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C C Wang

Publications and source records attributed to C C Wang.

At least 343 records · Page 19Linked to original sources
PubMed1995

Sequences at four termini of the giardiavirus double-stranded RNA.

The linear double-stranded RNA (dsRNA) genome of giardiavirus (GLV) was estimated to be 6100 nucleotides (nt) [Wang et al., Proc. Natl. Acad. Sci. USA 90 (1993) 8585-8599]. As the 5'- and 3'-untranslated regions (UTR) of viral genomes are known to contain critical information for viral replication, we reexamine the sequences at all four termini of GLV dsRNA by (i) direct RNA sequencing with RT, (ii) tailing GLV dsRNA with UTP or CTP in addition to ATP in 3' rapid amplification of cDNA ends (RACE) and (iii) adding poly(dG) to the products of primer extension in 5'-RACE. The results confirmed the reported sequence for the 5'-terminus of the GLV sense strand RNA, but uncovered an additional 177 nt at the 3'-terminus. The new study also showed conclusively that there are no protruding overhangs at either terminus of GLV dsRNA.

Base Sequence↗
PubMed1995

Rat antizyme inhibits the activity but does not promote the degradation of mouse ornithine decarboxylase in Trypanosoma brucei.

Ornithine decarboxylase (ODC) of African trypanosomes is an important target for anti-trypanosomal chemotherapy because of its remarkable stability in vivo. The in vivo activity and stability of mammalian ODC are regulated by polyamines. Polyamines induce antizyme, which inactivates ODC by tight association and promotes degradation of ODC by the mammalian 26 S proteasome. Here we found, in contrast to mammalian cells, that polyamines caused no reduction of ODC activity in Trypanosoma brucei. Mouse ODC expressed in T. brucei was also unaffected by exogenous polyamines, suggesting that a mammalian antizyme equivalent may be absent in T. brucei. The rat antizyme expressed in T. brucei was found capable of inhibiting mouse ODC activity by the formation of rat antizyme-mouse ODC complex. However, complex formation did not lead to degradation of mouse ODC in T. brucei. Further in vitro experiments suggested the presence of an inhibitory factor(s) in trypanosome, which interferes with the degradation of mouse ODC. We also demonstrated the presence of proteasomes in T. brucei. But the mobility of the trypanosomal proteasome on native gel is different from that of the mammalian proteasome. Thus, the absence of antizyme, the presence of inhibitory factor(s), and the differences between trypanosomal and mammalian proteasome may account for the stability of mouse ODC in T. brucei cells.

Amino Acid Sequence↗
PubMed1995

Orphon spliced-leader sequences form part of a repetitive element in Angiostrongylus cantonensis.

In nematodes, the 22 nucleotide (nt) spliced leader (SL) is normally encoded by a multi-copy, tandemly reiterated SL gene and is trans-spliced from SL-RNA onto the 5' end of a subset of mRNAs. We have found that the SL is also encoded at multiple (> 100) orphon genomic sites in the parasitic nematode Angiostrongylus cantonensis. At these sites the sequence forms part of a 198 bp repetitive element (designated con-198). Transcription from two genomic elements that contain the con-198 sequence has been characterised. At one element (G-2) an approximately 850 nt RNA with an internal SL is transcribed. At the other (G-1), transcription takes place 3 kb downstream of the con-198 sequence.

Angiostrongylus cantonensis↗
PubMed1995

Consistent presence of isochromosome 7q in hepatosplenic T gamma/delta lymphoma: a new cytogenetic-clinicopathologic entity.

Peripheral T-cell lymphoma (PTL), which is characterized by hepatosplenic presentation and the gamma/delta T-cell receptor (TCR) phenotype on the malignant cells, is a rare but distinct subtype of non-Hodgkin's lymphomas. Little is known about the chromosomal changes in these lymphomas. We report the cytogenetic analysis of three patients who had neoplastic proliferation of T gamma/delta cells in the spleen, bone marrow, and liver, but not in lymph nodes or skin. Isochromosome 7q and trisomy 8 were observed in all three patients. Isochromosome 7q as the sole abnormality has been previously reported in one patient with similar clinicopathologic features. It is suggested that i(7q) is a primary, nonrandom chromosomal abnormality in hepatosplenic T gamma/delta PTL.

Adult↗
PubMed1995

Unbiased usage of T-cell receptor beta variable region genes in peripheral blood cells of hepatitis C patients: no correlation with superantigen effect.

