Biomedical subjects

W Tsai

Publications and source records attributed to W Tsai.

At least 19 recordsLinked to original sources

PubMed2003

Hepatosplenic fungal infection in patients with acute leukemia in Taiwan: incidence, treatment, and prognosis.

Nosocomial fungal infection increases gradually and has become the leading pathogen at National Taiwan University Hospital since 1993. From January 1995 through May 2002, hepatosplenic fungal infection (HSF) was diagnosed in 37 (7.4%) of the 500 adult patients with acute leukemia who received chemotherapy at this hospital. There was no significant difference in the incidence of HSF between the patients with acute myeloid leukemia and those with acute lymphoblastic leukemia, or between the patients treated with high-dose chemotherapy and those with conventional or low-dose chemotherapy. Candida tropicalis was the leading pathogen, followed by Candida albicans. The computed tomography scan showed multiple hypodense lesions in the liver (89%), spleen (70%), and kidney (27%). Eighteen patients were initially treated with fluconazole and 19 with amphotericin B. Nineteen patients received the planned chemotherapy after the diagnosis of HSF. Among them, eight patients underwent hematopoietic stem cell transplantation and seven patients survived more than 100 days post-transplantation; none of these patients had relapse of prior HSF. Twenty-three patients (62%) died during a median follow up of 10 months, but only seven died due to HSF. In conclusion, a substantial percentage of patients with acute leukemia acquired HSF after chemotherapy and carried high mortality. However, HSF itself is not a contraindication for subsequent chemotherapy and hematopoietic stem cell transplantation.

Acute Disease↗

PubMed2002

Ontogeny, tissue distribution, and hormonal regulation of insulin-like growth factor binding protein-2 (IGFBP-2) in a marine fish, Sparus aurata.

In this study, we have cloned insulin-like growth factor binding protein (IGFBP)-2 from a marine hermaphroditic fish species, the gilthead sea bream (Sparus aurata), and determined its structure, ontogeny, tissue distribution, and hormonal regulation. The sea bream IGFBP-2 precursor consists of 286 amino acids, including a putative signal peptide of 22 residues and a mature protein of 264 residues. The overall sequence of sea bream preIGFBP-2 is 52-39% identical to that of zebrafish, chick, mouse, rat, pig, sheep, bovine, and human preIGFBP-2. The cysteine-rich N- and C-terminal domains, which are believed to be important for IGF binding, show a greater degree of conservation with a sequence identity of 73-56% and 53-50%, respectively. Sea bream IGFBP-2 contains 18 cysteine residues and their alignment is identical to those of other vertebrate IGFBP-2s. The Arg-Gly-Asp (RGD) sequence, present in the C-terminal domain of all known IGFBP-2, is also present in the sea bream. Northern blot analysis of RNA samples extracted from adult liver and developing larvae, revealed two transcripts of about 1.3 and 2.5 kb. RT-PCR analysis showed that IGFBP-2 was expressed in all adult tissues studied, with the highest levels found in liver and skin. IGFBP-2 transcripts were detected in gonad during reproductive cycle of S. aurata. Highest levels of IGFBP-2 mRNA were found in bisexual young gonads, and the levels decreased with gonad development and relatively high levels of IGFBP-2 mRNA were found in the ovary during spawning. By contrast, testicular IGFBP-2 mRNA levels were very low. RT-PCR detected IGFBP-2 mRNA throughout development in unfertilized eggs, embryos, and larvae, with highest levels observed after day 3 post-hatching, suggesting that this mRNA is the product of both the maternal and embryonic genomes. High variability in steady-state levels of hepatic IGFBP-2 mRNA was noted in adult fish, resulting in a statistically insignificant response to growth hormone treatment. These results suggest that the structure of IGFBP-2 is conserved in sea bream and that IGFBP-2 mRNA is expressed during early development and in gonad during the reproductive cycle, suggesting that it may play a role in gilthead sea bream development and reproduction.

