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Biomedical subjects

R P Wang

Publications and source records attributed to R P Wang.

17 recordsLinked to original sources
PubMed2000

[Effect of sodium nitroprusside on the spontaneous and induced responses of rat spinal cord dorsal horn neurons].

The present study was performed on anesthetized and paralyzed Wistar rats. Extracellular recordings were made from the lumbar enlargement of the spinal cord using carbon filament electrodes. NO donor sodium nitroprusside (SNP) was locally microdialyzed to the spinal cord, and effects of NO on spontaneous response and the responses induced by mechanical stimulation of the hind-foot were observed. After dialyzing for 10~20 min, SNP at a concentration of 1 micromol/L increased the induced responses to innocuous mechanical stimulation and decreased those to noxoius mechanical stimulation. The induced responses to both innocuous and noxious mechanical stimulations were all decreased after dialyzing the same dose of SNP for 20~30 min. The decrease was shown within 7~15 min dialysis when SNP was used at a concentration of 20 micromol/L. SNP at both concentrations of 1 and 20 micromol/L increased spontaneous spikes of the recorded spinal units. These effects were shown preferentially in deep units. The data suggest that in the deeper neurons of the spinal cord NO of different concentrations has different effects on transmission of nociceptive and nonnociceptive signals induced by mechanical stimulation, and No is involved in the antinociception in spinal neurons and facilitates their spontaneous activity.

Animals↗
PubMed2000

[Interaction between pelvic and pudendal afferent inputs in lumbar-sacral spinal cord in rats].

Time-dependent inhibition is one of the means to study the interactions between peripheral inputs. The present study was performed on anesthetized and paralyzed Wistar rats using the technique of conditioning-testing stimulation. Electric stimulation (1.5 3 folds of the threshold intensity) was given to pelvic and pudendal nerves. Extracellular recordings were made from convergent neurons at L(6)-S(1) segments of the spinal dorsal horn. Stimulus intervals between conditioning and testing stimulation were measured when a half of the testing responses were inhibited by conditioning responses or the inhibition just began to occur. The time-dependent inhibition was seen in neurons situated deeper than 300 m beneath the dorsal surface of the spinal cord, and not in more superficial neurons. The inhibition intervals were in the range of 1 360 ms and became longer when conditioning stimulation was given to the pelvic nerve. The inhibition intervals were 1 3 ms in superficial neurons (<300 micrometer) and no apparent time-dependent inhibition occurred. The inhibition in deeper neurons was partially reduced by cold block conducted at segments C(5-6) and the blockage was more significant when conditioning stimulation was applied to the pelvic nerve. These findings suggest that the inputs from the pelvic and pudendal nerves may converge on single neurons at deeper lumbosacral dorsal horn and the pudendal nerve induced responses are more likely to be inhibited in these neurons, which may be further strengthened by superspinal structures.

Afferent Pathways↗
PubMed1997

Molecular cloning and functional expression of a human cDNA encoding the antimutator enzyme 8-hydroxyguanine-DNA glycosylase.

The major mutagenic base lesion in DNA caused by exposure to reactive oxygen species is 8-hydroxyguanine (8-oxo-7, 8-dihydroguanine). In bacteria and Saccharomyces cerevisiae, this damaged base is excised by a DNA glycosylase with an associated lyase activity for chain cleavage. We have cloned, sequenced, and expressed a human cDNA with partial sequence homology to the relevant yeast gene. The encoded 47-kDa human enzyme releases free 8-hydroxyguanine from oxidized DNA and introduces a chain break in a double-stranded oligonucleotide specifically at an 8-hydroxyguanine residue base paired with cytosine. Expression of the human protein in a DNA repair-deficient E. coli mutM mutY strain partly suppresses its spontaneous mutator phenotype. The gene encoding the human enzyme maps to chromosome 3p25. These results show that human cells have an enzyme that can initiate base excision repair at mutagenic DNA lesions caused by active oxygen.

Amino Acid Sequence↗
PubMed1996

Putative human blue-light photoreceptors hCRY1 and hCRY2 are flavoproteins.

