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Biomedical subjects

H Wang

Publications and source records attributed to H Wang.

At least 1,801 records · Page 100Linked to original sources
PubMed1996

Salmonella detection by the polymyxin-cloth enzyme immunoassay using polyclonal and monoclonal detector antibodies.

Several commercially available O-antigen polyclonal antisera and a monoclonal antibody to the core region of lipopolysaccharide (LPS) were examined as sources of detector antibodies in a polymyxin-cloth enzyme immunoassay (polymyxin-CEIA) for Salmonella. In this assay, polymyxin-coated polyester cloth captured the LPS antigens from Salmonella broth cultures, followed by immunoenzymatic detection of the captured LPS using specific antibodies. Pools of polyvalent antisera reacted with all of the Salmonella strains tested, but also gave cross-reactions with some non-Salmonella bacteria. On the other hand, the monoclonal antibody gave positive reactions with all of the Salmonella tested except serogroup O-strains, but did not react with any of the non-Salmonella bacteria. The monoclonal antibody supplemented with a single factor serogroup O:35 rabbit antiserum was able to detect the serogroup O-strains without causing any cross-reactions with the non-Salmonella bacteria. As an example of the applicability of this assay system, low levels of Salmonella cells spiked into various food samples were successfully detected after an overnight enrichment in broth.

Animals↗
PubMed1996

Kinetics of HPV11 DNA replication after infection of keratinocytes with virions.

Human papillomavirus type 11 (HPV11) normally infects keratinocytes of stratified epithelia and replicates as an episome. To untangle early events in the development of a papilloma, we have infected human keratinocytes with human papillomavirus type 11 virions and monitored replication using density labeling with bromodeoxyuridine and subsequent density centrifugation. We show that only a portion of the virus reaching the nucleus undergoes replication and that continued replication occurs without recruitment from the nonreplicating pool of virus suggesting that HPV11 replication remains confined to a small number of cells. Increasing inoculum size leads to more viral DNA reaching the nucleus but not a corresponding increase in viral replication. Subsequent papilloma development must, therefore, occur within a small subset of cells.

3T3 Cells↗
PubMed1996

Identification and DNA typing of two Cw7 alleles (Cw*0702 and Cw*0704) in Japanese, with the corrected sequence of Cw*0702.

Two alleles encoding HLA-Cw7 antigens, tentatively called C7J1 and C7J2, have been identified in Japanese using a PCR-SSCP method. The nucleotide sequence of full-length C7J1 cDNAs showed a high degree of homology to the reported Cw*0702 sequence except in exon 1. We then resequenced the allele carried by the cell line JY in which Cw*0702 was first identified, according to a request from the WHO Nomenclature Committee. The results revealed complete identity between the corrected Cw*0702 sequence and the C7J1 sequence. On the other hand, the C7J2 sequence was completely identical to the reported Cw*0704 sequence. Sequences specific for Cw*0702 and Cw*0704 were confirmed using PCR-RFLP and PCR-SSO methods. Moreover, association analysis with other HLA locus alleles showed positive associations of Cw*0702 with HLA-B7, -B39, and -B67 and of Cw*0704 with HLA-B70 in Japanese.

Alleles↗
PubMed1996

Presence of neuronal nitric oxide synthase in the suprachiasmatic nuclei of mouse and rat.

Nitric oxide has recently been identified as a major neural regulator. It is synthesized by the enzyme nitric oxide synthase. Whilst considerable functional evidence has pointed to an involvement of nitric oxide in circadian regulation, all previous morphological studies have failed to demonstrate the presence of nitric oxide synthase in the mammalian suprachiasmatic nucleus. By use of an antibody directed against whole recombinant rat neuronal nitric oxide synthase we have identified the presence of immunoreactivity for this enzyme in the suprachiasmatic nucleus of mouse and rat, to provide the first detailed report of this enzyme in the suprachiasmatic nucleus for any species of mammal. Immunoreactivity for neuronal nitric oxide synthase was found in neurons throughout the suprachiasmatic nucleus of mice and no difference could be detected between the ventrolateral and dorsomedial parts of the nucleus in terms of the optical density of the immunostaining. Very small, rounded neuronal cell bodies were immunopositive. Electron microscopy revealed that these neurons had relatively large nuclei and scant cytoplasm containing relatively few organelles, which sometimes included some 100 nm dense-cored peptidergic vesicles. Only about 5% of such neurons were not detectably immunoreactive. By contrast, in the rat suprachiasmatic nucleus, a much smaller number of neurons were immunopositive and these cells were aggregated in the ventrolateral part of the nucleus. The immunoreactive neurons were bipolar cells with scanty cytoplasm. Electron microscopy revealed diffuse immunoreactivity in the cytosol, but not within any organelles. In the surrounding neuropil immunoreactive dendrites and axons mingled with much larger numbers of immunonegative processes, but immunoreactive boutons were only identified just outside the dorsal margin of the nucleus. Astrocytes, oligodendrocytes and endothelial cells in the suprachiasmatic nucleus were immunonegative. NADPH-diaphorase activity was not detectable in the suprachiasmatic nucleus of either mouse or rat. This morphological evidence for nitric oxide synthase-immunoreactive cells in the suprachiasmatic nucleus supports the existing functional evidence for an involvement of nitric oxide in the transmission of light-induced signals to the suprachiasmatic nucleus in mammals.

