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Biomedical subjects

R Wang

Publications and source records attributed to R Wang.

At least 559 records · Page 31Linked to original sources
PubMed1994

Effects of three fragments of parathyroid hormone on calcium channel currents in neonatal rat ventricular cells.

The effects of different fragments of bovine parathyroid hormone, bPTH-(1-34), bPTH-(1-84) and bPTH-(3-34), on two types of calcium channel currents in neonatal rat ventricular cells were compared in the present study. bPTH-(1-34) increased the amplitude of L channel currents, but not of the T channel currents. This effect of bPTH-(1-34) was sustained after a complete washout of the peptide from the bath. The intact PTH molecule, bPTH-(1-84), also increased L channel currents but not affecting T channel currents. While bPTH-(3-34) did not affect the amplitudes of either L or T channel currents by itself, pretreatment of cells with bPTH-(3-34) abolished the effects of both bPTH-(1-34) and bPTH-(1-84) on L channel currents. Moreover, the kinetics of L channel currents in the presence of bPTH-(1-34) or bPTH-(3-34) were different. bPTH-(1-34) increased the time constant of activation, but not of inactivation, of L channel currents from 1.8 to 2.5 ms (P < 0.05). In contrast, bPTH-(3-34) decreased the time constant of inactivation, but not of activation, of L channel currents from 159 to 117 ms (P < 0.05). These results indicate that different fragments of PTH exert different effects on the amplitudes or kinetics of cardiac calcium channel currents.

Animals↗
PubMed1994

Direct fluorescence analysis of genetic polymorphisms by hybridization with oligonucleotide arrays on glass supports.

A simple and rapid method for the analysis of genetic polymorphisms has been developed using allele-specific oligonucleotide arrays bound to glass supports. Allele-specific oligonucleotides are covalently immobilized on glass slides in arrays of 3 mm spots. Genomic DNA is amplified by PCR using one fluorescently tagged primer oligonucleotide and one biotinylated primer oligonucleotide. The two complementary DNA strands are separated, the fluorescently tagged strand is hybridized to the support-bound oligonucleotide array, and the hybridization pattern is detected by fluorescence scanning. Multiple polymorphisms present in the PCR product may be detected in parallel. The effect of spacer length, surface density and hybridization conditions were evaluated, as was the relative efficacy of hybridization with single or double-stranded PCR products. The utility of the method was demonstrated in the parallel analysis of 5 point mutations from exon 4 of the human tyrosinase gene.

Alleles↗
PubMed1994

Purification and characterization of a calcium-independent acidic phospholipase A2 from rat lung.

Several phospholipase A2 (PLA2) activities have been identified in rat lung homogenate and shown to be important in metabolism of lung phospholipids. One PLA2 activity is Ca(2+)-independent, active in vitro at pH 4, and inhibited by a substrate analogue, 1-hexadecyl-3-trifluoroethylglycero-sn-2-phosphomethanol (MJ33). Purification of this rat lung PLA2 by approx. 550-fold was carried out by sequential column chromatographies using DE-52, Sephacryl-100, heparin-Sepharose, and phenyl-Sepharose columns. The purified activity had an acidic pH optimum, was Ca(2+)-independent, was inhibited by MJ33 in a dose-dependent manner (50% inhibition at 3 mol%), was unaffected by treatment with p-bromophenacyl bromide or mercaptoethanol, and had a unique N-terminal amino acid sequence. The apparent molecular mass was 15 kDa on gel electrophoresis and activity was recovered in part by renaturation of protein from the gel. The properties of this enzyme suggest a new class of PLA2.

Amino Acid Sequence↗
PubMed1994

The human BAT3 ortholog in rodents is predominantly and developmentally expressed in testis.