Recently, a human cDNA clone with high sequence homology to the photolyase/blue-light photoreceptor family was identified. The putative protein encoded by this gene exhibited a strikingly high (48% identity) degree of homology to the Drosophila melanogaster (6-4) photolyase [Todo et al. (1996) Science 272, 109-112]. We have now identified a second human gene whose amino acid sequence displays 73% identity to the first one and have named the two genes CRY1 and CRY2, respectively. The corresponding proteins hCRY1 and hCRY2 were purified and characterized as maltose-binding fusion proteins. Similar to other members of the photolyase/blue-light photoreceptor family, both proteins were found to contain FAD and a pterin cofactor. Like the plant blue-light photoreceptors, both hCRY1 and hCRY2 lacked photolyase activity on the cyclobutane pyrimidine dimer and the (6-4) photoproduct. We conclude that these newly discovered members of the photolyase/photoreceptor family are not photolyases and instead may function as blue-light photoreceptors in humans.

Amino Acid Sequence↗
PubMed1996

Molecular cloning and functional analysis of a human cDNA encoding an Escherichia coli AlkB homolog, a protein involved in DNA alkylation damage repair.

The Escherichia coli AlkB protein is involved in protecting cells against mutation and cell death induced specifically by SN2-type alkylating agents such as methyl methanesulfonate (MMS). A human cDNA encoding a polypeptide homologous to E.coli AlkB was discovered by searching a database of expressed sequence tags (ESTs) derived from high throughput cDNA sequencing. The full-length human AlkB homolog (hABH) cDNA clone contains a 924 bp open reading frame encoding a 34 kDa protein which is 52% similar and 23% identical to E.coli AlkB. The hABH gene, which maps to chromosome 14q24, was ubiquitously expressed in 16 human tissues examined. When hABH was expressed in E.coli alkB mutant cells partial rescue of the cells from MMS-induced cell death occurred. Under the conditions used expression of hABH in skin fibroblasts was not regulated by treatment with MMS. Our findings show that the AlkB protein is structurally and functionally conserved from bacteria to human, but its regulation may have diverged during evolution.

AlkB Homolog 1, Histone H2a Dioxygenase↗
PubMed1995

Molecular cloning and expression of human cDNAs encoding a novel DNA ligase IV and DNA ligase III, an enzyme active in DNA repair and recombination.

Three distinct DNA ligases, I to III, have been found previously in mammalian cells, but a cloned cDNA has been identified only for DNA ligase I, an essential enzyme active in DNA replication. A short peptide sequence conserved close to the C terminus of all known eukaryotic DNA ligases was used to search for additional homologous sequences in human cDNA libraries. Two different incomplete cDNA clones that showed partial homology to the conserved peptide were identified. Full-length cDNAs were obtained and expressed by in vitro transcription and translation. The 103-kDa product of one cDNA clone formed a characteristic complex with the XRCC1 DNA repair protein and was identical with the previously described DNA ligase III. DNA ligase III appears closely related to the smaller DNA ligase II. The 96-kDa in vitro translation product of the second cDNA clone was also shown to be an ATP-dependent DNA ligase. A fourth DNA ligase (DNA ligase IV) has been purified from human cells and shown to be identical to the 96-kDa DNA ligase by unique agreement between mass spectrometry data on tryptic peptides from the purified enzyme and the predicted open reading frame of the cloned cDNA. The amino acid sequences of DNA ligases III and IV share a related active-site motif and several short regions of homology with DNA ligase I, other DNA ligases, and RNA capping enzymes. DNA ligases III and IV are encoded by distinct genes located on human chromosomes 17q11.2-12 and 13q33-34, respectively.

Amino Acid Sequence↗
PubMed1991

Colonic Cl channel blockade by three classes of compounds.

