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Biomedical subjects

J Wu

Publications and source records attributed to J Wu.

At least 1,243 records · Page 69Linked to original sources
PubMed1994

Morin: a wood pigment that protects three types of human cells in the cardiovascular system against oxyradical damage.

Morin is a yellowish pigment extractable from the wood of Chlorophora tinctoria. In the present study, we have determined that morin protects three types of human cells--ventricular myocytes, saphenous vein endothelial cells, and erythrocytes--against damage by oxyradicals generated in situ. In myocytes and endothelial cells, morin prolonged substantially and in a concentration-dependent manner the survival of cells exposed to either xanthine oxidase-generated oxyradicals or superoxide radicals produced with menadione. Morin protected erythrocytes from lytic attack by peroxyl radicals generated with 2,2'-azo-bis (2-amidinopropane) dihydrochloride. In all three types of human cells, the protective effect of morin clearly excelled that displayed by Trolox (a vitamin E analog), ascorbate, or mannitol, which are water-soluble antioxidants of similar molecular size. Chemically, we verified that morin behaves as an antioxidant by diminishing markedly the amount of malondialdehyde (lipid peroxidation product) found in human cardiocytes despite their exposure to oxyradicals. In agreement with related reports, we also observed that morin is non-toxic in rats even when used at concentrations 2-3 orders of magnitude higher than those in our in vitro studies. Thus, morin acts as a broad-spectrum and non-toxic antioxidant.

Ascorbic Acid↗
PubMed1994

Correction of accelerated autoimmune disease by early replacement of the mutated lpr gene with the normal Fas apoptosis gene in the T cells of transgenic MRL-lpr/lpr mice.

MRL-lpr/lpr mice develop a generalized autoimmune disease which includes increased autoantibody production, glomerulonephritis, and development of lymphadenopathy. The lpr genetic defect has been identified as a mutation in the Fas apoptosis gene that results in low expression of Fas mRNA. To determine the significance of the lpr mutation and T cells in the development of the autoimmune disease, we constructed transgenic MRL-lpr/lpr mice using a full-length murine Fas cDNA under the regulation of the T-cell-specific CD2 promoter and enhancer. Here we show that the early correction of the lpr gene defect in T cells eliminates glomerulonephritis and development of lymphadenopathy and decreases the levels of autoantibodies. In this model, early correction of the lpr defect in T cells is sufficient to eliminate the acceleration of autoimmune disease even in the presence of B cells and other cells that express the mutant lpr gene.

Animals↗
PubMed1994

Rapamycin-induced long-term allograft survival depends on persistence of alloantigen.

In this study the mechanism of rapamycin-induced long-term allograft tolerance was investigated in a rat model. We have demonstrated that the tolerance is strain specific, but is not organ specific. The tolerized rats failed to generate high levels of donor-specific cytotoxic Ab and cytotoxic cells in vivo. Removal of the alloantigen from the tolerized rats with or without concomitant thymectomy could break down the status of tolerance, and the rats regained the capability to reject the grafts and to develop specific cytotoxic Ab and cytotoxic cells. These results clearly indicate that the maintenance of the rapamycin-induced long-term tolerance to allografts depends on the persistence of alloantigens. Mechanistically, we have shown that the reduced IL-2 production and the reduced antigenicity of the graft in the tolerized rat contribute to, but are not solely responsile for, the tolerance. The results of adoptive transfer experiments suggest that regulatory cells or suppressive serum factors are not involved in the tolerance. The fact that the removal of the alloantigen in the thymectomized rat could reverse the tolerance, indicates that there is no clonal deletion. We propose that chronic desensitization due to prolonged engagement of TCR by persistent alloantigens is the major mechanism at the late stage of the rapamycin-induced allograft tolerance.

Animals↗
PubMed1994

Fetal isoform of human retinoic acid receptor beta expressed in small cell lung cancer lines.

