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H Wang

Publications and source records attributed to H Wang.

At least 1,585 records · Page 88Linked to original sources
PubMed1997

Mechanisms of loss of functional dendritic cells in HIV-1 infection.

Dendritic cells (DC) are lost from blood and skin during injection with HIV-1; those remaining show a reduced capacity to stimulate T cell proliferation [S. C. Knight, AIDS 10, 807-817]. Our recent studies investigate mechanisms underlying these effects. DC exposed to HIV-1 vitro can act as targets for cytotoxic T cells, although optimal killing was not obtained until DC were exposed to HIV-1 for 3 days. This cytotoxicity may provide a feedback mechanism by which DC that have presented antigens are removed. However, this effect could also contribute to the reduction in DC during persistent infection. We have also investigated the effect of exposure to HIV-1 on DC function. DC exposed to HIV-1 IIIB virus for 2 h stimulated primary proliferative and cytotoxic T cell responses in vitro; these effects may be similar to those occurring during the early activation of protective antiviral immunity in vivo. After exposure of DC to virus for 5 days, stimulation of allogeneic T cells was reduced. However, a different situation applied when using DC developed from CD34+ cord blood stem cells under the influence of granulocyte-macrophage colony-stimulating factor and tumor necrosis factor alpha that were exposed at 24 h to the same virus. These DC showed low levels of infection similar to peripheral blood DC but in contrast stimulated normal allogeneic T cell proliferation. The capacity of DC exposed to HIV-1 to stimulate T cell proliferation or to show a blocked stimulatory capacity may thus depend not only on the length of the exposure to virus but also on the maturational state of the DC. Loss in DC numbers and function on exposure to HIV-1 may result in lower levels of stimulation of T cells, which in turn may be instrumental in reduction of T cell numbers.

Cell Differentiation↗
PubMed1997

Protein heteronuclear NMR assignments using mean-field simulated annealing.

A computational method for the assignment of the NMR spectra of larger (21 kDa) proteins using a set of six of the most sensitive heteronuclear multidimensional nuclear magnetic resonance experiments is described. Connectivity data obtained from HNC alpha, HN(CO)C alpha, HN(C alpha)H alpha, and H alpha (C alpha CO)NH and spin-system identification data obtained from CP-(H)CCH-TOCSY and CP-(H)C(C alpha CO)NH-TOCSY were used to perform sequence-specific assignments using a mean-field formalism and simulated annealing. This mean-field method reports the resonance assignments in a probabilistic fashion, displaying the certainty of assignments in an unambiguous and quantitative manner. This technique was applied to the NMR data of the 172-residue peptide-binding domain of the E. coli heat-shock protein, DnaK. The method is demonstrated to be robust to significant amounts of missing, spurious, noisy, extraneous, and erroneous data.

Computer Simulation↗
PubMed1997

Multiplex PCR for avian pathogenic mycoplasmas.

Mycoplasma infections are of great concern in avian medicine, because they cause economic losses in commercial poultry production. A multiplex polymerase chain reaction (PCR) was optimized to simultaneously detect four pathogenic species of avian mycoplasmas. Four sets of oligonucleotide primers specific for Mycoplasma gallisepticum (MG), M. synoviae (MS), M. meleagridis (MM) and M. iowae (MI) were used in the test. By using agarose gel electrophoreses for detection of the PCR-amplified DNA products, the sensitivity of detection was between 1 pg for MG, 1 pg for MS, 100 fg for MM and 100 pg for MI after 35 cycles of PCR. Similar sensitivity of these primers was achieved with broth cultures of these four organisms.

Animals↗
PubMed1997

Lipids, lipoproteins and apolipoproteins abnormalities in patients undergoing dialysis.

Twenty hemodialysis (HD) patients and 20 patients on continuous ambulatory peritoneal dialysis (CAPD) were investigated for lipids, lipoproteins and apolipoproteins abnormalities. HD patients had elevated serum triglyceride, decreased high-density lipoprotein cholesterol (HDL-C) and apolipoprotein A-I (Apo A-I), while CAPD patients had elevated total cholesterol, triglyceride, low-density lipoprotein cholesterol (LDL-C), Apolipoprotein B (Apo B), Apo B/Apo A-I ratio, and decreased HDL-C, Apo A-I. Because of the molecular sieving effects of peritoneum, CAPD have a negative effect on these abnormalities. CAPD patients might be at greater risk of developing coronary artery disease than HD patients who are also at increased risk as compared with normals.