Amino Acid Sequence↗

PubMed2001

Poliovirus 3C protease-mediated degradation of transcriptional activator p53 requires a cellular activity.

Infection of HeLa cells with poliovirus leads to rapid shut-off of host cell transcription by RNA polymerase II. Previous results have suggested that both the basal transcription factor TBP (TATA-binding protein) and transcription activator proteins such as CREB (cyclic AMP-responsive element-binding protein) and Oct-1 (the octamer-binding factor) are cleaved by the viral-encoded protease, 3C(Pro). Here we demonstrate that the transcriptional activator (and tumor suppressor) p53 is degraded by the viral protease 3C both in vivo and in vitro. Unlike other transcription factors that are directly cleaved by 3C(pro), degradation of p53 requires a HeLa cell activity in addition to 3C(Pro). The degradation of p53 by 3C(Pro) does not appear to involve the ubiquitin pathway of protein degradation. Vaccinia virus infection of HeLa cells leads to inactivation of the cellular activity required for 3C(Pro)-mediated degradation of p53. The vaccinia-encoded protein (CrmA) is known to inhibit caspase I (ICE protease) that converts inactive IL-1beta to an active secreted form. Incubation of HeLa cells with caspase I inhibitor Z-VAD-fmk does not interfere with 3C(Pro)-mediated degradation of p53. The cellular activity present in extracts of HeLa cells can be fractionated through phosphocellulose. A partially purified fraction that elutes at 0.6 M KCl from phosphocellulose contains the activity that degrades p53 in a 3C(Pro)-dependent manner. These results suggest that both poliovirus-encoded protease 3C(Pro) and a cellular activity are required for the degradation of p53 observed in cells infected with poliovirus.

3C Viral Proteases↗

PubMed2001

Interaction of poliovirus-encoded 2C/2BC polypeptides with the 3' terminus negative-strand cloverleaf requires an intact stem-loop b.

The poliovirus-encoded, membrane-associated polypeptide 2C and its precursor, 2BC, is believed to be required for initiation and elongation of viral RNA synthesis. Previous studies have shown that the 2C polypeptide specifically interacts with the 3'-terminal sequence of poliovirus negative-strand RNA. This interaction is facilitated by the presence of the conserved sequence UGUUUU in the "stem a" within the 3'-terminal cloverleaf structure. We show that the 2C precursor, 2BC, also interacts with the 3'-terminal cloverleaf of the negative-strand RNA. We also demonstrate here that interaction of 2C/2BC with the negative strand 3'-terminal sequence not only depends on an intact "stem a" containing the UGUUUU sequence but is equally influenced by the presence of an intact "stem b" within the negative-strand cloverleaf. The results presented here suggest that the spatial configuration of stem a UGUUUU sequence with respect to an intact stem-b is crucial for 2C/2BC interaction with the 3'-terminal negative-strand cloverleaf structure.

Base Sequence↗

PubMed2001

Philadelphia chromosome-positive acute lymphoblastic leukemia in Taiwan.