We compared the potency and inhibitory actions of three different classes of organic acids on a Cl channel derived from colonic enterocyte plasma membrane vesicles. Chloride channels were incorporated into planar lipid bilayer membranes to examine the effects of the anthranilic acids, diphenylamine 2-carboxylic acid (DPC) and 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), the indanyl alkanoic acids, 2-[(2-cyclopentyl-6,7-dichloro-2,3-dihydro-2-methyl-1-oxo-1H-inden -5-yl)oxy] acetic acid (IAA-94) and its stereoenantiomer IAA-95, and the disulfonic stilbene, 4,4'-dinitro-stilbene-2,2'-disulfonic acid (DNDS). Except for DNDS, each of the blockers was equipotent from both the outer membrane and the cytoplasmic side of the channel protein. The potency order from the outmembrane side was DNDS greater than IAA-94 = IAA-95 greater than NPPB much greater than DPC. In contrast, the potency order from the cytoplasmic side was IAA-94 = IAA-95 greater than NPPB greater than DNDS much greater than DPC. DPC and NPPB caused a concentration-dependent decrease in the single-channel conductance (fast block). DNDS, IAA-94, and IAA-95 caused a flickery-type block and a concentration-dependent decrease in open-channel probability. Kinetic analysis revealed that blockade could be explained by a linear closed-opened-blocked kinetic scheme. Similarities in the electrostatic potential maps of these open-channel blockers suggest they may bind to a single shared binding site within the channel protein.

Animals↗
PubMed1991

Atrial natriuretic peptide inhibits sympathetic outflow in NaCl-sensitive spontaneously hypertensive rats.

The current study tested the hypothesis that circulating atrial natriuretic peptide (ANP) inhibits sympathetic outflow, as reflected in lumbar sympathetic nerve activity (LSNA), in NaCl-sensitive spontaneously hypertensive rats (SHR-S) and that this effect is exaggerated by high NaCl feeding. NaCl-resistant SHR (SHR-R) and Wistar-Kyoto (WKY) rats maintained on basal and high-NaCl diets were used as controls. Intravenous administration of ANP to conscious, freely moving rats with intact baroreflexes decreased blood pressure and LSNA in SHR-S, SHR-R and WKY rats maintained on basal or high-NaCl diets for 2-3 weeks. The depressor response to intravenous ANP was greater in 8% NaCl-fed SHR-S than in any other group; the LSNA response was greater in SHR-S on either diet than in any other group. Intracerebroventricular administration of ANP evoked small, transient sympatholytic responses in SHR-S on both diets and minimal responses in SHR-R and WKY rats; these responses could not be attributed to leakage of ANP into the peripheral circulation. Thus, circulating ANP has a sympatholytic effect in SHR-S that is not amplified by high-NaCl feeding and can be only partially accounted for by a central action.

Animals↗
PubMed1983

Effects of aging on changing arterial compliance and left ventricular load in a northern Chinese urban community.

Pulse wave velocity (PWV) was measured by means of transcutaneous Doppler techniques in the aorta, right arm, and right leg of 480 normal subjects of both sexes in urban Beijing, China (age range 3 to 89 years, mean age 41 +/- 20.8 SD); supine blood pressure was recorded in the brachial artery of each subject with standard sphygmomanometric procedures. Serum cholesterol was determined in a subgroup of 79 subjects (age 17 to 85 years, mean 47 +/- 26 SD). PWV (y in cm/sec) was found to vary with age (x, years) at each of the three locations according to the following regression equations: aorta, y = 9.2x + 615, r = .673 (p less than .001); right arm, y = 4.8x + 998, r = .453 (p less than .001); right leg, y = 5.6x + 791, r = .630 (p less than .001). Systolic, diastolic, mean, and pulse pressures were found to increase with age. PWV also increased with mean supine blood pressure but was not related to serum cholesterol (average 4.49 +/- 0.11 [SEM], mmol/l). Compared with that of Western populations, serum cholesterol tended to be lower at all age groups, systolic pressure higher at ages over 35 years, and PWV higher at all ages. Because change in PWV is directly related to change in arterial compliance, these results indicate that aging and not concomitant atherosclerosis (known to be rare in Asian populations) is the dominant factor associated with reduced arterial compliance and increased left ventricular load in these subjects.

Adolescent↗