Aged↗
PubMed1997

Sequence-based association analysis of HLA class I and II alleles in Japanese supports conservation of common haplotypes.

Alleles of HLA-A, B, C, DRB1, DQB1, and DPB1 loci were fully determined in 117 healthy Japanese. A*2402, A*3303, A*1101, A*0201, B*4403, B*5201, Cw*0102, Cw*1403, Cw*0304, Cw*0702, Cw*0801, and Cw*1202 showed frequencies of over 10%. Multi-locus haplotype frequencies were estimated by the maximum likelihood method. Strength of association between C and B loci was comparable with that between DRB1 and DQB1 loci. Alleles unidentified by a serological method and having very similar nucleotide sequences (A2: A*0201, A*0206, A*0207, B61: B*4002, B*4006) were carried by different haplotypes. Several frequent five-locus haplotypes were identified including A*3303-Cw*1403-B*4403-DRB1(*)1302-DQB1(*)0604, and A*2402-Cw*1202-B*5201-DRB1(*)1502-DQB1(*)0601. These sequence-based haplotypes corresponded to serology-based common haplotypes which have already been described in Japanese. These findings indicate that common HLA haplotypes consist of particular sets of HLA alleles and that these haplotypes have been conserved through recent human evolution.

Alleles↗
PubMed1997

Site-directed mutagenesis of a fungal beta-1,4-endoglucanase increases the minimum size required for the substrate.

Conserved regions within a family-5 beta-1,4,-endoglucanase (Eg11), produced by the fungus Macrophomina phaseolina, have been modified through site-directed mutagenesis, resulting in production of an enzyme with novel substrate requirements. The engineered form was generated through mutagenesis of D232A, which lies within the substrate-binding cleft of Eg11. Wild-type Eg11 requires a minimum substrate size of five glucosyl units, while the engineered form requires a minimum of six glucosyl units. Screening was facilitated by the unique ability to obtain functional expression of the fungal endoglucanase in Escherichia coli. Wild-type and mutated Eg11 have equivalent activity on cellohexaose, and both release cellobiose from the reducing end of the cellodextrin. This is the first example of protein engineering of an endoglucanase that results in a novel minimum substrate requirement for cellohexaose. This substrate specificity has not been reported for any native endoglucanases, thus the modified Eg11 may prove useful in applications requiring specific hydrolysis of complex carbohydrates such as beta-glucans.

Amino Acid Sequence↗
PubMed1997

Characterization of the Na+/H+ exchanger in human choriocarcinoma (BeWo) cells.

We examined the expression and activity of the Na+/H+ exchanger in the human choriocarcinoma BeWo cell line. When treated with methotrexate, these cells differentiated from cytotrophoblast-like cells to enlarged multinucleate syncytiotrophoblast-like cells. There was no change in the apparent Km for Na+ between undifferentiated and differentiated cells. However, differentiated cells could transport more than five times the proton flux of undifferentiated cells. There was no difference in the Hill coefficient between undifferentiated and differentiated cells. However, the maximal flux (Jmax) for undifferentiated cells was higher than that for differentiated cells. Inhibition of Na+/H+ exchange activity by an amiloride analog and Hoe694 revealed a sensitive and a resistant component in both differentiated and undifferentiated cells. Northern blot analysis and immunocytochemistry suggested that the sensitive component was due to the NHE1 isoform of the protein while the resistant component was due to the NHE3 isoform. The NHE1 isoform was localized to the brush border membrane of BeWo cells and Western blot analysis showed that the NHE1 protein was more abundant in brush border membranes from differentiated BeWo cells compared to undifferentiated cells. The results show that BeWo cells contain the NHE1 and NHE3 isoforms of the Na+/H+ exchanger and that the NHE1 isoform is primarily localized to the brush border membrane.