The retinoic acid receptor type beta (RAR beta) complementary DNA from a small cell tumor line was amplified, sequenced, and found to be homologous to the murine RAR beta 1. Seventeen lung tumor lines were analyzed. Five of seven small cell lung carcinoma lines expressed RAR beta 1, but only one other line (epidermoid) expressed the isoform, and this was at trace levels. Two other epidermoid lines, as well as three adenocarcinoma, two adenosquamous, and two large cell-derived lines did not express RAR beta 1. Nine adult human tissues, including lung, were analyzed, and in contrast to what has been reported for the mouse, undetectable or barely detectable levels were observed. On the other hand, a total of 13 different fetal tissues, at three different developmental stages, all expressed RAR beta 1. RAR beta 1 may be a master developmental gene in humans, and the remarkably specific association with small cell lung carcinoma suggests a molecular link between this type of cancer and development.

Amino Acid Sequence↗
PubMed1994

Autoimmune disease. A problem of defective apoptosis.

Human autoimmune diseases share the common feature of an imbalance between the production and destruction of various cell types including lymphocytes (SLE), synovial cells (RA), and fibroblasts (scleroderma). Patients with SLE have increased levels of soluble Fas that inhibit proper apoptosis of lymphocytes. In animal models of autoimmune diseases, mutations of genes involved in apoptosis including Fas, Fas ligand, and the hematopoietic cell phosphatase gene have been identified. Oncogenes, including bcl-2, p53, and myc, that regulate apoptosis are also expressed abnormally. Potent inducers of apoptosis including steroids, azathioprine, cyclophosphamide, and methotrexate are the most efficacious therapies for autoimmune disease currently known. Specific therapies that induce apoptosis without incurring side effects should improve treatment of autoimmune disease.

Antigens, Surface↗
PubMed1994

13C-labeled oligodeoxyribonucleotides: a solution study of a CCAAT-containing sequence at the nuclear factor I recognition site of human adenovirus.

The solution behavior of the single-stranded CCAAT-containing octamer 1, d(AGCCAATA), that comprises part of the nuclear factor I (NF-I) recognition site at the origin of replication of human adenovirus has been studied by nmr spectroscopy at 500 and 600 MHz. Proton resonance assignments for 1 were aided by selective 13C enrichment at C1' of A1 or A5. High-resolution 13C-1H heteronuclear multiple-bond coherence spectra of the 13C-labeled oligomers permitted the selective detection of furanosyl ring protons within each labeled residue due to short- and long-range 13C-1H couplings to the enriched C1'. The resulting assignments provided firm starting points in the interpretation of double quantum filtered correlated spectra, yielding information supplemented by total correlated spectroscopy (TOCSY) and rotating frame nuclear Overhauser effect spectroscopic data to completely assign the 1H-nmr spectrum of 1 and extract 3JHH values for furanose conformational analysis. Several 13C-1H spin-coupling constants within the 13C-enriched A1 or A5 residues were measured from cross-peak shifts in TOCSY spectra, and their signs determined by inspection of the relative orientations of these shifts. 1H-1H and 13C-1H spin-couplings both indicate a preference (> 75%) for south (C2'-endo) conformations by the furanosyl rings of 1.

Adenoviruses, Human↗
PubMed1994

The effect of rapamycin on T cell development in mice.

Rapamycin (RAPA) is a strong immunosuppressant with a chemical structure similar to that of FK506, although it acts by a mechanism different from both FK506 and cyclosporin A. The effect of RAPA on T cell development in mice was investigated in this study. RAPA caused significant thymic atrophy. The major histological change in the RAPA-treated thymus was thinning of the cortex. No other apparent damage in the cortex or medulla was observed. Consistent with these histological findings, in vivo thymocyte cycling was blocked by RAPA before the S phase, and the ex vivo and in vitro proliferation of the thymocytes was also strongly repressed by the drug. According to electron microscopy and DNA fragmentation assays, RAPA did not induce apoptosis. These results indicate that the repressed thymocyte proliferation is a major mechanism causing RAPA-induced thymic atrophy. Further, RAPA had no effect on thymocyte apoptosis induced by anti-CD3 or ionomycin, and the drug did not interfere with deletion of CD4+8+ thymocytes or peripheral V beta 6+ T cells induced by anti-CD3 or Mls-1a, respectively. These data suggest that RAPA does not hamper the negative selection. There was a relative increase in the CD3- fraction of the de novo developing CD4 and CD8 double-positive (DP) thymocytes in the RAPA-treated mice. Moreover, there were relative increases of the CD3-fractions of the CD4 or CD8 single-positive (SP) cells in both the thymi and periphery. The generation of the CD3-SP under the influence of RAPA could be used as a useful model for further study of the function and signal transduction of these CD3-defective SP cells.