A partial cDNA clone, RLC34, was isolated from a rat brain cDNA library. Its sequence exhibits high identity with BAT3 (88.4% and 94.9% for DNA and the deduced amino acid sequence, respectively), a gene located within the region of human major histocompatibility complex III (MCHIII region). RLC34 detected a transcript the same size in human and rat, similar to that reported for BAT3. Southern blot analysis of RLC34 showed similar restriction patterns as those of the human BAT3 gene. A panel of rodent tissue samples were examined and the RLC34 was found to be predominantly expressed in the germ cells of rodent testes. The expression is developmentally regulated with increased transcripts seen at 17-20 days after birth. Its testicular expression, its association with spermatogenesis, and its location in MCHIII suggest a correlation of RLC34 with the growth-reproduction complex (grc). This finding may also provide a clue to study the function of other genes localized in this area of the MCHIII region.

Adult↗
PubMed1994

Characterization and nucleotide sequence of the cardiac alpha-myosin heavy chain gene from Syrian hamster.

In small mammals, the cardiac alpha-myosin heavy chain (MyHC) gene is predominantly expressed in the adult ventricle, while the beta-MyHC gene is predominantly expressed in the fetal ventricle. In order to perform comparative studies of these genes, we sequenced the Syrian hamster alpha-MyHC gene by determining a continuous 32,415 base pair (bp) sequence. It contained 39 exons encoding a predicted polypeptide of 1939 amino acid residues (aa) with a molecular weight of 223,644 Da. Sequence comparison revealed that the coding sequence is highly identical to that of the human alpha-MyHC gene. A marked feature of the Syrian hamster alpha-MyHC gene was the abundance of reiterated sequences within introns. Eleven B1 and four B2 retroposons were identified, a variety of di-, tri-, and tetra-nucleotide repeats were present, and three longer direct repeats were found. Some of them appear to be associated with genetic recombination events.

Amino Acid Sequence↗
PubMed1994

Histamine-evoked Ca2+ oscillations in HeLa cells are sensitive to methylxanthines but insensitive to ryanodine.

The relative contribution of inositol-trisphosphate(InsP3)-sensitive and InsP3-insensitive Ca2+ stores to the agonist-evoked oscillatory release of Ca2+ in HeLa cells was investigated using fura-2 cytosolic Ca2+ measurements and whole-cell recordings of Ca(2+)-activated K+ currents [K(Ca2+)]. The experimental approach chosen consisted in studying the effects on Ca2+ oscillations of a variety of pharmacological agents such as ryanodine, ruthenium red, caffeine and theophylline, which are known to affect the Ca2+ channels responsible for Ca(2+)-induced Ca2+ release (CICR) in excitable cells. The results obtained essentially indicate (a) that neither ryanodine nor ruthenium red affects the generation of periodic K(Ca2+) current pulses in whole-cell experiments, and (b) that histamine-induced Ca2+ oscillations are inhibited by caffeine and theophylline in a dose-dependent manner. However, these methylxanthines were unable, at concentrations ranging from 0.1 mM to 10 mM, either to mobilize Ca2+ from internal stores or to block the initial Ca2+ rise evoked by histamine. In addition, both methylxanthines showed at high concentrations (10-20 mM) a moderate inhibitory action on the production of InsP3 induced by histamine. This effect was not essential to the action of caffeine on the oscillatory release of Ca2+, since an inhibition by caffeine of InsP3-induced Ca2+ oscillations was still observed in whole-cell experiments where the InsP3 concentration was kept constant. The results also show (c) that the application of either caffeine or theophylline during histamine stimulation leads systematically to an increased Ca2+ sequestration in InsP3-sensitive Ca2+ pools, the effect observed with theophylline being stronger than that resulting from the application of caffeine, and finally (d) that the action of caffeine and theophylline is not related to an increase in cAMP concentration since neither forskolin (10-50 microM) nor 8-Br-cAMP (1 mM) caused an inhibition of the InsP3-induced Ca2+ oscillations. It is concluded on the basis of these results that the agonist-evoked Ca2+ oscillations in HeLa cells do not involve directly or indirectly a ryanodine-sensitive Ca(2+)-release channel with CICR properties, but rather arise from a control by Ca2+ of the InsP3 Ca(2+)-release process.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1994

Cardiovascular effects of Buthus martensii (Karsch) scorpion venom.