From January 1986 to December 1998, 26 (23%) of 114 acute lymphoblastic leukemia (ALL) patients older than 15 years were found to have Philadelphia (Ph) chromosome. They accounted for 28% (26 of 94) of the patients with B-lineage ALL. Compared with the other 88 ALL patients, the leukemic cells from all but one Ph-positive ALL patients were early pre-B cells with a higher rate of CD34 expression (92% vs 50%, P<0.05). At presentation, the Ph-positive adult ALL patients had higher circulating blasts (mean 21.4 vs 3.66x10(4)/microl, P<0.05) and lower initial platelet counts (mean 4.47 vs 7.34x10(4)/microl, P<0.01) and showed a trend toward higher frequency of initial central nerve system (CNS) involvement (25% vs 11%, P=0.098) than the others. Among patients with adequate chemotherapy, 16 (64%) of the 25 Ph-positive ALL patients achieved complete remission (CR), an incidence marginally lower than that of Ph-negative ALL (62 of 76, 82%, P=0.06) and significantly lower than that of Ph-negative B-lineage ALL (50 of 58, 86%, P=0.0037). However, all patients relapsed except for those who received allogeneic hemopoietic stem cell transplantation (allo-HSCT). The probabilities of 5-year continuous CR and 5-year survival for Ph-positive adult ALL patients were significantly inferior to those for Ph-negative adult ALL patients (0% vs 12%, P=0.0001, and 7% vs 19%, P=0.047, respectively), and those for Ph-negative B-lineage ALL (0% vs 14%, P<0.0001, and 7% vs 23%, P=0.002, respectively). Prognostic factors were analyzed among the Ph-positive ALL patients including the 26 adults mentioned above and an additional 11 children. No clinical or biological characteristics such as age, sex, initial circulating blast count, extramedullary involvement, or CD34 expression had an impact on the disease outcome. Allo-HSCT in first CR may improve the probability of 5-year continuous CR (100% vs. 0% for those without allo-HSCT, P=0.0091) although only three patients received it in this study. In conclusion, Ph-positive ALL patients tended to have a poor prognosis, regardless of whether other possible risk factors were present or not. Aggressive treatment, such as high-dose chemotherapy with allo-HSCT, should be considered for these patients to improve survival.

Adolescent↗

PubMed2000

The prevalence of C677T mutation in the methylenetetrahydrofolate reductase gene and its association with venous thrombophilia in Taiwanese Chinese.

C677T mutation of the methylenetetrahydrofolate reductase gene remains a controversial risk factor for venous thrombosis in Whites. The prevalence of methylenetetrahydrofolate reductase C677T genotype and its association with vascular thrombosis are not well established in Chinese population. We conducted a case-control study to investigate the prevalence of methylenetetrahydrofolate reductase C677T gene mutation and its association with venous thrombophilia in Taiwanese Chinese. The subjects consisted of 112 venous thrombophilic patients and 125 healthy controls, with similar age (p=0.08) and sex (p=0.58). The prevalent rates of C/T heterozygote were 32.8 and 44.6%; whereas those of T/T homozygote were 6.4 and 8.0% in the controls and patients, respectively. Neither C/T heterozygote (odds ratio, 1.7; 95% confidence interval, 1.0-3.0, p=0.05] nor T/T homozygote (odds ratio, 1.4; 95% confidence interval, 0.5-4.0, p=0.5) was significantly associated with venous thrombosis. Even when only subjects (52 patients and 107 controls) with normal inhibitor protein levels were analyzed, the association of T/T homozygote with venous thrombosis remained insignificant (p=0.06) with an odds ratio (95% confidence interval) of 3.4 (0.99-11.7). We concluded that, in Taiwanese Chinese, methylenetetrahydrofolate reductase C677T mutation is a common genetic mutation, but T/T homozygote is not a significant risk factor for venous thrombophilia.

Adult↗

PubMed2000

Evaluation of the practice of routine culturing of native valves during valve replacement surgery.

BACKGROUND: We routinely cultured native heart valves removed during valve replacement surgery even when infected carditis (IE) was not suspected. Several probable contaminated cultures prompted us to evaluate this practice. METHODS: The medical records of all patients who had positive valve cultures from 1995 to 1997 were reviewed for admission diagnoses, operative surgery, pathology and microbiology report, postoperative infections, and antibiotic use. Cases were excluded only for incomplete medical records or preoperative suspicion of IE. Long-term outcome for the cases was obtained from review of outpatient records and phone contact with the patient or physician. RESULTS: Thirty-two of 222 (14.4%) evaluable patients had positive valve cultures. Coagulase-negative Staphylococcus was the most common isolate. IE was not suggested in any of these cases based upon the surgical or the pathology report. Only 1 of 32 (3%) developed postoperative prosthetic valve endocarditis (PVE). Three patients died of unrelated causes, and the 28 surviving patients showed no sign of PVE, with a mean follow-up of 23 months. CONCLUSIONS: The incidence of false-positive native valve cultures is high. Positive cultures did not predict the occurrence of PVE sufficiently to justify obtaining them. Treating patients who had positive native valve cultures would have been unwarranted and poses an unnecessary risk.

