Biomedical subjects

M C Chen

Publications and source records attributed to M C Chen.

At least 55 records · Page 3Linked to original sources

PubMed2000

A safe and effective regimen without heparin therapy after successful primary coronary stenting in patients with acute myocardial infarction.

Short-term heparin therapy has been administered routinely after primary coronary stenting. However. heparin therapy results in a significantly higher incidence of bleeding and vascular complications. A new therapeutic regimen of ticlopidine and aspirin without further heparin after coronary stenting in patients without AMI has been shown to be safe and reduce the incidence of stent thrombosis. The aim of this study was to evaluate whether a new therapeutic regimen of aspirin and ticlopidine without heparin is safe and effective in patients with acute myocardial infarction (AMI) who have undergone primary coronary stenting and have Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow in the infarct-related artery. Between January 1997 and September 1999, one hundred and fifty two consecutive patients with AMI on Killip score 1 or 2 who underwent primary coronary stenting resulting in TIMI grade 3 flow were enrolled and divided into two groups: Group 1 (n = 95 patients) received aspirin, ticlopidine and further intravenous heparin infusion for 48 hours following primary coronary stenting; Group 2 (n = 57 patients) received only aspirin and ticlopidine without further heparin therapy following primary coronary stenting. No in-hospital major cardiac events were observed in either group. However, the combined incidence of bleeding and vascular complications (27.4% vs 12.3%, p = 0.029) and the need for blood transfusions (9.5% vs 0%, p = 0.013) were significantly higher in Group I patients. Furthermore, hospital stay was also longer in Group I patients (5.8+/-2.4 vs 4.7+/-1.7 days, p = 0.0003). At the 30-day follow-up, there were no differences (1.05% vs 0%, p = 0.63) in the combined incidence of vascular complications and the major cardiac events were similar (1.05% vs 1.75%, p = 0.71) between the groups. The results suggest that further heparin therapy following primary coronary stenting increases the combined incidence of bleeding and vascular complications as well as the need for blood transfusions and prolongs the length of hospital stay without further benefit to those patients with coronary flow restored to TIMI 3 grade flow.

Aged↗

PubMed2000

Cytokines induce apoptosis in beta-cells isolated from mice lacking the inducible isoform of nitric oxide synthase (iNOS-/-).

Prolonged exposure of rodent beta-cells to combinations of cytokines induces the inducible form of nitric oxide synthase (iNOS) expression and Fas expression, nitric oxide (NO) production, and cell death. It also induces the expression of potential "defense" genes, such as manganese superoxide dismutase (MnSOD) and heat shock protein (hsp) 70. NO is a radical with multifaceted actions. Recent studies have shown that NO, in addition to having cytotoxic actions, may regulate gene transcription. It remains unclear whether NO mediates cytokine-induced gene expression and subsequent beta-cell death. Previous studies using NO synthase blockers yielded conflicting results, which may be due to nonspecific effects of these agents. In this study, we examined the effects of cytokines on gene expression, determined by reverse transcriptase-polymerase chain reaction (RT-PCR), and viability, determined by nuclear dyes, of pancreatic islets or fluorescence-activated cell sorter (FACS)-purified beta-cells isolated from iNOS knockout mice (iNOS-/-, background C57BL/6x129SvEv) or their respective controls (C57BL/6x129SvEv). The combination of cytokines used was interleukin-1beta (50 U/ml) plus gamma-interferon (1,000 U/ml) plus tumor necrosis factor-alpha (1,000 U/ml). The lack of cytokine-induced iNOS activity in the iNOS-/- islet cells was confirmed by RT-PCR and nitrite determination. Cytokines induced a >3-fold increase in Fas and MnSOD mRNA expression in wild-type (WT) and iNOS-/- islets. On the other hand, hsp 70 was induced in WT but not in iNOS-/- islets. Prolonged (6-9 days) exposure of WT islets to cytokines leads to an 80-90% decrease in islet cell viability, whereas viability decreased by only 10-30% in iNOS-/- islet cells. To determine the mode of cytokine-induced cell death, FACS-purified beta-cells were exposed to the same cytokines. After 9 days, the apoptosis index was similarly increased in WT (39 +/- 3%) and iNOS4-/- (33 +/- 4%) beta-cells. On the other hand, cytokines increased necrosis in WT (20 +/- 4%) but not in iNOS-/- (7 +/- 3%) beta-cells. From these data, we concluded that 1) NO is required for cytokine-induced hsp 70 mRNA expression but not for Fas and MnSOD expression, 2) cytokines induce both apoptosis and necrosis in mouse beta-cells, and 3) cytokine-induced apoptosis is mostly NO-independent, whereas necrosis requires NO formation.

