. The slides were then post-treated with 70% ethanol-HCl pH 1.0, or with phosphotungstic acid for 0.5-30 min. The integrated optical density of cell nuclei was measured with a VIDAS image analyzer. In the material stained with the Husain and Watts procedure, some Gallocyanin chromalum was removed from the nuclei in the early phase (5 min) of all the post-treatment steps, followed by a plateau phase where the integrated optical density remained constant for 30 min. In this phase, the nuclear absorbance was highly reproducible and of the same size regardless of the post-treatment. Both the Husain and Watts procedure and the Feulgen-reaction gave quantitative staining of DNA. The Gallocyanin chromalum stain after Husain and Watts is a quick staining procedure for quantitative evaluation of DNA in cytological material. Proper rinsing of the slides is necessary for a good reproducibility of results.

Blood Cells↗

Na,K-ATPase in resting and stimulated submandibular salivary glands in cats, studied by means of ouabain-sensitive, K(+)-dependent p-nitrophenylphosphatase activity.

Sections from normal resting submandibular glands were incubated in a p-nitrophenylphosphate substrate mixture to demonstrate Na,K-ATPase activity by light and electron microscopy. Dense deposits of reaction product occurred within the basal and lateral infoldings of striated duct cells. A similar, less heavy deposition occurred in demilune cells. Only scattered, infrequent pockets of staining occurred in association with the basal plasma membrane of central acinar cells, supporting the belief that these cells do not contribute greatly to the secretion of fluid. No reaction was detected on luminal membranes. Glands tested after sympathetic or parasympathetic nerve stimulation showed no obvious differences from resting glands. These results contrast with the previously reported paucity of immunostaining in resting demilunes with an antibody to Na,K-ATPase and the large increase in the immunostaining by these cells after parasympathetic stimulation. It is inferred that there must be an inaccessibility to immunobinding sites of Na,K-ATPase in resting glands, and that the increase in binding which occurs after parasympathetic stimulation represents a conformational change that did not affect its enzyme catalytic reaction with this substrate.

4-Nitrophenylphosphatase↗

Interaction of a new fluorescent reagent with sulfhydryl groups of the (Na+ + K+)-stimulate ATPase.

The (Na+ + K+)-ATPase enzyme of rat brain microsomes can be reversibly inhibited by a new fluorescent sulfhydryl (SH) probe, dimethylaminoaphthalenecysteine-Hg+ (Dn-cys-Hg+). This reagent is more reactive than N-ethylamaleimide (MalNEt) toward membrane sulfhydryl groups. A procedure using the two SH reagents sequentially seems to permit a more selective labelling of the SH groups involved in the (Na+ + K+)-ATPase than is possible by using MalNEt alone. Brain microsomes treated by this method incorporate the fluorescent label within or near the active site of the enzyme. When the probe was bound a blue shift of its fluorescence emission maximum (from 540 to 495 nm) and a 20-fold increase in relative fluorescence occurred. This indicates that the Dn moiety is within a very non-polar region of the membrane.

Adenosine Triphosphatases↗

Comparison of two bond strength testing methodologies for bilayered all-ceramics.