Hepatitis C virus (HCV) infection frequently causes chronic hepatitis and lack of virus clearance in these patients. In addition, many patients infected by HCV also present with hypergammaglobulinemia in the early stage of chronic infection. These observations raise a possible viral superantigen effect induced by HCV, because viral superantigen found in human immunodeficiency virus (HIV) or in replication of defective murine leukemia virus (MuLV) is associated with T-cell dysfunction and polyclonal activation of B cells. The possibility was investigated of whether HCV encodes any superantigen by analyzing the usage of T-cell receptor (TCR) from the peripheral blood lymphocytes (PBL) of patients with chronic hepatitis C. Two groups, one with hypergammaglobulinemia and the other without hypergammaglobulinemia, were studied for the usage of TCR beta chain by reverse transcription-polymerase chain reaction (RT-PCR) analysis. It was found that all genes of V beta variable chain were used in the PBL of these patients. Furthermore, there was no significant difference of the TCR expression pattern between these two groups, nor a complete deletion of a particular T-cell subset in either group. These results do not provide evidence for HCV superantigen, but indicate that the TCR usage in the patients was neither defective nor biased.

Adult↗
PubMed1995

Establishment and characterization of TSGH9201, a human gastric carcinoma cell line that is growth inhibited by epidermal growth factor.

A human signet ring gastric carcinoma cell line TSGH9201 was established in vitro. The cells grew in vitro as a monolayer with polygonal morphology and had a population doubling time of 34 hours. The cells secreted tumor markers CEA and CA 125. They were, however, not tumorigenic in athymic nude mice. Karyotypic analysis demonstrated a near tetraploidy with a modal chromosome number of 98. Northern blotting and immunocytochemical analysis revealed the expression of both transforming growth factor alpha and high levels of epidermal growth factor receptor. Cell growth was inhibited by the epidermal growth factor in vitro. The cell line may be a useful tool to study autocrine growth regulation through the epidermal growth factor receptor.

Animals↗
PubMed1995

Preliminary X-ray crystallographic analysis of Tritrichomonas foetus inosine-5'-monophosphate dehydrogenase.

Inosine-5'-monophosphate dehydrogenase (IMPDH) from the protozoan parasite Tritrichomonas foetus has been expressed in E. coli and crystallized. Crystals were grown to 0.1 mm in each dimension in 18 to 72 h using ammonium sulfate and low-molecular-weight polyethylene glycols. The crystals belong to the cubic space group P432 with unit cell edge = 157.25 A. The enzyme is a homotetramer with each monomer having a molecular weight of 55,534 Da. There is one monomer per asymmetric unit, based on a volume/mass ratio of 2.7 A3/Da and self-rotation analysis. The crystals are adequately stable to allow a complete data set to be collected from a single crystal. Complete native data sets have been collected to 2.3 A resolution at 4 degrees C using synchrotron radiation. High-quality complete data extending to 3.0 A resolution have been collected from crystals of four putative derivatives, and the data appear to be isomorphous with that of the native crystals in each case. Efforts to solve the derivatives for use in MIR phasing are underway.

Animals↗
PubMed1995

Dietary copper supplementation increases the catecholamine levels in genetically obese (ob/ob) mice.

The interactive relationship between Cu deficiency and depressed synthesis of certain neurotransmitters has been recognized. To investigate the effects of dietary Cu supplementation on the catecholamine levels in genetically obese mice, male obese (ob/ob) mice and their lean (+/?) counterparts were administered either a control diet (4.0 mg/kg) or a Cu-supplemented diet (50 mg/kg) for 4 wk. The ob/ob mice that were fed a control diet showed lower liver and higher plasma levels of Cu. Depressed levels of plasma and brain catecholamines were also found in ob/ob mice that were fed the control diet. The ob/ob mice that received a Cu-supplemented diet showed significant increases in the levels of catecholamine in the plasma and brain. This study showed that catecholamine levels in ob/ob mice can be increased by dietary Cu supplementation. However, the interaction between Cu and sympathetic nervous activity in obesity was not elucidated in this study.

Animals↗
PubMed1995

Hepatocellular carcinoma in a pregnant woman detected by routine screening of maternal alpha-fetoprotein.

A 26-year-old pregnant woman was diagnosed as having hepatocellular carcinoma at 16 weeks of gestation when extremely high levels of alpha-fetoprotein were found on a routine screening examination. The pregnancy was terminated, and a right hepatectomy was performed. Although such cases are rare, this one suggests that early measurement of maternal serum alpha-fetoprotein in a pregnant woman with hepatitis B surface antigenemia may allow the timely detection of a hepatocellular carcinoma.