Animals↗
PubMed1996

Comparison of MSX-1 and MSX-2 suggests a molecular basis for functional redundancy.

This study examines the biochemical properties of two members of the murine MSX family, MSX-1 and MSX-2, which have been implicated to have partially overlapping functions during embryogenesis. Our analyses show that MSX-1 and MSX-2 share many features in common including their DNA binding and transcriptional properties. In particular, MSX-1 and MSX-2 interact with a common consensus DNA site, and exhibit similar DNA binding site preferences. However, MSX-2 has a higher apparent affinity for DNA, and the distinction between MSX-1 and MSX-2 resides in their differing sequences N-terminal to the homeodomain. With respect to their transcriptional properties, both MSX-1 and MSX-2 function as repressors and share the distinct property that they do so independently of their consensus DNA binding sites. However, MSX-1 is a more potent repressor, and the difference between these proteins also maps to their N-terminal regions. Similarly, the expression patterns of Msx-1 and Msx-2 as examined by whole mount in situ hybridization are related but not identical. Thus, Msx-1 and Msx-2 are co-expressed in the limbs, neural tube, and branchial arches; however, Msx-1 has a broader expression pattern overall and is expressed uniquely in certain embryonic regions. These features suggest that these members of the Msx family are 'equivalent but not equal' and that their proposed redundancy may be achieved via distinct biochemical mechanisms that yield a similar functional outcome.

Amino Acid Sequence↗
PubMed1996

Screening potential corneal donors for HIV-1 by polymerase chain reaction and a colorimetric microwell hybridization assay.

PURPOSE: Current screening of potential corneal donors for human immunodeficiency virus type 1 (HIV-1) involves serologic detection of antibodies to the virus. However, this approach cannot detect infection during the seronegative window period of the disease. We therefore evaluated the polymerase chain reaction (PCR) assay for viral nucleic acid as a possible alternative to screening cadaveric blood for HIV-1. METHODS: Blood specimens from cadavers diagnosed at autopsy with acquired immunodeficiency syndrome (AIDS) (n = 21), at high risk for HIV-1 infection (n = 47), and at no known risk (n = 350) were screened by PCR for HIV-1 proviral DNA and human leukocyte antigen (HLA)-DQ alpha sequences, and for HIV antibodies. RESULTS: All AIDS group samples were seropositive; of these, 18 (86%) and 20 (95%) of 21 were positive for HIV by PCR of proteinase K- and Chelex-extracted pellets, respectively. The seropositive samples negative by PCR testing were shown to inhibit PCR amplification. Nine (19%) of 47 high-risk specimens were HIV-positive. The no-known-risk group yielded negative results. The overall sensitivities for PCR in the proteinase K- and Chelex-treated groups were 90% and 97%, respectively, compared with Western blot reactivity. If PCR-inhibitory samples and HLA-DQ alpha-negative samples had been eliminated, sensitivity would have been 100%. Specificity was 100% for each group. CONCLUSIONS: Screening cadaveric blood by PCR may be feasible, but further refinement of the assay and blood specimen collection practices will be necessary for it to become routine. Future studies should focus on optimizing specimen procurement and preparation to reduce or eliminate specimens that inhibit PCR.

AIDS Serodiagnosis↗
PubMed1996

A VISUAL BASIC program to pre-process MRI data for finite element modeling.