Blotting, Northern↗
PubMed1997

Excretory/secretory products of plerocercoids of Spirometra erinaceieuropaei induce the expression of inducible nitric oxide synthase mRNA in murine hepatocytes.

In this study, we observed the level of normal murine hepatocyte inducible NOS (iNOS) mRNA by semi-quantitative polymerase chain reaction (SQ-PCR) analysis after stimulation with ES products (ESP) and/or ESP fractions from the plerocercoids. We found that ESP are able to induce the expression of iNOS gene in a dose-dependent fashion. Treatment of ESP with polymyxin B did not affect their ability to induce the expression of iNOS gene, suggesting that bacterial lipopolysaccharide (LPS) is not involved. The iNOS-inducing factor (a) is soluble, and may be a component whose molecular mass exceeds 94 kDa as analyzed by a combination of SDS-PAGE and SQ-PCR. The peak of iNOS mRNA level was detected 3 h after stimulation with ESP; the mRNA level decreased sharply from 9 h. Dexamethasone inhibited the induction of mRNA for hepatocyte iNOS. In contrast, cycloheximide stimulated the induction; this suggests that de nova protein synthesis is important in the regulation of the ESP-induced expression of iNOS mRNA. Actinomycin D blocked the induction. In addition, the results of Northern blot analysis showed that ESP suppressed the LPS (10 micrograms/ml) and interferon-gamma (IFN-gamma, 100 U/ml)-induced hepatocyte iNOS mRNA expression in a dose-dependent fashion and the suppressing effect was more marked when hepatocytes were exposed to ESP 3 h prior to LPS and IFN-gamma. These results demonstrate that the soluble factor(s) of ESP is capable of inducing murine iNOS gene expression in hepatocytes in the absence of added cytokines.

Animals↗
PubMed1997

Expression of syndecan-1 in rabbit neointima following de-endothelialization by a balloon catheter.

Enrichment of proteoglycans is prominent in early atherogenesis, contributing not only to SMC migration and proliferation, but also to low density lipoprotein retention. A family of integral cell membrane proteoglycans termed syndecans has recently been recognized. Among syndecans, syndecan-1, the first isolated member, has received most research attention. In this study, we examined the expression of syndecan-1 in rabbit aorta and aortic neointima, developed in response to a balloon catheter-induced de-endothelialization. The tissues were processed for Northern blot analysis, in situ hybridization, immunohistochemical staining and immunoblotting. Our results indicate that in normal aorta, the signal for syndecan-1 is weak. However, arterial injury induces syndecan-1 expression at both mRNA and protein levels. The presence of syndecan-1 in the neointimal tissue is persistent, prominent even at the 12th week after injury. Syndecan positive cells are distributed in the whole layer of the neointima, but are not visible in the underlying media. The presence of syndecan-1 in arterial neointima suggests a novel means of mediating interactions between neointimal cells and various agents, including extracellular matrix components, growth factors and lipoproteins.

Animals↗
PubMed1997

The normal stress effect and equilibrium friction coefficient of articular cartilage under steady frictional shear.

During creep or stress relaxation, articular cartilage exhibits a time-dependent friction coefficient which has been shown to reach an equilibrium value, mu eq, as the tissue deformation equilibrates. This study investigates the frictional properties of articular cartilage explants under steady frictional shear and constant compressive strain after the tissue reaches stress-relaxation equilibrium. The two parameters measured are the normal force and frictional torque, from which the friction coefficient was then calculated. It is shown in this experimental study that: (1) Under a prescribed infinitesimal compressive strain, cartilage supports higher compressive normal stress under steady shear than it does in the absence of frictional shear. Furthermore, the normal stress increases with increasing sliding velocity, resulting in a velocity-dependent value of mu eq. The observed normal stress effectively increases the compressive stiffness of cartilage by a factor up to 3.1. (2) Under a prescribed steady frictional shear both the normal stress and frictional shear stress increase, though not proportionally, with increasing compressive strain, producing a decreasing friction coefficient. (3) This velocity-dependent normal stress effect is also shown to result, at least partly, from intrinsic properties of cartilage. The normal stress effect has not been previously reported for articular cartilage, and represents an intriguing mechanical response not commonly encountered in solids, though common in non-Newtonian fluids.