Adult↗

PubMed2000

TT virus infection among hemodialysis patients at a medical center in Taiwan.

Although the association between TT virus (TTV) infection and hepatitis is controversial, the high prevalence of TTV infection in healthy blood donors and even higher rate among frequently transfused patients poses a potential threat to public health and clinical care. In addition, there is a lack of data concerning the prevalence and mode of transmission of TTV infection in different subpopulations in Taiwan. In the present study, we investigated the prevalence of TTV infection in 111 uremic patients receiving regular hemodialysis in a single hospital in Taiwan. Blood samples were collected and analyzed using a seminested polymerase chain reaction (PCR) designed to amplify a 271 base-pair DNA fragment. The results show that the overall TTV positive rate in uremic patients in our hospital was 61% (68/111), which was much higher than the reported TTV prevalence rate among the normal population (ranging from 1%-12%). The results of analysis of the demographic and clinical characteristics of the patients indicate that blood transfusion may play an important role in TTV transmission (p < 0.05). In addition, the hepatitis B positive rate was significantly lower in TTV positive patients. However, liver function tests were not significantly different between TTV positive and TTV negative patients. The results of the present study suggest that blood transfusion plays an important role in TTV transmission in uremic patients.

Adult↗

PubMed2000

Costimulatory molecules expression and cytokine profiles of cord blood mononuclear cells in newborns with low and high risk of developing atopic diseases.

This study sought to determine predictors of atopic diseases in newborns. We evaluated the levels of expression of costimulatory molecules (CD80 and CD86) and the production of cytokines [interleukin (IL)-12, interferon (IFN)-gamma, IL-4, IL-10] in the cord blood of mononuclear cells in high risk newborns (n = 17), and compared them with those in low risk newborns (n = 25). Fluorescence-activated cell sorter (FACScan) analysis was performed to determine the expressions of CD80 and CD86 on activated B cells and monocytes of both groups. The levels of IL-10, IL-12p40 and IL-12p70 in the supernatant were assayed by enzyme-linked immunosorbent assay (ELISA), and also the mRNA levels by reverse transcription-polymerase chain reaction (RT-PCR). Intracellular staining of IL-4 and IFN-gamma in stimulated mononuclear cells was also performed as well. The expressions of CD80 and CD86 on B cells showed no significant difference between the high and low risk group. There was greater expression of CD86 on the monocytes of low risk newborns as compared to high risk newborns (p < 0.05). When B cells and monocytes isolated from the cord blood of both groups were stimulated with mitogens, the production of IL-10, IL-12p40, and IL-12p70 in the supernatants was not significantly different. The expressions of mRNA of IL-10, IL-12p35, and IL-12p40, and the intracellular staining of IL-4 and IFN-gamma in stimulated mononuclear cells were not significantly different between the two groups. These findings suggested that cytokine profiles in the cord blood cannot predict the development of atopic diseases. Determination of whether preferential expression of costimulatory molecules is of predictive value or not will require further study.

Antigens, CD↗

PubMed2000

No sex and age influence on the expression pattern and activities of human gastric alcohol and aldehyde dehydrogenases.