Adult↗

PubMed2000

Genetic epidemiology of type 1 diabetes mellitus in Taiwan.

Some environmental and genetic factors play important roles in etiopathogenesis of type 1 or insulin-dependent diabetes mellitus (IDDM). HLA genes, the IDDM1 locus located the human chromosome 6, were found to be associated with insulin-dependent diabetes mellitus. However, the incidence of IDDM varied greatly among various populations. To evaluate the pathogenetic factors contributing to the development of IDDM in Taiwan, HLA typing was performed in a group of IDDM unrelated individuals and IDDM pedigrees along with the normal controls from the northern Taiwan. DNA genotypes of class II HLA were done by polymerase-chain-reaction based oligotyping techniques. We confirmed that class II HLA genes were significantly associated with IDDM in Taiwan. To study detailed molecular structure of class II HLA molecules and disease association, we examined several amino acid residues on DQalpha and DQbeta chains and the molecular mechanisms to explain the heterozygotic effect of the DR3/DR4 and DR3/DR9 in the Chinese population. Linkage analysis in our pedigrees confirmed the association between HLA and IDDM in population association studies. Among the several class II alleles, a closer segregation of HLA-DQB1*0401 to the affected persons might suggest that HLA-DQB1*0401 itself or an allele closely linked to the DQB1 locus was the IDDM-predisposing allele in Taiwanese. For IDDM2 (INS) region, association with IDDM was not found due to that more than 90% of the population carried class I alleles. In our collection of IDDM, we found few cases (2.4%) carried mitochondrial DNA mutation. Our studies in Taiwanese confirm a multigenetic nature for IDDM.

Autoantibodies↗

PubMed2000

Heterozygous loss of Six5 in mice is sufficient to cause ocular cataracts.

Myotonic dystrophy (DM) is an autosomal dominant disorder characterized by skeletal muscle wasting, myotonia, cardiac arrhythmia, hyperinsulinaemia, mental retardation and ocular cataracts. The genetic defect in DM is a CTG repeat expansion located in the 3' untranslated region of DMPK and 5' of a homeodomain-encoding gene, SIX5 (formerly DMAHP; refs 2-5). There are three mechanisms by which CTG expansion can result in DM. First, repeat expansion may alter the processing or transport of the mutant DMPK mRNA and consequently reduce DMPK levels. Second, CTG expansion may establish a region of heterochromatin 3' of the repeat sequence and decrease SIX5 transcription. Third, toxic effects of the repeat expansion may be intrinsic to the repeated elements at the level of DNA or RNA (refs 10,11). Previous studies have demonstrated that a dose-dependent loss of Dm15 (the mouse DMPK homologue) in mice produces a partial DM phenotype characterized by decreased development of skeletal muscle force and cardiac conduction disorders. To test the role of Six5 loss in DM, we have analysed a strain of mice in which Six5 was deleted. Our results demonstrate that the rate and severity of cataract formation is inversely related to Six5 dosage and is temporally progressive. Six5+/- and Six5-/- mice show increased steady-state levels of the Na+/K+-ATPase alpha-1 subunit and decreased Dm15 mRNA levels. Thus, altered ion homeostasis within the lens may contribute to cataract formation. As ocular cataracts are a characteristic feature of DM, these results demonstrate that decreased SIX5 transcription is important in the aetiology of DM. Our data support the hypothesis that DM is a contiguous gene syndrome associated with the partial loss of both DMPK and SIX5.

Animals↗

PubMed2000

Kainic acid-induced neurotrophic activities in developing cortical neurons.

