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Biomedical subjects

S Zheng

Publications and source records attributed to S Zheng.

At least 19 recordsLinked to original sources
PubMed1996

Naturally processed T cell epitopes from human glutamic acid decarboxylase identified using mice transgenic for the type 1 diabetes-associated human MHC class II allele, DRB1*0401.

The identification of class II binding peptide epitopes from autoimmune disease-related antigens is an essential step in the development of antigen-specific immune modulation therapy. In the case of type 1 diabetes, T cell and B cell reactivity to the autoantigen glutamic acid decarboxylase 65 (GAD65) is associated with disease development in humans and in nonobese diabetic (NOD) mice. In this study, we identify two DRB1*0401-restricted T cell epitopes from human GAD65, 274-286, and 115-127. Both peptides are immunogenic in transgenic mice expressing functional DRB1*0401 MHC class II molecules but not in nontransgenic littermates. Processing of GAD65 by antigen presenting cells (APC) resulted in the formation of DRB1*0401 complexes loaded with either the 274-286 or 115-127 epitopes, suggesting that these naturally derived epitopes may be displayed on APC recruited into pancreatic islets. The presentation of these two T cell epitopes in the islets of DRB1*0401 individuals who are at risk for type 1 diabetes may allow for antigen-specific recruitment of regulatory cells to the islets following peptide immunization.

Alleles↗
PubMed1996

APC mutations in colorectal tumors with mismatch repair deficiency.

We have investigated the influence of genetic instability [replication error (RER) phenotype] on APC (adenomatous polyposis coli), a gene thought to initiate colorectal tumorigenesis. The prevalence of APC mutations was similar in RER and non-RER tumors, indicating that both tumor types share this step in neoplastic transformation. However, in a total of 101 sequenced mutations, we noted a substantial excess of APC frameshift mutations in the RER cases (70% in RER tumors versus 47% in non-RER tumors, P < 0.04). These frameshifts were characteristic of mutations arising in cells deficient in DNA mismatch repair, with a predilection for mononucleotide repeats in the RER tumors (P < 0.0002), particularly (A)n tracts (P < 0.00007). These findings suggest that the genetic instability that is reflected by the RER phenotype precedes, and is responsible for, APC mutation in RER large bowel tumors and have important implications for understanding the very earliest stages of neoplasia in patients with tumors deficient in mismatch repair.

Adenomatous Polyposis Coli↗
PubMed1996

An autosomal dominant, qualitative platelet disorder associated with multimerin deficiency, abnormalities in platelet factor V, thrombospondin, von Willebrand factor, and fibrinogen and an epinephrine aggregation defect.

Multimerin is a massive soluble, multimeric protein found in platelets and endothelial cells. Recent studies identified multimerin as a specific coagulation factor V binding protein, complexed with platelet, but not plasma, factor V. These findings led us to investigate individuals with inherited factor V deficiencies for possible multimerin abnormalities. Platelet proteins were evaluated using immunoassays, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, immunoprecipitation, and direct binding studies. Patients with factor V Quebec, a disorder with abnormal platelet factor V, had a quantitative deficiency in multimerin (n = 11 tested; mean, 12.5%; range, 5% to 27% of the normal pool; normal range, 45% to 214%) with a normal multimer pattern. Quantitative and qualitative abnormalities were detected in their platelet factor V. An unrelated patient who was deficient in platelet and plasma factor V had normal platelet multimerin. The levels of platelet beta-thromboglobulin, von Willebrand factor, thrombospondin, and fibrinogen antigen were normal in the factor V Quebec patients. However, proteins with abnormal mobility were detected in their platelet lysate and releasate, and their platelet thrombospondin, von Willebrand factor, and fibrinogen showed evidence of proteolytic degradation. Platelet counts of the factor V Quebec patients ranged from mildly thrombocytopenic to low normal (mean, 159 x 10(9)/L; range, 104 to 198 x 10(9)/L). In addition, their platelets failed to aggregate in response to 6 to 10 micromol/L epinephrine despite normal numbers of platelet alpha 2-adrenergic receptors. These data indicate that patients with factor V Quebec have an inherited bleeding disorder distinct from other platelet disorders and associated with multiple abnormalities, including multimerin deficiency, abnormal platelet factor V, thrombospondin, von Willebrand factor, and fibrinogen, and an epinephrine aggregation defect.