Investigators use non-invasive imaging to collect geometric data for finite element models. A preprocessor is described to facilitate model generation of anatomical regions from serial Magnetic Resonance Imaging (MRI) data stored in a bitmap format. The MRI Data Transfer System is a stand-alone Windows-based program developed in VISUAL BASIC 3.0 which generates a NASTRAN input file. The program can be modified to generate input files for other solvers. The software will executive on any IBM-compatible computer which runs Windows Version 3.1 or higher. To demonstrate the software, model generation of a portion of a tibia is described.

Computer Graphics↗
PubMed1996

Clinical neuroimmunology.

Clinical research has focused on autoimmune disease (AID) for a couple of decades. More sensitive and specific methods have been developed for neuroimmunological research. Gamma fraction bands (bands separated by electrophoresis and visualized by amino black staining) and IgG fraction bands (bands separated by iso-electric focusing and visualized by immunostaining) are used instead of oligoclonal bands. Myasthenia gravis (MG) mainly involves acetylcholine receptors of the postsynaptic membrane at the neuromuscular junction. Myasthenia gravis has been considered to be a generalized AID, because 7% of patients with myasthenia gravis associate with other AIDs and more than one autoimmune antibody is detected in 52.5% patients with myasthenia gravis. Pyramidal signs in myasthenia gravis patients are described; the possible mechanism may at least be partly due to the acetylcholine receptor antibody. P2 protein and its antibody are studied in patients with acute and chronic inflammatory demyelinating polyneuropathy.

Adolescent↗
PubMed1996

DNA vaccines against malaria: immunogenicity and protection in a rodent model.

Since the first demonstration of the technology a few years ago, DNA vaccines have emerged as a promising method of vaccination. In a variety of experimental systems, DNA vaccines have been shown not only to induce potent immune responses, but also to offer many advantages in terms of ease of construction, testing, and production. In this article we summarize the progress achieved in development of DNA vaccines that can protect mice from infection by the rodent malaria parasite Plasmodium yoelii, describe initial studies of immunogenicity of a malaria DNA vaccine in a primate model, and outline the strategies being employed to design the next generation of malaria DNA vaccines.

Animals↗
PubMed1996

Characterization of platelet-reactive antibodies in children with varicella-associated acute immune thrombocytopenic purpura (ITP).

Biochemical analyses were performed on blood samples obtained from two children (P1, P2) who presented with acute immune thrombocytopenic purpura (ITP) following a recent varicella zoster virus (VZV) infection. Patient sera had antibodies that were reactive with normal blood-group O platelets as measured by flow-cytometric assay. Western blot analysis of electrophoretically separated normal blood-group O platelets under reducing and non-reducing conditions demonstrated that these sera were reactive with platelet antigens of approximately 50 and approximately 110 kD, respectively. These 50/110 kD antigens were not reactive with seven sera from acute ITP patients whose illness was not preceded by VZV infection, with serum from a patient with a prior history of VZV and no thrombocytopenia, nor with normal healthy control sera. VZV antibodies (IgG and IgM), isolated from patient sera by affinity chromatography using immobilized purified VZV glycoproteins, were found to bind to gel-filtered autologous platelets and with normal blood-group O platelets, as analysed by flow cytometry. No binding was observed using antibodies similarly prepared from healthy volunteer sera. To investigate their ability to sensitize platelets to complement activation, affinity-purified VZV antibodies were incubated with platelets and then with purified complement components C1 and 125 I-labelled C4. Platelets reacted with VZV-specific antibodies from the two patients and showed increases of 2.3-2.4-fold of platelet-surface deposition of 125 I-C4b, compared to controls. These data provide evidence that virus-specific antibodies occurring in children with varicella-associated acute ITP cross-react with normal platelet antigens, and may contribute to platelet clearance.

Antibodies, Viral↗
PubMed1996

Characterization of the CheAS/CheZ complex: a specific interaction resulting in enhanced dephosphorylating activity on CheY-phosphate.