Animals↗
PubMed1994

Ligand-induced conformational changes in the lactose permease of Escherichia coli: evidence for two binding sites.

By using a lactose permease mutant containing a single Cys residue in place of Val 331 (helix X), conformational changes induced by ligand binding were studied. With right-side-out membrane vesicles containing Val 331-->Cys permease, lactose transport is inactivated by either N-ethylmaleimide (NEM) or 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM). Remarkably, beta,D-galactopyranosyl 1-thio-beta,D-galactopyranoside (TDG) enhances the rate of inactivation by CPM, a hydrophobic sulfhydryl reagent, whereas NEM inactivation is attenuated by the ligand. Val 331-->Cys permease was then purified and studied in dodecyl-beta,D-maltoside by site-directed fluorescence spectroscopy. The reactivity of Val 331-->Cys permease with 2-(4'-maleimidylanilino)-naphthalene-6-sulfonic acid (MIANS) is not changed over a low range of TDG concentrations (< 0.8 mM), but the fluorescence of the MIANS-labeled protein is quenched in a saturable manner (apparent Kd approximately equal to 0.12 mM) without a change in emission maximum. In contrast, over a higher range of TDG concentrations (1-10 mM), the reactivity of Val 331-->Cys permease with MIANS is enhanced and the emission maximum of MIANS-labeled permease is blue shifted by 3-7 nm. Furthermore, the fluorescence of MIANS-labeled Val 331 -->Cys permease is quenched by both acrylamide and iodide, but the former is considerably more effective. A low concentration of TDG (0.2 mM) does not alter quenching by either compound, whereas a higher concentration of ligand (10 mM) decreases the quenching constant for iodide by about 50% and for acrylamide by about 20%.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1994

CGRP in the serum of endotoxin-treated rats suppresses lymphoproliferation.

Mouse mesenteric lymph node cells were incubated with concanavalin A (Con A) with or without the rat form of calcitonin gene-related peptide (rCGRP) (0.1 fM-1 microM) +/- human (h)CGRP8-37 (1 microM) for 48 h. DNA synthesis was assessed by [3H]thymidine incorporation. Con A-stimulated DNA synthesis was suppressed by 13, 20, and 30% at 10 fM, 1 pM, and 100 pM of rCGRP, respectively. hCGRP8-37 (1 microM), a selective blocker of CGRP1 receptor, completely inhibited the suppression of DNA synthesis by rCGRP (10 fM-100 pM). rCGRP caused concentration-dependent elevations of cAMP levels, which were potentiated by pretreatment with 3-isobutyl-1-methylxanthine (0.3 mM, 10 min), an inhibitor of cAMP-phosphodiesterase. hCGRP8-37 (1 microM) significantly inhibited cAMP elevations induced by rCGRP at the lower concentrations, but not at the highest concentrations of rCGRP. These data suggest that rCGRP, at circulating levels (1-10 pM), appears to directly interact with receptors on mouse mesenteric lymph node cells that are coupled to cAMP generation, ultimately inhibiting lymphocyte proliferation. To test the involvement of CGRP in suppression of lymphocyte proliferation by serum from endotoxin-treated rats, mouse mesenteric lymph node cells were stimulated with Con A with or without dilutions of endotoxin treated rat serum. At a 1:20 dilution, DNA synthesis was suppressed 30%, at a 1:40 dilution, DNA synthesis was suppressed by 34%, and at a 1:80 dilution, DNA synthesis was suppressed 25%. At all serum dilutions, coincubation with hCGRP8-37 (1 microM) significantly inhibited the suppressive effect of the endotoxin treated rat serum. These data suggest that the immunosuppression observed during endotoxin shock may be due, at least in part, to CGRP in serum.

