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Biomedical subjects

W C Yang

Publications and source records attributed to W C Yang.

At least 91 records · Page 5Linked to original sources

Characterization of the soybean early nodulin cDNA clone GmENOD55.

Two cDNA clones of the soybean early nodulin GmENOD55 were characterized. These clones may represent two members of the soybean early nodulin gene family GmENOD55. GmENOD55 has an N-terminal signal peptide and it contains an internal domain consisting of proline and serine residues. Analyses of nodules lacking infection threads and intracellular bacteria suggest that the GmENOD55 gene is first expressed after release of Bradyrhizobium japonicum in plant cells. This conclusion is supported by in situ hybridization studies showing that the expression is restricted to the infected cell type.

Amino Acid Sequence↗

Immunostimulatory factor(s) in the porcine uterine fluids.

The effects of porcine uterine fluid filtrate (PUF) on lymphocyte activity were examined. PUF was added to porcine lymphocyte cultures in the presence or absence of phytohemagglutinin (PHA), concanavalin A (Con A), or pokeweed mitogen (PWM). Lymphocyte response was determined by 3H-thymidine incorporation. Results indicated that PUF enhanced PHA-, Con A-, and PWM-induced lymphocyte mitogenesis by 222%, 207%, and 261%, respectively, as compared to control. The stimulatory effect of PUF was unlikely to be due to the presence of interleukin-2 in PUF, by evidence that PUF alone did not support cytotoxic T-lymphocyte proliferation, nor was the stimulatory effect due to microbial contamination, because treatment of PUF through PyroBind columns to remove endotoxin did not affect its activity. Incubation of PUF at 56C for 40 min did not alter the enhancement effect. Treatment of PUF with trypsin or pronase for up to 24 hr did not change its activity. In mixed lymphocytes reaction, PUF augmented 3H-thymidine uptake by 242%. The molecular (mol) mass of the immunostimulatory activity in PUF was examined by size-exclusion HPLC, Sephadex G-50, and G-15 gel filtration, and it was found to be 0.8 kDa and < 0.5 kDa. Extracts of porcine endometrium also enhanced PHA-induced lymphocyte mitogenesis, suggesting that the factor(s) may be of uterine origin. In conclusion, we have identified an factor(s) in PUF capable of stimulating lymphocyte mitogenesis and mixed lymphocyte reaction.

Adjuvants, Immunologic↗

Differential effects of IFN-gamma on kidney cell expression of MHC class II molecules, kidney cell associated molecules and their stimulatory capacity in mixed lymphocyte kidney cell culture.

Mixed cell co-cultures of lymphocytes responding to kidney cells (MLKC), islets of Langerhans (MLIC) and mixed lymphocyte culture (MLC), were used to clarify mechanisms in allogeneic and autoimmune (tissue-associated) antigen presentation. Fresh kidney cortical tubular cells (KC), Madin-Darby canine kidney (MDCK), and dog kidney (6247) (DK) cell lines were used in MLKC reactions, and islets of Langerhans were used in the MLIC as putative antigen presenting cells (APC). The stimulating cells were treated with purified or recombinant dog interferon-gamma (IFN-gamma), and the detection of class II MHC molecule expression was assessed by a moneclonal antibody (mAb) (B1F6). Transcription of MHC class II mRNA and IFN-gamma mRNA was measured by semiquantitative polymerase chain reaction, and detection of a kidney cell (tissue-associated) antigen molecule was assessed by the mAb I1F6 that recognizes 72 and 150 kDa tubular cell protein(s) (KT1). The MDCK cell line constitutively expressed low levels of MHC class II molecules and KT1. The steady-state level of the MHC class II mRNA transcription was virtually unaltered by treatment with IFN-gamma (400 units) for 48 hours; however, the MHC cell surface protein expression was enhanced. The KC and DK cell lines constitutively expressed KT1, but not MHC class II molecules; these cells required a minimum of 4000 units, and a 62-hour incubation with IFN-gamma was needed to upregulate both surface MHC class II molecules and the transcription of corresponding specific mRNA. In the MLKC reaction both the MDCK and DK cell lines, as well as fresh KC cells, could serve as lymphocyte activators. This could be amplified by exogenous IFN-gamma. The removal of APC from the responding T cell population did not reduce the IFN-gamma effect. This indicates that IFN-gamma treatment allows for the expression of all of the co-stimulating factors and/or adhesion molecules necessary for these cells to serve as (surrogate) APC (direct as opposed to indirect antigen presentation). The requirements for purified IFN-gamma to increase this amplification was greater in the MLKC reactions with kidney cells than in the MLC reactions. The mAbs anti-IFN-gamma and I1F6 differed in their ability to inhibit lymphocyte proliferation depending on the different cell types involved. The I1F6 inhibited the MDCK and DK cell-driven MLKC (in the absence of exogenous IFN-gamma).(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗

Characterization of GmENOD40, a gene showing novel patterns of cell-specific expression during soybean nodule development.

In this paper, the soybean 'early nodulin' clone pGmENOD40 is characterized. The GmENOD40 encoded protein does not contain methionine and does not show homology to proteins identified so far. In situ hybridizations showed that this gene has a complex expression pattern during development of determinate soybean nodules. At early stages of development transcription is induced in dividing root cortical cells, the nodule primordium and the pericycle of the root vascular bundle. In mature soybean nodules, the gene is expressed in the uninfected cells of the central tissue and in the pericycle of the nodule vascular bundles. Studies on nodules devoid of intracellular bacteria and infection threads, showed that the expression of the gene in the nodule primordium is induced in these empty nodules, while the induction of the GmENOD40 gene in the nodule vascular bundle requires the presence of intracellular bacteria or infection threads. A pea cDNA clone homologous to GmENOD40 was isolated to enable in situ hybridization studies on indeterminate nodules. The expression patterns in both determinate and indeterminate nodules suggests that the ENOD40 protein might have a transport function.

Amino Acid Sequence↗

Production and characterization of recombinant canine interferon-gamma from Escherichia coli.

We have used the recently cloned cDNA for canine interferon-gamma (IFN-gamma) to engineer bacteria to produce large amounts of the recombinant cytokine. The resulting protein can be recognized by monoclonal and polyclonal antibodies largely species specific for canine IFN-gamma. The purified recombinant IFN-gamma (rIFN-gamma) also had biological activity in vitro in three assay systems: (i) vesicular stomatitis virus plaque inhibition, (ii) class II major histocompatibility complex antigen upregulation on canine kidney parenchymal cells, and (iii) amplification of in vitro tissue-associated lymphoproliferation, all known to be effected by native IFN-gamma (nIFN-gamma). The availability of large amounts of active canine rIFN-gamma will be an important tool in studies of the role of this cytokine in the widely used experimental canine organ transplant model and also will be of diagnostic and therapeutic veterinary interest.

Amino Acid Sequence↗

Suppressive effect of serum from pigs and dogs fed a diet deficient in vitamin E and selenium on lymphocyte proliferation.

The effect of sera collected from either pigs or dogs previously fed a vitamin E (vit E)- and selenium (Se)-deficient diet on in vitro lymphocyte blastogenesis response to mitogens was studied. Porcine sera were obtained from pigs used in 2 different trials. In I trial, 4-wk-old pigs received either a basal diet deficient in vit E and Se or the basal diet supplemented with Vit E, Se or Vit E and Se. Pigs were maintained on their respective diet for 25 d. Canine sera were collected from pups maintained on a deficient diet for 8 wk. Four dogs and 4 pigs maintained on a commercial diet were used as donors of peripheral blood lymphocytes (PBL). The addition of sera from pigs or dogs maintained on a vit E- and Se-deficient diet markedly suppressed both porcine and canine PBL response to mitogens. Porcine PBL blastogenesis was also suppressed when porcine or canine sera were added 8, 24 or 48 h after the beginning of the incubation period to culture containing 1% of fetal bovine serum (FBS). However, the suppressive effect caused by porcine sera was less severe than the one due to canine sera. Addition of 1% FBS in the cultures was sufficient to eliminate the suppression caused by the presence of sera from pigs fed a vit E- and Se-deficient diet. Other attempts to restore the lymphocyte response to mitogens by the addition of indomethacin, diethylcarbamazine or eicosatetraynoic acid, inhibitors of prostaglandin and/or leukotriene synthesis, were not successful. Because of the severe suppression caused by sera from animals maintained on a vit E- and Se-deficient diet on the in vitro response of lymphocytes to mitogenic stimulations, it is very important to take precautions to avoid such deficiency. In vivo suppression of immunocompetent cells to antigenic stimulations may impair the capacity of the host to control infections.