Afibrinogenemia↗
PubMed1996

Structural heterogeneity of phospholipase D in 10 dicots.

The occurrence of multiple forms of phospholipase D (EC 3.1.4.4) was investigated in different tissues of castor bean (Ricinus communis) and in other plant species. Phospholipase D variants were resolved by nondenaturing and isoelectric focusing polyacrylamide gel electrophoresis and detected by immunoblotting using anti-phospholipase D antibodies and by enzyme activity assay. Three phospholipase D variants were produced differentially in the roots, endosperm, cotyledons, and hypocotyl of 5-day postgermination seedlings of castor bean. Furthermore, different patterns of phospholipase D variants were found in the different regions of hypocotyl (elongated and hook). Multiple phospholipase D forms were found in florets of cauliflower and broccoli, leaves of cabbage, celery, tomato, and potato, and alfalfa sprouts, suggesting that structural heterogeneity of phospholipase D occurs widely in plants.

Electrophoresis, Polyacrylamide Gel↗
PubMed1996

Increased endothelin expression in a rat cardiac allograft model of chronic vascular rejection.

Endothelins (ET) are potent vasoconstrictors that are directly mitogenic for vascular smooth muscle cells and fibroblasts. It is possible that the vasoconstrictor and mitogenic effects of ET could play a significant role in the vascular remodeling process that occurs in chronic vascular rejection (CVR). We have previously shown that cardiac allografts in the indefinitely surviving major histocompatibility complex identical WF.1L (RT1(1)) to Lewis (LEW) (RT1(1)) inbred rat strain combination provide a highly reproducible model of progressive CVR. The objective of this investigation was to measure endothelin-1 ventricular content of WF.1L-LEW cardiac allografts and to determine the immunohistochemical patterns of ET cellular reactivity at well defined posttransplant time periods. Data were compared with those obtained in similar studies of LEW-LEW syngeneic: heart grafts as well as all recipients' own hearts. The ventricular ET-1 content of the WF.1L cardiac allografts was markedly higher (4.3-, 7.0-, and 4.8-fold at 20, 40, and 60 days, respectively) than in corresponding recipients' hearts. Also, the increase in ventricular ET-1 levels as compared with the recipients' hearts rose significantly only in the allograft group. No comparable differences were observed in the syngeneic heart graft controls. Allografts consistently showed ET staining of intimal myocytes at sites of occlusive and subocclusive intimal proliferation associated with CVR. Allografts also showed ET cellular staining in areas of reparative fibrosis associated with indolent interstitial rejection and ischemic myocardial damage. The results of this study strongly suggest that ET may play a significant role in the pathogenesis of CVR.

Animals↗
PubMed1996

The induction of ret by Wnt-1 in PC12 cells is atypically dependent on continual Wnt-1 expression.

Wild type PC12 pheochromocytoma cells that had been infected with a Wnt-1-carrying virus and thus express Wnt-1 (PC12/Wnt-1) are known to acquire the same flat cell phenotype as that of spontaneously occurring PC12 flat cell variants except that the latter do not presently express Wnt-1. Flat cell variants of PC12 cells exhibit markedly altered morphology and gene expression. In order to assess the possibility that the spontaneously occurring flat cell variants could have been induced in wild type PC12 cells by previous transient expression of the cell's endogenous Wnt-1, we have isolated PC12/Wnt-1 cells expressing little or no Wnt-1. In spite of absent Wnt-1 expression, they retained their flat cell morphology, glutamate/aspartate transporter activity, increased neu mRNA levels and lack of both norepinephrine transporter activity and nerve growth factor-induced differentiation. Thus, Wnt-1 expression is not required to maintain the flat cell phenotype. However, we identified one gene, ret, whose mRNA level in PC12 was not only increased by Wnt-1 expression, but whose increased mRNA level was also dependent on continual Wnt-1 expression. This finding suggests that the induction of ret by Wnt-1 can be used to elucidate the Wnt-induced signalling pathway in mammalian cells.