BACKGROUND: Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are the principal enzymes responsible for ethanol metabolism in humans. The stomach is involved in the metabolism of alcohol during absorption. Conflicting reports exist with regard to the influence of sex and age on the activity of ADH in the human gastric mucosa. The purpose of the present study was to determine the effects of age and sex on the expression pattern and activities of stomach ADH and ALDH. METHODS: A total of 115 endoscopic gastric biopsy specimens were investigated from Han Chinese men (n = 70) and women (n = 45) aged 20-79 years with approximately even distribution among 10-year age intervals. The expression patterns of ADH and ALDH were identified by isoelectric focusing, and the activities were assayed spectrophotometrically. RESULTS: The expression patterns of gastric ADH and ALDH remained unchanged with respect to sex and age. At 33 mM or 500 mM ethanol, pH 7.5, the ADH activities did not differ significantly among the various age groups or between men and women. At 200 microM or 20 mM acetaldehyde, the ALDH activities did not differ significantly in relation to sex and age. No correlations were found between the ADH or ALDH activities at both the high and low substrate concentrations and the ages in men and women. CONCLUSIONS: The results indicate that there is no significant effect of either sex or age on the expression pattern and activity of ADH and ALDH in the human gastric mucosa. The stomach ADH seems unlikely to account for possible variations in the first-pass metabolism of alcohol with regard to sex and age.

Adult↗

PubMed1999

Incidence of highly genetically diversified Vibrio parahaemolyticus in seafood imported from Asian countries.

Vibrio parahaemolyticus is an important pathogen of humans and aquacultured animals, especially in Asian countries. In this study, we examined 686 samples of seafood imported from Hong Kong, Indonesia, Thailand and Vietnam for V. parahaemolyticus. V. parahaemolyticus was recovered from 315 (45.9%) samples. The incidence of V. parahaemolyticus in products from Hong Kong and Thailand was markedly higher than the incidence in products from Indonesia and Vietnam. The incidence rates in shrimp, crab, snail, lobster, sand crab, fish and crawfish were 75.8, 73.3, 44.3, 44.1, 32.5, 29.3 and 21.1%, respectively. None of the isolates possessed the hemolysin genes (tdh, trh). The chromosomal DNA of 121 randomly selected imported isolates and three local environmental strains was digested with SfiI and analyzed by pulsed-field gel electrophoresis (PFGE). These isolates were then grouped into 96 different but mostly unique PFGE patterns. After hierarchical cluster analysis, these patterns could be arbitrarily grouped into twenty-two PFGE types (type A to V). A wide range of PFGE types were identified in isolates from different origin. Moreover, the PFGE types were not specifically associated with the origin or kind of seafood. These results reveal the high genetic diversity in V. parahaemolyticus isolated from seafood.

Animals↗

PubMed1999

Inhibition of human pancreatic islet insulin release by receptor-selective somatostatin analogs directed to somatostatin receptor subtype 5.

Somatostatin (SS)-14 and SS28 are produced by pancreatic D cells and gut mucosa and inhibit pancreatic islet insulin and glucagon release. There are five distinct SS receptor (SSTR) subtypes, namely SSTR1-5, which show different affinities for SS14 and SS28. In order to identify the subtype responsible for inhibition of insulin release by human B cells, SSTR-selective SS analogs were tested in isolated human islets. Glucose-stimulated insulin secretion in human islets incubated for 1 hr at 20 mM glucose, and in islets cultured for 24 hr at a near-physiological (6.1 mM) glucose concentration, was inhibited (<50% of the control) by SSTR5-specific analogs and by SS14 and SS28. SS14, SS28, and different SSTR5 preferential analogs also inhibited islet amyloid polypeptide release during the 24-hr culture. On the other hand, a group of SSTR2-selective analogs failed to inhibit insulin release. Analysis by reverse transcription-polymerase chain reaction indicated that human islets express similar amounts of SSTR2 and SSTR5 mRNAs, while human pancreatic ductal cells express much lower levels of these mRNAs. In conclusion, our data suggest that SSTR5 is an important mediator of the insulin inhibitory action of SS in cultured human islets.

Amyloid↗

PubMed1999

Induction of p21(CIP1/Waf1) and activation of p34(cdc2) involved in retinoic acid-induced apoptosis in human hepatoma Hep3B cells.