The cheA gene encodes two overlapping polypeptides with a common carboxyl terminus: CheAL and CheAS. CheAL plays a central role in the Escherichia coli chemotaxis signalling pathway by autophosphorylation and transferring the phosphate to both CheY and CheB. On the other hand, the physiological functions of CheAS remain unknown. We have observed that overproduction of CheAS in wild-type cells increased counterclockwise-biased flagellar rotation, and this effect is dependent on the presence of CheZ. CheZ specifically facilitates CheY-phosphate (CheY-P) dephosphorylation and generates a smooth swimming signal. A physical interaction was detected between CheZ and CheAS in wild-type cell lysates by immunoprecipitation. The CheAS/CheZ interaction does not require other chemotaxis components, as we could form the complex using purified CheAS and CheZ proteins. The ability of CheAS to bind to CheZ depends on its being in the reduced state. We found that under non-reducing conditions, CheAS appears to form intermolecular disulphide bonds and loses the ability to bind to CheZ. Finally, the CheAS/CheZ complex formed in vitro shows a greater dephosphorylating activity on CheY-P than does free CheZ.

Bacterial Proteins↗
PubMed1996

Pollination induces mRNA poly(A) tail-shortening and cell deterioration in flower transmitting tissue.

Pollination induces many physiological responses in the flower, including deterioration and death in specific pistil cell types. It is shown here that within the style of tobacco, pollination-induced cell deterioration was restricted to the transmitting tissue while the surrounding cortical tissue was not affected. It was distinct from general senescence since exogenously applying the senescence-inducing hormone ethylene, or its precursor aminocyclopropane-1-carboxylic acid (ACC), to the flower or the pistil induced overall deterioration in the entire flower. Furthermore, both pollen tube growth and ethylene action were needed for the entire spectrum of cellular changes associated with this pollination-induced transmitting tissue deterioration process. It is also shown that pollination-induced mRNA poly(A) tail-shortening for at least three major classes of transmitting tissue-specific mRNAs. As is commonly observed for poly(A) tail-shortened mRNAs, the levels of two of these three mRNA classes decline after pollination. On the other hand, the third class of mRNAs, transmitting tissue-specific (TTS) mRNAs, was maintained at a very high level subsequent to pollination, even after substantial poly(A)-tail shortening. TTS mRNAs encode a pollen tube growth-promoting and -attracting protein needed for optimal in vivo pollen tube growth. The specific preservation of TTS mRNAs in the deteriorating transmitting tissue cells suggests that these cells can distinguish molecules needed in the pollinated styles from those that are dispensable, and protect them from degradation. It is suggested that the pollination-induced mRNA poly(A) tail-shortening and cell death are programmed processes suited to the post-pollination transmitting tissue environment. Results showing that ACC is a candidate signal molecule for the pollination-induced mRNA-shortening which is accentuated by ethylene and mediated via a protein phosphorylation-dependent signal transduction pathway are also presented.

Amino Acids↗
PubMed1996

Radiation-modifying effect of oxygen in synchronized cells pre-treated with acute or prolonged hypoxia.

The objective of the present study was to measure the sensitizing effect of oxygen following acute as well as protracted hypoxia in cell cultures of maximum homogeneity. Human cells of the established line NHIK 3025 were synchronized by the method of mitotic selection and irradiated while in late G1 (aerobic conditions) or in the oxygen-sensitive restriction point near the G1/S border. Four different conditions of oxygen treatment were used: (1) Cells were irradiated under aerobic conditions 5 h after mitotic selection (G1 lasts for 6.5 h in these cells). (2) Cells were irradiated under extremely hypoxic conditions (i.e. < 4 ppm O2) following an acute (30 min) deoxygenation starting 5 h after mitotic selection. (3) Cells were irradiated under aerobic conditions immediately following 20 h of extreme hypoxia which was started 5 h following mitotic selection. (4) Cells were irradiated under extremely hypoxic conditions after 20 h of extreme hypoxia starting 5 h after mitotic selection. In addition, asynchronous cells were irradiated, either without any pretreatment, or after reoxygenation, following 20 h of extreme hypoxia. The data indicate that the sensitizing effect of oxygen (after protracted hypoxia) is strictly dose-modifying for cells rendered hypoxic while in late G1 and arrested in the oxygen-sensitive restriction point in late G1. There is a non-dose-modifying sensitization when synchronized control cells, irradiated under aerobic conditions, are compared with synchronized cells irradiated during acute hypoxia. In this case, however, one is comparing cells in two different biological states, i.e. cells in an ordinary G1 are compared with cells arrested in the oxygen-sensitive restriction point in G1. The non-dose-modifying nature of the oxygen sensitization as observed in this case may, therefore, reflect biological differences between the cell cultures that are compared rather than differences with respect to the radiochemical sensitization as induced by oxygen at small and large radiation doses. The present data indicate, however, that if cancer cells in human tumours are best represented by a mixture of aerobic cells and chronic hypoxic cells, oxygen may appear to exert no effect, or perhaps even a net protective effect, at low radiation doses.