5,8,11,14-Eicosatetraynoic Acid↗

The 16S rRNA gene of Streptomyces lividans TK64 contains internal promoters.

A 632-bp Sau3AI fragment of Streptomyces lividans TK64 genome was found to confer promoter activity in Streptomyces and Escherichia coli. This fragment showed almost identical sequence (97.8%) to the S. coelicolor 16S rRNA segment encompassing from nucleotide 706 to 1337 region. The transcription start points of this fragment were identified by the primer extension method. Analysis of the nucleotide sequence upstream the transcription start points revealed two putative E. coli-like promoters resided within this fragment. The occurrence of internal promoters active in Streptomyces and E. coli was also confirmed in the 16S rRNA gene of rrnE operon from TK64.

Base Sequence↗

Neurogenic bladder in acquired immune deficiency syndrome (AIDS).

This is the first report of incidence of neurogenic bladder in patients with acquired immune deficiency syndrome (AIDS) and emphasizes that it is a significant cause of various voiding dysfunctions in these patients. Neurologic disease occurs in about one third of patients with AIDS. Both central and peripheral nervous systems may be involved. Urodynamic evaluation is necessary for assessment and management of neurogenic voiding disorders in this group of patients.

Acquired Immunodeficiency Syndrome↗

Cross-species reactivity of the anti-idiotype anti-OKT3 cascade between mice and humans.

The administration of murine mAb specific for the CD3 epsilon subunit of the TCR complex (OKT3) has been demonstrated to engender in humans an anti-OKT3 idiotypic cascade. This study used murine-derived anti-OKT3 (Ab2) as a bioreagent to determine whether this Ab2 and polyclonal anti-(anti-OKT3) (Ab3) generated in some human kidney transplant patients are idiotypically connected. Two anti-OKT3 mAbs G-880 (IgG1) and M-12 (IgM) were derived by immunizing BALB/c mice with the OKT3-secreting hybridoma. The two mAbs exhibited specificity for OKT3 F(ab)'2 idiotypic determinants. Both mAbs were tested for their ability to inhibit OKT3 induced mitogenesis and to block FITC-OKT3 binding to cell surface CD3 epsilon chain. The M-12 mAb inhibited OKT3-induced mitogenesis and blocked (approximately 60%) the binding of OKT3 to peripheral blood (PBL) T-cell CD3 epsilon chain in flow cytometry. In contrast, the G-880 mAb did not inhibit mitogenesis and only weakly blocked OKT3 binding to CD3 epsilon chain (approximately 12%). Sera of kidney transplant recipients who received OKT3 antirejection therapy and who developed antiidiotypic anti-OKT3 antibodies could be divided into two subgroups exhibiting anti-OKT3 activity: (a) those who had similar specificity as M-12 and failed to enhance the M-12 inhibition of OKT3 binding to PBL T-cell CD3 epsilon chain when added as a third component (n = 3), and (b) those with anti-OKT3 antibodies with idiotype specificity dissimilar to M-12 and who were able to increase the (maximum 60%) inhibition obtained with M-12 in the OKT3 to T-cell CD3-binding assay (n = 4). From these observations, we conclude that M-12 had the characteristics of an Ab2 beta and G-880 that of an Ab2 alpha. Additionally, there was an idiotypic connectivity of mouse-derived M-12 anti-OKT3 (Ab2) and OKT3-engendered human polyclonal anti-(anti-OKT3) (Ab3), in that three of seven patients examined had human serum IgG antibodies that specifically recognized M-12 idiotypic determinants as demonstrated in ELISA.

Animals↗

Cerebral infarction in young men with nephrotic syndrome.