Using primary cultured cortical neurons from embryonic rat brains, we elucidated an alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)/kainic acid (KA) receptor-mediated neuroprotective mechanism through actions of nerve growth factor (NGF) in developing neurons. Neurotoxicity of KA in early days in vitro neurons was quite low compared with the mature neurons. However, pretreatment with anti-NGF antibody or TrkA inhibitor AG-879 profoundly raised KA toxicity. Furthermore, KA stimulation resulted in an increase of TrkA expression and phosphorylation, which was blocked not only by the AMPA/KA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione and AG-879, but also by the phospholipase C inhibitor U73122 and the intracellular calcium chelator BAPTA. A study of polyphosphoinositide turnover showed that KA-stimulated phospholipase C (PLC) activity was directly triggered by the AMPA/KA receptor activity, but not by the activity of TrkA or other excitatory amino acid receptor subtypes. Sources of KA-increased intracellular calcium levels were contributed by both extracellular calcium influx and intracellular calcium release and were partially sensitive to guanosine 5'-O-(2-thiodiphosphate). These results indicate that in developing cortical neurons, activation of AMPA/KA receptors by KA may induce expression, followed by activation of TrkA via PLC signaling and intracellular calcium elevation and hence increase reception of NGF on KA-challenged neurons. A G protein-coupled AMPA/KA receptor may be involved in these metabotropic events for neuronal protection.

2-Amino-5-phosphonovalerate↗

PubMed2000

Induction of growth hormone (GH) mRNA by pulsatile GH-releasing hormone in rats is pattern specific.

Growth hormone-releasing hormone (GHRH) is a main inducer of growth hormone (GH) pulses in most species studied to date. There is no information regarding the pattern of GHRH secretion as a regulator of GH gene expression. We investigated the roles of the parameters of exogenous GHRH administration (frequency, amplitude, and total amount) upon induction of pituitary GH mRNA, GH content, and somatic growth in the female rat. Continuous GHRH infusions were ineffective in altering GH mRNA levels, GH stores, or weight gain. Changing GHRH pulse amplitude between 4, 8, and 16 microg/kg at a constant frequency (Q3.0 h) was only moderately effective in augmenting GH mRNA levels, whereas the 8 microg/kg and 16 microg/kg dosages stimulated weight gain by as much as 60%. When given at a 1.5-h frequency, GHRH doubled the amount of GH mRNA, elevated pituitary GH stores, and stimulated body weight gain. In the rat model, pulsatile but not continuous GHRH administration is effective in inducing pituitary GH mRNA and GH content as well as somatic growth. These studies suggest that the greater growth rate, pituitary mRNA levels, and GH stores seen in male compared with female rats are likely mediated, in part, by the endogenous episodic GHRH secretory pattern present in males.

Animals↗

PubMed1999

Insulin-like growth factor binding protein 2 is a growth inhibitory protein conserved in zebrafish.

Fish serum contains several specific binding proteins for insulin-like growth factors (IGFBPs). The structure and physiological function of these fish IGFBPs are unknown. Here we report the complete primary sequence of a zebrafish IGFBP deduced from cDNA clones isolated by library screening and rapid amplification of cDNA ends. The full-length 1,757-bp cDNA encodes a protein of 276 aa, which contains a putative 22-residue signal peptide and a 254-residue mature protein. The mature zebrafish IGFBP has a predicted molecular size of 28,440 Da and shows high sequence identity with human IGFBP-2 (52%). The sequence identities with other human IGFBPs are <37%. Chinese hamster ovary cells stably transfected with the zebrafish IGFBP-2 cDNA secreted a 31-kDa protein, which bound to IGF-I and IGF-II with high affinity, but did not bind to Des(1-3)IGF-I or insulin. Northern blot analyses revealed that the zebrafish IGFBP-2 transcript is a 1.8-kb band expressed in many embryonic and adult tissues. In adult zebrafish, IGFBP-2 mRNA levels were greatly reduced by growth hormone treatment but increased by prolonged fasting. When overexpressed or added to cultured zebrafish and mammalian cells, the zebrafish IGFBP-2 significantly inhibited IGF-I-stimulated cell proliferation and DNA synthesis. These results indicate that zebrafish IGFBP-2 is a negative growth regulator acting downstream in the growth hormone-IGF-I axis.