Abortion, Induced↗
PubMed1995

Differentiation of a culture-adapted mutant bloodstream form of Trypanosoma brucei into the procyclic form results in growth arrest of the cells.

The bloodstream forms of Trypanosoma brucei monomorphic strain 427 serially passaged in rats can differentiate in vitro equally well in HMI-9, HMI-10, SDM-79 or Cunningham's medium into the insect (procyclic) forms by a simple temperature shift from 37 to 26 degrees C in the presence of citrate and cis-aconitate. The procyclic forms thus generated can also continue to multiply at 26 degrees C without replacing the culture medium. The same strain of T. brucei pre-adapted to grow as bloodstream forms in HMI-10 medium at 37 degrees C is also capable of differentiating showing a similar rate of variant surface glycoprotein (VSG) disappearance and appearance of phosphoenolpyruvate carboxykinase (PEPCK) under the same experimental conditions. However, appearance of both procyclin mRNA and procyclin protein is much delayed and the resulting procyclic forms cannot multiply. The culture-adapted bloodstream forms are capable of infecting rats, and the cells thus harvested from the rats can differentiate but cannot multiply in the same manner as the original culture-adapted bloodstream forms. Apparently, a certain variant has been selected during the adaptation of T. brucei bloodstream forms from rat blood to the culture medium. This variant could be a useful tool for identifying the genes involved in differentiation of T. brucei and multiplication of the procyclic forms. Comparison of the protein profiles between the wild-type and the variant during differentiation showed that a major protein band of about 70 kDa remained in the non-dividing variant procyclic forms but vanished in the rapidly dividing wild type procyclic forms. N-terminal determinations indicated that the 70-kDa protein band consists of bovine serum albumin and fetuin. Presumably these two serum proteins are actively taken up by the bloodstream forms via endocytosis. Since the procyclic forms are incapable of endocytosis, the serum proteins may be rapidly diluted in the growing wild type procyclic cells but remain unchanged in the non-dividing procyclic cells of the variant. Further studies are underway in trying to identify the key distinctions between these two lines of cells at the molecular level.

Animals↗
PubMed1995

Progression to complete atrioventricular block in a patient with bundle branch re-entry tachycardia.

A 38-year-old man with no significant structural heart disease suffered from one episode of wide QRS tachycardia. The electrocardiogram showed a PR interval of 0.20 second and a QRS duration of 0.10 second. His bundle recording revealed an HV interval of 90-100 ms. The tachycardia was inducible with programmed stimulation and displayed a QRS morphology of complete left bundle branch block. It was characterized by an atrioventricular dissociation, a cycle length of 280 ms, and an H deflection preceding each QRS complex. Pacing from the right ventricular apex at a cycle length of 270 ms entrained the tachycardia, while at a cycle length of 260 ms, the tachycardia was terminated. Four years later, the patient presented with complete atrioventricular block with a wide QRS escape rhythm. An electrophysiologic study conducted while he was in 1:1 atrioventricular conduction showed an HV interval of 100 ms. Second-degree infrahisian block developed at an atrial paced cycle length of 700 ms. There was no induction of tachycardia with programmed stimulation before or after isoproterenol. The patient was treated with an implantation of a permanent pacemaker.

Adult↗
PubMed1995

Atrial pacing leads: the clinical contribution of steroid elution.

Although the original atrial pacing leads were passive fixation and J shaped for right atrial appendage placement, the subsequent development of the active fixation screw-in lead found favor because of a perceived low incidence of lead dislodgment and a wider selection of atrial pacing sites. A bipolar atrial lead study was undertaken to compare the long-term atrial implant data in 215 patients. Study leads comprised one passive fixation, steroid-eluting lead (Medtronic CapSure SP, 119 patients) and three nonsteroid-eluting leads; two active fixation (Medtronic BISPING model 4058, 30 patients; and Telectronics ACCUFIX model 330-801, 44 patients) and one passive fixation (Telectronics ENCOR model 330-854, 22 patients). Bipolar atrial voltage stimulation thresholds and electrograms were measured using the Telectronics META DDDR immediately postimplantation, and at 1-, 3-, 6-, 12-, and 18-month follow-up. There were 135 males and the mean age 68 years. The incidence of lead dislodgment was 4% for active fixation and 2% for passive fixation. All nonsteroid leads showed a typical rise in stimulation threshold with the highest being the ACCUFIX followed by the BISPING and ENCOR. The steroid-eluting CapSure SP, however, demonstrated a flat response with 98% of leads at 18 months having a value < or = 1.3 volts allowing voltage programming to 2.5 volts (2:1 safety ratio). Telemeted electrograms showed no differences for all leads at all visits. For low voltage atrial pacing with a low incidence of dislodgment and satisfactory atrial sensing, the steroid-eluting passive fixation lead is superior to all nonsteroid-eluting leads.