Animals↗
PubMed1997

An antiserum to idazoxan recognizes an immunoreactive substance in human serum and cerebral spinal fluid which is not agmatine.

A polyclonal antibody was generated in rabbits to an idazoxan-albumin antigen. The anti-idazoxan antiserum had high affinity for unconjugated 3H-idazoxan (Kd of 19.8 nM) in a radio-immunoassay (RIA). Of various drugs and native molecules only idazoxan potently (Ki of 24 nM) inhibited 3H-idazoxan binding to the anti-idazoxan antibody. A few drugs weakly inhibited 3H-idazoxan binding (IC50 > 605 microM) with rank order of UK 14304 > guanabenz > cirazoline > amiloride > naphazoline. Neither agmatine, an endogenous clonidine displacing substance (CDS), catecholamines or imidazoles inhibited the binding of 3H-idazoxan to the anti-idazoxan antibody. The anti-idazoxan RIA was 4-6 fold more sensitive than an antibody to para-amino clonidine. The CDS detected by ligand displacement from bovine brain dose-dependently inhibited 3H-idazoxan binding. This immunoreactive (ir-) CDS activity was present in human (0.9-4.1 U/ml) and rat sera (1-2 U/ml) and in the cerebro-spinal fluid of eight patients with serious disease of the central nervous system, but not in controls. We conclude: (1) an anti-idazoxan RIA is a sensitive, selective and clinically applicable RIA for measuring ir-CDS; (2) ir-CDS is not agmatine; (3) CDS represents a family of endogenous ligands for imidazoline receptors including ir-CDS and agmatine.

Agmatine↗
PubMed1997

Ultrastructural immunocytochemical localization of mu-opioid receptors in dendritic targets of dopaminergic terminals in the rat caudate-putamen nucleus.

Many motor effects of opiates acting at mu-opioid receptors are thought to reflect functional interactions with dopaminergic inputs to the caudate-putamen nucleus. We examined the cellular and subcellular bases for this interaction in the rat caudate-putamen nucleus by dual immunocytochemical labelling for mu-opioid receptors and tyrosine hydroxylase, a marker mainly for dopamine in this region. mu-Opioid receptor-like immunoreactivity showed a patchy distribution by light microscopy. Within the patches, electron microscopy revealed that immunogold labelling for mu-opioid receptors was mainly distributed along extrasynaptic plasma membranes of medium spiny neurons. In contrast, immunoperoxidase labelling for tyrosine hydroxylase was exclusively located in axons and axon terminals without detectable mu-opioid receptor-like immunoreactivity. Forty-six percent of the total mu-opioid receptor-labelled neuronal profiles (n = 1441) were in contact with tyrosine hydroxylase-immunoreactive axons and terminals. These contacts were characterized by closely apposed parallel plasma membrane segments, without well-defined synaptic junctions, or with punctate symmetric specializations. From 639 noted appositions, over 90% were between mu-opioid receptor-labelled dendrites and/or dendritic spines and tyrosine hydroxylase-containing terminals. The dendritic spines containing mu-opioid receptor-like immunoreactivity often received asymmetric synapses characteristics of excitatory inputs from unlabelled terminals. Axon terminals containing mu-opioid receptor-like immunoreactivity formed asymmetric synapses with dendritic spines, or apposed tyrosine hydroxylase-labelled terminals. Our results suggest that, in striatal patch compartments, mu-agonists and dopamine dually modulate the activity of single spiny neurons mainly through changes in their postsynaptic responses to excitatory inputs. In addition, our findings implicate mu-opioid receptors and dopamine in the presynaptic regulation of excitatory neurotransmitter release within the striatal patch compartments.

Animals↗
PubMed1997

Telomerase expression in primary neuroblastomas.

Maintenance of chromosomal telomeres is necessary for continued cell growth, and this is carried out in germline tissues by telomerase. In contrast to most somatic tissues, many tumours have telomerase activity. The RNA component of human telomerase (hTR) was measured by Northern analyses of 150 primary untreated neuroblastomas and compared with clinical stage at diagnosis. hTR expression > 33 (relative to cell line control = 100) was seen in 41% of all tumours and the frequency of hTR > 33 increased with stage of disease. Expression of hTR may be involved in progression of neuroblastoma.