ATP-Binding Cassette Transporters↗
PubMed1996

A glutamate/glutamine/aspartate/asparagine transport operon in Rhodobacter capsulatus.

A mutant of Rhodobacter capsulatus was identified in which an operon encoding a binding-protein-dependent transporter was interrupted by Tn5 transposition. Cloning and sequence analysis of the wild-type operon revealed a four-gene cluster with similarities to genes encoding periplasmic binding proteins (BztA), integral membrane proteins (BztB and BztC), and ATP-binding proteins (BztD). To assess the function of this putative binding-protein-dependent transport system, a mutant was constructed in which most of the bztABCD operon was deleted and replaced by an antibiotic-resistance marker. The deletion mutant grew more slowly than the wild type in NH4(+)-free medium supplemented by glutamate, glutamine, aspartate or asparagine; it was resistant to toxic analogues of Glu, Asp, and Asn at concentrations that inhibited growth of the wild type; and it was defective in the uptake of Glu, Gln, and Asp. A complementing plasmid containing the wildtype copy of bztABCD was able to rescue all the mutant phenotypes. Taken together, these results indicate that the proteins encoded by bztABCD are active in the uptake of Glu, Gln, Asp, and Asn. In addition, competition experiments, in which the ability of each of the four amino acids to compete for the transport of one another was examined, demonstrated that all four substrates share at least one component of this transport system.

Amino Acid Sequence↗
PubMed1995

Expression cloning of a protective Leishmania antigen.

Parasite-specific CD4+ T cells have been shown to transfer protection against Leishmania major in susceptible BALB/c mice. An epitope-tagged expression library was used to identify the antigen recognized by a protective CD4+ T cell clone. The expression library allowed recombinant proteins made in bacteria to be captured by macrophages for presentation to T cells restricted to major histocompatibility complex class II. A conserved 36-kilodalton member of the tryptophan-aspartic acid repeat family of proteins was identified that was expressed in both stages of the parasite life cycle. A 24-kilodalton portion of this antigen protected susceptible mice when administered as a vaccine with interleukin-12 before infection.

Amino Acid Sequence↗
PubMed1995

Mutational analysis of patients with X-linked adrenoleukodystrophy.

Adrenoleukodystrophy (ALD) is an X-linked neurodegenerative disorder characterized by elevated very long chain fatty acid (VLCFA) levels, reduced activity of peroxisomal VLCFA-CoA ligase, and variable phenotypic expression. A putative gene for ALD was recently identified and surprisingly encodes a protein (ALDP) that belongs to a family of transmembrane transporters regulated or activated by ATP (the ABC proteins). We have examined genomic DNA from ALD probands for mutations in the putative ALD gene. We detected large deletions of the carboxyl-terminal portion of the gene in 4 of 112 probands. Twenty-five of the ALD probands whose ALD genes appeared normal by Southern blot analysis were surveyed for mutations by Single Strand Conformation Polymorphism (SSCP) procedures and DNA sequence analysis. SSCP variants were detected in 22 probands and none in 60 X-chromosomes from normal individuals. Mutations were detected in all of the ALD probands. The mutations were distributed throughout the gene and did not correlate with phenotype. Approximately half were non-recurrent missense mutations of which 64% occurred in CpG dinucleotides. There was a cluster of frameshift mutations in a small region of exon 5, including an identical AG deletion in 7 unrelated probands. These data strongly support the supposition that mutations in the putative ALD gene result in ALD.