OBJECTIVES: This study compared the shear bond strength (SBS) and microtensile (MTBS) testing methodologies for core and veneering ceramics in four types of all-ceramic systems. METHODS: Four different ceramic veneer/core combinations, three of which were feldspathic and the other a fluor-apatite to their respectively corresponding cores, namely leucite-reinforced ceramic ((IPS)Empress, Ivoclar), low leucite-reinforced ceramic (Finesse, Ceramco), glass-infiltrated alumina (In-Ceram Alumina, Vita) and lithium disilicate ((IPS)Empress 2, Ivoclar) were used for SBS and MTBS tests. Ceramic cores (N=40, n=10/group for SBS test method, N=5 blocks/group for MTBS test method) were fabricated according to the manufacturers' instructions (for SBS: thickness, 3mm; diameter, 5mm and for MTBS: 10 mm x 10 mm x 2 mm) and ultrasonically cleaned. The veneering ceramics (thickness: 2mm) were vibrated and condensed in stainless steel moulds and fired onto the core ceramic materials. After trying the specimens in the mould for minor adjustments, they were again ultrasonically cleaned and embedded in PMMA. The specimens were stored in distilled water at 37 degrees C for 1 week and bond strength tests were performed in universal testing machines (cross-head speed: 1mm/min). The bond strengths (MPa+/-S.D.) and modes of failures were recorded. RESULTS: Significant difference between the two test methods and all-ceramic types were observed (P<0.05) (2-way ANOVA, Tukey's test and Bonferroni). The mean SBS values for veneering ceramic to lithium disilicate was significantly higher (41+/-8 MPa) than those to low leucite (28+/-4 MPa), glass-infiltrated (26+/-4 MPa) and leucite-reinforced (23+/-3 MPa) ceramics, while the mean MTBS for low leucite ceramic was significantly higher (15+/-2 MPa) than those of leucite (12+/-2 MPa), glass-infiltrated (9+/-1 MPa) and lithium disilicate ceramic (9+/-1 MPa) (ANOVA, P<0.05). SIGNIFICANCE: Both the testing methodology and the differences in chemical compositions of the core and veneering ceramics influenced the bond strength between the core and veneering ceramic in bilayered all-ceramic systems.

Aluminum Silicates↗

Examination of the jarosite-alunite precipitate addition in the raw meal for the production of sulfoaluminate cement clinker.

The aim of the present research work was to investigate the possibility of adding a jarosite-alunite chemical precipitate, a waste product of a new hydrometallurgical process developed to treat economically low-grade nickel oxides ores, in the raw meal for the production of sulfoaluminate cement clinker. For that reason, two samples of raw meals were prepared, one contained 20% gypsum, as a reference sample ((SAC)Ref) and another with 11.31% jarosite-alunite precipitate ((SAC)J/A). Both raw meals were sintered at 1300 degrees C. The results of chemical and mineralogical analyses as well as the microscopic examination showed that the use of the jarosite-alunite precipitate did not affect the mineralogical characteristics of the so produced sulfoaluminate cement clinker and there was confirmed the formation of the sulfoaluminate phase (C4A3S), the most typical phase of this cement type. Furthermore, both clinkers were tested by determining the grindability, setting time, compressive strength and expansibility. The hydration products were examined by XRD analysis at 2, 7, 28 and 90 days. The results of the physico-mechanical tests showed that the addition of jarosite-alunite precipitate did not negatively affect the quality of the produced cement.

Aluminum Compounds↗

The higher toxicity of cereulide relative to valinomycin is due to its higher affinity for potassium at physiological plasma concentration.

Valinomycin and cereulide are bacterial toxins with closely similar chemical structure and properties but different toxic effects. Emetic poisoning is induced by cereulide but not by valinomycin. Both are specific potassium ionophores. Such compounds may affect mitochondrial functions. Both compounds cause a potassium-dependent drop in the transmembrane inner membrane potential due to the uptake of K+ as positively charged ionophore complex. Valinomycin is more potent than cereulide at high [K+] (>80 mM), whereas cereulide in contrast to valinomycin is active already at <1 mM. With cereulide, there is a substantial lag, while valinomycin acts without lag. Both ionophores induce mitochondrial swelling in the presence of K+, in the case of cereulide with a lag. These toxins strongly inhibited respiration at the level of complex IV when used at higher concentrations than that used for detection of ionophoretic transport of K+. At high [KCl] (120 mM), valinomycin was more potent than cereulide both as ionophore and inhibitor, but at low [KCl] (2.5 mM), cereulide was much more potent. Thus, valinomycin needed 20-30 mM KCl for substantial effects, cereulide only 1-3 mM K+, which is close to its level in blood serum. This explains the higher toxicity of cereulide at low concentrations with the positively charged potassium complex being accumulated in the cell by transport through the plasma membrane driven by the membrane potential. Furthermore, with similar concentrations, the final concentration of cereulide in the cells may become higher than that of valinomycin.