Amino Acid Sequence↗

PubMed1999

Adsorption properties and breakthrough model of 1,1-dichloro-1-fluoroethane on activated carbons.

Hydrochlorofluorocarbons (HCFCs), among the major replacements for chlorofluorocarbons (CFCs) and chlorinated solvents, are now considered to the prime contribution to the stratospheric ozone depletion. This paper reports the adsorption equilibrium of 1,1-dichloro-1-fluoroethane (HCFC-141b) vapor in air streams on the commercial activated carbons PCB and BPL, which were made from coconut shell and bituminous coal, respectively, at 283, 293, 303, and 313 K. The experimental results show that within the experimental conditions, the adsorption capacities of the two adsorbents for HCFC-141b in dry air streams can be well-fitted by the Langmuir, Freundlich, and Dubinin-Radushkevich (D-R) adsorption equations. The physical properties of the adsorbents are consistent with the isotherm parameters obtained from the adsorption results. In addition, available experimental data of adsorption breakthrough at 283 K were used in conjunction with the Yoon and Nelson model to generate theoretical breakthrough curves based on the obtained values of parameters. In each case, calculated theoretical breakthrough curves are in agreement with corresponding experimental data.

Journal Article↗

PubMed1999

Receptor protein tyrosine phosphatase alpha participates in the m1 muscarinic acetylcholine receptor-dependent regulation of Kv1.2 channel activity.

The phosphorylation state of a given tyrosine residue is determined by both protein tyrosine kinase (PTK) and protein tyrosine phosphatase (PTP) activities. However, little is known about the functional interaction of these opposing activities at the level of an identified effector molecule. G protein-coupled receptors (GPCRs), including the m1 muscarinic acetylcholine receptor (mAChR), regulate a tyrosine kinase activity that phosphorylates and suppresses current generated by the Kv1.2 potassium channel. We examined the possibility that PTPs also participate in this signaling pathway since the tyrosine phosphatase inhibitor vanadate increases the extent of both Kv1.2 phosphorylation and suppression. We show that an endogenous transmembrane tyrosine phosphatase, receptor tyrosine phosphatase alpha (RPTPalpha), becomes tyrosine phosphorylated and co-immunoprecipitates with Kv1.2 in a manner dependent on m1 receptor activation. The N- and C-termini of Kv1.2 are shown to bind RPTPalpha in vitro. Overexpression of RPTPalpha in Xenopus oocytes increases resting Kv1.2 current. Biochemical and electrophysiological analysis reveals that recruiting RPTPalpha to Kv1.2 functionally reverses the tyrosine kinase-induced phosphorylation and suppression of Kv1.2 current in mammalian cells. Taken together, these results identify RPTPalpha as a new target of m1 mAChR signaling and reveal a novel regulatory mechanism whereby GPCR-mediated suppression of a potassium channel depends on the coordinate and parallel regulation of PTK and PTP activities.

Animals↗

PubMed1999

Thrombin activates the growth, cell-cycle kinetics, and clustering of human dental pulp cells.

Thrombin is activated during vascular injury and inflammation of the dental pulp. In the present study, we found that thrombin can stimulate the proliferation of pulp cells in a dose-dependent manner as analyzed by modified MTT assay. The cell number increased by 1.6, 1.77, and 2.14-fold over that of control after exposure to 5, 10, and 20 units/ml of thrombin for 5 days. Flow cytometry studies also found that thrombin (10 units/ml) can induce the cell cycle progression of pulp cells after 24 h of incubation, as revealed by increasing the proportion of cells in the S phase and the G2/M phase from 29 to 72%. Moreover, exposure to thrombin (> 5 units/ml) for 3 days led to marked clustering of pulp cells. We concluded that thrombin can regulate the growth, cell cycle progression, and functional reorganization of the pulp tissue during pulp healing and inflammatory processes.