ATP Binding Cassette Transporter, Subfamily D, Mem↗
PubMed1995

Analysis of Pichinde arenavirus transcription and replication in human THP-1 monocytic cells.

Human promonocytic THP-1 cells were previously shown to be nonpermissive for Pichinde virus (PV) replication unless the cells were induced to differentiate to macrophages by stimulation with phorbol ester (PMA) (J. Virol. 65, 3575, 1991). The restriction did not involve receptor modulation, virus binding, nor internalization of virus but a requirement for a host cell function in PV replication was observed in that the phorbol ester effect required protein kinase C activation and was inhibited by actinomycin D. In this report we demonstrate that PV S RNA genomes, antigenomes, GPC mRNA and NP mRNA are expressed at high levels in PMA treated THP-1 cells but at significantly lower levels or not at all in untreated cells. We have also determined that degradation of input viral S RNA does not account for decreased PV RNA synthesis in the undifferentiated cells. This suggests that the restriction of PV replication in THP-1 cells is a post-penetration event which precedes transcription of viral mRNAs and replication of viral genomes and supports a role for differentiation-specific host cell factors early in PV replication.

Cell Line↗
PubMed1995

5' termini of Pichinde arenavirus S RNAs and mRNAs contain nontemplated nucleotides.

Primer extension of Pichinde arenavirus purified virion RNA suggests that genomes have at least a single nontemplated base at the 5' end which is a G in all cDNA clones having one such single base. On the other hand, the predominant products of primer extension on total virus-infected-cell RNA are at positions -1 and -2. The primer extension product at position -2 is not represented in virion RNA, and neither of these products is proportionally represented in mRNA. mRNA is predominantly 3 or 4 bases longer than genomes and antigenomes, but primer extension products as long as 7 bases were observed. The sequence of nontemplated bases reported here is unambiguous with respect to the 5'-terminal base and supports the view that there is a sequence preference for a G at the 5' termini of mRNAs. Assessment of our sequence data in the context of the sequences of Tacaribe and lymphocytic choriomeningitis viruses suggests that the mechanism of initiation of arenavirus transcription is fundamentally different from that of members of the families Orthomyxoviridae and Bunyaviridae.

Animals↗
PubMed1995

Influence of steric stabilization of liposome-encapsulated hemoglobin on Listeria monocytogenes host defense.

Liposome-encapsulated hemoglobin (LEH) products are being investigated as potential blood substitutes. To determine if changes in LEH composition can modify the immune response, red blood cell substitutes based on conventional lipids containing phosphatidylinositol (LEH1) and sterically stabilized lipid vesicles containing polyethylene glycol phosphatidylethanolamine (LEH2) were tested for effects on host resistance. On Day 0, groups of 18 to 20 female CD-1 mice were given an intravenous (i.v.) infectious challenge with a 20% lethal dose of Listeria monocytogenes. Mice received a single i.v. dose of LEH1, LEH2, or albumin vehicle on Day +1 or Day -3 relative to infectious challenge. Mice dosed with LEH1 and LEH2 on Day +1 died rapidly from Listeria infection; but mice dosed with LEH2 lived significantly longer than did mice receiving LEH1. By contrast, when administered on Day -3, LEH1 had no significant effect on host immunity, while LEH2 increased susceptibility to Listeria infection. In addition, LEH1 and LEH2 both caused significant reduction of phagocytic activity as measured by rat alveolar macrophage (AM) ingestion of latex microspheres. AM incubated 4 hr with either LEH1 or LEH2 prior to addition of microspheres ingested fewer beads in a dose-dependent manner. No difference in in vitro phagocytic activity was observed between LEH1 or LEH2. The inability to differentiate LEH formulations based on in vitro phagocytic activity suggests that the in vivo Listeria infection model may be more relevant in discerning the immunotoxicity of the LEH formulations tested.

Animals↗
PubMed1995

[Clinical significance of urodynamic study in benign prostatic hyperplasia].