Animals↗

Ultrasound assisted PTC catalyzed saponification of vegetable oils using aqueous alkali.

A few vegetable oils were saponified using aqueous KOH and different PTCs at room temperature in the presence of ultrasound. The extent of saponification was studied using the saponification value as a reference. Optimizations of various parameters such as time, selection of PTC, quantity of PTC, quantity of KOH and quantity of water were carried out using soyabean oil as a sample oil under sonication with stirring. To study the effect of ultrasound, the saponification was also carried out at 35 +/- 2 degrees C under different conditions, namely stirring, sonication, stirring and sonication, and heating at 100 degrees C. It was found that the heterogeneous liquid-liquid phase saponification of different vegetable oils using aq. KOH/CTAB was remarkably accelerated at 35 +/- 2 degrees C in the presence of ultrasound along with stirring.

Catalysis↗

Carbon monoxide production from degradation of desflurane, enflurane, isoflurane, halothane, and sevoflurane by soda lime and Baralyme.

Anecdotal reports suggest that soda lime and Baralyme brand absorbent can degrade inhaled anesthetics to carbon monoxide (CO). We examined the factors that govern CO production and found that these include: 1) The anesthetic used: for a given minimum alveolar anesthetic concentration (MAC)-multiple, the magnitude of CO production (greatest to least) is desflurane > or = enflurane > isoflurane >> halothane = sevoflurane. 2) The absorbent dryness: completely dry soda lime produces much more CO than absorbent with just 1.4% water content, and soda lime containing 4.8% or more water (standard soda lime contains 15% water) generates no CO. In contrast, both completely dry Baralyme and Baralyme with 1.6% water produce high concentrations of CO, and Baralyme containing 4.7% water produces concentrations equaling those produced by soda lime containing 1.4% water. Baralyme containing 9.7% or more water and standard Baralyme (13% water) do not generate CO.3) The type of absorbent: at a given water content, Baralyme produces more CO than does soda lime. 4) The temperature: an increased temperature increases CO production. 5) The anesthetic concentration: more CO is produced from higher anesthetic concentrations. These results suggest that CO generation can be avoided for all anesthetics by using soda lime with 4.8% (or more) water or Baralyme with 9.7% (or more) water, and by using inflow rates of less than 2-3 L/min. Such inflow rates are low enough to ensure that the absorbent does not dry out.

Adsorption↗

Synaptosomes from rat brain: morphology, compartmentation, and transmembrane pH and electrical gradients.

Morphological studies of synaptosomes isolated from rat brains show that approximately 68% of the synaptosomes in these preparations contain synaptic vesicles (range, 62--72.5%). Approximately 30% of the synaptosomes contain mitochondria, and only less than 20% of the total mitochondria in good preparations are free and not enclosed in synaptic structures. The mitochondrial volume percent calculated on the basis of the measured cytochrome c content is 5% for synaptosomes isolated from anesthetized animals and 11% for synaptosomes isolated from unanesthetized animals. These numbers bracket the value of 8.7% obtained from electron micrographs. The volume percent of intrasynaptic vesicles is 1.5% as calculated from electron micrographs. The pH gradient between the extracellular pH and the mean intracellular pH is --0.45, as measured by equilibrium distributions of methylamine and dimethylamine, and --0.05, as determined by equilibrium distributions of 5,5-dimethyloxazolidine-2,4-dione and trimethylacetic acid. Analysis of these data shows that there cannot be a large pH gradient (alkaline inside) across the mitochondria, nor can the synaptic vesicle compartment be very large (less than 1.85%). Equilibrium distribution of [3H]triphenylmethylphosphonium ion in synaptosomal preparations gives a calculated apparent potential of --85 mV, in agreement with our previous value. Analysis of these data using the measured volumes of mitochondrial and intrasynaptic vesicular compartments (8.7 and 1.5%, respectively) gives a maximum possible transmitochondrial membrane potential of --59 mV.