Cell Cycle↗
PubMed1996

Differentiation modulates the balance of positive and negative Jun/AP-1 DNA binding activities to regulate cellular proliferative potential: different effects in nontransformed and transformed cells.

Differentiation of 3T3T cells into adipocytes results in the progressive repression of growth factor responsiveness. This is associated with the transcriptional repression of the inducibility of c-jun and junB expression by serum. In contrast, differentiation of SV-40 large T antigen-transformed 3T3T cells (CSV3-1) does not repress growth factor responsiveness nor c-jun or junB inducibility even though CSV3-1 cells can differentiate into adipocytes. To better explain these observations, we have studied compositional changes in AP-1 DNA binding activity attributed to c-Jun, JunB, and JunD during the differentiation process in 3T3T and CSV3-1 cells. The results show that in nontransformed 3T3T cells, differentiation represses AP-1 DNA binding activity via a proportionate downregulation of c-Jun, JunB, and JunD. In contrast, in CSV3-1 cells, AP-1 DNA binding activity increases twofold during differentiation, which is accounted for by an increase in JunD with no change in c-Jun and JunB. If c-Jun and JunB serve as positive regulators and JunD serves as a negative regulator for cell proliferation as suggested by previous studies, the repression of JunD expression in differentiating CSV3-1 cells should be mitogenic because decreasing JunD/AP-1 DNA binding activity would allow c-Jun/AP-1 and JunB/AP-1 DNA binding activities to be dominant. The results confirm this prediction showing that antisense junD oligodeoxyribonucleotides are mitogenic for differentiating CSV3-1 cells whereas antisense c-jun and junB inhibit mitogenesis. These data support the conclusion that differentiation can regulate cellular proliferative potential by modulating the balance of positive and negative Jun/AP-1 DNA binding activities in distinct ways in nontransformed and transformed cells.

3T3 Cells↗
PubMed1996

Pixel driven implementation of filtered backprojection for reconstruction of fan beam SPECT data using a position dependent effective projection bin length.

Filtered backprojection is commonly implemented as a pixel driven algorithm in which the density is reconstructed on an array of grid points that are usually associated with the centres of square image pixels. In fan beam geometry, this conventional pixel driven approach using two-bin linear interpolation leads to an inefficient use of the projection data due to magnification at the projection line. For typical count limited SPECT data, this results in increased reconstructed image noise. We propose an alternative type of pixel driven algorithm that makes efficient use of projection data by averaging over all bins within a position dependent "effective' projection bin interval. The effective bin interval is equivalent to the image pixel length magnified on the projection line. This heuristic method leads to more complete use of the projection data and results in reconstructed images with superior noise properties compared with the conventional method while presenting similar spatial resolution characteristics.

Algorithms↗
PubMed1996

Overexpression of cyclin D1 and cyclin E in N-nitrosomethylbenzylamine-induced rat esophageal tumorigenesis.

Dysregulation of G1 cyclins has been implicated in several human malignancies. To further investigate the role of G1 cyclins in chemical carcinogenesis, the expression of cyclin D1 and cyclin E was analyzed by RT-PCR and immunohistochemical studies in N-nitrosomethylbenzylamine (NMBA)-induced rat esophageal tumorigenesis. Cyclin D1 mRNA levels were increased 2.8-fold in 25 week (P < 0.05) and 6.8-fold in 45 week (P < 0.01) papillomas induced by NMBA, when compared with normal rat esophageal epithelium. Cyclin E mRNA levels were increased 6.2-fold in 25 week (P < 0.01) and 6.9-fold in 45 week (P < 0.01) papillomas. Immunohistochemical staining revealed exclusive nuclear staining of both cyclin D1 and cyclin E. Furthermore, there was a sequential increase in cyclin D1- and cyclin E-positive cells from normal epithelium, to preneoplastic lesions, to papillomas. These findings suggest that overexpression of cyclin D1 and cyclin E occur relatively early in rat esophageal tumorigenesis and participate in tumor progression in this model.

Animals↗

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