The biological activity of retinoic acid (RA) was examined in human hepatoma Hep3B cells. Under serum-deprived conditions, RA induced S/M-phase elevation and mitotic index increase within 24 h, followed by apoptosis. This RA-induced apoptosis was accompanied by p53-independent up-regulation of endogenous p21(CIPI/Waf1) and Bax proteins, as well as activation of p34(cdc2) kinase, and increase of Rb2 protein level and phosphorylation pattern. In addition, RA had no effect on the levels of Bcl-XL; Bcl-XS; cyclins A, B, D1, D3, or E; or Rb1 expression but markedly down-modulated Cdk2 kinase activity and reduced Cdk4 expression. RA also slightly delayed p27(Kip1) expression. Olomoucine, a potent p34(cdc2) and Cdk2 inhibitor, effectively blocked RA-mediated p34(cdc2) kinase activation and prevented RA-induced apoptosis. Furthermore, antisense oligonucleotide complementary to p21(CIP2/Waf1) and p34(cdc2) mRNA significantly rescued RA-induced apoptosis. Our data indicate that p21(CIP2/Waf1) overexpression may not be the only regulatory factor necessary for RA-induced apoptosis in human hepatoma Hep3B cells. RA treatment leads to Rb2 hyperphosphorylation, and p34(cdc2) kinase activation is coincident with an aberrant mitotic progression, followed by appearance of abnormal nucleus. This aberrant cell cycle progression appeared requisite for RA-induced cell death. These findings suggest that inappropriate regulation of the cell cycle regulators p21(CIP2/Waf1) and p34(cdc2) is coupled with induction of Bax and involved in cell death with apoptosis when Hep3B cells are exposed to RA.

Apoptosis↗

PubMed1999

Short- and long-term outcomes after percutaneous transluminal coronary angioplasty in chronic hemodialysis patients.

The aim of this study was to obtain data on the outcomes of chronic hemodialysis patients who underwent percutaneous transluminal coronary angioplasty (PTCA). A retrospective chart analysis identified 31 such patients between August 1992 and October 1996. The mean follow-up period was 12.4 +/- 11.7 months. Angiographic success was achieved in 39 of 41 (95.1%) stenoses attempted. There were three in-hospital deaths. Clinical success was achieved in 28 of 31 patients (90%). Two of the 28 survivors were lost to follow-up. Recurrent angina developed within 6 months in 14 of 26 patients (53.8%). Eleven and 17 of the 26 patients (42.3% and 65.4%) died within 6 and 14 months, respectively, after the PTCA procedure. Ten of the 17 deaths (58.8%) were due to cardiovascular events. Our study suggests that PTCA is technically feasible with high angiographic success rate in chronic hemodialysis patients. In-hospital mortality rate and rate of recurrent angina are high. Long-term prognosis is poor.

Aged↗

PubMed1999

Interleukin 1beta increases arginine accumulation and activates the citrulline-NO cycle in rat pancreatic beta cells.

Nitric oxide (NO) may contribute to pancreatic beta cell damage during the development of type 1 diabetes. Its formation can be triggered by cytokines which induce the expression of the inducible form of nitric oxide synthase (iNOS) in pancreatic islets. In the iNOS-catalyzed reaction, arginine is converted into citrulline and NO. Cellular NO formation may be regulated by the availability of arginine. Arginine can be provided extracellularly, entering the cell mainly through the cationic amino acid transporter system y+CAT, and intracellularly, by protein degradation or synthesis from citrulline (the citrulline-NO cycle). This study demonstrates for the first time that the citrulline-NO cycle is induced in FACS-purified rat beta cells exposed to interleukin-1beta(IL-1beta) and that extracellular arginine or citrulline is required for NO production by beta cells. Moreover, the accumulation of arginine was higher in IL-1beta-treated beta cells than in control cells.beta cells expressed mRNAs for the two y+CAT transporters CAT-2A and CAT-2B with no change in transporter expression after exposure to IL-1beta. It is concluded that the activation of the citrulline-NO cycle and an increase in arginine accumulation may be adaptive responses in cytokine-exposed beta-cells to assure an adequate arginine supply for continuous NO production in the presence of low extracellular arginine levels which may prevail during insulitis.

Amino Acid Transport Systems↗

PubMed1999

IL-1beta induces serine protease inhibitor 3 (SPI-3) gene expression in rat pancreatic beta-cells. Detection by differential display of messenger RNA.