Blotting, Northern↗
PubMed1997

Comparison of synovial fluid cartilage marker concentrations and chondral damage assessed arthroscopically in acute knee injury.

OBJECTIVE: To examine the correlation between synovial fluid cartilage markers and degree of cartilage damage determined by arthroscopic evaluation in subjects with acute knee injury. DESIGN: Chondral damage was quantified using a validated arthroscopic scoring system in 20 subjects with effusive acute knee injuries of less then 4 months duration and no history or radiographic evidence of joint pathology. Levels of synovial fluid 3B3(-) neoepitope, 3B3(+) chondroitinase generated epitope of proteoglycan, keratan sulfate (KS) and hyaluronic acid (HA) were measured by competitive enzyme-linked immunosorbent assays using monoclonal antibodies 3B3 and 5D4. Total sulfated glycosaminoglycan (GAG) was measured by 1,9-dimethylmethylene blue colorimetric dye-binding assay. RESULTS: We found a dramatic decrease in levels of 3B3(-) (rs = -0.62, P = 0.004), and GAG (rs = -0.49, P = 0.03) with increasing chondral damage score; but no correlation of damage score with 3B3(+), KS or HA levels. CONCLUSION: These data reveal a change in cartilage metabolism within the first 4 months of symptomatic knee injury evinced by a significant inverse correlation of 3B3(-) and GAG levels to chondral lesion severity. These results suggest that serial measurement of these synovial fluid markers in the setting of acute knee injury could predict chondral lesion severity and aid in the decision to intervene surgically.

Acute Disease↗
PubMed1997

Anserinones A and B: new antifungal and antibacterial benzoquinones from the coprophilous fungus Podospora anserina.

Two new benzoquinones with antifungal, antibacterial, and cytotoxic activities have been isolated from liquid cultures of the coprophilous fungus Podospora anserina. The structures of anserinones A (1) and B (2) were assigned on the basis of MS and NMR results, and the absolute stereochemistry of 2 was deduced by analysis of 1H-NMR data for its (R)- and (S)-2-phenylbutyryl ester derivatives.

Anti-Bacterial Agents↗
PubMed1997

Zrp2: a novel maize gene whose mRNA accumulates in the root cortex and mature stems.

A near full-length cDNA clone (pZRP2) was isolated from a cDNA library constructed from maize root mRNAs. The predicted polypeptide has a calculated molecular mass of 66,975 Da, is largely hydrophilic, and contains 26 repeats of a motif the consensus sequence of which is RKATTSYG[S][D/E][D/E][D/E][D/E][P]. The function of the putative protein remains to be elucidated. The ZRP2 mRNA accumulates to the highest levels in young roots, and is also present in mature roots and stems of maize. Further analysis of young roots indicates that the lowest level of ZRP2 mRNA is near the root tip, with relatively high levels throughout the remainder of the root. In situ hybridization reveals that ZRP2 mRNA accumulates predominantly in the cortical parenchyma cells of the root. In vitro nuclear run-on transcription experiments indicate a dramatically higher level of zrp2 gene transcription in 3-day old roots than in 5-day old leaves. A zrp2 genomic clone, which includes the transcribed region and 4.7 kb of upstream sequence, was isolated and characterized.

Amino Acid Sequence↗
PubMed1997

Expression of a synthetic antifreeze protein in potato reduces electrolyte release at freezing temperatures.

A synthetic antifreeze protein gene was expressed in plants and reduced electrolyte leakage from the leaves at freezing temperatures. The synthetic AFP was expressed as a fusion to a signal peptide, directing it to the extracytoplasmic space where ice crystallization first occurs. The gene was introduced to Solanum tuberosum L. cv. Russet Burbank by Agrobacterium-mediated transformation. Transformants were identified by PCR screening and expression of the introduced protein was verified by immunoblot. Electrolyte-release analysis of transgenic plant leaves established a correlation between the level of transgenic protein expression and degree of tolerance to freezing. This is the first identification of a phenotype associated with antifreeze protein expression in plant tissue.

Amino Acid Sequence↗