Buthus martensii (Karsch) (BMK) scorpion envenomation is a common medical problem in China and BMK scorpion has been widely used in traditional Chinese medicine. However, the cardiovascular effects of this venom have not been systematically investigated. In the present study, i.v. BMK venom injection significantly increased the blood pressure in conscious rats in a concentration-dependent manner (ED50 = 59 +/- 12 micrograms/kg). The increase in blood pressure occurred within 1 min of injection of the venom and was sustained for more than 50 min. Heart rate was not changed by the venom in conscious rats. In vitro studies with BMK venom revealed the increase in the force of contraction, without modification of the contraction frequency (within 20 min) of isolated atrial strips. Contractions of isolated arterial strips from aorta, renal and vertebral arteries were also enhanced by BMK venom with a time lag of 8 min between the application of the venom and the initiation of the contraction. Furthermore, BMK venom-induced rises in blood pressure in vivo and increased contraction of isolated vessel strips were inhibited by prazosin and tolazoline, respectively, two alpha 1-adrenergic antagonists. BMK venom alone did not alter intracellular calcium concentrations, [Ca2+]i, in cultured vascular smooth muscle cells and endothelial cells. However, BMK venom significantly increased the metabolism of InsP3 in dispersed cardiac myocytes, indicating a direct effect on cardiac myocytes. These results demonstrate the significant cardiovascular effects of BMK venom, which may be mediated by an alteration in InsP3 in cardiac myocytes but not by [Ca2+]i in vascular smooth muscle cells.

Animals↗
PubMed1994

Enhanced calcium influx by parathyroid hormone in identified Helisoma trivolvis snail neurons.

The modulation of [Ca2+]i by parathyroid hormone (PTH) has been extensively studied in vertebrates. The present study examined the effects of PTH on [Ca2+]i in isolated invertebrate neurons B5 from buccal ganglia of the pond snail, Helisoma trivolvis, utilizing the Fura-2 fluorescence technique. Bovine PTH, bPTH-(1-84), induced a slow and sustained increase in [Ca2+]i in neurons B5. In contrast, the elevation of extracellular K+ concentration from 1.7 mM to 15 mM induced a rapid and transient increase in [Ca2+]i. Simultaneous application of 15 mM KCl and bPTH-(1-84), or application of 15 mM KCl in the presence of bPTH-(1-84) additively increased [Ca2+]i in neurons B5. An increase in [Ca2+]i in neurons B5 was also induced by a PTH agonist [bPTH-(1-34)], but not by a PTH antagonist [bPTH-(3-34)]. The absence of calcium, or the presence of lanthanum (2 mM) or omega-conotoxin (10 microM), in the bath solution abolished the effect of bPTH-(1-84) on [Ca2+]i. Furthermore, our results demonstrated that the effect of PTH on [Ca2+]i in neurons B5 was not due to the hormonal modulation of voltage-dependent Na+ or K+ channels or a Na+/Ca2+ exchanger. The results from this study show that PTH can modulate [Ca2+]i in an identified invertebrate neuron mainly by promoting extracellular calcium influx via the N-like voltage-dependent calcium channel.

Animals↗
PubMed1994

Effects of descending inhibitory systems on the c-Fos expression in the rat spinal cord during formalin-induced noxious stimulation.

This study provides morphological evidence for the activation of the descending modulatory control by nociceptive afferent pathways. Fos-like immunoreactivity in the spinal dorsal horn is used as an indicator of efficacy of transmission of noxious inputs at this level. Wistar rats were anesthetized with nembutal and the spinal dorsolateral funiculus was transected unilaterally at the level of T11,12. Two days later, an equal volume (0.2 ml) of formalin (5% in saline) was injected into the plantar aspect of two hindpaws. After 1 h of injection, rats were deeply anesthetized and killed for the immunocytochemical examination of Fos-like protein product by using an immunocytochemical technique. The results show that the mean number of Fos protein-like immunoreactive neurons is significantly lower in the superficial laminae and in the neck of the dorsal horn on the side ipsilateral to the intact dorsolateral funiculus than that on the opposite side (i.e. 18.4 +/- 1.0 vs 30.0 +/- 1.3 and 33.9 +/- 0.2 vs 56.8 +/- 1.7, respectively). We conclude that the peripheral noxious inputs (which ascend via the ventral half of the spinal cord in this study) can activate the supraspinal descending inhibitory systems, which in turn suppress the synthesis of Fos-like protein in the related dorsal horn neurons.