Immune-mediated beta-cell damage induces diverse intracellular signals, leading to transcription of different genes which may either contribute to beta-cell repair and/or defence or lead to cell death. The cytokine interleukin-1beta (IL-1) is a potential mediator of beta-cell dysfunction and damage in type 1 diabetes mellitus. To understand the molecular actions of this cytokine upon beta-cells, this study aimed at the cloning of genes induced in FACS-purified rat pancreatic beta-cells by a 6- or 24-h exposure to IL-1 by using differential display of mRNA with reverse transcription-polymerase chain reaction (DDRT-PCR). Among these cytokine-induced genes, a gene encoding for rat serine protease inhibitor (SPI-3) was isolated. SPI-3 may be involved in cellular defence responses against inflammatory stress. RT-PCR analysis confirmed that SPI-3 mRNA expression in rat beta-cells is increased by IL-1 at an early stage (2 h), with maximal accumulation during 6-12 h and decline after 24 h. Similar observations were made in mouse pancreatic islets and in the rat insulinoma cell line RINm5F. IFN-gamma neither increased SPI-3 gene expression nor potentiated its induction by IL-1 in rat beta-cells. The stimulatory effects of IL-1 on SPI-3 mRNA expression were decreased by co-incubation with an inhibitor of gene transcription (actinomycin D), an inhibitor of protein synthesis (cycloheximide) or an inhibitor of NF-kappaB activation (PDTC). On the other hand, a blocker of inducible nitric oxide synthase (iNOS) activity (N(G)-methyl-L-arginine) did not prevent IL-1-induced SPI-3 expression. Thus, SPI-3 mRNA expression following IL-1 exposure depends on gene transcription, protein synthesis and activation of the nuclear transcription factor NF-kappaB, but it is independent of NO formation.

Acute-Phase Proteins↗

PubMed1999

Identification of IL-1beta-induced messenger RNAs in rat pancreatic beta cells by differential display of messenger RNA.

AIMS/HYPOTHESIS: Interleukin-1beta is a putative mediator of pancreatic beta-cell dysfunction and damage in Type I (insulin-dependent) diabetes mellitus. To better understand the molecular mechanisms involved in IL-1beta effects, we carried out a differential display of mRNA by RT-PCR to identify novel cytokine-regulated genes. METHODS: Fluorescence activated cell sorting-purified rat pancreatic beta-cells were exposed for 6 or 24 h to IL-1beta. Differentially expressed cDNA bands were cloned and then identified by comparing their sequences with data from the GenBank. Differential gene expression was confirmed by RT-PCR using specific primers. RESULTS: Interleukin-1beta increased the expression of adenine nucleotide translocator-1, phospholipase D-1 and cytokine-induced neutrophil chemoattractant-1 and decreased expression of the protein tyrosine phosphatase-like protein IA-2. Interleukin-1beta-induced differential expression of these genes in beta cells was confirmed by RT-PCR. In additional studies, IL-1beta was shown to induce chemokines other than cytokine-induced neutrophil chemoattractant-1, including cytokine-induced neutrophil chemoattractant-3 and monocyte chemotactic protein-1. CONCLUSION/INTERPRETATION: Our observations indicate that IL-1beta modifies the expression of several genes in pancreatic beta cells. These genes may affect both function, viability and beta-cell recognition by the immune system. Functional characterization of the mRNAs which have been identified could facilitate a better understanding of the mechanisms leading to beta-cell destruction in Type I diabetes. [Diabetologia (1999) 42: 1199-1203]

Animals↗

PubMed1999

Therapeutic benefits of infrared (810-nm) diode laser macular grid photocoagulation in prophylactic treatment of nonexudative age-related macular degeneration: two-year results of a randomized pilot study.