Urodynamic study, including UFR, cystometry, UPP, and EMG was carried out in 74 patients with benign prostatic hyperplasia (BPH) during the past 5 years. The correlations of prostate volume (57.3 +/- 6.3 ml), prostate length (4.94 +/- 1.23 cm) and detrusor pressure (6.54 +/- 4.73 kPa) were analysed. The results showed that urodynamic study plays an important role in indicating the time and method of operative treatment and predicting the prognosis of BPH operation. The fistularization of the bladder should be performed on the patients with the bladder disfunction. The measurement of the urethral resistance is of value in the diagnosis of posterior urethra obstruction than that of UFR.

Aged↗
PubMed1995

[Reduced intracellular drug accumulation related to overexpression of P-glycoprotein cannot explain the complete drug resistance in MDR cell variant K562/Dox].

The purpose of this study was to determine what proportion of drug resistance of MDR cell variant K562/Dox was attributed to the reduced steady state intracellular drug accumulation related to overexpression of P-glycoprotein. K562/Dox was derived from repeated exposure of human erythroleukaemic cell line K562 cell to doxorubicin. As assessed with MTT assay, the resistance of K562/Dox to 7 cytotoxic drugs increased variably in the range between 1200 and 11 folds. K562/Dox cells were positively stained with anti-human p-glycoprotein monoclonal antibody JSB-1, indicating overexpression of P-gp. Intracellular drug accumulation in K562/Dox, though significantly reduced as compared with that in K562 cells, was maximally restored by concurrent exposure of cells to 6 mumol/L verapamil and 1.72 mumol/L either of the doxorubicin, epirubicin or daunorubicin. Similar results were obtained by exposure of cells to 12 mumol/L verapamil and 8.62 mumol/L drug, indicating that restoration of intracellular drug accumulation in MDR cells was dependent on the relative concentrations of verapamil to drug. However, the resistances of K562/Dox cells to epirubicin and daunorubicin still remained for about 5.6 and > 6.6 folds, respectively, even at verapamil concentration of 6 mumol/L, suggesting at least a relatively big fraction of drug resistance was not directly related to the altered cellular pharmacokinetics associated with overexpression of P-glycoprotein.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1995

[Inhibition of colorectal carcinoma induced by 1, 2-dimethylhydrazine in mice with tea polyphenols].

Colorectal cancer was induced by subcutaneous injection of 1,2-dimethylhydrazine (DMH) in mice, and the animals were administered orally with 0.4% of tea polyphenols (TP) simultaneously for 20 weeks to study its preventive effects. Results showed incidence of colorectal cancer in mice administered with TP was significantly lower than in positive controls (P < 0.05), and average number of tumor foci and proportion of adenocarcinoma were significantly fewer in TP group than in positive controls (P < 0.01). Level of cytochrome P450 (CP450) in liver microsome of the mice administered with TP was lower than that both in negative and positive controls (P < 0.01), and activities of superoxide dismutase (SOD) in liver tissue were higher than those in positive controls (P < 0.01), but lower than in negative controls (P < 0.01). Proliferative index of epithelial cells in mice was not obviously influenced by TP. It suggested that TP could have preventive effects on experimental colorectal cancer with a possible mechanism of lowering CP450 and increasing the activity of SOD in the liver.

1,2-Dimethylhydrazine↗
PubMed1995

[Synthesis of substituted benzeneseleninic acid and its antineoplastic activity].

In view of the negative relationship between selenium and cancer, we designed and synthesized eleven substituted benzeneselenic acids (IVa-k) The antineoplastic activity of the title compounds was evaluated via the tests of inhibition of acetylcholinesterase, HL-60 and K562 in vitro, and Ehrlich carcinoma in mice. The result showed that the compounds IVa and IVb had distinct antineoplastic activity. The life span increase rate of IVb on mice bearing Ehrlich carcinoma was 20.30% (20mg/kg), which was higher than the increase rate of sodium selenite (10.65%, 0.5mg/kg).

Animals↗