Afferent Pathways↗
PubMed1994

Norepinephrine and serotonin-induced antinociception are blocked by naloxone with different dosages.

The effects of intrathecally (IT) administered naloxone (Nal) on the antinociception produced by IT norepinephrine (NE), serotonin (5-HT), or morphine (Mor) were observed and compared in rats using the tail-flick (TF) assay. The results show that: a) NE, 5-HT, and Mor in doses of 1 nmol, 240 nmol, and 0.5 nmol, respectively, produce similar increases in amplitude and time in TF latency (TFL); b) Nal treatment of 240 and 360 nmol has no effects on TFL; c) the antinociception produced by NE (1 nmol) can be blocked by Nal (240 nmol); d) antinociception produced by Mor (0.5 nmol) can also be blocked by Nal (240 nmol); e) 240 nmol of Nal does not affect the 5-HT (120 nmol)-produced antinociception, while 360 nmol of Nal show a delayed blockade to the 5-HT (120 nmol)-produced antinociception. The results suggest that endogenous opiate-like substances may be involved in both NE- or 5-HT-produced antinociception at the spinal level, and these effects may be mediated through different types of opiate receptors.

Animals↗
PubMed1994

Effect of sodium nitroprusside on sperm motility, viability, and lipid peroxidation.

OBJECTIVE: To analyze the effect of sodium nitroprusside, a nitric oxide releaser, on sperm motion and lipid peroxidation-induced membrane damage in cryopreserved human sperm. DESIGN: Post-thaw, cryopreserved, human sperm samples were washed and divided into three aliquots. Each aliquot was incubated with either 0, 50, or 100 nM sodium nitroprusside. INTERVENTIONS: Samples were analyzed for lipid peroxidation (measured by malonaldehyde-thiobarbituric acid reactivity) at 3 hours post-thaw. MAIN OUTCOME MEASURES: Percent viability and motion parameters were assessed at 0, 10, and 30 minutes and 2, 3, 5, and 6 hours post-thaw. RESULTS: All results represent a mean +/- SEM, n = 10. Lipid peroxidation in samples incubated with 50 nM sodium nitroprusside (15.1 +/- 2.1 nM malonaldehyde/10(8) sperm) or 100 nM sodium nitroprusside (13.2 +/- 2.1 nM malonaldehyde/10(8) sperm) was significantly lower than in controls (22.7 +/- 3.1 nM malonaldehyde/10(8) sperm). Percent viability was significantly reduced from 0 minutes (60.6% +/- 3.5%) to 6 hours post-thaw in controls (38.0% +/- 5.1%) but not in 50 nM (46.8% +/- 10.4%) or 100 nM (48.8% +/- 6.5%) sodium nitroprusside-treated samples. Compared with controls (18.3% +/- 3.4%), maintenance of percent motility at 3 hours post-thaw was significantly improved in 50 nM (24.5% +/- 2.9%) and in 100 nM (26.3% +/- 3.2%) sodium nitroprusside-treated samples. Straight line velocity maintenance was significantly improved in 50 nM (37.3 +/- 1.3) and in 100 nM (37.0 +/- 1) sodium nitroprusside-treated samples as compared with controls (30.5 +/- 1.7). Significant improvements in curvilinear velocity maintenance compared with controls (56.3 +/- 2.9) also were observed in 50 nM (65.9 +/- 2.1) and 100 nM (72.1 +/- 4.1) sodium nitroprusside-treated samples. Significant differences in the motion parameters of sodium nitroprusside-treated samples were maintained at 5 and 6 hours post-thaw in comparison to controls. CONCLUSION: These results suggest that sodium nitroprusside is beneficial to the maintenance of post-thaw sperm motion and viability for up to 6 hours and that reduction of lipid peroxidative damage to sperm membranes may be the mechanism for these benefits.

Cell Survival↗