OBJECTIVE: This pilot study collected preliminary information on the effectiveness and safety of infrared (810-nm) diode laser macular grid photocoagulation in patients with nonexudative age-related macular degeneration (AMD). Results from this pilot study were used in designing a larger, multicenter, randomized clinical trial. DESIGN: A multicenter, randomized, controlled, clinical trial. PARTICIPANTS: A total of 229 eyes of 152 patients with AMD were enrolled in the pilot study. Seventy-five patients with 1 eye eligible (75 eyes) were enrolled in the unilateral arm of the study; 77 patients with both eyes eligible (154 eyes) were enrolled in the bilateral arm of the study. In the unilateral study arm, 32 eyes were randomized to the observation group, 27 eyes were treated with visible endpoint burns, and 16 eyes were treated with invisible endpoint (subthreshold) lesions. In the bilateral study arm, 77 eyes were in the observation group, 36 eyes were treated with visible burns, and 41 eyes were treated with subthreshold (invisible) lesions. INTERVENTION: Eyes were treated with infrared (810-nm) diode laser macular grid photocoagulation using either visible burns or subthreshold (invisible) lesions and compared to eyes receiving no treatment. MAIN OUTCOME MEASURES: Reduction of drusen, change in visual acuity, and rate of choroidal neovascularization (CNV) membrane formation. RESULTS: At 12 months after treatment, 62% of eyes treated with visible burns had a clinically significant reduction in drusen, whereas this proportion (65%) was reached in 18 months for eyes treated with subthreshold lesions. At 24 months' follow-up, treated eyes had a significant reduction in drusen compared to observation eyes (P < 0.0001). Visual acuity was significantly improved in treated eyes at 12, 18, and 24 months compared to observation eyes (P < 0.001). Choroidal neovascularization formation was similar in treated and observation eyes through 24 months' follow-up. Complications included CNV associated with six eyes treated with visible burns and a juxtafoveal laser scar in one eye treated with visible burns. CONCLUSIONS: Infrared (810-nm) diode laser macular grid photocoagulation in patients with nonexudative AMD significantly reduces drusen levels (P < 0.0001) and significantly improves visual acuity (P < 0.001) when either visible endpoint burns or subthreshold endpoint lesions are used. Complications were fewer using subthreshold endpoint lesions. A larger, multicenter, prospective clinical trial with longer follow-up is needed to determine the efficacy of treatment in reducing the rate of CNV formation. Data from this clinical pilot study have been used to design the Prophylactic Treatment of AMD Trial (PTAMD), a multicenter, randomized, prospective clinical trial currently in progress comparing subthreshold (invisible) treatment to observation in eyes with nonexudative AMD.

Aged↗

PubMed1999

Cultured composite autografts as coverage for an extensive body surface area burn: case report and review of the technology.

Cultured epithelial autografts (CEA) have been used as an adjunct in the surgical management of extensive thermal burns. Unfortunately, the lack of a dermal matrix makes CEA susceptible to infection, shearing forces and limits their incorporation into the burn wound. A cultured composite autograft (CCA) has been developed in which autologous keratinocytes and fibroblasts are surgically harvested from the burn patient's normal skin. These components are proliferated and then combined to form an epidermal and dermal matrix, grown to confluence then applied. Standard wound coverage techniques as well as CCA technology were utilized for successful wound closure in a 12 yr-old female with an 81% third degree burn. After fascial excision and allograft coverage, autografts were placed on her posterior burns and then 7500 cm2 of CCA was placed onto her anterior thorax, abdomen and lower extremities. Sixty percent of the burn was covered with CCA resulting in a success rate of 40%. No evidence of infection was noted, even in areas where CCA failed, although in those areas random epithelialization appeared to occur which then seemed to facilitate autograft placement. Early debridement and allografting followed by conventional autografts and CCA placement may provide an effective skin coverage strategy in patients with extensive deep burns.

Burns↗

PubMed1999

Junctional tachycardia during radiofrequency ablation of the slow pathway in patients with AV nodal reentrant tachycardia: effects of autonomic blockade.

INTRODUCTION: The autonomic nervous system richly innervates the peri-AV nodal region and may be activated during radiofrequency (RF) ablation for AV nodal reentrant tachycardia, resulting in the generation of junctional tachycardia. The purpose of this prospective study was to determine the role of the autonomic nervous system in the genesis of junctional tachycardia. METHODS AND RESULTS: We compared the characteristics of junctional tachycardia in patients with (n = 10) and without (n = 10) autonomic blockade undergoing RF ablation for AV nodal reentrant tachycardia. Intravenous administration of atropine (0.04 mg/kg) and propranolol (0.2 mg/kg) were used to block the autonomic nervous system. There were no differences in clinical variables and baseline electrophysiologic characteristics between the two groups except for slightly longer effective refractory periods of the fast pathway and of the atrium in the autonomic blockade group. The autonomic blockade shortened the baseline sinus cycle length and effective refractory period of the ventricle only but not other electrophysiologic characteristics of the AV node. The junctional tachycardia was observed during ablation in each patient, but its occurrence and cycle length, as well as numbers of consecutive junctional beats, were not altered by the autonomic blockade. CONCLUSION: Our results indicate that the muscarinic and beta-adrenergic components of the autonomic nervous system play no role in the genesis of junctional tachycardia.