BACKGROUND AND PURPOSE: Thrombosis is one of the main complications of nephrotic syndrome; however, cerebral infarction associated with nephrotic syndrome has been rarely reported. SUMMARY OF REPORT: We describe acute cerebral infarction in two young men with nephrotic syndrome. Both had a hypercoagulable state based on hemostatic studies. By retrospectively reviewing the medical records of the past 10 years at our hospital, we found an additional five cases of cerebral infarction with nephrotic syndrome. Two of the patients were found to have nephrotic syndrome during admission for stroke. CONCLUSIONS: Hypercoagulability may be the major contributing factor of cerebral infarction in patients with nephrotic syndrome.

Acute Disease↗

Methotrexate, vinblastine, adriamycin and cisplatin (M-VAC) for advanced TCC of urothelium.

After the introduction of methotrexate, vinblastine, adriamycin and cisplatin combination (M-VAC) chemotherapy for transitional cell carcinoma (TCC) of the urothelium, the reported response and survival rates improved when compared with the previous regimens. From July 1989 to March 1991, 43 consecutive cases of invasive or metastatic TCC that had received at least one course of M-VAC treatment were collected by a computer-assisted search and analyzed. The overall response rate, including complete response (CR) and partial response (PR), was 44% (CR, 16%; PR, 28%). Tumors originating from the renal pelvis or bladder responded better than those from the ureter (kidney, 57%; bladder, 42%; and ureter 30%). Lymph node, soft tissue, lung metastases and local recurrent tumors had a greater chance of response than bone or liver metastases. Although patients from the blackfoot disease endemic area seemed to respond better than those from the nonendemic area, there was no statistically significant survival benefit. The duration of follow-up ranged from nine to 38 months. The length of survival of nonresponders ranged from one month to 19 months (median, eight months; mean, seven months). Ten of the 19 responders relapsed, and the others were still responding. Three patients (7%) survived two years, disease-free. Although the response rate improved, the chance of long-term survival was still unsatisfactory in our study.

Adult↗

Immunochemical and biochemical characterization of purified canine interferon-gamma. Production of a monoclonal antibody, affinity purification, and its effect on mixed lymphocyte culture and mixed lymphocyte kidney culture reactions.

We have advanced the hypothesis that the primary autolymphoproliferative response of dog T cells in mixed lymphocyte kidney cultures (MLKC) results from their recognition of tissue-specific (kidney-associated) antigen(s) presented in conjunction with class II MHC antigens. Lymphocyte culture-derived supernatants had been found previously to upregulate class II antigen expression on kidney cells and enhance T cell activation. In the present study we have isolated and characterized dog IFN-gamma, a class II-inducing substance that is secreted in the culture supernatant of activated T lymphocytes. Dog IFN-gamma was induced with A-23187 and PMA and purified stepwise using controlled-pore glass, Mono Q anion exchange chromatography, and Superose 6-gel filtration on FPLC. The purification resulted in two molecules of 42 Kd and 31 Kd molecular weights. An IgG1 monoclonal antibody was engendered to these molecules. With this mAb reagent, in immunochemical experiments, we have developed a sensitive ELISA and a method for purifying dog IFN-gamma by affinity chromatography. Species specificity studies indicated that purified dog IFN-gamma reacted with a polyclonal rabbit antihuman IFN-gamma, but not with a mAb to human IFN-gamma. However, the antidog IFN-gamma mAb that was generated also reacted with recombinant human IFN-gamma. In in vitro biological studies, the purified IFN-gamma (two mol. wt. species) upregulated the expression of canine class II MHC molecules on dog tubular epithelial cells and the dog kidney epithelial cell line (MDCK). The antidog IFN-gamma mAb blocked T cell proliferative response to kidney cell and, by inference, the interaction between endogenously released IFN-gamma in vitro with its cell surface receptor, thus inhibiting the induced upregulation of class II. Interestingly, although antidog IFN-gamma markedly blocked the MLKC (10 micrograms mAb/well), there was no effect on the allogeneic MLC. This observation indicates that the cytokine IFN-gamma may be a uniquely key substance amplifying the immune response of T cells to tissue-associated antigens on surrogate antigen-presenting cells that require induced upregulation of class II MHC antigen expression (MLKC), in contrast to reactions in which these antigens are already constitutively expressed on the antigen-presenting cells (mixed lymphocyte culture).