Aged↗
PubMed1995

Comparative complement selection in bacteria enables screening for lead compounds targeted to a purine salvage enzyme of parasites.

Expression plasmids encoding the hypoxanthine phosphoribosyltransferases (HPRTs) of Plasmodium falciparum, Schistosoma mansoni, Tritrichomonas foetus, and Homo sapiens were subcloned into genetically deficient Escherichia coli that requires complementation by the activity of a recombinant HPRT for growth on semidefined medium. Fifty-nine purine analogs were screened for their abilities to inhibit the growth of these bacteria. Several compounds that selectively altered the growth of the bacteria complemented by the malarial, schistosomal, or tritrichomonal HPRT compared with the growth of bacteria expressing the human enzyme were identified. These results demonstrate that the recombinant approach to screening compounds by complement selection in a comparative manner provides a rapid and efficient method for the identification of new lead compounds selectively targeted to the purine salvage enzymes of parasites.

Animals↗
PubMed1995

Maturation of giardiavirus capsid protein involves posttranslational proteolytic processing by a cysteine protease.

The double-stranded RNA genome of giardiavirus (GLV) has only two large open reading frame (ORFs). The 100-kDa capsid polypeptide (p100) is encoded by ORF1, whereas the only other viral polypeptide, the 190-kDa GLV RNA-dependent RNA polymerase (p190), is synthesized as an ORF1-ORF2 fusion protein by a (-1) ribosomal frameshifting. Edman degradation revealed that p100 was N-terminally blocked except for 2 to 5% of it that showed free N terminus starting from amino acid residue 33 of ORF1. Studies using antiserum targeted against amino acid residues 6 to 27 indicated that this region (NT) is absent from viral p100 and p190, while pulse-labelling experiments showed that NT is present in nascent p100 synthesized in GLV-infected Giardia lamblia but removed subsequently. In contrast, this region was retained in the two viral proteins synthesized in vitro, and it was not removed upon prolonged incubation or inclusion of microsomal fraction in the in vitro translation reaction mixtures. These results suggest that endoplasmic reticulum is not involved in the protein processing and that the precursors of p100 and p190 are incapable of cleaving themselves or each other. This specific cleavage was reproduced when lysates from GLV-infected G. lamblia were added, but not those from uninfected cells. The cleavage activity was relatively insensitive to phenylmethylsulfonyl fluoride, but it was inhibitable by leupeptin or E-64, two known specific inhibitors of cysteine protease. The possible origin of this processing activity is discussed.

Amino Acid Sequence↗
PubMed1995

Virus-mediated expression of firefly luciferase in the parasitic protozoan Giardia lamblia.

Giardia lamblia, a prevalent human pathogen and one of the lineages that branched earliest from prokaryotes, can be infected with a double-stranded RNA virus, giardiavirus (GLV). The 6,277-bp viral genome has been previously cloned (A.L. Wang, H.-M. Yang, K.A. Shen, and C.C. Wang, Proc. Natl. Acad. Sci. USA 90:8595-8599, 1993; C.-H. Wu, C.C. Wang, H.M. Yang, and A.L. Wang, Gene, in press) and was converted to a transfection vector for G. lamblia in the present study. By flanking the firefly luciferase gene with the 5' and 3' untranslated regions (UTRs) of the GLV genome, transcript of the construct was synthesized in vitro with T7 polymerase and used to transfect G. lamblia WB trophozoites already infected with GLV (WBI). Optimal electroporation conditions used for the transfection were set at 1,000 V/cm and 500 microF, which resulted in expression of significant luciferase activity up to 120 h after electroporation. Furthermore, the mRNA and the antisense RNA of the luciferase gene were both detected by reverse transcription and PCR from 6 to 120 h postelectroporation, whereas no antisense RNA of luciferase was observed in the electroporated virus-free Giardia WB trophozoites. The mRNA of luciferase was detectable in the virus-free trophozoites by reverse transcription and PCR only up to 20 h after the electroporation, indicating that the introduced mRNA was replicated only by the viral RNA-dependent RNA polymerase inside the WBI cells. This expression of luciferase was dependent on the presence of UTRs on both ends of the viral genome transcript, including a putative packaging site that was apparently indispensable for luciferase expression. This is the first time that a viral vector in the form of mRNA URTs has been successfully used in transfecting a protozoan.

Animals↗