Atrioventricular Node↗

PubMed1999

Membrane permeabilization by small hydrophobic nonstructural proteins of Japanese encephalitis virus.

Infection with Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, may cause acute encephalitis in humans and induce severe cytopathic effects in various types of cultured cells. We observed that JEV replication rendered infected baby hamster kidney (BHK-21) cells sensitive to the translational inhibitor hygromycin B or alpha-sarcine, to which mock-infected cells were insensitive. However, little is known about whether any JEV nonstructural (NS) proteins contribute to virus-induced changes in membrane permeability. Using an inducible Escherichia coli system, we investigated which parts of JEV NS1 to NS4 are capable of modifying membrane penetrability. We found that overexpression of NS2B-NS3, the JEV protease, permeabilized bacterial cells to hygromycin B whereas NS1 expression failed to do so. When expressed separately, NS2B alone, but not NS3, was sufficient to alter bacterial membrane permeability. Similarly, expression of NS4A or NS4B also rendered bacteria susceptible to hygromycin B inhibition. Examination of the effect of NS1 to NS4 expression on bacterial growth rate showed that NS2B exhibited the greatest inhibitory capability, followed by a modest repression from NS2A and NS4A, whereas NS1, NS3, and NS4B had only trivial influence with respect to the vector control. Furthermore, when cotransfected with a reporter gene luciferase or beta-galactosidase, transient expression of NS2A, NS2B, and NS4B markedly reduced the reporter activity in BHK-21 cells. Together, our results suggest that upon JEV infection, these four small hydrophobic NS proteins have various modification effects on host cell membrane permeability, thereby contributing in part to virus-induced cytopathic effects in infected cells.

Animals↗

PubMed1999

Effects of growth factors and trefoil peptides on migration and replication in primary oxyntic cultures.

Restitution, the lateral migration of cells over an intact basement membrane, maintains mucosal integrity. We studied the regulation of migration and proliferation of enzyme-dispersed canine oxyntic mucosa cells in primary culture. Confluent monolayers were wounded and cultured in serum-free medium, and cells migrating into the wound were counted. [3H]thymidine incorporation into DNA was studied using subconfluent cultures. Considerable migration occurred in untreated monolayers; however, epidermal growth factor (EGF), transforming growth factor (TGF)-alpha, basic fibroblast growth factor (bFGF), insulin-like growth factor I (IGF-I), two trefoil peptides, and interleukin (IL)-1beta further enhanced migration. The specific EGF receptor (EGFR) monoclonal antibody, MAb-528, inhibited both basal and TGF-alpha- or IL-1beta-stimulated migration, but not the response to trefoil peptide, bFGF, or IGF-I. Exogenous TGF-beta inhibited cell proliferation but did not alter migration. Immunoneutralization with anti-TGF-beta blocked the response to exogenous TGF-beta and produced a small enhancement of basal thymidine incorporation but did not attenuate basal or TGF-alpha-stimulated migration. In conclusion, endogenous EGFR ligands regulate proliferation and migration. TGF-beta inhibits mitogenesis; it did not upregulate migration in these cultures. Although bFGF, IGF-I, and IL-1beta enhance gastric epithelial migration, only IL-1beta acted in a TGF-alpha-dependent fashion.

Animals↗

PubMed1999

Feline immunodeficiency virus subtype C is prevalent in northern part of Taiwan.

The seroepidemiological survey of cats conducted in northern part of Taiwan in 1998 revealed that the positive rate of feline immunodeficiency virus (FIV)-infection was 21.9% (7/32) and the rate was much higher than those of previous reports. We succeeded in isolation of three strains of FIV from peripheral blood mononuclear cells of the blood samples. Nucleotide sequences of the env variable V3 to V5 region of the strains revealed that the isolates from distinct areas belong to subtype C. These data together with our previous report (Inada et al. 1997. Arch. Virol., 142: 1459-1467) indicate that FIV subtype C is prevalent in northern part of Taiwan.

Amino Acid Sequence↗