Animals↗
PubMed1994

Defective clonal deletion and anergy induction in TCR transgenic lpr/lpr mice.

Although autoreactive T cells are thought to play a prominent role in autoimmune disease in MRL-lpr/lpr mice, it has been difficult to directly determine if autoreactive T cells escape from the thymus and react with self-antigens in the periphery. To identify a possible defect in clonal deletion or clonal anergy induction of auto-specific T cells, we have studied C57BL/6-lpr/lpr transgenic mouse expressing TCR genes that recognize a known self-antigen, the male H-Y antigen and analyzed clonal deletion and tolerance induction after neonatal tolerance induction with the class II MHC reactive superantigen staphylococcal enterotoxin B (SEB) in V beta 8 TCR transgenic and non-transgenic MRL-lpr/lpr mice. In lpr/lpr mice, the main defect was a thymic-dependent loss of self-tolerance by auto-specific T cells and also a small defect of clonal deletion of autoreactive thymocytes. Defective expression of the Fas apoptosis antigen results from the insertion of the ETn retrotransposon. The fas defect can be partially corrected in TCR-beta TCR transgenic mice in which accelerated T cell development prevents the lymphoproliferative disease and in CD2-fas transgenic mice in which fas expression is corrected in T cells. These results suggest that expression of the TCR-beta, fas and retrovirus genes are co-regulated during early thymocyte development, most likely by common enhancer transcription factors.

Animals↗
PubMed1994

Donor's and recipient's antigen presenting cells co-operatively enhance the allo-reactive proliferation of peripheral blood lymphocytes.

Antigen-presenting cells (APC) play important roles in the allo-reactive proliferation of T cells in vitro and in the allo-graft rejection in vivo. This work was aimed at establishing whether a co-operation of any kind occurs between donor's and recipient's APC during allo-reactions, and whether certain cell surface molecules, such as beta 2-microglobulin (beta 2m) and human leucocyte antigens (HLA), are involved in such an interaction. The data herein reported show that the allo-reactive proliferation of peripheral blood lymphocytes (PBL) is markedly intensified by a positive co-operation between donor's and recipient's APC. Such a synergistic co-stimulatory action can be significantly hindered by a monoclonal antibody aimed against beta 2m while being not changed by monoclonal antibodies binding to either HLA-A-B-C or HLA-DR molecules. Hence, our preliminary results indicate that a positive interaction between donor's and recipient's APC may be of importance for allo-graft rejection.

Antigen-Presenting Cells↗
PubMed1994

The opposing effects of an inhibitor of nitric oxide synthesis and of a donor of nitric oxide in rabbits undergoing myocardial ischemia reperfusion.

We observed that N-nitro-L-arginine (NOLA), a nitric oxide biosynthesis inhibitor, exacerbated necrosis in the rabbit heart during ischemia-reperfusion while 3-morpholino-sydnonimine-hydrochloride (SIN-1) (a nitric oxide donor) reduced myocardial damage in the same model. In rabbits undergoing 1-h ligation of the anterior ventricular coronary artery, a single bolus injection of NOLA (30 mg/kg) or continuous infusion of SIN-1 (3 mg/kg) were introduced into the post-ischemic heart immediately before 4-h reperfusion. Against negligible necrosis in 6 sham-operated control animals, and 33.8 (SD 13.5)% necrosis in the area at risk for the saline control group (n = 8), the NOLA-treated group (n = 8) had a necrosis of 44.3 (SD 8.6)% whereas the SIN-1-treated group (n = 10) showed a necrosis of 16.8 (SD 4.9)% (both with p < 0.05 vs saline control group). The pressure-rate index increased in the NOLA-treated group but decreased in the SIN-1-treated group. These data support the contention that a nitric oxide donor is an effective cardioprotector during ischemia-reperfusion in vivo.