Animals↗

Methods for distinguishing gram-positive from gram-negative bacteria.

Lysis by KOH and hydrolysis of L-alanine-4-nitroanilide were compared with the Gram reaction of aerobic, microaerophilic, and anaerobic bacteria. Both tests correlated well with the Gram reaction with nonfermentative bacilli and Bacillus species, whereas they did not correlate with nonsporulating anaerobes. Only campylobacteria were KOH positive and L-alanine-4-nitroanilide and gram negative.

Aniline Compounds↗

Ethanol has thyrotropin-like activity in cultured porcine thyroid follicles.

We describe TSH-like activity of ethanol for thyroid hormone formation in the physiological culture system. Porcine thyroid follicles were preincubated with 0-100 mM (0-0.58%) ethanol in the presence of 0-1280 microU/ml bovine TSH for 24 h; these follicles were then incubated with the mixture of Na125I and NaI to measure iodide uptake, iodine organification, and de novo thyroid hormone formation. Ethanol stimulated iodide uptake in a dose-response manner in TSH-free medium. Ethanol augmented the effect of TSH on iodide uptake, iodide organification, and thyroid hormone formation in the presence of 20-80 microU/ml TSH. When TSH concentration was 320 microU/ml or greater, ethanol no longer stimulated iodide uptake and thyroid hormone formation. Ethanol mediated iodide uptake and iodine organification were inhibited by potassium perchlorate and propylthiouracil respectively. The effect of ethanol on the thyroid follicle was reversible 24 h after removal of ethanol from the medium. The mechanism of TSH-like activity of ethanol was studied by measuring cAMP generation and Na+K+ATPase activity, a sodium pump necessary for iodide transport, in the presence of 0-1280 microU/ml TSH. Ethanol increased cAMP production in TSH-free medium; the increment of cAMP by ethanol was more prominent when 20-80 microU/ml TSH were present. Ethanol also augmented (Bu)2cAMP-mediated iodide uptake and TSH-mediated thyroid Na+K+ATPase activity. Thus, TSH-like activity of ethanol for thyroid hormone formation can be explained by activation of the cAMP system and Na+K+ATPase activity. Our results indicate that ethanol concentrations equivalent to the blood level of moderate to heavy alcohol drinkers exert TSH-like activity in the thyroid follicle.

Animals↗

Hyperpolarization/depolarization on human spermatozoa.

The accumulation of the lipophilic cation radiolabeled triphenylmethylphosphonium (TPMP+) was utilized to determine the resting membrane potential across the plasma membrane (psi) of human sperm. Washed sperm were suspended and incubated in low-K+ and high-K+ medium and allowed to take up the cation to a steady state (20 min at 37 degrees C). By using this differential, the value obtained was inserted in the Nernst equation and the value yielded a psi of -69 +/- 2 mV. When the Na+ or K+ concentration is high in the medium, the accumulation of TPMP+ in the membrane sperm cells was increased or decreased, respectively, inducing hyperpolarization and depolarization of the membrane 20% and 85%, respectively. The presence of divalent cations Zn++ and Mg++ in the incubation medium both induced a hyperpolarization of 10% and 8.6%, respectively. The addition of specific reagents such as p-chloromercuribenzenosulfonate and ethylenediaminetetraacetic acid sodium salt both decreased the psi 35% and 58%, respectively. The agents acting on the sperm cell membrane, such as dithiothreitol and progesterone, both induced hyperpolarization and depolarization of the membrane 16% and 40%, respectively. The presence of propranolol and L-alpha-lysophosphatidylcholine, which affect the ionic gradients present across the plasma membrane, both induced a depolarization from 43% and 92%, respectively. Finally the psi was glucose-dependent. The result of these studies was that, by the use of agents causing hyperpolarization or depolarization, we obtained changes in the psi of -83.4 +/- 2.2 mV, until -6 +/- 0.6 mV changes of -76.8 +/- 2 mV translated across the sperm cell membrane.

Cations↗