Cell Aggregation↗

PubMed1998

A longitudinal study of immunological status in Chinese haemophiliacs: importance of the heat viral inactivation of factor concentrates. II. Improvements of CD4/CD8 ratio after treatments with heat-inactivated factor concentrates.

Screened and heated clotting factor concentrates of intermediate purity have been used in Taiwanese haemophiliacs since the end of 1986. A significant improvement of CD4/CD8 ratio during the years 1987-1989 as compared with those during the years 1984-1986 was observed in haemophilia A patients [mean +/- SD (median), 1.191 +/- 0.495 (1.163) vs. 0.880 +/- 0.325 (0.838), P = 0.0020] who were seronegative for human immunodeficiency virus. Almost all patients received an increased amount of factor VIII concentrates and total plasma products since 1987. Multiple linear regression analysis for the association of CD4/CD8 ratio with changes in dosage of plasma products revealed that there was a significant positive association of CD4/CD8 ratio measured during 1987-1989 with dosage of factor VIII concentrate administered during 1984-1986 (P = 0.0230), which is an indicator for changes in viral load, but not with changes in dosage of plasma products, which are indicators for changes in plasma protein intake. Our data indicate that immunological abnormalities after replacement therapy observed in haemophiliacs are mainly attributed to virus infection through infusion of factor concentrates, not to allogeneic proteins existing in plasma products.

Adolescent↗

PubMed1998

A small yeast RNA blocks hepatitis C virus internal ribosome entry site (HCV IRES)-mediated translation and inhibits replication of a chimeric poliovirus under translational control of the HCV IRES element.

Hepatitis C virus (HCV) infection frequently leads to chronic hepatitis and cirrhosis of the liver and has been linked to development of hepatocellular carcinoma. We previously identified a small yeast RNA (IRNA) capable of specifically inhibiting poliovirus (PV) internal ribosome entry site (IRES)-mediated translation. Here we report that IRNA specifically inhibits HCV IRES-mediated translation both in vivo and in vitro. A number of human hepatoma (Huh-7) cell lines expressing IRNA were prepared and characterized. Constitutive expression of IRNA was not detrimental to cell growth. HCV IRES-mediated cap-independent translation was markedly inhibited in cells constitutively expressing IRNA compared to control hepatoma cells. However, cap-dependent translation was not significantly affected in these cell lines. Additionally, Huh-7 cells constitutively expressing IRNA became refractory to infection by a PV-HCV chimera in which the PV IRES is replaced by the HCV IRES. In contrast, replication of a PV-encephalomyocarditis virus (EMCV) chimera containing the EMCV IRES element was not affected significantly in the IRNA-producing cell line. Finally, the binding of the La autoantigen to the HCV IRES element was specifically and efficiently competed by IRNA. These results provide a basis for development of novel drugs effective against HCV infection.

Autoantigens↗

PubMed1997

The m1 muscarinic acetylcholine receptor transactivates the EGF receptor to modulate ion channel activity.

Intracellular tyrosine kinases link the G protein-coupled m1 muscarinic acetylcholine receptor (mAChR) to multiple cellular responses. However, the mechanisms by which m1 mAChRs stimulate tyrosine kinase activity and the identity of the kinases within particular signaling pathways remain largely unknown. We show that the epidermal growth factor receptor (EGFR), a single transmembrane receptor tyrosine kinase, becomes catalytically active and dimerized through an m1 mAChR-regulated pathway that requires protein kinase C, but is independent of EGF. Finally, we demonstrate that transactivation of the EGFR plays a major role in a pathway linking m1 mAChRs to modulation of the Kv1.2 potassium channel. These results demonstrate a ligand-independent mechanism of EGFR transactivation by m1 mAChRs and reveal a novel role for these growth factor receptors in the regulation of ion channels by G protein-coupled receptors.

Carbachol↗