Animals↗

Antibodies to hepatitis C virus in uremic patients on continuous ambulatory peritoneal dialysis.

The prevalence of antibody to hepatitis C virus (anti-HCV) among 101 uremic patients receiving continuous ambulatory peritoneal dialysis (CAPD) was evaluated using a synthetic peptide-based HCV antibodies enzyme immunoassay. Thirty (29.7%) were found anti-HCV positive. This is significantly higher than 500 unselected paid blood donors (4.2%, P less than 0.0001). Among CAPD patients, anti-HCV positivity was found more frequently in patients who had received frequent and longer duration of hemodialysis previously (40.4% vs. 20.4%, P less than 0.05). These findings suggest that hemodialysis patients have a higher risk of HCV infection. At present, CAPD may be a suitable way to reduce the incidence of HCV infection in uremic patients.

Adolescent↗

Comparison of nonspecific radioimmunoassay, high-performance liquid chromatography, and fluorescence polarization immunoassay for cyclosporine monitoring in renal transplantation.

Three methods, i.e., nonspecific radioimmunoassay (RIA; Incstar), fluorescence polarization immunoassay (FPIA; TDx Abbott), and high-performance liquid chromatography (HPLC), have been used for monitoring cyclosporine blood levels in renal transplantation patients. The levels obtained from 135 samples showed a modest correlation between RIA and HPLC, FPIA and HPLC, RIA and FRIA. The mean ratios of RIA to HPLC, FPIA to HPLC, and RIA to FPIA were 2.96, 4.14, and 0.73. The significant variations in cyclosporine levels result from the cross-reaction of antibody with some cyclosporine metabolites, by which these two methods often overestimate the true blood cyclosporine level. HPLC is a more effective and reliable method for pharmacokinetic studies and blood level monitoring of cyclosporine in clinical practice.

Adult↗

Evidence that antibodies to cytomegalovirus and the T cell receptor (TCR)/CD3 complex may have common ligands.

Immunoglobulins derived from sera containing anti-antiidiotypic antibodies (Ab2) generated in renal transplant recipients after OKT3 monoclonal antibody therapy, as reported in our previous study (1), have now been proved to bind to several bands of T cell membrane lysates (TCML) in immunoblotting analyses ranging in molecular weight from 40 to 55 KD. These sera also blocked the expression of the ligand binding to WT31 in flow cytometry. WT31 is a MAb that recognizes a common determinant on the T cell receptor (TCR). Immunoglobulins from these sera suppressed the activation of normal peripheral blood T lymphocytes (PBT) induced by OKT3. All patients (7/32) who developed this Ab2 had distinct culture-proved cytomegaloviral infections. In further immunoblotting studies, alpha F1, another MAb recognizing the framework of the TCR alpha chain, more deeply inserted in the T cell membrane, also showed binding to protein bands of cytomegalovirus pellet lysates derived from virus-infected embryonic fibroblasts. In addition, alpha F1 showed positive binding to several ligands in the membrane lysate of CMV-infected, but not noninfected MRC-5 cells. An anti-CMV MAb recognizing late nuclear antigen (LAb), also strongly bound to a approximately 50 KD band of TCML and several bands (approximately 34, approximately 40, and approximately 50 KD) of H33HJAJ1 (human T leukemia) cell lysate. Furthermore, alpha F1 immunoprecipitated a approximately 96 KD ligand of CMV-infected MRC5 lysate that had the same electrophoretic mobility as one of the proteins precipitable with LAb. Both LAb and alpha F1 also showed positive binding to paraformaldehyde-fixed and Triton X-100-permeabilized PBT in flow cytometry. Sera containing Ab2 blocked alpha F1 binding to acetone-fixed cytofuged PBT preparations on slides. Moreover, both alpha F1 and LAb inhibited mitogen-stimulated lymphocyte activation in vitro. These data support the notion that T cell functional abnormalities associated with CMV infection observed after treatment of transplant recipients with anti-T cell monoclonals might be caused by binding to T cell ligands by a variety of crossreacting human Igs operative in a regulatory network. Confirmatory evidence is the effect of MAbs generated against CMV virion epitopes crossreacting with T cell ligands, and vice versa.

Antibodies, Monoclonal↗