Analysis of Variance↗
PubMed1994

Purpurogallin: in vivo evidence of a novel and effective cardioprotector.

We observed that purpurogallin (PPG) which is a flavonoid markedly protects the rabbit against myocardial ischemia-reperfusion injury. In rabbits undergoing 1-h ligation of the anterior ventricular coronary artery, a bolus infusion of PPG was introduced into the post-ischemic heart immediately before 3-h reperfusion. Against negligible necrosis in 6 sham-operated controls, and 41.7 (SD 11.3)% necrosis in the area at risk for the placebo control group (n = 14 animals), the PPG-treated groups (n = 6, 6, 14) had a necrosis of 26.8 (6.4)%, 10.8 (3.5)%, and 11.7 (5.2)% at doses of 2.5, 5, and 10 mumol/kg, respectively (each with p < 0.01 vs control value). By comparison, infusion of Trolox (a vitamin E analogue) at 5 mumol/kg produced a higher necrosis of 17.7 +/- 7.2% (n = 6, p < 0.05 vs value obtained from 5 mumol/kg PPG-treated group) in the same model. Note that myocardial necrosis was estimated by tetrazolium-based histochemistry and confirmed by light and transmission electron microscopies. These data support our contention that PPG is an effective cardioprotector, whose mechanism of action will be reported separately.

Analysis of Variance↗
PubMed1994

An extended cellular space method for simulating autocatalytic oligonucleotides.

Self-replicating nucleotides and other self-replicating molecules are an active area of study today by organic chemists. Such studies are important for improving our understanding of the origins of life. Computational studies of self-replicating molecules could increase our insight into their properties, but existing computational techniques have been limited in their usefulness for such reactions (numerical simulation of differential equations requires reaction rate constants that are difficult to obtain, cellular automata models are too restrictive for modeling molecular movements and bindings, etc.). We have thus developed an efficient modified cellular automata method that supports the study of self-replicating oligonucleotides. We explain this method and illustrate its use with a specific self-replicating (autocatalytic) deoxyribohexanucleotide.

Algorithms↗
PubMed1994

Quantitative polymerase chain reaction for hepatitis B virus DNA.

To monitor and compare the effect of drugs on hepatitis B virus (HBV) replication in hepatitis patients, an accurate quantitative method with high sensitivity is needed. Polymerase chain reaction (PCR) is the most sensitive method for the detection of HBV DNA, but because of sample to sample variability in amplification efficiency, the quantification of target nucleic acids by conventional PCR is inaccurate. Therefore, we developed a competitive PCR method with an internal standard which uses the same primers as the target HBV DNA. The standard was generated from the HBV S gene by PCR site-directed mutagenesis and differed from the native S gene by a single base which introduced an internal restriction enzyme site. For practical application, this method was used to quantitate the effect of the carbocyclic analogue 2'-deoxyguanosine (2'-CDG) on HBV DNA replication in 2.2.15 cells, an HBV transfected HepG2 cell line. A decrease of HBV DNA level in the attomole range was demonstrated by the assay in the media of 2.2.15 cells treated with 2'-CDG. The results were verified by conventional HBV DNA slot blot analysis followed by densitometric scanning. Competitive PCR proved to be a sensitive, accurate and reliable method for HBV quantitation.

Antiviral Agents↗
PubMed1994

Effect of acoustic streaming on ultrasonic heating.

It was found in in vitro experiments performed on tissue phantoms that acoustic streaming in low attenuating fluid may dramatically reduce the temperature rise generated by ultrasound at the surface of bone, if the ultrasound path contains a low-attenuating-fluid/thin-soft-tissue/bone structure. The temperature rise at the surface of bone generated by a 3.5 MHz focused ultrasound beam of 180 mW source acoustic power and an average acoustic intensity at the focus of 5.8 W/cm2 was measured to be only 18% of the value predicted by an existing model, when the bone was in contact with the low attenuating fluid. When a bone surface that is covered by 1 mm thick soft tissue is exposed to the same ultrasound beam, the measured temperature rise at the tissue-bone surface is only 60% of the calculated value.

Acoustics↗